VIOLIN: Vaccine Investigation and Online Information Network

Bacillus anthracis

Table of Contents

General Information
Vaccine Related Pathogen Genes
1. capsule biosynthesis protein (Other)
2. EF (Other)
3. Lef (Protective antigen)
4. Lef from B. anthracis str. A2012 (Protective antigen)
5. LF (Protective antigen)
6. NEAT (Protective antigen)
7. PA (Protective antigen)
8. PA63 (Protective antigen)
9. PA63-LAMP1 (Protective antigen)
10. PA83 (Protective antigen)
11. pagA (Protective antigen)
12. PagA from B. anthracis str. 'Ames Ancestor' (Protective antigen)
13. S-layer protein (Protective antigen)
14. TPA-PA63 (Protective antigen)
15. TPA-PA63-LAMP1 (Protective antigen)
16. UQ-PA63 (Protective antigen)
17. mntA (Virmugen)
Vaccine Related Host Genes
Vaccine Information
References

I. General Information

1. NCBI Taxonomy ID:

1392

2. Disease:

Anthrax

3. Introduction

The pathology of anthrax is almost entirely mediated by expression of two secreted toxins: lethal toxin (LeTx) and edema toxin (EdTx). The natural form of anthrax is extremely rare in the United States, with only 244 cases reported between 1944 and 1999. Natural infection of humans occurs through direct exposure to spores from infected animals or their products, such as hides or wool. Anthrax is primarily a disease of animals and is probably propagated in the environment through spores present in soil at sites of infected carcasses. The causative agent of anthrax is B. anthracis, which produces transmissible and infectious spores. The vegetative anthrax bacillus is not readily transmissible, but the spores are environmentally robust for years and can be easily transmitted to humans. This phenomenon is the core of anthrax biowarfare and bioterrorism concern, since infective spores can be obtained from fermentation cultures and purified in large quantities in a pure non-aggregable form suitable for aerosol dissemination. This could potentially result in the intentional dissemination of spores to cause human infection (Brey, 2005).

4. Microbial Pathogenesis

In general, spores are phagocytosed by macrophages and germinate within phagolysosomes. Vegetative bacteria release many toxins leading to macrophage death. Lethal toxin acts on host macrophages and induces the release of proinflammatory cytokines responsible for inducing sudden and fatal shock (Hanna et al., 1999). Edema toxin causes localized edema and systemic shock (Hirsh et al, 2004). Other virulence factors allow for survival within phagolysosomes and on mucosal surfaces (Inh and MprF), escape from phagolysosomes and phagocytic cells (anthrolysins), iron acquiring products (Dlp), and regulation of cellular products (AtxA and AcpA) (Hirsh et al, 2004).

Link to pathogenesis of Bacillus anthracis in HazARD.

5. Host Ranges and Animal Models

Host ranges include the following: livestock or other herbivores (eg, cattle, sheep, goats, pigs, bison, water buffalo) acquire infection by consuming contaminated soil or feed; spores are infectious agents that can enter the human body through skin lesions, ingestion, or inhalation; and laboratory animal models include Guinea pigs, Syrian hamsters, and various mouse models (PathPort).

6. Host Protective Immunity

Since protection against anthrax is induced by vaccines containing PA as the major immunogen, with minor amounts of EF and LF, antibodies against PA and other toxin components are essential in the protection against anthrax. Serum therapy has been used in the past for the treatment of human anthrax with some success. PA is a key immunogen for antianthrax vaccine development since it induces the production of toxin-neutralizing Abs. However, vital, anti-PA Abs are not the only, or completely sufficient, means for an immune host to impede the development of anthrax (Glomski et al., 2007). Immune serum containing antibodies to PA can be effective in the therapy of established experimental infection in guinea pigs. However, The identification of anti-toxic immunity as the most important means for protection against B. anthracis has been complicated by lack of an entirely well-accepted animal model for evaluating immunity to LeTx and to spores of different anthrax isolates, due to varying susceptibility of animal models to spores of different origin (Brey, 2005).

Anti-capsule antibodies may also be important in controlling the outgrowth phase of anthrax infection, since they would be presumed to fix complement and kill vegetative cells. However, antibodies to the poly γ-d-glutamate capsule have not been well studied, because the capsule is poorly immunogenic and is a T cell-independent antigen. Recently, a series of murine monoclonal antibodies to the capsule has been obtained by immunizing mice with an anthrax capsule isoform isolated from B. licheniformis. The capsule is a major virulence factor in mice. Although there is a definitive role for anti-toxin antibodies in protection against anthrax, it is not yet clear what levels of antibodies will be required to protect humans against anthrax after vaccination or passive injection of protective antibodies. This consideration is important since challenge studies cannot be performed in humans, and correlates of immunity have to be extrapolated from animal studies (Brey, 2005).

Humoral immunity does not protect from nontoxinogenic capsulated bacteria; however, a cellular immune response by IFN-{gamma}-producing CD4 T lymphocytes protect mice. These results provide evidence of protective cellular immunity against capsulated B. anthracis and suggest that future antianthrax vaccines should strive to augment cellular adaptive immunity (Glomski et al., 2007).

II. Vaccine Related Pathogen Genes

1. capsule biosynthesis protein

Gene Name : capsule biosynthesis protein
Sequence Strain (Species/Organism) : Bacillus anthracis
NCBI Protein GI : WP_000409765
Other Database IDs : CDD:311953
CDD:332959
CDD:282847
CDD:99910
Taxonomy ID : 86661
Gene Strand (Orientation) : ?
Protein Name : capsule synthesis positive regulator AcpB
Protein pI : 8.69
Protein Weight : 56188.45
Protein Length : 586
Protein Note : M protein trans-acting positive regulator (MGA) HTH domain; pfam08280

Protein Sequence : Show Sequence

>WP_000409765.1 MULTISPECIES: capsule synthesis positive regulator AcpB [Bacillus cereus group]
MEKDIKRQIQILEIITSEEKWFTTIEISKILRCCNKTIMKDISFIKDFLPEDWHIKIKKGKGVRIYLPYN
KHRNEITFLLFRESLTFRILQHLFERETKTIATLAERLYIQVPSILPALKRVENYLKKFGLKLRKKPLRL
EGDEVRIMIMYLDLYLKSYNDTEWPFEKLKKEVIFQYLGTLEESLGISLHVVSKRHLSFFIAILLKRKQQ
GYKVQLNRKFLYFNTETPDYVKIGRIFEKLEREFGVSLTVQDKILLTISIKSSKYVYKDINKEKEESVQY
FKEGNLSIYELVKDFINSLEEKLKVDLISDEEFIFALVDYFKRTIYHLQYLCMFERPQKQTIQYMQTEHS
ETFSAVKEVYTEFVKKNEIADYVSVEEIAKVTMYIEASRLRYTSNYKKVLLVTGESESWAEYLAATLAKR
FGDKIQISTVFFAKKSDHDVNADFIISTIPLDLGSTPIICINSIPTERDYTNIQYYLDLQDG

Molecule Role : Other

2. EF

Gene Name : EF
Sequence Strain (Species/Organism) : Bacillus anthracis
NCBI Protein GI : 142813
Other Database IDs : CDD:149028
CDD:202671
Taxonomy ID : 1392
Gene Strand (Orientation) : ?
Protein Name : edema factor
Protein Length : 800
Protein Note : Anthrax toxin lethal factor, N- and C-terminal domain; pfam07737

Protein Sequence : Show Sequence

>gi|142813|gb|AAA79215.1| edema factor [Bacillus anthracis]
MTRNKFIPNKFSIISFSVLLFAISSSQAIEVNAMNEHYTESDIKRNHKTEKNKTEKEKFKDSINNLVKTE
FTNETLDKIQQTQDLLKKIPKDVLEIYSELGGEIYFTDIDLVEHKELQDLSEEEKNSMNSRGEKVPFASR
FVFEKKRETPKLIINIKDYAINSEQSKEVYYEIGKGISLDIISKDKSLDPEFLNLIKSLSDDSDSSDLLF
SQKFKEKLELNNKSIDINFIKENLTEFQHAFSLAFSYYFAPDHRTVLELYAPDMFEYMNKLEKGGFEKIS
ESLKKEGVEKDRIDVLKGEKALKASGLVPEHADAFKKIARELNTYILFRPVNKLATNLIKSGVATKGLNE
HGKSSDWGPVAGYIPFDQDLSKKHGQQLAVEKGNLENKKSITEHEGEIGKIPLKLDHLRIEELKENGIIL
KGKKEIDNGKKYYLLESNNQVYEFRISDENNEVQYKTKEGKITVLGEKFNWRNIEVMAKNVEGVLKPLTA
DYDLFALAPSLTEIKKQIPTKRMDKVVNTPNSLEKQKGVTNLLIKYGIERKPDSTKGTLSNWQKQMLDRL
NEAVKYTGYTGGDVVNHGTEQDNEEFPEKDNEIFIINPEGEFILTKNWEMTGRFIEKNITGKDYLYYFNR
SYNKIAPGNKAYIEWTDPITKAKINTIPTSAEFIKNLSSIRRSSNVGVYKDSGDKDEFAKKESVKKIAGY
LSDYYNSANHIFSQEKKRKISIFRGIQAYNEIENVLKSKQIAPEYKNYFQYLKERITNQVQLLLTHQKSN
IEFKLLYKQLNFTENETDNFEVFQKIIDEK

Molecule Role : Other

3. Lef

Gene Name : Lef
Sequence Strain (Species/Organism) : Bacillus anthracis str. 'Ames Ancestor'
VO ID : VO_0010873
NCBI Gene ID : 2820148
NCBI Protein GI : 47566484
Locus Tag : GBAA_pXO1_0172
Genbank Accession : AE017336
Protein Accession : YP_016503
3D structure: PDB ID : 1J7N
Other Database IDs : CDD:285037
CDD:286266
CDD:238144
Taxonomy ID : 261594
Plasmid No : pXO1
Gene Starting Position : 149356
Gene Ending Position : 151785
Gene Strand (Orientation) : -
Protein Name : lethal factor
Protein pI : 5.69
Protein Weight : 89799.18
Protein Length : 809
Protein Note : Anthrax toxin lethal factor, N- and C-terminal domain; pfam07737

DNA Sequence : Show Sequence

>NC_007322.2:149356-151785 Bacillus anthracis str. 'Ames Ancestor' plasmid pXO1, complete sequence
CTTATGAGTTAATAATGAACTTAATCTGATCGTTAATAAATTGGAAAGTTTTCGGAGCATTTTTTTGAAC
TTTTAAACGTTCAGCATGGTCCGTAGAATGCATTAACCTAAAGGCTTCTGCAAAAAATTCCGCTTCATTT
GTTCTCCCATACGAAGTTAAATTACTCCCTTCTTCCTTAAAAATATCAATGAATTTTTTAGAATTTGTAA
CTAAATCAGATTGGTTCTTATCTAATAGATATCCAGCATAATCATCCACAGCATGTCCAAATTCGTGTAT
AAAACCCTCACTATCATTCCTTAATTCTACACCTTTTGAAGGTCCATGGAGTAATATAGAACGGGATTCT
GGAACATATAACCCTTTTGAATGAACTTGCTCATATATCTCATCTTGATGTGTATATTGTTCAGCTATAT
TAGGGAGAGTAATATCGGTAAAAACAAATCTTCCATTACCATCAACTAAGTAATTTGTTACCTTTTTTAT
AAGATCACTTTGAATATTATTTTTCCATTCATTCAATATTAAATAAGCACTTTCTACAATATTGGATGCA
TATCTATTATGCACGTTGAATGTAATAAGCTTTGTATATTTTGGTAACCCTAATGCTTTATTCCATTCCT
GATTTATATTTAACTGTGCTTCTTGAATTTTTGTATCTATTTTACTCTTTGGCACTACTTTCGCATCAAT
CCTTATATATTCTTTTTCGGATTGCTTAATTATTTGTACATCCTTTATTTCCAGACCGATGTTTCTTTGT
AATATAAGCTTTCCATTTTCTAAATATCCTGCTCGAGTATCTGGTGATAATTGGATTCTCCATTTCAAAC
GCTCATTATCTAATGCAGGCCTTTCATTTATATCAACAATCATATAGTTACTAGAAATACTATATTTGAA
ATTTTTTTTGAATTCATTGAAAATACCTCTATTAATTTTAGTATTATCAGTGGAATCAACTAAATCCGCA
CCTAGGGTTGCTGTAAGGTTATTGATATTCATATTTTCATACAAATAAATTTTATTGTACAAGGTACTTC
CAATGGATTGATGTAATAAAGCATCAATATTTTGAATATCCCTTTTATACTGCTTTCTTACATCAAGATT
AATTGACGGACTATCAATTAACCCTCCTGTATCTTGCAACCTTTGATTAATATCATATGGTTGAATATCA
AGTTTCAGCTTTTTTAAAAACTCTTTTTCTTTTTCAGATAAAGGATTACTACTATCCACCTGTATTCTAT
TTAAAAGCTCTTTTTCTTCTTCAGATAAAGAATCACGAATATCAATTTGTAGCTTTTTTAAAAACTCTTT
TTCCTCAGTAGATAAAAAATCACTACTATCAATTTGTATTCTTTTTAGAAGCTCTTTTTCTTCTTGAGAT
AAAGAATGAATTATGTCATCTTTCTTTGGCTCAATAGGAATCTGCAGCTTTTTTAAAAGTCCTCTTCCTT
CTTCAGATAAAGAATCGCTCCAGTGTTGATAGTGCTGTTTTATCTTTTCCCATTTTTCATATCTTGCCAG
CATCCGTTGATCTTTAAGTTCTTCCAAGGATAGATTTATTTCTTGTTCGTTAAATTTATCCATGTAATTA
AAAGCTTCCGGTGCATAAAGCTGTAAAACATCACGATGCTGTGGCTCGATATAATATGCAAAAGCTTTCG
CAAATACTTCTTGTACCTCATTGCTATTTTGTTCCAAGAATTCTACAGAAAAGTCTGTGGGATGTTCCTT
AAGCTGATTAGTAAATAAAAGATCTTGTCCATCTGAATCAGATGCATTTTTAATGGTATTTAATACATCT
AAAAATTTCTGATATGGTTGATTAATTTTACTTAAAATATCCCTTGATAATATCTTACCTATTTCATAAT
AAACGTTCAGTGCCTTTTCAGTATTTTCTACATAATCTTCCGAAGATTGGATTACAAGTACGGGTTCATA
TCCTTCTTTTGCATATACATAATGTTCATGTAATAAAGCATCTTTCCCATAAATGTCTTTTATTTTTTTC
TTATCTTCAGATAATGCTTCTAAAGATATATGTTTTGTAATATCACCATCCACAATATATATCTTTCCTC
CAATTGCTTTATACATCTCTAAAACATCAGATGGTACTTTCTCAAGTAGCTTTTCTGCTGCCTCTTTTTT
AACAGCTTCCTCCCCTTTTACTTCTATTTTTACAATGTGTTTCATGATTTCCTTTAAATGCTCTTCCTGT
GTTTTATTTCGTTCTTCATCTTTTCTCTTATTCTCATCTTTATTTTTCTCTTTCTCTTTTACGTGCATAC
CTACATCACCATGACCGCCCGCCCCCTGTACAAGGGGGATAAAGACGGGACCACTCAAAGTAATTGCTGT
TACTAAACATGACATACTAATTACTTTTATAAATTCTTTTTTTATATTCA

Protein Sequence : Show Sequence

>YP_016503.2 lethal factor (plasmid) [Bacillus anthracis str. 'Ames Ancestor']
MNIKKEFIKVISMSCLVTAITLSGPVFIPLVQGAGGHGDVGMHVKEKEKNKDENKRKDEERNKTQEEHLK
EIMKHIVKIEVKGEEAVKKEAAEKLLEKVPSDVLEMYKAIGGKIYIVDGDITKHISLEALSEDKKKIKDI
YGKDALLHEHYVYAKEGYEPVLVIQSSEDYVENTEKALNVYYEIGKILSRDILSKINQPYQKFLDVLNTI
KNASDSDGQDLLFTNQLKEHPTDFSVEFLEQNSNEVQEVFAKAFAYYIEPQHRDVLQLYAPEAFNYMDKF
NEQEINLSLEELKDQRMLARYEKWEKIKQHYQHWSDSLSEEGRGLLKKLQIPIEPKKDDIIHSLSQEEKE
LLKRIQIDSSDFLSTEEKEFLKKLQIDIRDSLSEEEKELLNRIQVDSSNPLSEKEKEFLKKLKLDIQPYD
INQRLQDTGGLIDSPSINLDVRKQYKRDIQNIDALLHQSIGSTLYNKIYLYENMNINNLTATLGADLVDS
TDNTKINRGIFNEFKKNFKYSISSNYMIVDINERPALDNERLKWRIQLSPDTRAGYLENGKLILQRNIGL
EIKDVQIIKQSEKEYIRIDAKVVPKSKIDTKIQEAQLNINQEWNKALGLPKYTKLITFNVHNRYASNIVE
SAYLILNEWKNNIQSDLIKKVTNYLVDGNGRFVFTDITLPNIAEQYTHQDEIYEQVHSKGLYVPESRSIL
LHGPSKGVELRNDSEGFIHEFGHAVDDYAGYLLDKNQSDLVTNSKKFIDIFKEEGSNLTSYGRTNEAEFF
AEAFRLMHSTDHAERLKVQKNAPKTFQFINDQIKFIINS

