VIOLIN: Vaccine Investigation and Online Information Network

Leishmania amazonensis

Table of Contents

General Information
Vaccine Related Pathogen Genes
1. CP cysteine proteinase (Protective antigen)
2. Gp46 (Protective antigen)
3. hypothetical protein [Leishmania braziliensis] (Protective antigen)
4. M2 (Protective antigen)
5. P4 (Protective antigen)
6. WD (Protective antigen)
Vaccine Information
References

I. General Information

1. NCBI Taxonomy ID:

5659

2. Disease:

Leishmaniasis

3. Introduction

The different Leishmania species cause a broad spectrum of human diseases. L. amazonensis is known to be associated with cutaneous, diffuse cutaneous, and visceral leishmaniasis in South and Central America. The pathological mechanisms responsible for the variable outcomes of infection in humans are not fully understood; however, it is generally agreed that long-lasting immunity against reinfection can be developed in cutaneous leishmaniasis patients. Several vaccination trials have demonstrated that killed L. amazonensis can induce protection from natural infection. However, the efficacy of heat-killed vaccines against Leishmania has been extremely low or highly variable within the same study. Live parasites have been used as a vaccine strategy, and although they are highly effective in inducing immunity, this strategy has been virtually abandoned due to safety issues associated with injecting virulent organisms (Campbell et al., 2003).

4. Host Protective Immunity

Protective mechanisms in L. amazonensis are unclear. During L. amazonensis infection, BALB/c mice display a mixed profile of both Th1 and Th2 responses, although the implications of this mixed profile are ambiguous. However, the necessity for vaccines to induce a Th1-dominant response for these species of Leishmania appears to be consequential for protection (Campbell et al., 2004). The immune responses induced against L. (L.) amazonensis have showed a distinct pattern from that described for L. (L.) major. While mice that are resistant or susceptible to infection with L. (L.) major exhibit immune responses polarized to Th1 and Th2, respectively, susceptibility to L. (L.) amazonensis can not be correlated to an increased Th2 response (Fedeli et al., 2010).

II. Vaccine Related Pathogen Genes

1. CP cysteine proteinase

Gene Name : CP cysteine proteinase
Sequence Strain (Species/Organism) : Leishmania amazonensis
VO ID : VO_0011295
NCBI Nucleotide GI : 30142041
NCBI Protein GI : 30142042
Protein Accession : AAN34826.1
Other Database IDs : CDD:185513
CDD:214853
CDD:278538
CDD:239068
Taxonomy ID : 5659
Gene Strand (Orientation) : ?
Protein Name : cysteine proteinase
Protein pI : 6.56
Protein Weight : 36075.3
Protein Length : 418
Protein Note : forma: amastigote

DNA Sequence : Show Sequence

>gi|30142041|gb|AY141759.1| Leishmania mexicana amazonensis clone 2A1 cysteine proteinase mRNA, complete cds
CCGTCACGCCCTTCTCGTGCGCACGTTGCAGAACCAGGATCGGCAAAGATGGCGCGCCGCAACCCCCTTT
TGTTTGCGATAGTGGTGACGATCCTGTTCGTGGTGTGCTACGGTTCCGCTCTCATCGCCCAGACACCCCC
CGCCGTCGACAACTTCGTTGCCTCAGCGCATTACGGAAGTTTTAAGAAGCGCCACAGTAAGGCCTTCGGC
GGGGACGCCGAGGAGGGTCACCGCTTCAATGCCTTCAAGCAGAACATGCAGACAGCCTACTTCCTCAACA
CGCAGAACCCCCACGCGCACTACGACGTGTCCGGCAAGTTCGCGGACCTCACCCCGCAGGAGTTCGCCAA
GCTGTACCTGAATCCCGACTACTACACGAGCCACCTCAAGGATCACAAGGAGGACGTGCACGTCGACGAC
AGCGCCCCCAGTGGTGTGATGTCGGTGGACTGGCGTGATAAGGGTGCCGTGACGCCGGTGAAGAACCAGG
GATTATGCGGCTCGTGCTGGGCCTTCTCCGCCATTGGCAACATCGAAGGTCAGTGGGCTGCAAGCGGCCA
CTCGTTGGTTTCACTGTCAGAACAGATGCTCGTGTCGTGCGACAACGTCGATGAAGGGTGCAACGGCGGG
CTGATGGACCAGGCAATGAACTGGATCATGCAGAGTCACAACGGCAGTGTGTTCACGGAGGCCAGCTATC
CCTACACCTCTGGCGGCGGCACCAGACCGCCGTGCCATGACGAAGGTGAAGTTGGCGCTAAAATCACCGG
TTTCCTCTCCCTGCCGCACGACGAGGAGCGGATCGCGGACTGGGTGGAGAAGAGAGGCCCCGTCGCCGTC
GCCGTCGACGCGACAACCTGGCAGCTGTACTTTGGCGGTGTGGTCTCCCTCTGCCTCGCGTGGTCGCTCA
ACCACGGTGTGCTCATTGTCGGCTTCAACAAAAACGCGAAACCGCCGTACTGGATCGTGAAGAACTCGTG
GGGCTCCTCGTGGGGTGAGAAGGGGTACATCCGCCTTGCCATGGGCAGCAACCAGTGCATGCTCAAGAAT
TACCCCGTGTCGGCCACGGTTGAAAGCCCCCACACCCCACACGTGCCGACGACAACGGCCTAGCTGGAGC
CCAGGCCCACGGCGCCTGGGCGAATGAGCTGCTTTTTTTTTCGAGTGGGTGCTCGAGGCTGTGCACAAGC
ACGACCTATCCGACCTGTGTGTGCCTGAAGCGCCAGAATGGCGGCTCTGCCCAGGTGACCTGTGGCAAAC
CCGAGACGGCGGAGCTTCTCTACAACTGGCCGGGTTGCTCCGAGGCTGCCACCGAGGCCTGCATGCCGCT
GAACAATATGTATGCGCAGCTACGCCGGCTACCTCCAGAACGTCTGCACTGCGTTCGTCGCTTGCACTGA
GAACCAACGACGCCGTCTCTGACTTGACCAAGCCAACCATCTTGGGCCAACCACACGTCCCGTACGCGAG
GCACGCGGGGATCCAGGACTGGCAGGATCCACTGCGCACTATACGCCTCAGCTGCACCGATGCGGATCCC
TCCCTCAAAAAAAAAAAAAAAAAAAAAA

Protein Sequence : Show Sequence

>AAN34826.1 cysteine proteinase [Leishmania amazonensis]
MARRNPLLFAIVVTILFVVCYGSALIAQTPPAVDNFVASAHYGSFKKRHSKAFGGDAEEGHRFNAFKQNM
QTAYFLNTQNPHAHYDVSGKFADLTPQEFAKLYLNPDYYTSHLKDHKEDVHVDDSAPSGVMSVDWRDKGA
VTPVKNQGLCGSCWAFSAIGNIEGQWAASGHSLVSLSEQMLVSCDNVDEGCNGGLMDQAMNWIMQSHNGS
VFTEASYPYTSGGGTRPPCHDEGEVGAKITGFLSLPHDEERIADWVEKRGPVAVAVDATTWQLYFGGVVS
LCLAWSLNHGVLIVGFNKNAKPPYWIVKNSWGSSWGEKGYIRLAMGSNQCMLKNYPVSATVESPHTPHVP
TTTA

Molecule Role : Protective antigen
Molecule Role