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA vaccine LF pDNA encoding LF , pCLF4

4. Lef from B. anthracis str. A2012

Gene Name : Lef from B. anthracis str. A2012
Sequence Strain (Species/Organism) : Bacillus anthracis str. A2012
VO ID : VO_0010857
NCBI Gene ID : 1158731
NCBI Protein GI : 21392848
Locus Tag : BXA0172
Genbank Accession : AE011190
Protein Accession : NP_652928
3D structure: PDB ID : 1J7N
Taxonomy ID : 191218
Gene Starting Position : 149356
Gene Ending Position : 151785
Gene Strand (Orientation) : -
Protein Name : lethal factor
Protein pI : 5.69
Protein Weight : 89799.18
Protein Length : 809

DNA Sequence : Show Sequence

>NC_003980.1:149356-151785 Bacillus anthracis str. A2012 plasmid pXO1, complete sequence
CTTATGAGTTAATAATGAACTTAATCTGATCGTTAATAAATTGGAAAGTTTTCGGAGCATTTTTTTGAAC
TTTTAAACGTTCAGCATGGTCCGTAGAATGCATTAACCTAAAGGCTTCTGCAAAAAATTCCGCTTCATTT
GTTCTCCCATACGAAGTTAAATTACTCCCTTCTTCCTTAAAAATATCAATGAATTTTTTAGAATTTGTAA
CTAAATCAGATTGGTTCTTATCTAATAGATATCCAGCATAATCATCCACAGCATGTCCAAATTCGTGTAT
AAAACCCTCACTATCATTCCTTAATTCTACACCTTTTGAAGGTCCATGGAGTAATATAGAACGGGATTCT
GGAACATATAACCCTTTTGAATGAACTTGCTCATATATCTCATCTTGATGTGTATATTGTTCAGCTATAT
TAGGGAGAGTAATATCGGTAAAAACAAATCTTCCATTACCATCAACTAAGTAATTTGTTACCTTTTTTAT
AAGATCACTTTGAATATTATTTTTCCATTCATTCAATATTAAATAAGCACTTTCTACAATATTGGATGCA
TATCTATTATGCACGTTGAATGTAATAAGCTTTGTATATTTTGGTAACCCTAATGCTTTATTCCATTCCT
GATTTATATTTAACTGTGCTTCTTGAATTTTTGTATCTATTTTACTCTTTGGCACTACTTTCGCATCAAT
CCTTATATATTCTTTTTCGGATTGCTTAATTATTTGTACATCCTTTATTTCCAGACCGATGTTTCTTTGT
AATATAAGCTTTCCATTTTCTAAATATCCTGCTCGAGTATCTGGTGATAATTGGATTCTCCATTTCAAAC
GCTCATTATCTAATGCAGGCCTTTCATTTATATCAACAATCATATAGTTACTAGAAATACTATATTTGAA
ATTTTTTTTGAATTCATTGAAAATACCTCTATTAATTTTAGTATTATCAGTGGAATCAACTAAATCCGCA
CCTAGGGTTGCTGTAAGGTTATTGATATTCATATTTTCATACAAATAAATTTTATTGTACAAGGTACTTC
CAATGGATTGATGTAATAAAGCATCAATATTTTGAATATCCCTTTTATACTGCTTTCTTACATCAAGATT
AATTGACGGACTATCAATTAACCCTCCTGTATCTTGCAACCTTTGATTAATATCATATGGTTGAATATCA
AGTTTCAGCTTTTTTAAAAACTCTTTTTCTTTTTCAGATAAAGGATTACTACTATCCACCTGTATTCTAT
TTAAAAGCTCTTTTTCTTCTTCAGATAAAGAATCACGAATATCAATTTGTAGCTTTTTTAAAAACTCTTT
TTCCTCAGTAGATAAAAAATCACTACTATCAATTTGTATTCTTTTTAGAAGCTCTTTTTCTTCTTGAGAT
AAAGAATGAATTATGTCATCTTTCTTTGGCTCAATAGGAATCTGCAGCTTTTTTAAAAGTCCTCTTCCTT
CTTCAGATAAAGAATCGCTCCAGTGTTGATAGTGCTGTTTTATCTTTTCCCATTTTTCATATCTTGCCAG
CATCCGTTGATCTTTAAGTTCTTCCAAGGATAGATTTATTTCTTGTTCGTTAAATTTATCCATGTAATTA
AAAGCTTCCGGTGCATAAAGCTGTAAAACATCACGATGCTGTGGCTCGATATAATATGCAAAAGCTTTCG
CAAATACTTCTTGTACCTCATTGCTATTTTGTTCCAAGAATTCTACAGAAAAGTCTGTGGGATGTTCCTT
AAGCTGATTAGTAAATAAAAGATCTTGTCCATCTGAATCAGATGCATTTTTAATGGTATTTAATACATCT
AAAAATTTCTGATATGGTTGATTAATTTTACTTAAAATATCCCTTGATAATATCTTACCTATTTCATAAT
AAACGTTCAGTGCCTTTTCAGTATTTTCTACATAATCTTCCGAAGATTGGATTACAAGTACGGGTTCATA
TCCTTCTTTTGCATATACATAATGTTCATGTAATAAAGCATCTTTCCCATAAATGTCTTTTATTTTTTTC
TTATCTTCAGATAATGCTTCTAAAGATATATGTTTTGTAATATCACCATCCACAATATATATCTTTCCTC
CAATTGCTTTATACATCTCTAAAACATCAGATGGTACTTTCTCAAGTAGCTTTTCTGCTGCCTCTTTTTT
AACAGCTTCCTCCCCTTTTACTTCTATTTTTACAATGTGTTTCATGATTTCCTTTAAATGCTCTTCCTGT
GTTTTATTTCGTTCTTCATCTTTTCTCTTATTCTCATCTTTATTTTTCTCTTTCTCTTTTACGTGCATAC
CTACATCACCATGACCGCCCGCCCCCTGTACAAGGGGGATAAAGACGGGACCACTCAAAGTAATTGCTGT
TACTAAACATGACATACTAATTACTTTTATAAATTCTTTTTTTATATTCA

Protein Sequence : Show Sequence

>NP_652928.1 lethal factor (plasmid) [Bacillus anthracis str. A2012]
MNIKKEFIKVISMSCLVTAITLSGPVFIPLVQGAGGHGDVGMHVKEKEKNKDENKRKDEERNKTQEEHLK
EIMKHIVKIEVKGEEAVKKEAAEKLLEKVPSDVLEMYKAIGGKIYIVDGDITKHISLEALSEDKKKIKDI
YGKDALLHEHYVYAKEGYEPVLVIQSSEDYVENTEKALNVYYEIGKILSRDILSKINQPYQKFLDVLNTI
KNASDSDGQDLLFTNQLKEHPTDFSVEFLEQNSNEVQEVFAKAFAYYIEPQHRDVLQLYAPEAFNYMDKF
NEQEINLSLEELKDQRMLARYEKWEKIKQHYQHWSDSLSEEGRGLLKKLQIPIEPKKDDIIHSLSQEEKE
LLKRIQIDSSDFLSTEEKEFLKKLQIDIRDSLSEEEKELLNRIQVDSSNPLSEKEKEFLKKLKLDIQPYD
INQRLQDTGGLIDSPSINLDVRKQYKRDIQNIDALLHQSIGSTLYNKIYLYENMNINNLTATLGADLVDS
TDNTKINRGIFNEFKKNFKYSISSNYMIVDINERPALDNERLKWRIQLSPDTRAGYLENGKLILQRNIGL
EIKDVQIIKQSEKEYIRIDAKVVPKSKIDTKIQEAQLNINQEWNKALGLPKYTKLITFNVHNRYASNIVE
SAYLILNEWKNNIQSDLIKKVTNYLVDGNGRFVFTDITLPNIAEQYTHQDEIYEQVHSKGLYVPESRSIL
LHGPSKGVELRNDSEGFIHEFGHAVDDYAGYLLDKNQSDLVTNSKKFIDIFKEEGSNLTSYGRTNEAEFF
AEAFRLMHSTDHAERLKVQKNAPKTFQFINDQIKFIINS

Molecule Role : Protective antigen
Related Vaccine(s): DNA vaccine encoding PA83 and LF

5. LF

Gene Name : LF
Sequence Strain (Species/Organism) : synthetic construct
NCBI Protein GI : 40646955
Other Database IDs : CDD:285037
CDD:294115
CDD:238144
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : lethal factor domains I-III
Protein pI : 5.34
Protein Weight : 64562.77
Protein Length : 645
Protein Note : Homo sapiens tissue plasminogen activator signal peptide

Protein Sequence : Show Sequence

>AAR88322.1 lethal factor domains I-III [synthetic construct]
MDAMKRGLCCVLLLCGAVFVSPSAGGHGDVGMHVKEKEKNKDENKRKDEERNKTQEEHLKEIMKHIVKIE
VKGEEAVKKEAAEKLLEKVPSDVLEMYKAIGGKIYIVDGDITKHISLEALSEDKKKIKDIYGKDALLHEH
YVYAKEGYEPVLVIQSSEDYVENTEKALNVYYEIGKILSRDILSKINQPYQKFLDVLNTIKNASDSDGQD
LLFTNQLKEHPTDFSVEFLEQNSNEVQEVFAKAFAYYIEPQHRDVLQLYAPEAFNYMDKFNEQEINLSLE
ELKDQRMLSRYEKWEKIKQHYQHWSDSLSEEGRGLLKKLQIPIEPKKDDIIHSLSQEEKELLKRIQIDSS
DFLSTEEKEFLKKLQIDIRDSLSEEEKELLNRIQVDSSNPLSEKEKEFLKKLKLDIQPYDINQRLQDTGG
LIDSPSINLDVRKQYKRDIQNIDALLHQSIGSTLYNKIYLYENMNINNLTATLGADLVDSTDNTKINRGI
FNEFKKNFKYSISSNYMIVDINERPALDNERLKWRIQLSPDTRAGYLENGKLILQRNIGLEIKDVQIIKQ
SEKEYIRIDAKVV

Molecule Role : Protective antigen

6. mntA

Gene Name : mntA
Sequence Strain (Species/Organism) : Bacillus anthracis str. A0248
NCBI Gene ID : 7848141
NCBI Protein GI : 229600559
Locus Tag : BAA_3238
Genbank Accession : CP001598
Protein Accession : YP_002867390
Taxonomy ID : 592021
Gene Starting Position : 2936644
Gene Ending Position : 2937579
Gene Strand (Orientation) : -
Protein Name : manganese ABC transporter, manganese-binding protein
Protein pI : 5.78
Protein Weight : 32608.66
Protein Length : 311
Protein Note : identified by match to protein family HMM PF01297

DNA Sequence : Show Sequence

>gi|229599883:2936644-2937579 Bacillus anthracis str. A0248, complete genome
TTTACTTTTGTAACCCATTAATAATGGTATCTATATTCCATTTCATCATTTTTAAATACGTATCTCCATC
CTCACCTGATTTACCTAGTGAATCTGTAAAAATTGTACCTGCAATTGGCACGTTTGTTTCTTTTGAAACA
GTTTCCATGCTGCGTCGATCTACACTAGTTTCTACAAATAGAGCCGGAACTTTATTTGTTTGAATTACAC
TTACAACATCTCGAATTTGATCTGGTGTACCTTGATTTTCTGAGTTAATTTCCCAAATGTATCCCGTTTT
AATATCATATGCTTTTCCAAAGTATTTAAAAGCACCTTCACTAGAGATTAAGAACCGTTTCTCCTCAGGG
ATTTGATGAATTCTGTTTACTGTCTCATCATGTAACTTTTGAAGTTCTGCTACATAGTTGTCAGCATTTT
TAGTATAGAACTCTTTATTTTTAGGGTCTTCTTTAATTAATGCCTTTTTCACATTTTCAGCATATAAAAT
ACCATTTTTAATGTTCATCCATGCATGCGGGTCTGGTTCTTTTTCTAATCCTTTTGTCTCTAAATAAATA
GCTTCTACGCCTTCACTTACTTTATAAACCGGTGCATCTTTCTCTGATTTATTTGCCGTTTTTAATAGCT
TTTTAAACCACGCTCCACCTTCTTCTAGGTTCAATCCATTGTAAAGAACCATATCTGCATCTGTCATTTT
CATAACATCTTTTGGTAGTGGATCATATTCATGCGGGTTAGCTCCAATTGGAACAAGACTATGAATCTCA
ACTTTCTCTCCGCCAATTTGCTTCACCATATCATATATAATGGAGTATGTAGTTACAACTTTTAATTTTC
CACTGCCCTCTTCTTTTCCATTTGTGTTACTAGAACACGCTGTTAATGCAAATACGAAAATACAAAGTAT
CGATAATACAACATTTTTAAATTTCA

Protein Sequence : Show Sequence

>gi|229600559|ref|YP_002867390.1| manganese ABC transporter, manganese-binding protein [Bacillus anthracis str. A0248]
MKFKNVVLSILCIFVFALTACSSNTNGKEEGSGKLKVVTTYSIIYDMVKQIGGEKVEIHSLVPIGANPHE
YDPLPKDVMKMTDADMVLYNGLNLEEGGAWFKKLLKTANKSEKDAPVYKVSEGVEAIYLETKGLEKEPDP
HAWMNIKNGILYAENVKKALIKEDPKNKEFYTKNADNYVAELQKLHDETVNRIHQIPEEKRFLISSEGAF
KYFGKAYDIKTGYIWEINSENQGTPDQIRDVVSVIQTNKVPALFVETSVDRRSMETVSKETNVPIAGTIF
TDSLGKSGEDGDTYLKMMKWNIDTIINGLQK

Molecule Role : Virmugen
Molecule Role Annotation : An mntA deletion mutant, generated by allelic replacement, in Bacillus anthracis was attenuated and provided protection for guinea pigs against lethal spore dose (60 LD50) of the virulent B. anthracis Vollum strain (Gat et al., 2005).
Related Vaccine(s): Bacillus anthracis mntA deletion mutant vaccine

7. NEAT

Gene Name : NEAT
Sequence Strain (Species/Organism) : Bacillus anthracis
NCBI Protein GI : WP_080489610
Other Database IDs : CDD:309946
CDD:132993
CDD:227223
CDD:275380
CDD:275378
CDD:306825
Taxonomy ID : 1392
Gene Strand (Orientation) : ?
Protein Name : NEAT domain-containing leucine-rich repeat protein
Protein pI : 5.8
Protein Weight : 81601.3
Protein Length : 872
Protein Note : Iron Transport-associated domain; pfam05031

Protein Sequence : Show Sequence

>WP_080489610.1 NEAT domain-containing leucine-rich repeat protein [Bacillus anthracis]
MIGDIILKKIYMKALVVATTLAIPFAAYSTPALAAIKIEANQSVAASDRTYDTEIKIYKDQKDEPSMVSQ
YIKDPKVAIVAGKKIVTVTMQDSDYFQYLRIEDRNQPGVFHDVKVLSEDKRKNGTKVIQFEIGEFEKKHN
MQMHILIPAIGYDHKYQVQFEIKDPTVGDKETEKPDDNSNSGNTETDKPVDNQNMITDNKLRELVNKKVF
NRKDVNTPITKEELLQVKNLFLNTNEILDYSALKYMPNLKSLTVANAKIKDPSFFANLKQLNHLALRGNE
FSDVTPLVKMDHLDSLDLSNNKITNVAPLIEMKNVKSLYLSGNQIEDVTALAKMEQLDYLNLANNKITNV
APLSALKNVTYLTLAGNQIEDIKPLYSLPLTDLVLTRNKVKDLSGIEQMKQLEELWIGKNEIKDVTPLSK
MTQLKQLHLPNNELKDITPLSSLVNLQKLDLEANYISDLTPASNLKKLVFLSFVANEIRDVRPVIELSKT
AYINVQNQKVFLEETEVNKEVKVPIYEKDGKISTKIRLKDEGGTYSNDAVKWSTPGEKVYEFGVKDPFAD
TGIFFTGSVIQNVVESKADNISKEDNTSKEDAKVEVVEFKDVPKGHWSEEAIHYLAKENIFKGYGNGQFG
FGDSITRGQVASLVQRYLKLENKVEQKERFTDTKGHMFEQDIATVAQAGIMQGDGTGEFRPDGVLTRYEM
SVVLYKVFQLKEDGNNKVNFKDVPTGHWAEGYVKALVDNNISKGDGKERFLGDDFVTREQYAQFLYNAIT
K

Molecule Role : Protective antigen
Molecule Role Annotation : (Balderas et al., 2016)

8. PA

Gene Name : PA
Sequence Strain (Species/Organism) : Bacillus anthracis str. H9401
NCBI Gene ID : 12796252
NCBI Protein GI : 386733821
Locus Tag : H9401_5741
Genbank Accession : CP002093
Protein Accession : YP_006204206
Taxonomy ID : 768494
Plasmid No : BAP2
Gene Starting Position : 46551
Gene Ending Position : 46985
Gene Strand (Orientation) : -
Protein Name : Phosphoesterase PA-phosphatase related protein
Protein pI : 9.43
Protein Weight : 14561.26
Protein Length : 144

DNA Sequence : Show Sequence

>NC_017727.1:46551-46985 Bacillus anthracis str. H9401 plasmid BAP2, complete sequence
ATTATAATTCCTTCGATTTACTTTTTAATGGTAAAACATATTGCTCTACTCTTTCATAAAGATTAAGTAT
TTGTGTAATAAATTTAATTTTTGGTACTATCCAATATGAAAATAAAGCTGAAATACTCCCTATTAAGGCG
CCTGTTGCAACATCAAAAGGATAATGAACCCCGACCCAAATACGAGAGATACCCACACAGAATGCAATTA
CAAGCAATAACAACCCAGCCTTTTTATGAACGAGCTTAACTGAAACACAAATTGAAAAAAATAGAATCGT
GTGATCACTAGGAAATGAATTATCCACTACATGATCAGCGAGTTTATTAACATCAGGTAATACTGCAAAT
GGTTGATAATTCAAATGCAATTTCCCTAGTAGCTTTCCAATTATCTCAGCAGTCATGAAAGCAAACACCG
CTTGTATAATCATCA

Protein Sequence : Show Sequence

>YP_006204206.1 Phosphoesterase PA-phosphatase related protein (plasmid) [Bacillus anthracis str. H9401]
MMIIQAVFAFMTAEIIGKLLGKLHLNYQPFAVLPDVNKLADHVVDNSFPSDHTILFFSICVSVKLVHKKA
GLLLLVIAFCVGISRIWVGVHYPFDVATGALIGSISALFSYWIVPKIKFITQILNLYERVEQYVLPLKSK
SKEL

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA Vaccine expressing PA , Typhi strain Ty21a-PA (Bacillus anthracis)

9. PA63

Gene Name : PA63
Sequence Strain (Species/Organism) : synthetic construct derived from Bacillus anthracis
NCBI Protein GI : 193873769
Other Database IDs : CDD:281492
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : PA63
Protein pI : 5.74
Protein Weight : 60404.36
Protein Length : 613
Protein Note : derived from Bacillus anthracis

Protein Sequence : Show Sequence

>ACF23542.1 PA63 [synthetic construct]
MPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTG
RIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVHGNAEVHA
SFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTT
SLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLD
TDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARISFNGKDLNLVERRIAAVNPSDPLETTKPGM
TLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLNAKMNILI
RDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKEVINDRYD
MLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKG
YEIG

Molecule Role : Protective antigen

10. PA63-LAMP1

Gene Name : PA63-LAMP1
Sequence Strain (Species/Organism) : synthetic construct derived from Bacillus anthracis
NCBI Protein GI : 193873761
Other Database IDs : CDD:281492
CDD:213052
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : PA63-LAMP1
Protein pI : 6.22
Protein Weight : 64720.28
Protein Length : 659
Protein Note : Clostridial binary toxin B/anthrax toxin PA; pfam03495

Protein Sequence : Show Sequence

>ACF23538.1 PA63-LAMP1 [synthetic construct]
MPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTG
RIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVHGNAEVHA
SFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTT
SLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLD
TDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPLETTKPDM
TLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLNAKMNILI
RDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKISSGYIVEIEDTEGLKEVINDRYD
MLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKG
YEIGGLNNMLIPIAVGGALAGLVLIVLIAYLIGRKRSHAGYQTI

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA vaccine pLAMP1-PA63

11. PA83

Gene Name : PA83
Sequence Strain (Species/Organism) : synthetic construct
NCBI Protein GI : 40646953
Protein Accession : AAR88321.1
Other Database IDs : CDD:284994
CDD:281492
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : protective antigen PA83
Protein pI : 5.32
Protein Weight : 80526.18
Protein Length : 822
Protein Note : Homo sapiens tissue plasminogen activator signal peptide

Protein Sequence : Show Sequence

>AAR88321.1 protective antigen PA83 [synthetic construct]
MDAMKRGLCCVLLLCGAVFVSPSEVKQENRLLNESESSSQGLLGYYFSDLNFQAPMVVTSSTTGDLSIPS
SELENIPSENQYFQSAIWSGFIKVKKSDEYTFATSADNHVTMWVDDQEVINKASNSNKIRLEKGRLYQIK
IQYQRENPTEKGLDFKLYWTDSQNKKEVISSDNLQLPELKQKSSNTSAGPTVPDRDNDGIPDSLEVEGYT
VDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEARHPLVAAYPIVH
VDMENIILSKNEDQSTQNTDSETRTISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVSAGFSNSNSSTVA
IDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTTSLVLGKNQTLATIKAKENQLSQ
ILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLDTDQVYGNIATYNFENGRVRVDT
GSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPLETTKPDMTLKEALKIAFGFNEPNGNLQYQ
GKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLNAKMNILIRDKRFHYDRNNIAVGADESVVK
EAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKEVINDRYDMLNISSLRQDGKTFIDFKKYND
KLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKGYEIG

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA vaccine PA83 furin , B. anthracis DNA vaccine pDNA encoding PA

12. pagA

Gene Name : pagA
Sequence Strain (Species/Organism) : Bacillus anthracis
VO ID : VO_0011029
NCBI Protein GI : 9280533
3D structure: PDB ID : 1ACC
Other Database IDs : ATCC:14185
CDD:311565
CDD:281492
Taxonomy ID : 1392
Gene Strand (Orientation) : ?
Protein Name : protective antigen
Protein pI : 6.09
Protein Weight : 81760.71
Protein Length : 838
Protein Note : PA14 domain; pfam07691

Protein Sequence : Show Sequence

>AAF86457.1 protective antigen (plasmid) [Bacillus anthracis]
MKKRKVLIPLMALSTILVSSTGNLEVIQAEVKQENRLLNESESSSQGLLGYYFSDLNFQAPMVVTSSTTG
DLSIPSSELENIPSENQYFQSAIWSGFIKVKKSDEYTFATSADNHVTMWVDDQEVINKASNSNKIRLEKG
RLYQIKIQYQRENPTEKGLDFKLYWTDSQNKKEVISSDNLQLPELKQKSSNSRKKRSTSAGPTVPDRDND
GIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEA
RHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVS
AGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTTSLVLGKNQTL
ATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLDTDQVYGNIAT
YNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPLETTKPDMTLKEALKIAF
GFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNVTNIYTVLDKIKLNAKMNILIRDKRFHYDRN
NIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKEVINDRYDMLNISSLRQD
GKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKGYEIG

Molecule Role : Protective antigen
Molecule Role Annotation : Study described the construction and evaluation of an adenovirus vaccine expressing domain 4 of Bacillus anthracis protective antigen, Ad.D4. Ad.D4 elicited antibodies to protective antigen 14 days after a single intramuscular injection, which were further increased upon boosting. Furthermore, two doses of Ad.D4 4 weeks apart were sufficient to protect 67% of mice from toxin challenge (McConnell et al., 2006).
Related Vaccine(s): B. anthracis DNA Vaccine expressing PA , B. anthracis DNA vaccine PA , B. anthracis DNA vaccine pCPA , B. anthracis DNA vaccine pIMS-120 encoding PA , pCMV/ER-PA83 , rLAG- PA-DCpep

13. PagA from B. anthracis str. 'Ames Ancestor'

Gene Name : PagA from B. anthracis str. 'Ames Ancestor'
Sequence Strain (Species/Organism) : Bacillus anthracis str. 'Ames Ancestor'
VO ID : VO_0010872
NCBI Gene ID : 2820165
NCBI Protein GI : 47566476
Locus Tag : GBAA_pXO1_0164
Genbank Accession : AE017336
Protein Accession : YP_016495
3D structure: PDB ID : 1ACC
Taxonomy ID : 261594
Plasmid No : pXO1
Gene Starting Position : 143778
Gene Ending Position : 146072
Gene Strand (Orientation) : +
Protein Name : protective antigen
Protein pI : 6.09
Protein Weight : 81542.47
Protein Length : 764
Protein Note : similar to SP:P13423; identified by sequence similarity; putative
Protein Annotation : Protective antigen (PA) is the dominant antigen in both natural and vaccine-induced immunity to anthrax infection and consists of four distinct and functionally independent domains. Domain 1 is divided into domains 1a (consisting of amino acids 1-167) and 1b (consisting of aa 168-258). Domain 2 comprises aa 259-487 and domain 3 aa 488-595. After the antigen binds to the host cell receptor via the binding site of domain 4, the N-terminal amino acids (1-167, i.e. domain 1a) of domain 1, which contain a furin protease cleavage site, are cleaved off, exposing the LF or EF binding site located in domain 1b and the adjacent domain 3. Domains 2 and 3 then form part of a heptameric pore on the cell surface, the LF or EF binds to its receptor, and the whole toxin complex undergoes receptor-mediated endocytosis into the cell. After acidification of the endosome, the toxin is translocated into the cell cytosol where it exerts its cytotoxic effect. Therefore, inhibition of the binding and entry of the toxin complex, particularly lethal toxin, into the host cell is clearly important for the prevention of infection (Flick-Smith et al., 2002).

DNA Sequence : Show Sequence

>NC_007322.2:143778-146072 Bacillus anthracis str. 'Ames Ancestor' plasmid pXO1, complete sequence
TATGAAAAAACGAAAAGTGTTAATACCATTAATGGCATTGTCTACGATATTAGTTTCAAGCACAGGTAAT
TTAGAGGTGATTCAGGCAGAAGTTAAACAGGAGAACCGGTTATTAAATGAATCAGAATCAAGTTCCCAGG
GGTTACTAGGATACTATTTTAGTGATTTGAATTTTCAAGCACCCATGGTGGTTACCTCTTCTACTACAGG
GGATTTATCTATTCCTAGTTCTGAGTTAGAAAATATTCCATCGGAAAACCAATATTTTCAATCTGCTATT
TGGTCAGGATTTATCAAAGTTAAGAAGAGTGATGAATATACATTTGCTACTTCCGCTGATAATCATGTAA
CAATGTGGGTAGATGACCAAGAAGTGATTAATAAAGCTTCTAATTCTAACAAAATCAGATTAGAAAAAGG
AAGATTATATCAAATAAAAATTCAATATCAACGAGAAAATCCTACTGAAAAAGGATTGGATTTCAAGTTG
TACTGGACCGATTCTCAAAATAAAAAAGAAGTGATTTCTAGTGATAACTTACAATTGCCAGAATTAAAAC
AAAAATCTTCGAACTCAAGAAAAAAGCGAAGTACAAGTGCTGGACCTACGGTTCCAGACCGTGACAATGA
TGGAATCCCTGATTCATTAGAGGTAGAAGGATATACGGTTGATGTCAAAAATAAAAGAACTTTTCTTTCA
CCATGGATTTCTAATATTCATGAAAAGAAAGGATTAACCAAATATAAATCATCTCCTGAAAAATGGAGCA
CGGCTTCTGATCCGTACAGTGATTTCGAAAAGGTTACAGGACGGATTGATAAGAATGTATCACCAGAGGC
AAGACACCCCCTTGTGGCAGCTTATCCGATTGTACATGTAGATATGGAGAATATTATTCTCTCAAAAAAT
GAGGATCAATCCACACAGAATACTGATAGTCAAACGAGAACAATAAGTAAAAATACTTCTACAAGTAGGA
CACATACTAGTGAAGTACATGGAAATGCAGAAGTGCATGCGTCGTTCTTTGATATTGGTGGGAGTGTATC
TGCAGGATTTAGTAATTCGAATTCAAGTACGGTCGCAATTGATCATTCACTATCTCTAGCAGGGGAAAGA
ACTTGGGCTGAAACAATGGGTTTAAATACCGCTGATACAGCAAGATTAAATGCCAATATTAGATATGTAA
ATACTGGGACGGCTCCAATCTACAACGTGTTACCAACGACTTCGTTAGTGTTAGGAAAAAATCAAACACT
CGCGACAATTAAAGCTAAGGAAAACCAATTAAGTCAAATACTTGCACCTAATAATTATTATCCTTCTAAA
AACTTGGCGCCAATCGCATTAAATGCACAAGACGATTTCAGTTCTACTCCAATTACAATGAATTACAATC
AATTTCTTGAGTTAGAAAAAACGAAACAATTAAGATTAGATACGGATCAAGTATATGGGAATATAGCAAC
ATACAATTTTGAAAATGGAAGAGTGAGGGTGGATACAGGCTCGAACTGGAGTGAAGTGTTACCGCAAATT
CAAGAAACAACTGCACGTATCATTTTTAATGGAAAAGATTTAAATCTGGTAGAAAGGCGGATAGCGGCGG
TTAATCCTAGTGATCCATTAGAAACGACTAAACCGGATATGACATTAAAAGAAGCCCTTAAAATAGCATT
TGGATTTAACGAACCGAATGGAAACTTACAATATCAAGGGAAAGACATAACCGAATTTGATTTTAATTTC
GATCAACAAACATCTCAAAATATCAAGAATCAGTTAGCGGAATTAAACGCAACTAACATATATACTGTAT
TAGATAAAATCAAATTAAATGCAAAAATGAATATTTTAATAAGAGATAAACGTTTTCATTATGATAGAAA
TAACATAGCAGTTGGGGCGGATGAGTCAGTAGTTAAGGAGGCTCATAGAGAAGTAATTAATTCGTCAACA
GAGGGATTATTGTTAAATATTGATAAGGATATAAGAAAAATATTATCAGGTTATATTGTAGAAATTGAAG
ATACTGAAGGGCTTAAAGAAGTTATAAATGACAGATATGATATGTTGAATATTTCTAGTTTACGGCAAGA
TGGAAAAACATTTATAGATTTTAAAAAATATAATGATAAATTACCGTTATATATAAGTAATCCCAATTAT
AAGGTAAATGTATATGCTGTTACTAAAGAAAACACTATTATTAATCCTAGTGAGAATGGGGATACTAGTA
CCAACGGGATCAAGAAAATTTTAATCTTTTCTAAAAAAGGCTATGAGATAGGATA

Protein Sequence : Show Sequence

>YP_016495.2 protective antigen (plasmid) [Bacillus anthracis str. 'Ames Ancestor']
MKKRKVLIPLMALSTILVSSTGNLEVIQAEVKQENRLLNESESSSQGLLGYYFSDLNFQAPMVVTSSTTG
DLSIPSSELENIPSENQYFQSAIWSGFIKVKKSDEYTFATSADNHVTMWVDDQEVINKASNSNKIRLEKG
RLYQIKIQYQRENPTEKGLDFKLYWTDSQNKKEVISSDNLQLPELKQKSSNSRKKRSTSAGPTVPDRDND
GIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEA
RHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVS
AGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTTSLVLGKNQTL
ATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLDTDQVYGNIAT
YNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPLETTKPDMTLKEALKIAF
GFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLNAKMNILIRDKRFHYDRN
NIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKEVINDRYDMLNISSLRQD
GKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKGYEIG

Molecule Role : Protective antigen
Related Vaccine(s): Anthrax Spore Vaccine , DAAV using PA and PGA , DNA vaccine encoding PA (PA63) , DNA vaccine encoding PA83 and LF , pCLF4 , Recombinant PA domain 4

14. S-layer protein

Gene Name : S-layer protein
Sequence Strain (Species/Organism) : Bacillus anthracis
NCBI Protein GI : WP_047401514
Other Database IDs : CDD:278807
CDD:224259
CDD:302596
CDD:302792
Taxonomy ID : 1392
Protein Name : S-layer protein
Protein pI : 9.61
Protein Weight : 73443.67
Protein Length : 711
Protein Note : S-layer homology domain; pfam00395

Protein Sequence : Show Sequence

>WP_047401514.1 S-layer protein [Bacillus anthracis]
MKAITSFTLIGSILFGGNINSTQAESDLKKIVPDQMVPAFLDVPSNHWANVAINNLLKKEIIVGYGNNKF
GLGDSVTREQVAAVIYRIKFPEKEVEAEAQNPYRDVNKSSTMFLDEILTLTKMGIFSGDEKGNFRPKSPI
SRAEMTQVIKNAFNIPVVGKHNFRDVAKDYWANDAISALQSNQLVSGTGNGLFEPYKSVTREEYAQFIFN
VLNFEDLVNKEKQLDELNKRRDVINQKIDEFDKLNSQRKDLERMLEELNQKLSQLKQQSPQLQDLKNKLK
ESQSRLLELNKKDSNRLELNSEIKKLNDRKAELLSLIMELIKQQSEFDKKIKNEKDDLNKKREDLINRIA
ESKELAKKKAELNTKLVELFKVQEALNKKSGQYLYYINKLDNELRELADKYKNSDNKISRLKNHIGEYNK
QLEKIENELEECNKKIDNTKKQLAEFDKSNKKQQELESELVQLNKKIDELGKRHKHRQELEASQKKALDE
AKEINKKLAEKDPERQHINDKLKKLNNDWRLTNNELNRLNSELQKVNTKLLGYSQEHQYYNALNKERTEL
SKKVDEIYQKQSEIDKNYIELNRQLDELSNPKPNEQVQLLKNQINELNSSINQILNHRKNEINNITQRKD
HIIDQLNQFPKLRIPNS

Molecule Role : Protective antigen
Molecule Role Annotation : (Mikshis et al., 2014)

15. TPA-PA63

Gene Name : TPA-PA63
Sequence Strain (Species/Organism) : synthetic construct derived from Bacillus anthracis
NCBI Protein GI : 193873765
Other Database IDs : CDD:281492
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : TPA-PA63
Protein pI : 5.74
Protein Weight : 62856.12
Protein Length : 640
Protein Note : Clostridial binary toxin B/anthrax toxin PA; pfam03495

Protein Sequence : Show Sequence

>ACF23540.1 TPA-PA63 [synthetic construct]
MDAMKRGLCCVLLLCGAVFVSPSMPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTK
YKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRT
ISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTA
RLNANIRYVNTGTAPIYNVLPTTSLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFS
STPITMNYNQFLELEKTKQLRLDTDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDL
NLVERRIAAVNPSDPLETTKPDMTLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAE
LNATNIYTVLDKIKLNAKMNILIRDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKI
LSGYIVEIEDTEGLKEVINDRYDMLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTII
NPSENGDTSTNGIKKILIFSKKGYEIG

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA vaccine pTPA-PA63

16. TPA-PA63-LAMP1

Gene Name : TPA-PA63-LAMP1
Sequence Strain (Species/Organism) : synthetic construct derived from Bacillus anthracis
NCBI Protein GI : 193873763
Other Database IDs : CDD:281492
CDD:213052
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : TPA-PA63-LAMP1
Protein pI : 6.22
Protein Weight : 67115.01
Protein Length : 686
Protein Note : Clostridial binary toxin B/anthrax toxin PA; pfam03495

Protein Sequence : Show Sequence

>ACF23539.1 TPA-PA63-LAMP1 [synthetic construct]
MDAMKRGLCCVLLLCGAVFVSPSMPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTK
YKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRT
ISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTA
RLDANIRYVNTGTAPIYNVLPTTSLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFS
STPITMNYNQFLELEKTKQLRLDTDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDL
NLVERRIAAVNPSDPLETTKPDMTLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAE
LNATNIYTVLDKIKLNAKMNILIRDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKI
LSGYIVEIEDTEGLKEVINDRYDMLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTII
NPSENGDTSTNGIKKILIFSKKGYEIGGLNNMLIPIAVGGALAGLVLIVLIAYLIGRKRSHAGYQTI

Molecule Role : Protective antigen
Related Vaccine(s): B. anthracis DNA vaccine pTPA-P

17. UQ-PA63

Gene Name : UQ-PA63
Sequence Strain (Species/Organism) : synthetic construct derived from Bacillus anthracis
NCBI Protein GI : 193873767
Other Database IDs : CDD:176398
CDD:214563
CDD:281492
Taxonomy ID : 32630
Gene Strand (Orientation) : ?
Protein Name : UQ-PA63
Protein pI : 5.91
Protein Weight : 68767.41
Protein Length : 694
Protein Note : Ubiquitin; cd01803

Protein Sequence : Show Sequence

>ACF23541.1 UQ-PA63 [synthetic construct]
MQIFVKTLTGKTTTLGVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLV
LRLRGAAMPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYS
DFEKVTGRIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVH
GNAEVHASFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPI
YNVLPTTSLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEK
TKQLRLDTDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPL
ETTKPDMTLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLN
AKMNILIRDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKE
VINDRYDMLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKI
LIFSKKGYEIG

Molecule Role : Protective antigen

III. Vaccine Related Host Genes

1. Ifng (Interferon gamma)

Gene Name : Ifng (Interferon gamma)
Sequence Strain (Species/Organism) : Mouse
NCBI Gene ID : 15978
NCBI Protein GI : 33468859
Genbank Accession : NM_008337
Protein Accession : NP_032363.1
Other Database IDs : MGI:107656; UniProt: P01580
Taxonomy ID : 10090
Gene Strand (Orientation) : ?

DNA Sequence : Show Sequence

>gi|145966741|ref|NM_008337.3| Mus musculus interferon gamma (Ifng), mRNA
ATAGCTGCCATCGGCTGACCTAGAGAAGACACATCAGCTGATCCTTTGGACCCTCTGACTTGAGACAGAA
GTTCTGGGCTTCTCCTCCTGCGGCCTAGCTCTGAGACAATGAACGCTACACACTGCATCTTGGCTTTGCA
GCTCTTCCTCATGGCTGTTTCTGGCTGTTACTGCCACGGCACAGTCATTGAAAGCCTAGAAAGTCTGAAT
AACTATTTTAACTCAAGTGGCATAGATGTGGAAGAAAAGAGTCTCTTCTTGGATATCTGGAGGAACTGGC
AAAAGGATGGTGACATGAAAATCCTGCAGAGCCAGATTATCTCTTTCTACCTCAGACTCTTTGAAGTCTT
GAAAGACAATCAGGCCATCAGCAACAACATAAGCGTCATTGAATCACACCTGATTACTACCTTCTTCAGC
AACAGCAAGGCGAAAAAGGATGCATTCATGAGTATTGCCAAGTTTGAGGTCAACAACCCACAGGTCCAGC
GCCAAGCATTCAATGAGCTCATCCGAGTGGTCCACCAGCTGTTGCCGGAATCCAGCCTCAGGAAGCGGAA
AAGGAGTCGCTGCTGATTCGGGGTGGGGAAGAGATTGTCCCAATAAGAATAATTCTGCCAGCACTATTTG
AATTTTTAAATCTAAACCTATTTATTAATATTTAAAACTATTTATATGGAGAATCTATTTTAGATGCATC
AACCAAAGAAGTATTTATAGTAACAACTTATATGTGATAAGAGTGAATTCCTATTAATATATGTGTTATT
TATAATTTCTGTCTCCTCAACTATTTCTCTTTGACCAATTAATTATTCTTTCTGACTAATTAGCCAAGAC
TGTGATTGCGGGGTTGTATCTGGGGGTGGGGGACAGCCAAGCGGCTGACTGAACTCAGATTGTAGCTTGT
ACCTTTACTTCACTGACCAATAAGAAACATTCAGAGCTGCAGTGACCCCGGGAGGTGCTGCTGATGGGAG
GAGATGTCTACACTCCGGGCCAGCGCTTTAACAGCAGGCCAGACAGCACTCGAATGTGTCAGGTAGTAAC
AGGCTGTCCCTGAAAGAAAGCAGTGTCTCAAGAGACTTGACACCTGGTGCTTCCCTATACAGCTGAAAAC
TGTGACTACACCCGAATGACAAATAACTCGCTCATTTATAGTTTATCACTGTCTAATTGCATATGAATAA
AGTATACCTTTGCAACC

Protein Sequence : Show Sequence

>gi|33468859|ref|NP_032363.1| interferon gamma [Mus musculus]
MNATHCILALQLFLMAVSGCYCHGTVIESLESLNNYFNSSGIDVEEKSLFLDIWRNWQKDGDMKILQSQI
ISFYLRLFEVLKDNQAISNNISVIESHLITTFFSNSKAKKDAFMSIAKFEVNNPQVQRQAFNELIRVVHQ
LLPESSLRKRKRSRC

Molecule Role Annotation : IFN-gamma plays a critical role in Th1 type immune response. It is important for protection against infections by various viruses and intracellular bacteria.
Additional Molecule Role : Vaximmutor
Additional Molecule Role Annotation : The experimental data demonstrated that three time vaccinations with BCG in BALB/c mice induced strong TB Ag-specific IFN-gamma immune responses in splenocytes (Wang et al., 2009).
Related Vaccine(s): pSecTag-PA83

2. Ighg1

Gene Name : Ighg1
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16017
Genbank Accession : AC160982
Molecule Role : Vaximmutor
Related Vaccine(s): pSecTag-PA83

3. Il4 (interleukin 4)

Gene Name : Il4 (interleukin 4)
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16189
NCBI Protein GI : 10946584
Locus Tag : RP23-188H3.4
Genbank Accession : NM_021283.1
Protein Accession : NM_021283.1
Taxonomy ID : 10090
Gene Strand (Orientation) : ?
Protein Name : IL-4, Interleukin 4

DNA Sequence : Show Sequence

>gi|10946583|ref|NM_021283.1| Mus musculus interleukin 4 (Il4), mRNA
GGATCCCCGGGCAGAGCTGGGGGGGGATTTGTTAGCATCTCTTGATAAACTTAATTGTCTCTCGTCACTG
ACGGCACAGAGCTATTGATGGGTCTCAACCCCCAGCTAGTTGTCATCCTGCTCTTCTTTCTCGAATGTAC
CAGGAGCCATATCCACGGATGCGACAAAAATCACTTGAGAGAGATCATCGGCATTTTGAACGAGGTCACA
GGAGAAGGGACGCCATGCACGGAGATGGATGTGCCAAACGTCCTCACAGCAACGAAGAACACCACAGAGA
GTGAGCTCGTCTGTAGGGCTTCCAAGGTGCTTCGCATATTTTATTTAAAACATGGGAAAACTCCATGCTT
GAAGAAGAACTCTAGTGTTCTCATGGAGCTGCAGAGACTCTTTCGGGCTTTTCGATGCCTGGATTCATCG
ATAAGCTGCACCATGAATGAGTCCAAGTCCACATCACTGAAAGACTTCCTGGAAAGCCTAAAGAGCATCA
TGCAAATGGATTACTCGTAGTACTGAGCCACCATGCTTTAACTTATGAATTTTTAATGGTTTTATTTTAA
TATTTATATATTTATAATTCATAAAATAAAATATTTGTATAATGT

Protein Sequence : Show Sequence

>gi|10946584|ref|NP_067258.1| interleukin 4 [Mus musculus]
MGLNPQLVVILLFFLECTRSHIHGCDKNHLREIIGILNEVTGEGTPCTEMDVPNVLTATKNTTESELVCR
ASKVLRIFYLKHGKTPCLKKNSSVLMELQRLFRAFRCLDSSISCTMNESKSTSLKDFLESLKSIMQMDYS

Molecule Role : Vaximmutor
Molecule Role Annotation : IL-4 plays an important role in Th2 immune response.
Related Vaccine(s): pSecTag-PA83

4. Il5

Gene Name : Il5
Sequence Strain (Species/Organism) : Mus musculus
NCBI Gene ID : 16191
NCBI Protein GI : 6754336
Locus Tag : RP23-239O19.2
Genbank Accession : AC084392
Protein Accession : NP_034688
Taxonomy ID : 10090
Chromosome No : 11
Gene Starting Position : 53720793
Gene Ending Position : 53725102
Gene Strand (Orientation) : +
Protein Name : interleukin 5
Protein pI : 8.9
Protein Weight : 14616.09
Protein Length : 133
Protein Note : Also known as Il-5

DNA Sequence : Show Sequence

>gi|372099099:53720793-53725102 Mus musculus strain C57BL/6J chromosome 11, GRCm38 C57BL/6J
GCGCTCTTCCTTTGCTGAAGGCCAGCGCTGAAGACTTCAGAGTCATGAGAAGGATGCTTCTGCACTTGAG
TGTTCTGACTCTCAGCTGTGTCTGGGCCACTGCCATGGAGATTCCCATGAGCACAGTGGTGAAAGAGACC
TTGACACAGCTGTCCGCTCACCGAGCTCTGTTGACAAGCAATGAGGTAAAGTATAACTTATTCCTTCAGC
TTTGTTTTTAAGATCAGGACCTTGCTATACCGCTCTGACTGGCCTCAAACTTGCTATGTAGGGTAGGCTG
TCCTAACCCCTACCAGATCTCCTTACCTATGTCTCCCAAATACTAGGATTACAGACACATTACCTTGCCT
GACGCTATGGTTCTTCAGAATGCATAAATAGCTGCATTTGGCCTTTAATCCCAGAACTTGGGAGGCAGGG
TCAGGTGGATCTCTGTGAGTTCAAGGCCAGACTTGTCTACGTGGCCAGTTACAGGACAGCCAGAGCTAAA
GCAAGACCCTGATTCAAAATAATTTTTTTTCAAAACAAAAAAAAAAAACCCAAACCATTTGTGGCAATTC
ATTTCTAAACATAAAGATCTGCTTTAAATAGTGCAATTATGGCTTGTTCCCTTGCCTTCTTGCTCCCGTT
CTGTCCTCTTGTCCCACTCTCTCCCCATTCCACCCCCACCATGTGCTCATGGCCCGCATCTCTACTTCTC
TACTCTCTTTCTCTCCCTCTCCCCTCCTTCTTCCTTTCCCTCTCTCTCTCCCTCTTCTTCTCCTCCTCTC
TTTCTCTCTCTCTCCCTCTCTCTCTCTCTTTCTCTCTCTCTCTGCTTTTTTCTATCTCTACTACCCTCTC
AACTCCCCTCTCCATGCCCTGAATAAGCTCTATTCTATACTAAAAAAAAAAAGTGCAATTATGAATGTGT
TAGTGTTAATGCACAGGTGATAACCCTATCACCAGCAAGCATTGCATTAAAAAAGGCAACGGACTCTCTT
TAGGATGACCCTATGATGTTCTTTCCTTTGCAGACGATGAGGCTTCCTGTCCCTACTCATAAAAATGTAA
GTTATTCTTTACTGCCGTGCTTGCATGAGTAAGTCAGCTTCGCATACTAAGCTATAAGTCATCTGCATCT
AGCTTTCTGGTGTTGTGTGTGTCTGGGATGGGGACCTCTCTAGGTCTCAAGCTCCTGGGTTCAAGTGATT
CTCTTGCCTTGATAGAGCAGCTGGGACACAGGCCTGTGCCACCACACCCAGCAGAGCTTTTGATTTCAGT
TAAACTGTTTGACTTTCTTGGAAAAGAAAATTTATGTAGGTAGATATGAAAGTTTGTGCTTATAAATAAA
AAGAATATGAGAGTGGCAAATTATGTAATCCCAGTACTTGGGAGCCAAAGGCAGGGGTAGTCTGAGTCTA
GGGCCAGCTTAGATACATTGCCCTGTATGTATCAAAAGTAAATCCTATAAATAAATAAACAAAAACATTA
GAGGGCTGGAGATATAAGCTCTGTTGATAGATGGCCTAATATGCTGGGTTGACTCTTAGCACCCCATAAA
CTAAACATGGAAGTACCTGGCTGTAATCTCATGATGGTGAAATGGAGGCGGGAAGATCATAGGTTCAAGG
TCATCCTCAGCTACATTTTTGAGCTAGAGGCCAGCCTGGGCTATGAGACACGCAAAAACCACCAGCCAAT
TAATATTAGGAATGGCTTTGAGCTAGATCTGTTATGTAAGTGGCCAGCTGGAGCTGTCAGTCATACATCT
CACAGCCTCACAAGATTCTTTGCATGGCGAGAGGTCCTGCTGGGCTCCCTTTGGCTCTGTCCATGGCTCT
CTTCATCCTAGTGCCTCTCTTTGTTTTCCTTGTCTTATTTCTTACTGCTGAGGATCAAGCCCAGGGCCTT
CAGTGTGTGAAGTGAGCACTCTACCACTGAATTCCAGAGCCCGCCCACTCTAATGCCTTTCTGAAAGTAT
TAAGAGTTTAGGGTTATATATTCCTTTTGTTTATTTTATGTGTATGAGCATTTTGCCTGCATATATATAT
ATATATATATATATATATGTGTGTGTGTGTGTGTGTGTGTGTGTATATATATATGTATGTATGTATGTAT
GTATGTATGTATGTATATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTTCCACGTATGTGTCTATGTG
TCTGGTGTTCCTGAAGGCTAAAAGAAGGGCATCAGATCACCTGGGGCTGGATATGCAGATGGTTGTGAGC
CAACCATCTGGATGCTGGGAACTGCATCAAGTGTTCTTAACCACTGAGCCATCTCTCCCGCTCAGAGGGT
TATATTCTTAGGTAATGATAGAAAGACATAAAAATATCATGAATGCCTTTATTAATAATTTCTAAACAGT
TTAATGAATATGACTATGTAGTGATATTGTATACATTTCAATATTATCTTATTCTAGCGTAAAGTACATT
ATTTAACTTTTTCTAAATAGAAGAAAATTCATCAGCCTAAATTTCAAAAGAAAATATTAATATGGGTGTG
GTACCACTCACCTTTAATCCAGATGGTTGTGAGCCACCACAAGGGTGCTGGTAACTGAACCCAGGTCCTC
TGGAAGAGGACCCAGTGATCTTAACCACTGAGCCATCTCCCCAGCCCCAATCCTAACTTTGGGTTCATTT
TTTTGAAATGATCTCATGTAGCACTAGCTGGCCTCAAACTCTATGTATCAGAGGCTGGCCTTCAACTCCT
GATCCTCTTACCTCAACTTCCTGAATGCTGGCATTACAGATAAGCACCATCACATCTTGTATTGTCTGGG
GTTTTTTATTGATGCATTTAAATTGCATGTATTTATTGCATATGGCATGATATTTCAAAATATGTGTACG
TTGTGGGCAGTCTGATCTATTTGCTTCTTGATAATCTTCTTTCAGCACCAGCTATGCATTGGAGAAATCT
TTCAGGGGCTAGACATACTGAAGAATCAAACTGTCCGTGGGGGTACTGTGGAAATGCTATTCCAAAACCT
GTCATTAATAAAGAAATACATTGACCGCCAAAAAGTAAGTTCCCCAGGGACCCTGTGAATCCGGCTGCAG
CTGGTTCTCCAGGAGCCAACCTGACAGTCTGTTCTTTTCACAGGAGAAGTGTGGCGAGGAGAGACGGAGG
ACGAGGCAGTTCCTGGATTACCTGCAAGAGTTCCTTGGTGTGATGAGTACAGAGTGGGCAATGGAAGGCT
GAGGCTGAGCTGCTCCATGGTGACAGGACTTCACAATTTAAGTTAAATTGTCAACAGATGCAAAAACCCC
ACAAAACTGTGCAAATGCAAGGGATACCATATGCTGTTTCCATTTATATTTATGTCCTGTAGTCAGTTAA
ACCTATCTATGTCCATATATGCAAAGTGTTTAACCTTTTTGTATACGCATAAAAGAAATTCCTGTAGCGC
AGGCTGGCCTCAAACTGGTAATGTAGCCAAGGATAACCTTGAATTTCTGATCCTCCTGCCTCCTCTTCCT
GAAGGCTGAGGTTACAGACATGCACCATTGCCACTAGTTCATGAAGTGCTGGAGATGGAACCCAAGGCTT
TGTGCATGTTACCAACTGAGTTATACTCCCTCCCCCTCATCCTCTTCGTTGCATCAGGGTCTCAAGTATT
CCAGGCTGACTTTGAACTCAGTGTGTAGCCAAGGGTGACCCTGAACTCTTGGTCCAGATGGACGCAGGAG
GATCACATACCCAACCTTAGCATCCTTTCTCCTAGCCCCTTTAGATAGATGATACTTAATGACTCTCTTG
CTGAGGGATGCCACACCGGGGCTTCCTGCTCCTATCTAACTTCAATTTAATACCCACTAGTCAATCTCTC
CTCAACTCCCTGCTACTCTCCCCAAACTCTAGTAAGCCCACTTCTATTTCTTGGGGAGAGAGAAGGTTGA
CTTTTCTTATGTCCTATGTATGAATCAGACTGTGCCATGACTGTGCCTCTGTGCCTGGAGCAGCTGGATT
TTGGAAAAGAAAAGGGACATCTCCTTGCAGTGTGAATGAGAGCCAGCCACATGCTGGGCCTTACTTCTCC
GTGTAACTGAACTTAAGAAGCAAAGTAAATACCACAACCTTACTACCCCATGCCAACAGAAAGCATAAAA
TGGTTGGGATGTTATTCAGGTATCAGGGTCACTGGAGAAGCCTCCCCCAGTTTACTCCAGGAAAAACAGA
TGTATGCTTTTATTTAATTCTGTAAGATGTTCATATTATTTATGATGGATTCAGTAAGTTAATATTTATT
ACAACGTATATAATATTCTAATAAAGCAGAAGGGACAACT

Protein Sequence : Show Sequence

>gi|6754336|ref|NP_034688.1| interleukin-5 precursor [Mus musculus]
MRRMLLHLSVLTLSCVWATAMEIPMSTVVKETLTQLSAHRALLTSNETMRLPVPTHKNHQLCIGEIFQGL
DILKNQTVRGGTVEMLFQNLSLIKKYIDRQKEKCGEERRRTRQFLDYLQEFLGVMSTEWAMEG

Related Vaccine(s): pSecTag-PA83

IV. Vaccine Information

1. Anthrax Spore live culture Vaccine (USDA: 1011.00)

a. Manufacturer:

Colorado Serum Company

b. Vaccine Ontology ID:

VO_0001557

c. Type:

Live vaccine

d. Status:

Licensed

e. Location Licensed:

USA

f. Host Species for Licensed Use:

Cattle

2. Anthrax Spore Vaccine

a. Product Name:

B. anthracis Sterne strain spore vaccine

b. Manufacturer:

Colorado Serum Company

c. Vaccine Ontology ID:

VO_0000874

d. Type:

Live, attenuated vaccine

e. Status:

Licensed

f. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: Protein
Description:
Detailed Gene Information: Click here.

g. Preparation

Anthrax Spore Vaccine is prepared with a relatively nonpathogenic, noncapsulated variant strain of B. anthracis, originally developed at the Onderstepoort Laboratory, Pretoria, South Africa. Further work was conducted in England, India and in many other countries. Vaccine Strain is Sterne 34F2. The vaccine is a suspension of viable Bacillus anthracis spores in saponin (Spore vaccine).

h. Guinea pig Response

Host Strain: Female Hartley guinea pigs
Vaccination Protocol: Three 0.5-ml doses of the PA vaccine were administered at 2-week intervals. The commercial live veterinary Sterne strain spore vaccine was administered in three doses: 0.2, 0.3, and 0.5 ml i.m. at 2-week intervals. The stock spore vaccine contained 5 x 10^6 to 6 x 10^6 spores per ml (Little et al., 1986).
Immune Response: A high ELISA titer was obtained after immunization, as demonstrated by immunization with Sterne strain spores or PA vaccine + LF. However, it did not reflect the level of expected protection. This was demonstrated after challenge with a vaccine-resistant isolate. Comparison of the vaccine-resistant isolates with the Vollum cultures suggested that it is not the difference in the LD50s of the isolates that determines vaccine resistance but some other factors (Little et al., 1986).
Challenge Protocol: The animals were then challenged i.m. with 2,500 spores of Vollum 1B 2 weeks after immunization.
Efficacy: With Vollum and Vollum 1B, strains of B. anthracis killed 50% or more of the PA-immunized animals. The data confirm earlier findings: although guinea pigs were immunized effectively against a Vollum challenge, they were not protected against challenge with some isolates of B. anthracis. The study tested 4 of the10 strains used from the earlier study challenging their guinea pigs. In total 9 of 27 isolates tested were found to be resistant to immunization with the PA vaccine. Vaccination of guinea pigs with Sterne strain spores appears to provide broad protection against i.m. challenge with various anthrax isolates. A dose-response curve of the Sterne spore vaccine was obtained by injecting guinea pigs with 0.5 ml i.m. in a single dose or as two doses 14 days apart. The data indicate that excellent protection and antibody response to PA antigen can be achieved with two immunization doses of 10^6 Sterne spores (Little et al., 1986).
Description: Although various antigen preparations appear to provide a substantial degree of protection when immunized animals are challenged with the standard Vollum strain, earlier studies demonstrated that certain B. anthracis isolates were able to override this immunity in guinea pigs. This current study seeks an overall effort to evaluate and improve the PA vaccine presently used for humans and to confirm and expand upon those earlier studies (Little et al., 1986).

3. Anthrax Vaccine Adsorbed (AVA)

a. Product Name:

Anthrax Vaccine Adsorbed

b. Tradename:

Biothrax

c. Manufacturer:

BioPort Corp

d. Vaccine Ontology ID:

VO_0000014

e. CDC CVX code:

f. Type:

Subunit vaccine

g. Status:

Licensed

h. Location Licensed:

USA (License #1260)

i. Host Species for Licensed Use:

Human

j. Antigen

A cell-free filtrate of B. anthracis culture

k. Adjuvant:

VO ID: VO_0001241

l. Preservative:

benzethonium chloride,formaldehyde

m. Preparation

This vaccine is prepared by adsorbing filtered culture supernatants of an attenuated strain (V770-NP1-R) to aluminum hydroxide (Alhydrogel) as an adjuvant (Brey, 2005). The strain V770-NP1-R used for AVA preparation is a toxigenic, nonencapsulated strain. The filtrate contains a mix of cellular products including all three toxin components (LF, EF, and PA) (Amphogel, Wyeth Laboratories) (CDC, 2000).

n. Immunization Route

Intramuscular injection (i.m.)

o. Virulence

Most studies show that AVA only induces localized, minor, and self-limited adverse effects. No studies have definitively documented any occurrence of chronic diseases (e.g. cancer or infertility) following anthrax vaccination (CDC, 2000).

p. Storage

Vaccine should be stored at 2°C TO 8°C (36 TO 46°F). Do not freeze.

q . Approved Age for Licensed Use

Ages 18-65 (FDA: Anthrax Vaccine Adsorbed).

r. Contraindication

The use of BioThrax is contraindicated in subjects with a history of anaphylactic or anaphylactic-like reaction following a previous dose of BioThrax, or any of the vaccine components.

s. Description

AVA is the only licensed human anthrax vaccine in the United States. This vaccine was developed in the early 1950s and was licensed by the FDA in 1970. AVA has been shown to have a 92.5% efficacy for protection in both cutaneous and inhalational anthrax cases (Brey, 2005).

t. Human Response

Host Strain: Most of the study subjects were male (70.9%) and Caucasian (83.7%; 11.6% were African–American). The median age of study subjects was 33 years (range 19–61).
Vaccination Protocol: The vaccinations followed a minimal-risk protocol reviewed and administratively approved by the institutional review board at USAMRIID and the Human Subjects Research Review Board at the U.S. Army Surgeon General's office. Overall, AVA was given in a 6-dose series (subcutaneous injections at 0, 2, and 4 weeks and 6, 12, and 18 months with subsequent yearly boosters). Specifically, Vaccinations occurred within defined time intervals after receipt of the initial AVA injection (day 0 [dose #1], day 14 [range 11–21], day 28 [range 25–35], day 182 [range 154–216], day 364 [range 336–413], day 546 [range 518–609]) (Pittman et al., 2006).
Immune Response: A total of 671 sera were analyzed for IgG to PA. All subjects seroconverted after receiving AVA, as defined by a fourfold or greater increase over baseline in dilutional IgG to PA titer. The mean time after receipt of the initial AVA injection to seroconversion was 27.7 days (range = 14–63 days). The mean number of days after the first vaccine dose needed to reach a serum concentration of IgG to PA of 3 μg/mL was 24.2 days (Pittman et al., 2006).
Side Effects: No side effects were noted in this study (Pittman et al., 2006). In another independent report, AVA was linked to the development of adverse side effects including joint pain, gastrointestinal disorders, and pneumonia, leading many U.S. soldiers to refuse vaccination (Xie et al., 2005).
Efficacy: A serum concentration of IgG to PA ≥ 3 μg/mL was observed in all subjects after vaccination. This level of antibody was reached by 39.5% of subjects after the first injection, by a total of 96.5% after the second injection, and by 100% after the third injection. The analysis confirms that AVA is an effective immunogen, and significant increases in antibody concentration occurred after each injection, with peak responses achieved after the fourth (6-month) dose. No cases of anthrax disease have been observed among individuals receiving the 6-month dose of AVA (Pittman et al., 2006).
Description: The antibody profile during and after the six-dose primary vaccination series with anthrax vaccine adsorbed (AVA) was characterized in 86 human volunteers. The present study describes the kinetics of IgG antibodies to Bacillus anthracis protective antigen (PA) in AVA vaccinees receiving the entire six-dose primary series using sera obtained as part of the occupational health program and stored in the USAMRIID archive (Pittman et al., 2006).

u. Mouse Response

Host Strain: Male A/J mice obtained from the National Cancer Institute
Vaccination Protocol: Mice were immunized with AVA formulated with and without CpG ODN, PLG, or CpG ODN adsorbed onto PLG (CpG ODN-PLG). The mice were bled weekly, and their serum was stored at –20°C until use. Mice were challenged intraperitoneally with 3 x 102 to 9 x 103 50% lethal doses (LD50) of STI spores suspended in 0.5 ml of sterile phosphate-buffered saline (1 LD50 = 1.1 x 103 STI spores). Survival was monitored for 21 days (Xie et al., 2005).
Persistence: Immunized mice were protected from lethal anthrax challenge within 1 week of vaccination with CpG ODN-PLG plus AVA, with the level of protection correlating with serum immunoglobulin G anti-protective antigen titers (Xie et al., 2005).
Side Effects: No side effects noted.
Efficacy: Coadministering CpG ODN-PLG with 8 to 25 µl of AVA boosted the resultant IgG anti-PA antibody response by nearly 50-fold compared to AVA alone (Xie et al., 2005).
Description: This work examines the ability of immunostimulatory CpG oligodeoxynucleotides (ODN) adsorbed onto cationic polylactide-co-glycolide (PLG) microparticles (CpG ODN-PLG) to accelerate and boost the protective immunity elicited by AVA. The results indicate that coadministering CpG ODN-PLG with AVA induces a stronger and faster immunoglobulin G response against the protective antigen of anthrax than AVA alone (Xie et al., 2005).

v. Monkey Response

Host Strain: Rhesus Macaques
Vaccination Protocol: In the first experiment, rhesus macaques were immunized at 0 and 6 weeks with 0.5 ml of AVA plus 250 μg of an equimolar mixture of 3 CpG ODN, and then challenged with 105 Sterne strain anthrax spores when serum anti-PA titers returned to baseline at week 26.
In the second experiment, macaques were immunized with 0.5 ml of AVA plus 500 ug of ODN 7909 or the above mixture of 3 CpG ODN (Klinman et al., 2004).
Persistence: The results show that co-administering CpG ODN with AVA generates high levels of toxin neutralizing antibodies very rapidly, exceeding AVA alone by 17-fold at 11 days post-immunization (Klinman et al., 2004).
Side Effects: No serious local or systemic adverse reactions were observed in any of the macaques treated with CpG ODN plus AVA (Klinman et al., 2004).
Efficacy: Macaques immunized with AVA+ODN 7909 had on average a 17-fold higher toxin neutralizing titer than those immunized with AVA alone (Klinman et al., 2004).
Description: Synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs can improve the immune response to co-administered antigens. In mice, CpG ODN have been shown to boost the protective efficacy of vaccines against bacterial, viral and parasitic pathogens. However, due to evolutionary divergence in CpG recognition between species, ODNs that are highly active in rodents may be less efficacious in primates. Thus, pre-clinical studies to examine whether CpG ODN can accelerate and boost the immune response elictied by AVA must be conducted in a pertinent primate model. This study shows that co-administering GMP-grade CpG ODN with AVA to rhesus macaques does indeed increase rapidity, titer, affinity, and protective efficacy of their resultant IgG anti-PA response (Klinman et al., 2004).

w. Human Response

Vaccination Protocol: This study included 1,249 workers [379 received anthrax vaccine, 414 received placebo, 116 received incomplete inoculations (with either vaccine or placebo) and 340 were in the observational group (no treatment)] in four mills in the northeastern United States that processed imported animal hides (FDA: Anthrax Vaccine Adsorbed).
Immune Response: During the trial, there were 26 cases of anthrax reported across the four mills - five inhalation and 21 cutaneous.
Side Effects: Side effects after vaccination were mostly limited to local site reactions, fever, chills, nausea and general body aches.
Efficacy: In a comparison of anthrax cases between the placebo and vaccine groups, including only those who were completely vaccinated, the calculated vaccine efficacy level against all reported cases of anthrax combined was 92.5% (FDA: Anthrax Vaccine Adsorbed).

4. Anthrax vaccine adsorbed with Squalene adjuvant

a. Vaccine Ontology ID:

VO_0004246

b. Type:

Toxoid vaccine

c. Status:

Research

d. Adjuvant:

VO ID: VO_0000190

e. Immunization Route

Intramuscular injection (i.m.)

f. Mouse Response

Host Strain: Hartley
Vaccination Protocol: Mice were immunized intramuscularly (i.m.) at 0 and 4 weeks (Ivins et al., 1995).
Challenge Protocol: Animals were challenged at 10 weeks post vaccination with an aerosol of spores of B. anthracis Ames strain (Ivins et al., 1995).
Efficacy: The vaccine adjuvanted with SLT (squalene) was more protective than the vaccine without the squalene adjuvant (Ivins et al., 1995).

5. B. anthracis DNA Vaccine expressing PA

a. Vaccine Ontology ID:

VO_0004543

b. Type:

DNA vaccine

c. Status:

Research

d. Gene Engineering of pagA

Type: DNA vaccine construction
Description:
Detailed Gene Information: Click here.

e. Vector:

pIMS-120 (Livingston et al., 2010)

f. Immunization Route

Intramuscular injection (i.m.)

g. Macaque Response

Vaccination Protocol: Four groups of five animals each were assigned to the vaccine recipient groups; two animals were held as unimmunized controls. All experimental groups received intramuscular injection of vaccine at days 0 and 56 in the quadriceps. One group of rhesus macaques was immunized with 0.5 mL of Biothrax, the approved human dose. Another group was injected intramuscularly with 1.5 mg of pIMS-120. Two groups of animals were immunized with either 0.3 or 1.5 mg of pIMS-120 using an EP device referred to as the TriGrid Delivery System (TDS) (Livingston et al., 2010).
Immune Response: EP delivery effectively induced anti-PA neutralizing antibody responses in 100% of subjects at both dose levels (Livingston et al., 2010).
Challenge Protocol: One year after the first immunization, the rhesus macaques were exposed to a targeted dose of 100 LD50 spores of Ames isolate of B. anthracis (Livingston et al., 2010).
Efficacy: For the animals administered the pIMS-120 candidate by EP delivery, 40% (2/5) of the animals administered the 0.3 mg DNA dose and 80% (4/5) administered the 1.5 mg DNA dose survived the challenge (Livingston et al., 2010).

6. B. anthracis DNA vaccine LF pDNA encoding LF

a. Vaccine Ontology ID:

VO_0004417

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit

e. Gene Engineering of Lef

Type: DNA vaccine construction
Description: This DNA vaccine expressed lethal factor (LF) (Hermanson et al., 2004).
Detailed Gene Information: Click here.

f. Vector:

VR1012 (Hermanson et al., 2004)

g. Immunization Route

Intramuscular injection (i.m.)

h. Rabbit Response

Vaccine Immune Response Type: VO_0000286
Immune Response: High titers of anti-LF and neutralizing antibody to lethal toxin (Letx) were achieved in all rabbits (Hermanson et al., 2004).
Efficacy: Eight or nine animals in each group were challenged with 100x LD(50) of aerosolized anthrax spores 5 or 9 weeks after vaccination and 5 of 9 animals receiving LF pDNA survived. In addition, the time to death was significantly delayed in the others (Hermanson et al., 2004).

7. B. anthracis DNA vaccine PA

a. Vaccine Ontology ID:

VO_0004386

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit, guinea pig, mouse

e. Gene Engineering of pagA

Type: DNA vaccine construction
Description: Vector pWRG7079 expressed protective (PA) gene of B. anthracis (Riemenschneider et al., 2003).
Detailed Gene Information: Click here.

f. Vector:

pWRG7079 (Riemenschneider et al., 2003)

g. Immunization Route

Gene gun

h. Rabbit Response

Vaccine Immune Response Type: VO_0000286
Immune Response: All rabbits vaccinated with the DNA vaccine or with AVA vaccine developed antibody responses predictive of protective immunity to anthrax challenge (Riemenschneider et al., 2003).
Efficacy: Evaluating antibody responses by ELISA and TNA revealed that all rabbits vaccinated with the DNA vaccine developed antibody responses predictive of protective immunity to anthrax challenge. After a fourth vaccination, titers rebounded and the rabbits were challenged by subcutaneous injection of 100 LD50 of B. anthracis Ames strain heat-shocked spores - 9/10 rabbits given the PA DNA vaccine survived (Riemenschneider et al., 2003).

8. B. anthracis DNA vaccine PA83 furin

a. Vaccine Ontology ID:

VO_0004481

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit

e. Gene Engineering of PA83

Type: DNA vaccine construction
Description: Vector VR1012 expressed the PA construct that was chemically synthesized (Retrogen, San Diego) to include an amino terminal human tissue plasminogen activator (hTPA) leader peptide (replacing the Bacillus leader peptide) fused to a PA83 sequence (Hermanson et al., 2004).
Detailed Gene Information: Click here.

f. Vector:

VR1012 (Hermanson et al., 2004)

g. Immunization Route

Intramuscular injection (i.m.)

h. Rabbit Response

Vaccine Immune Response Type: VO_0000286
Efficacy: All animals receiving PA or PA plus LF pDNA vaccines were protected (Hermanson et al., 2004).

9. B. anthracis DNA vaccine pCPA

a. Vaccine Ontology ID:

VO_0004480

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Mouse

e. Gene Engineering of pagA

Type: DNA vaccine construction
Description: The gene fragment encoding amino acids 175 to 764 of a B. anthracis PA protein was PCR amplified using the forward primer 5′-ACA AGT CTC GAG ACC ATG GTT CCA GAC CGT GAC-3′ and the reverse primer 3′-CTC TAT CCT ATT CCA TTA AGA TCT ACT AAA-5′, with the pYS2 template. The PA gene fragment expressed corresponds to the biologically active, protease-cleaved PA63 fragment of the full-length 83-kDa protein. The PCR product was then digested with two restriction enzymes XhoI and XbaI and ligated into the eucaryotic expression plasmid pCI (Promega, Inc., Madison, Wis.) (Price et al., 2001).
Detailed Gene Information: Click here.

f. Immunization Route

Gene gun

g. Mouse Response

Vaccine Immune Response Type: VO_0000286
Efficacy: All mice immunized with pCLF4, pCPA, or the combination of both survived the challenge, whereas all unimmunized mice did not survive (Price et al., 2001).

10. B. anthracis DNA vaccine pDNA encoding PA

a. Vaccine Ontology ID:

VO_0004416

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit

e. Gene Engineering of PA83

Type: DNA vaccine construction
Description: This DNA vaccine expressed the protective antigen (PA) (Hermanson et al., 2004).
Detailed Gene Information: Click here.

f. Vector:

VR1012 (Hermanson et al., 2004)

g. Immunization Route

Intramuscular injection (i.m.)

h. Rabbit Response

Vaccine Immune Response Type: VO_0000286
Immune Response: High titers of anti-PA and neutralizing antibody to lethal toxin (Letx) were achieved in all rabbits (Hermanson et al., 2004).
Efficacy: Eight or nine animals in each group were challenged with 100x LD(50) of aerosolized anthrax spores 5 or 9 weeks after vaccination. An additional 10 animals vaccinated with PA pDNA were challenged >7 months postvaccination. All animals receiving PA pDNA vaccines were protected (Hermanson et al., 2004).

11. B. anthracis DNA vaccine pIMS-120 encoding PA

a. Vaccine Ontology ID:

VO_0004479

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Rabbit

e. Gene Engineering of pagA

Type: DNA vaccine construction
Description: Vector pVAX1 expressed the mature 83 kDa full-length PA protein (without the 29 aminoacid prokaryotic secretory signal sequence) (Luxembourg et al., 2008).
Detailed Gene Information: Click here.

f. Vector:

pVAX1 (Luxembourg et al., 2008)

g. Immunization Route

Intramuscular injection (i.m.)

h. Rabbit Response

Vaccine Immune Response Type: VO_0000286
Efficacy: Anthrax toxin neutralizing antibodies were also induced in rabbits immunized with electroporation with ED50 values comparable to those previously found to be protective in rabbits immunized with rPA (Luxembourg et al., 2008).

12. B. anthracis DNA vaccine pLAMP1-PA63

a. Vaccine Ontology ID:

VO_0004477

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Mouse

e. Gene Engineering of PA63-LAMP1

Type: DNA vaccine construction
Description: This DNA vaccine expressed C-terminal LAMP1 membrane anchor and 63 kDa mature protein (Midha and Bhatnagar, 2009).
Detailed Gene Information: Click here.

f. Immunization Route

Intramuscular injection (i.m.)

g. Mouse Response

Vaccine Immune Response Type: VO_0000286
Efficacy: Highest survival was elicited by all groups when they were challenged at week 12 and 14. Challenge was 100% fatal in control mice immunized with vector and PBS. Time-to-death analysis revealed that DNA vaccination with constructs pTPA-PA63, pPA63-LAMP1 and pTPA-PA63- LAMP1 was more protective than the native PA encoding construct (Midha and Bhatnagar, 2009).

13. B. anthracis DNA vaccine pTPA-P

a. Vaccine Ontology ID:

VO_0004478

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Mouse

e. Gene Engineering of TPA-PA63-LAMP1

Type: DNA vaccine construction
Description: This DNA vaccine expressed A63-LAMP1 N-terminal TPA signal, C-terminal LAMP1 membrane anchor and 63 kDa mature protein (Midha and Bhatnagar, 2009).
Detailed Gene Information: Click here.

f. Immunization Route

Intramuscular injection (i.m.)

g. Mouse Response

Vaccine Immune Response Type: VO_0000286
Efficacy: Highest survival was elicited by all groups when they were challenged at week 12 and 14. Challenge was 100% fatal in control mice immunized with vector and PBS (Midha and Bhatnagar, 2009).

14. B. anthracis DNA vaccine pTPA-PA63

a. Vaccine Ontology ID:

VO_0004476

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

Mouse

e. Gene Engineering of TPA-PA63

Type: DNA vaccine construction
Description: This DNA vaccine expressed the N-terminal TPA signal, and 63 kDa mature protein (Midha and Bhatnagar, 2009).
Detailed Gene Information: Click here.

f. Immunization Route

Intramuscular injection (i.m.)

g. Mouse Response

Vaccine Immune Response Type: VO_0000286
Efficacy: Highest survival was elicited by all groups when they were challenged at week 12 and 14. Challenge was 100% fatal in control mice immunized with vector and PBS. Time-to-death analysis revealed that DNA vaccination with constructs pTPA-PA63, pPA63-LAMP1 and pTPA-PA63- LAMP1 was more protective than the native PA encoding construct (Midha and Bhatnagar, 2009).

15. B. anthracis PA protein Vaccine with TMDP

a. Vaccine Ontology ID:

VO_0004270

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

B. anthracis PA protein (Ivins et al., 1992).

e. Adjuvant:

VO ID: VO_0001351

f. Immunization Route

Intramuscular injection (i.m.)

g. Guinea pig Response

Host Strain: Hartley
Vaccination Protocol: Several adjuvant preparations combined with PA were compared with each other and with MDPH-PA with respect to protection and elicitation of anti-PA antibody. In the first experiment, guinea pigs received one or more injections of each preparation (Ivins et al., 1992).
Challenge Protocol: Guinea pigs were challenged i.m. 10 weeks after the first immunization with 7,300 (73 LD50) of B. anthracis Ames spores (Ivins et al., 1992).
Efficacy: 12/18 guinea pigs survived challenge after immunization with 1 dose of PA + TMDP and 13/20 survived after 2 doses of PA + TMDP (Ivins et al., 1992).

16. B. anthracis rPA Vaccine with Rehydragel HPA adjuvant

a. Vaccine Ontology ID:

VO_0004223

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

purified rPA (Rhie et al., 2005).

e. Adjuvant:

VO ID: VO_0001311

f. Immunization Route

Intramuscular injection (i.m.)

g. Guinea pig Response

Host Strain: Hartley
Vaccination Protocol: Groups of female Hartley guinea pigs (Damul Science, Korea) weighing 300–320 g were immunized by intramuscular injection on days 0, 14, and 28 with 50 μg of the purified rPA. rPA was dissolved in 500 μl PBS containing Rehydragel HPA (alum hydroxide fluid gel, 250 μg; Reheis Inc., USA), according to the manufacturer's protocol. Phosphate-buffered saline (PBS) was used as a negative control (Rhie et al., 2005).
Challenge Protocol: Fourteen days after the third immunization, the guinea pigs were challenged with 100 LD50 of B. anthracis ATCC14578 spores by intramuscular injection. After injection with B. anthracis spores, guinea pigs were observed for a period of 14 days. Animals surviving for 14 days after the challenge were considered survivors (Rhie et al., 2005).
Efficacy: Guinea pigs immunized with rPA + Rehydragel HPA had 100% survival rate to challenge with B. anthracis ATCC14578 spores (Rhie et al., 2005).

17. Bacillus anthracis mntA deletion mutant vaccine

a. Vaccine Ontology ID:

VO_0002775

b. Type:

Live, attenuated vaccine

c. Status:

Licensed

d. Gene Engineering of mntA

Type: Gene mutation
Description:
Detailed Gene Information: Click here.

e. Preparation

An mntA deletion, generated by allelic replacement resulted in complete loss of MntA expression (Gat et al., 2005).

f. Immunization Route

Other

g. Guinea pig Response

Persistence: The mntA mutant resulted in severe attenuation; a 10(4)-fold drop in LD(50) in a guinea pig model (Gat et al., 2005).
Efficacy: All the guinea pigs were challenged with 60 LD50 of the virulent Vollum strain. All guinea pigs survived this challenge and exhibited antibody titers 103−105 of either anti-PA or anti-LF (Gat et al., 2005).

18. DAAV using PA and PGA

a. Vaccine Ontology ID:

VO_0000522

b. Type:

Conjugate vaccine

c. Antigen

Two antigens: PA-B and capsular poly-γ-d-glutamate. Both antigens are conjugated.

d. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: Protein
Description:
Detailed Gene Information: Click here.

e. Adjuvant:

VO ID: VO_0001241
Description: Anthrax involves a dual process of bacterial replication and toxin production. The dually active anthrax vaccine (DAAV) confers simultaneous protection against both bacilli and toxins was highly sought after through research. The weakly immunogenic and antiphagocytic PGA capsule disguises the bacilli from immune surveillance in a similar manner to the role of capsular polysaccharides in protecting pathogens, such as pneumococci and meningococci. Encapsulated B. anthracis strains grow unimpeded in the infected host, whereas isolates lacking the capsule are phagocytized and are virtually avirulent. Anthrax toxins are formed by PA, lethal factor (LF), and edema factor (EF), which are secreted separately as nontoxic monomers. The binding of LF or EF to PA results in the formation of active lethal toxin (LT) or edema toxin (ET), respectively. Because of its ability to elicit a protective immune response against both anthrax toxins, PA is the target antigen of existing anthrax vaccine. However, a vaccine based on both PGA and PA might allow direct targeting of bacillar growth, as well as inhibiting toxin activity, making it more effective than a vaccine based on PA alone. PGA is an attractive antigen because it consists of d-glutamic acid residues linked by γ peptide bonds, and thus bears no resemblance to mammalian host molecules (Rhie et al., 2003).

f. Preparation

This conjugate vaccine is constructed by conjugating two major virulence factors of B. anthracis, the capsular poly-γ-D-glutamic acid (PGA) and the essential toxin component and protective antigen (PA). This is a DAAV that confers simultaneous protection against both bacilli and toxins. Two sets of conjugates with 1:2 and 1:1 (wt/wt) PGA-to-PA ratios, designated DAAV-1 and DAAV-2, respectively (Rhie et al., 2003).

g. Virulence

(Rhie et al., 2003)

h. Description

Anthrax involves a dual process of bacterial replication and toxin production. The dually active anthrax vaccine (DAAV) confers simultaneous protection against both bacilli and toxins was highly sought after through research. The weakly immunogenic and antiphagocytic PGA capsule disguises the bacilli from immune surveillance in a similar manner to the role of capsular polysaccharides in protecting pathogens, such as pneumococci and meningococci. Encapsulated B. anthracis strains grow unimpeded in the infected host, whereas isolates lacking the capsule are phagocytized and are virtually avirulent. Anthrax toxins are formed by PA, lethal factor (LF), and edema factor (EF), which are secreted separately as nontoxic monomers. The binding of LF or EF to PA results in the formation of active lethal toxin (LT) or edema toxin (ET), respectively. Because of its ability to elicit a protective immune response against both anthrax toxins, PA is the target antigen of existing anthrax vaccine. However, a vaccine based on both PGA and PA might allow direct targeting of bacillar growth, as well as inhibiting toxin activity, making it more effective than a vaccine based on PA alone. PGA is an attractive antigen because it consists of d-glutamic acid residues linked by γ peptide bonds, and thus bears no resemblance to mammalian host molecules (Rhie et al., 2003).

i. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Groups of female BALB/c mice at 6–8 weeks of age were immunized by i.p. injection on days 0, 14, and 28. DAAV-1 was tested at 10- and 20-µg doses, and DAAV-2 was tested at 2-, 10-, and 20-µg doses (doses refer to PA content). Controls include PA and PGA at 20-µg doses, unconjugated PGA–PA mixture including 20 µg of PGA and 20 µg of PA. Each dose was dissolved in 50 µl of PBS and adsorbed to an equal volume of Al(OH)3 gel adjuvant (equivalent to 0.187 mg per dose). PBS/Al(OH)3 was used as a negative control (Rhie et al., 2003).
Persistence: (Rhie et al., 2003)
Side Effects: None were noted (Rhie et al., 2003).
Efficacy: After three immunizations in mice, DAAV-1 induced high levels of serum anti-PGA IgG, and booster injections significantly enhanced the IgG response. PGA-specific antibodies bound to encapsulated bacilli and promoted the killing of bacilli by complement. PA-specific antibodies neutralized toxin activity and protected immunized mice against lethal challenge with anthrax toxin. Thus, DAAV combines both antibacterial and antitoxic components in a single vaccine against anthrax (Rhie et al., 2003).
Description: PGA-specific antibodies bound to encapsulated bacilli and promoted the killing of bacilli by complement. PA-specific antibodies neutralized toxin activity and protected immunized mice against lethal challenge with anthrax toxin. Thus, DAAV combines both antibacterial and antitoxic components in a single vaccine against anthrax. DAAV introduces a vaccine design that may be widely applicable against infectious diseases and provides additional tools in medicine and biodefense (Rhie et al., 2003).

19. DNA vaccine encoding PA (PA63)

a. Vaccine Ontology ID:

VO_0000518

b. Type:

DNA vaccine

c. Antigen

B. anthracis PA (Gu et al., 1999)

d. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: Protein
Description:
Detailed Gene Information: Click here.

e. Vector:

pJW4303 (Gu et al., 1999)

f. Preparation

The gene fragment encoding AAs 173–764 of PA was PCR amplified. The PCR product was digested with NheI and BamHI and ligated into the pJW4303 vector, which was cut with the same two restriction enzymes. Both PA plasmid and control DNA were purified from E. coli DH5a using Endo-free plasmid preparation kits (Qiagen) and resuspended in PBS before use (Gu et al., 1999).

g. Virulence

Virulent strains of B. anthracis are characterized by their expression of a polyglutamic acid capsule and the production of a protein toxin. In vivo studies to determine whether cell mediated immunity provided protection against virulent B. anthracis could not be performed, since such studies require BL3 con-tainment facilities (Gu et al., 1999).

h. Storage

Not virulent.

i. Description

There have been many attempts to improve the safety and immunogenicity of the current licensed anthrax vaccine, including the formulation of PA in different adjuvants, the use of recombinant, mutant PA, expression of PA by attenuated salmonellae, and the generation of attenuated B. anthracis strains lacking one or more toxin components. Current studies have examined the possibility of inducing protection against anthrax toxin by immunizing with a DNA vaccine encoding PA. Studies in other model systems indicate that antigen-encoding DNA plasmids can stimulate strong cellular and humoral immune responses against proteins from pathogens (Gu et al., 1999).

j. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: BALB/c mice were immunized at 6–8 weeks of age by bilateral injection into the gastrocnemius muscle three times at 3-week intervals with 50 μg of purified plasmid in 50 μl of saline. Mice were bled two weeks after each vaccination. Some mice were challenged by tail vein injection of PA (60 μg/mouse) and LF (25–30 μg/mouse), a combination equivalent to approximately five LD50 (Gu et al., 1999).
Persistence: (Gu et al., 1999)
Side Effects: none (Gu et al., 1999)
Efficacy: The PA DNA vaccine protects against lethal challenge with a combination of anthrax PA + LF (Gu et al., 1999).
Description: Splenocytes from immunized BALB/c mice were stimulated to secrete IFNγ and IL-4 when exposed to PA in vitro. Immunized mice also mounted a humoral immune response dominated by IgG1 anti-PA antibody production. A 1:100 dilution of serum from these animals protected cells in vitro against cytotoxic concentrations of PA. Moreover, 7/8 mice immunized three times with the PA DNA vaccine were protected against lethal challenge with a combination of anthrax PA plus LF (Gu et al., 1999).

20. DNA vaccine encoding PA83 and LF

a. Vaccine Ontology ID:

VO_0000521

b. Type:

DNA vaccine

c. Antigen

B. anthracis PA and LF (Hermanson et al., 2004)

d. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: DNA vaccine construction
Description: The PA construct is chemically synthesized to include an amino terminal human tissue plasminogen activator (hTPA) leader peptide fused to a PA83 sequence (amino acids 30–764) with the furin cleavage site deleted (SRKKRS, amino acids 192–197). This construct, designated PA83 furin, is cloned into the mammalian expression vector VR1012 (Hermanson et al., 2004).
Detailed Gene Information: Click here.

e. Gene Engineering of Lef from B. anthracis str. A2012

Type: Protein
Description: contains Bacillus anthracis lethal factor x-ray crystal structure derived domain I; LF; vaccine construct
Detailed Gene Information: Click here.

f. Vector:

VR1012 (Hermanson et al., 2004)

g. Preparation

The PA construct is chemically synthesized to include an amino terminal human tissue plasminogen activator (hTPA) leader peptide fused to a PA83 sequence (amino acids 30–764) with the furin cleavage site deleted (SRKKRS, amino acids 192–197). This construct, designated PA83 furin, is cloned into the mammalian expression vector VR1012. The LF coding sequences are derived from the B. anthracis LF93 protein sequence, codon-optimized, and chemically synthesized as above to include the hTPA leader peptide. The LF domain I–III is PCR amplified from this clone by using a forward and reverse primer pair to amplify the 1,740-bp fragment encoding the hTPA leader peptide fused to LF amino acids 34–583. The LF domain I is also derived from the LF93 plasmid by PCR amplification using forward and reverse primer pairs to amplify an 876-bp fragment encoding an hTPA leader peptide fused to LF amino acids 34–295. Both LF genes are cloned into the VR1012 vector (Hermanson et al., 2004).

h. Virulence

The virulence of B. anthracis in rabbits, non-human primates, and humans is primarily the result of a multicomponent toxin secreted by the organism. The toxin consists of three separate gene products, designated protective antigen (PA), lethal factor (LF), and edema factor (EF), that are encoded on a 184-kb plasmid designated pXO1 (Hermanson et al., 2004).

i. Description

DNA vaccines provide an attractive technology platform against bioterrorism agents due to their safety record in humans and ease of construction, testing, and manufacture. Monovalent and bivalent anthrax plasmid DNA (pDNA) vaccines encoding genetically detoxified protective antigen (PA) and lethal factor (LF) proteins have been designed and tested for their immunogenicity and ability to protect rabbits from an aerosolized inhalation spore challenge. Immune responses after two or three injections of cationic lipid-formulated PA, PA + LF, or LF pDNAs were at least equivalent to two doses of anthrax vaccine adsorbed (AVA) (Hermanson et al., 2004).

j. Rabbit Response

Host Strain: New Zealand White
Vaccination Protocol: Plasmid DNA was prepared from overnight cultures of transformed XL-2 Blue bacteria in Terrific Broth plus 50 µg/ml kanamycin sulfate and processed by using Endo-free Giga kits. One milliliter of sterile water for irrigation (SWFI) was added to a vial containing a dried film of 3.75 µmol each of a 1:1 mixture of cationic lipid and colipid and vortex mixed for 5 minutes. The liposome suspension was diluted to 1.5 mM with SWFI and added to an equal volume of pDNA and vortex mixed briefly. The final molar ratio of all formulations was 4:1, DNA/cationic lipid (Hermanson et al., 2004). Two- to five-kilogram female New Zealand White rabbits were injected bilaterally in the quadriceps muscles with 1 ml of pDNA formulated with Vaxfectin or DMRIE/DOPE (0.5 ml per leg). Rabbits vaccinated with PA, LF, or vector received 1 mg of that pDNA whereas rabbits co-injected with PA + LF pDNAs received a mixture of 0.5 mg of each plasmid. Groups of rabbits receiving three doses were injected on days 0, 28, and 56; rabbits receiving only two doses were injected on study days 0 and 28. Rabbits immunized with AVA were injected unilaterally with 50 µl of AVA diluted to 0.5 ml in PBS on days 28 and 56. Prebleeds and biweekly postvaccination bleeds were taken for all groups for analysis of serum antibodies (Hermanson et al., 2004).
Persistence: Spore challenge induced a significant increase in the Letx neutralization titer in group 4 rabbits, suggesting that there was limited spore germination after challenge in these animals. This post-challenge increase in Letx neutralization titer, however, was smaller than the increase seen in AVA- or LF pDNA-vaccinated rabbits challenged at week 12 (Hermanson et al., 2004).
Immune Response: Both the PA and the LF pDNAs generate anti-PA and anti-LF antibody responses, respectively, when injected alone or co-injected. Furthermore, co-injection of PA and LF pDNAs does not cause detectable interference in the immunogenicity of either of the pDNAs (Hermanson et al., 2004). Immune responses after two or three injections of cationic lipid-formulated PA, PA + LF, or LF pDNAs were at least equivalent to two doses of anthrax vaccine adsorbed (AVA). High titers of anti-PA, anti-LF, and neutralizing antibody to lethal toxin were achieved in all rabbits.
Side Effects: none reported (Hermanson et al., 2004)
Challenge Protocol: Eight or nine animals in each group were challenged with 100x LD50 of aerosolized anthrax spores 5 or 9 weeks after vaccination. An additional 10 animals vaccinated with PA pDNA were challenged over 7 months post-vaccination.
Efficacy: All animals receiving PA or PA + LF pDNA vaccines were protected. In addition, 5/9 animals receiving LF pDNA survived, and the time to death was significantly delayed in the others. Groups receiving three immunizations with PA or PA + LF pDNA showed no increase in anti-PA, anti-LF, or Letx neutralizing antibody titers post-challenge, suggesting little or no spore germination (Hermanson et al., 2004).

21. pCLF4

a. Vaccine Ontology ID:

VO_0000880

b. Type:

DNA vaccine

c. Antigen

B. anthracis lethal factor (LF)

d. Gene Engineering of Lef

Type: Protein
Description:
Detailed Gene Information: Click here.

e. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: Protein
Description:
Detailed Gene Information: Click here.

f. Vector:

pCl (Price et al., 2001)

g. Preparation

Plasmid pCLF4 contains the N-terminal region (amino acids [aa] 10-254) of Bacillus anthracis LF cloned into the pCI expression plasmid. Plasmid pCPA contains a biologically active portion (aa 175-764) of B. anthracis PA cloned into the pCI expression vector. PA, LF, and LFE687C (LF7) were expressed and purified. LFE687C is the full-length enzymatically inactive LF protein containing the indicated aa substitution within the zinc-binding active site (Price et al., 2001).

h. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Micrometer-diameter gold particles were coated with plasmid pCLF4, pCPA, or a 1:1 mixture of both. Separate groups of female BALB/c mice at 4-5 weeks of age were immunized i.d. in the abdomen via biolistic particle injection on d 0, 14, and 28 with approximately 1 µg of plasmid DNA-coated gold particles for each injection. Immunization groups included mice injected with the same microparticles coated with pCPA, pCLF4, a 1:1 mixture of the pCPA and pCLF4 plasmids, or, as a vector control, the pCI plasmid. For the prime-boost immunization experiments, groups of BALB/c mice were first immunized twice with plasmid DNA as described above and then with a third and final boost of purified antigen emulsified in Freund's incomplete adjuvant. The protein immunizations were administered i.m. Blood samples were obtained 2 weeks following each vaccination, and the sera were pooled and stored at -20°C until analyzed (Price et al., 2001).
Immune Response: Titers of anti-LF antibody remain at high levels for much longer periods of time than do titers of anti-PA antibody. The LF antigen appears to be much more immunogenic and produces an immune response which lasts much longer than the response to the PA antigen. Co-administration of the pCPA and pCLF4 plasmids followed by a final protein booster immunization with the recombinant PA and LF7 antigens produced a substantially higher endpoint titer against either PA or LF at the same time-point than the antibody titers resulting from DNA-based immunization alone (Price et al., 2001).
Challenge Protocol: Plasmid-immunized BALB/c mice that had received a total of three injections were challenged with purified Letx 2 weeks following the third and final injection. The challenge was conducted by tail vein injection of a previously mixed combination of purified PA and LF proteins (60 μg of PA and 25 to 30 μg of LF per mouse), the equivalent of approximately 5 50% lethal doses (LD50) of Letx (Price et al., 2001).
Efficacy: All mice immunized with pCLF4, pCPA, or the combination of both survived the challenge, whereas all unimmunized mice did not survive. A significant antibody response is generated using DNA-based immunization alone and the levels of antibody produced are sufficient to protect animals against an Letx challenge that is 5 times the LD50. Also, co-administration of the pCPA and pCLF4 plasmids followed by a final protein booster immunization with the recombinant PA and LF7 antigens produced a substantially higher endpoint titer against either PA or LF at the same time point than the antibody titers resulting from DNA-based immunization alone (Price et al., 2001).

i. Rabbit Response

Host Strain: NZW
Vaccination Protocol: Groups of rabbits were immunized with various vaccine preparations. The first group was immunized (i.m.) twice using the needleless Biojector device with 500 ug of plasmid DNA (pCPA and/or pCLF4) resuspended in 0.5 ml of sterile phosphate buffered saline (PBS) at 4-week intervals. These animals were boosted by needle (i.m.) 4 weeks later with 200 ug of purified full-length rPA protein or full-length recombinant lethal factor (LF) protein LF7 with a point mutation resuspended in incomplete Freund’s adjuvant. The second group was immunized three times by gene gun with 10 ug plasmid DNA containing the PA63 gene fragment and/or the LF4 gene fragment bound to gold beads, at 4-week intervals. The third group of animals was immunized (s.c.) at 4-week intervals with AVA (lot FAV059), 800 ug rPA protein with Alum, or a mixture of 400 ug rPA and 400 ug rLF7 protein with Alum. Controls consisted of either non-immunized animals or a plasmid vector control not containing the PA and/or LF genes. All rabbits were aerosol challenged with B. anthracis spores, Ames strain, with an average dose of 50 LD50s with a range of 18-169 LD50s. Rabbit sera were collected prior to and following aerosol challenge and titrated for PA antibodies by indirect ELISA (Galloway et al., 2004).
Persistence: (Galloway et al., 2004)
Side Effects: None were noted (Galloway et al., 2004).
Efficacy: The results of this study indicate that DNA-based immunization against PA and LF followed by protein boosting induces significant protective immunity against aerosol challenge in the rabbit model and compares favorably with protein-based immunization (Galloway et al., 2004).
Description: None of the rabbits immunized with the DNA vaccines i.d. survived the challenge. Of the 5 vaccinated rabbits that survived, 2 were immunized i.m. with DNA followed with a protein boost and 3 were immunized subcutaneously (s.q.) with recombinant protein. DNA prime-boosted animals mount a protective response against aerosol challenge more than 1 year following the final immunization. Priming immunizations with plasmid DNA appear to set up a substantial memory response which is recalled upon protein boosting. A major factor predicting survival was the ability of the animal to mount a lasting antibody response to PA (Galloway et al., 2004).

22. pCMV/ER-PA83

a. Vaccine Ontology ID:

VO_0000876

b. Type:

DNA vaccine

c. Antigen

B. anthracis PA

d. Gene Engineering of pagA

Type: DNA vaccine construction
Description:
Detailed Gene Information: Click here.

e. Vector:

pCMV/myc/ER (Hahn et al., 2004)

f. Preparation

For the pCMV/ER PA83 construct, PA83 was cloned into the eukaryotic expression plasmid pCMV/myc/ER. A gene fragment coding for PA83 was amplified by PCR, and the resulting 2205-bp PCR fragment was digested with Pau I and Not I and ligated into the plasmid pCMV/myc/ER (Hahn et al., 2004).

g. Mouse Response

Host Strain: BALB/c and A/J
Vaccination Protocol: In the first vaccination trial, groups of 5 BALB/c mice were vaccinated on days 0, 14 and 28, with one of the three different PA-expressing plasmid constructs. A fourth group of five negative control mice received pCMV/ myc/ER vector DNA without insert. Each immunization consisted of a dose of approximately 1 mg plasmid DNA, precipitated onto 1.6 mm gold carriers per mouse. The DNA was applied to the shaved abdomen of anesthetized mice using a Helios gene gun. DNA-coated gold particles were discharged with 250 psi helium pressure. In the second vaccination trial, A/J mice were immunized according to the same vaccination protocol, except that the helium pressure for discharge of the DNA-coated gold carriers was increased to 400 psi. The treatment groups in this trial were pSecTag PA83 (13 mice), pCMV/ER PA83 (14 mice), and pCMV/myc/ER as a negative control group (10 mice). In the third vaccination trial, A/J mice were immunized with two shots per immunization using an increased DNA dose of 2.5 mg per shot, which were discharged with 400 psi. Six mice were vaccinated with pCMV/ER PA83 and eight with pSecTag PA83 (Hahn et al., 2004).
Immune Response: 26 d after the final immunization, the mice were killed, bled, and the serum samples analyzed for PA-specific antibody titers. All three plasmids induced anti-PA Ig and IgG1 antibody titers. Sera from mice vaccinated with pCMV/ER PA83 had significantly higher anti-PA total immunoglobulin titers than sera from mice vaccinated with pSecTag PA83, or pCMV/ER PA63. The GMT for PA-specific IgG1 of mice immunized with pCMV/ER PA83 was higher but not significantly different than PA-specific IgG1 responses of mice vaccinated with pSecTag PA83 or pCMV/ER PA63. There were no statistically significant differences between the titers of A/J mice immunized with pSecTag PA83 and A/J mice that received the pCMV/ER PA83 plasmid (Hahn et al., 2004).
Side Effects: Commercial anthrax vaccines can cause transient side effects, such as local pain and edema, which are probably due to trace amounts of LF and EF. None of these were noted in conjunction with the use of vaccine candidates studied here (Hahn et al., 2004).
Challenge Protocol: 14 A/J mice were vaccinated with pCMV/ER PA83, and 13 A/J mice with pSecTag PA83. The negative control group (10 A/J mice) received pCMV/myc/ER plasmid DNA without insert. Ten days after the third immunization, all mice were bled and their individual anti-PA titers were determined by ELISA. The mice of each treatment group were then marked to be challenged with either 10 LD50 of B. anthracis STI spores or 100 LD50 of STI spores. A/J mice immunized with the increased amount of plasmid DNA were all challenged with 100 LD50. After injection with B. anthracis spores, mice were observed for a period of 14 days.Surviving mice were killed and bled. The post-challenge serum samples were also analyzed by anti-PA ELISA and for toxin neutralization titers (Hahn et al., 2004).
Efficacy: Vaccination with either pSecTag PA83 or pCMV/ER PA83 showed significant protection of A/J mice against infection with B. anthracis STI spores (Hahn et al., 2004).

23. pSecTag-PA83

a. Vaccine Ontology ID:

VO_0000875

b. Type:

DNA vaccine

c. Status:

Research

d. Antigen

Protective antigen

e. Preparation

The vector pSecTag PA83, encoding the full-length PA protein, has a signal sequence for secretion of the expressed protein. For the pSecTag PA83 construct, DNA encoding the full-length B. anthracis PA83 was cloned into the eukaryotic expression plasmid pSecTag 2B. A gene fragment coding for the PA83 protein, without its own signal sequence, was amplified by PCR from DNA of a pXO2 strain of B. anthracis. The resulting 2204-bp PCR fragment was digested with Apa I and Kpn I and ligated into the vector pSecTag 2B (Hahn et al., 2004).

f. Immunization Route

Intramuscular injection (i.m.)

g. Mouse Response

Host Strain: BALB/c and A/J
Vaccination Protocol: In the first vaccination trial, groups of 5 BALB/c mice were vaccinated on days 0, 14 and 28, with one of the three different PA-expressing plasmid constructs. A fourth group of five negative control mice received pCMV/ myc/ER vector DNA without insert. Each immunization consisted of a dose of approximately 1 mg plasmid DNA, precipitated onto 1.6 mm gold carriers per mouse. The DNA was applied to the shaved abdomen of anesthetized mice using a Helios gene gun. DNA-coated gold particles were discharged with 250 psi helium pressure. In the second vaccination trial, A/J mice were immunized according to the same vaccination protocol, except that the helium pressure for discharge of the DNA-coated gold carriers was increased to 400 psi. The treatment groups in this trial were pSecTag PA83 (13 mice), pCMV/ER PA83 (14 mice), and pCMV/myc/ER as a negative control group (10 mice). In the third vaccination trial, A/J mice were immunized with two shots per immunization using an increased DNA dose of 2.5 mg per shot, which were discharged with 400 psi. Six mice were vaccinated with pCMV/ER PA83 and eight with pSecTag PA83 (Hahn et al., 2004).
Immune Response: 26 d after the final immunization, the mice were killed, bled, and the serum samples analyzed for PA-specific antibody titers. All three plasmids induced anti-PA Ig and IgG1 antibody titers. Sera from mice vaccinated with pCMV/ER PA83 had significantly higher anti-PA total immunoglobulin titers than sera from mice vaccinated with pSecTag PA83, or pCMV/ER PA63. The GMT for PA-specific IgG1 of mice immunized with pCMV/ER PA83 was higher but not significantly different than PA-specific IgG1 responses of mice vaccinated with pSecTag PA83 or pCMV/ER PA63. There were no statistically significant differences between the titers of A/J mice immunized with pSecTag PA83 and A/J mice that received the pCMV/ER PA83 plasmid (Hahn et al., 2004).
Side Effects: Commercial anthrax vaccines can cause transient side effects, such as local pain and edema, which are probably due to trace amounts of LF and EF. None of these were noted in conjunction with the use of vaccine candidates studied here (Hahn et al., 2004).
Challenge Protocol: 14 A/J mice were vaccinated with pCMV/ER PA83, and 13 A/J mice with pSecTag PA83. The negative control group (10 A/J mice) received pCMV/myc/ER plasmid DNA without insert. Ten days after the third immunization, all mice were bled and their individual anti-PA titers were determined by ELISA. The mice of each treatment group were then marked to be challenged with either 10 LD50 of B. anthracis STI spores or 100 LD50 of STI spores. A/J mice immunized with the increased amount of plasmid DNA were all challenged with 100 LD50. After injection with B. anthracis spores, mice were observed for a period of 14 days.Surviving mice were killed and bled. The post-challenge serum samples were also analyzed by anti-PA ELISA and for toxin neutralization titers (Hahn et al., 2004).
Efficacy: Vaccination with either pSecTag PA83 or pCMV/ER PA83 showed significant protection of A/J mice against infection with B. anthracis STI spores (Hahn et al., 2004).
Host Gene Response of Ifng (Interferon gamma)
- Gene Response: Spleen cells collected from plasmid-vaccinated BALB/c mice 26 days after the third immunization produced PA-specific interleukin-4, interleukin-5, and interferon-gamma in vitro. All levels were significantly higher than those from negative control mice immunized with pCMV/myc/ER, a eukaryotic expression plasmid without the insert (Hahn et al., 2004).
- Detailed Gene Information: Click here.
Host Gene Response of Ighg1
- Gene Response: pSecTag PA83 induced PA-specific humoral immune responses, predominantly IgG1 antibodies, in mice (Hahn et al., 2004).
- Detailed Gene Information: Click here.
Host Gene Response of Il4 (interleukin 4)
- Gene Response: Spleen cells collected from plasmid-vaccinated BALB/c mice 26 days after the third immunization produced PA-specific interleukin-4, interleukin-5, and interferon-gamma in vitro. All levels were significantly higher than those from negative control mice immunized with pCMV/myc/ER, a eukaryotic expression plasmid without the insert (Hahn et al., 2004).
- Detailed Gene Information: Click here.
Host Gene Response of Il5
- Gene Response: Spleen cells collected from plasmid-vaccinated BALB/c mice 26 days after the third immunization produced PA-specific interleukin-4, interleukin-5, and interferon-gamma in vitro. All levels were significantly higher than those from negative control mice immunized with pCMV/myc/ER, a eukaryotic expression plasmid without the insert (Hahn et al., 2004).
- Detailed Gene Information: Click here.

24. Recombinant PA domain 4

a. Vaccine Ontology ID:

VO_0000629

b. Type:

Subunit vaccine

c. Antigen

PA domain 4 from B. anthracis strain Sterne

d. Gene Engineering of PagA from B. anthracis str. 'Ames Ancestor'

Type: Protein
Description:
Detailed Gene Information: Click here.

e. Adjuvant:

VO ID: VO_0001241
Description: Domain 4 contains the dominant protective epitopes of PA and comprises amino acids 596-735 of the carboxy terminus of the PA polypeptide. Cell intoxication is thought to occur when full-length PA (PA83) binds to the cell surface receptor via domain 4, which contains the host cell receptor binding site. After binding to the host cell receptor, the N-terminal amino acids (1-167, i.e. domain 1a) of domain 1, which contains a furin protease cleavage site, are cleaved off, exposing the LF or EF binding site located in domain 1b and the adjacent domain 3. Domains 2 and 3 then form part of a heptameric pore on the cell surface, the LF or EF binds to its receptor, and the whole toxin complex undergoes receptor-mediated endocytosis into the cell. After acidification of the endosome, the toxin is translocates into the cell cytosol, where it exerts its cytotoxic effect. Therefore, inhibition of the binding and entry of the toxin complex, particularly lethal toxin, into the host cell is clearly important for the prevention of infection. The crystal structure of PA shows domain 4 to be more exposed than the other three domains, which are closely associated with each other. This structural arrangement may make the epitopes in domain 4 the most prominent for recognition by immune effector cells (Flick-Smith et al., 2002).

f. Preparation

DNA encoding the PA domains, which comprise amino acids 1-258, 168-487, 1-487, 168-595, 1-595, 259-735, 488-735, 596-735, and 1-735 (fusion proteins GST1, GST1b-2, GST1-2, GST1b-3, GST1-3, GST2-4, GST3-4, GST4, and GST1-4, respectively), was PCR amplified from B. anthracis strain Sterne DNA and cloned into the XhoI and BamHI sites of the expression vector pGEX-6-P3. Proteins produced by this system were expressed as fusion proteins with an N-terminal glutathione S-transferase (GST) protein. Immunization was done with rPA, with recombinant GST control protein, or with fusion proteins comprising domains 1, 4, and 1 to 4, which had the GST tag removed by incubation with PreScission Protease and removal of the GST on a glutathione Sepharose column (Flick-Smith et al., 2002).

g. Virulence

(Flick-Smith et al., 2002)

h. Description

Domain 4 contains the dominant protective epitopes of PA and comprises amino acids 596-735 of the carboxy terminus of the PA polypeptide. Cell intoxication is thought to occur when full-length PA (PA83) binds to the cell surface receptor via domain 4, which contains the host cell receptor binding site. After binding to the host cell receptor, the N-terminal amino acids (1-167, i.e. domain 1a) of domain 1, which contains a furin protease cleavage site, are cleaved off, exposing the LF or EF binding site located in domain 1b and the adjacent domain 3. Domains 2 and 3 then form part of a heptameric pore on the cell surface, the LF or EF binds to its receptor, and the whole toxin complex undergoes receptor-mediated endocytosis into the cell. After acidification of the endosome, the toxin is translocates into the cell cytosol, where it exerts its cytotoxic effect. Therefore, inhibition of the binding and entry of the toxin complex, particularly lethal toxin, into the host cell is clearly important for the prevention of infection. The crystal structure of PA shows domain 4 to be more exposed than the other three domains, which are closely associated with each other. This structural arrangement may make the epitopes in domain 4 the most prominent for recognition by immune effector cells (Flick-Smith et al., 2002).

i. Mouse Response

Host Strain: Female A/J mice
Vaccination Protocol: Mice were vaccinated with 10 µg of protein adsorbed to a 20% (vol/vol) solution of 1.3% Alhydrogel on days 1 and 28 of the study. Also included were groups of mice that were immunized with rPA (expressed and purified from B. subtilis), with recombinant GST control protein, or with fusion proteins comprising domains 1, 4, and 1-4, which had the GST tag removed by incubation with PreScission Protease and removal of the GST on a glutathione Sepharose column. Blood samples from mice were collected 37 days after primary immunization for serum antibody analysis by enzyme-linked immunosorbent assay. Mice were challenged i.p. with either 10^5 or 10^6 spores of the B. anthracis STI strain (equivalent to 10^2 or 10^3 minimum lethal doses [MLDs], respectively) on day 70 of the immunization regimen and were monitored for 14 days postchallenge to determine their protected status (Flick-Smith et al., 2002).
Persistence: (Flick-Smith et al., 2002)
Side Effects: No side effects noted (Flick-Smith et al., 2002).
Efficacy: At the lower challenge level of 10^2 MLDs, mice in the GST1-2-, GST4-, and cleaved 4-immunized groups were all fully protected. All mice in the groups immunized with fusion proteins containing domain 4 were fully protected against challenge with 10^3 MLDs of STI spores (Brey, 2005).

25. Recombinant PA with Poly(I:C) Adjuvant

a. Vaccine Ontology ID:

VO_0004255

b. Type:

Subunit vaccine

c. Status:

Research

d. Antigen

Recombinant PA (Sloat and Cui, 2006).

e. Adjuvant:

VO ID: VO_0001332

f. Immunization Route

intranasal immunization

g. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Mice were lightly anesthetized and given a total volume of 20 mL of rPA/pI:C solution in two 10-mL doses, with 10-15 min between each dose, half in each nare. As controls, mice (n = 5) were subcutaneously (s.c.) injected with rPA (5 mg/mouse) admixed with aluminum hydroxide gel, nasally dosed with rPA admixed with cholera toxin as a mucosal adjuvant, or left untreated. Mice were dosed on days 0, 7, and 14 (Sloat and Cui, 2006).
Immune Response: Mice nasally immunized with rPA adjuvanted with pI:C developed strong systemic and mucosal anti-PA responses with lethal toxin neutralization activity. These immune responses compared favorably to that induced by nasal immunization with rPA adjuvanted with cholera toxin. Poly(I:C) enhanced the proportion of DCs in local draining lymph nodes and stimulated DC maturation (Sloat and Cui, 2006).

26. rLAG- PA-DCpep

a. Vaccine Ontology ID:

VO_0004715

b. Type:

Recombinant vector vaccine

c. Status:

Research

d. Host Species for Licensed Use:

Mouse

e. Gene Engineering of pagA

Type: Recombinant vector construction
Description:
Detailed Gene Information: Click here.

f. Preparation

Targeted B. anthracis protective antigen (PA) genetically fused to a DC-binding peptide (DCpep) was delivered by Lactobacillus acidophilus (Mohamadzadeh et al., 2010)

g. Immunization Route

Intramuscular injection (i.m.)

h. Mouse Response

Vaccination Protocol: Groups of mice were orally vaccinated with 100 µl (108 CFU) L. gasseri expressing PA–DCpep, PA–Ctrlpep, or cells harboring the empty vector. Oral vaccination was administered four times on a weekly basis (Mohamadzadeh et al., 2010).
Vaccine Immune Response Type: VO_0003057
Challenge Protocol: The groups of mice were challenged intraperitoneally with B. anthracis Sterne pXO1+/pXO2- (5 × 10^4 CFU/mouse) (Mohamadzadeh et al., 2010).
Efficacy: L. gasseri expressing PA–DCpep fusion was 100% efficacious in protection of the mice compared with 30% survival when vaccinated with L. gasseri expressing PA–Ctrl pep (Figure 3A & B). Additionally, vaccinated mice with recombinant PA plus alhydrogel were fully protected from Sterne lethal challenge (Mohamadzadeh et al., 2010).

27. rPA with adjuvant Nanoemulsion

a. Vaccine Ontology ID:

VO_0000526

b. Type:

Toxoid vaccine

c. Antigen

For this vaccine, recombinant Bacillus anthracis protective antigen was used (Bielinska et al., 2007).

d. Adjuvant:

VO ID: VO_0001319

e. Preparation

The NE was manufactured by the emulsification of cetyl pyridum chloride, Tween 20, and ethanol in water with hot-pressed soybean oil, using a high-speed emulsifier. Every rPA-NE formulation was prepared by mixing rPA protein solution with NE, using saline as a diluent 30 to 60 min prior to immunization. For immunization with immunostimulants, 20 μg rPA was mixed with either 5 μg of MPL A or 10 μg CpG oligonucleotides in saline (Bielinska et al., 2007).

f. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Groups of mice were immunized intranasally with either one or two administrations of experimental vaccine 3 weeks apart. rPA-NE mixes were applied to the nares with a pipette tip administering 10 μl per nare, and the animals were then allowed to inhale the material (Bielinska et al., 2007).
Immune Response: rPA-NE immunization was effective in inducing both serum anti-PA IgG and bronchial anti-PA IgA and IgG antibodies after either one or two mucosal administrations. Serum anti-PA IgG2a and IgG2b antibodies and PA-specific cytokine induction after immunization indicate a Th1-polarized immune response. rPA-NE immunization also produced high titers of lethal-toxin-neutralizing serum antibodies in mice (Bielinska et al., 2007).
Challenge Protocol: The immune responses of mice were not challenged.

g. Guinea pig Response

Host Strain: Hartley
Vaccination Protocol: Hartley guinea pigs were vaccinated intranasally with one or two administrations of vaccine, each about 50 μl per nare, 4 weeks apart (Bielinska et al., 2007).
Immune Response: serum anti-PA immunoglobulin G and bronchial anti-PA IgA and IgG antibodies were produced following either one of two mucosal immunizations of rPA-NE. The anti-PA IgG2a and IgG2b antibodies and PA-specific cytokine induction found in the serum indicated a Th-1-polarized immune response. High titers of lethal-toxin-neatralizing antibodies were also found after rPA-NE immunization (Bielinska et al., 2007).
Challenge Protocol: The guinea pigs were challenged intradermaly with ~1,000 times the 50% lethal dose of B. anthracis Ames strain spores, which was about 1.38 × 10^3 spores (Bielinska et al., 2007).
Efficacy: Nasal immunization resulted in 70% and 40% survival rates against intranasal challenge with 1.2 × 10^6 and 1.2 × 10^7 Ames strain spores (Bielinska et al., 2007).

28. Typhi strain Ty21a-PA (Bacillus anthracis)

a. Vaccine Ontology ID:

VO_0004702

b. Type:

Recombinant vector vaccine

c. Status:

Research

d. Host Species for Licensed Use:

Mouse

e. Preparation

Anthrax protective antigen was delivered by Salmonella enterica serovar Typhi Ty21a (Osorio et al., 2009)

f. Immunization Route

Intramuscular injection (i.m.)

g. Mouse Response

Vaccination Protocol: Control mice received three doses of Ty21a alone. Mice that were immunized i.n. received 5 × 10^8 CFU per dose, and mice that were immunized i.p. received 5 × 10^7 CFU per dose. i.n. immunization was performed by administering 20 μl of a bacterial solution to the nares (Osorio et al., 2009).
Vaccine Immune Response Type: VO_0003057
Challenge Protocol: Mice were exposed for 90 min to aerosolized spores (5 × 10^9 spores per ml in deionized water with 0.01% Tween 80) prepared from B. anthracis strain 7702(pXO1+, pXO2−) (Osorio et al., 2009).
Efficacy: Vaccinated mice demonstrated 100% protection against a lethal intranasal challenge with aerosolized spores of B. anthracis 7702 (Osorio et al., 2009).

V. References

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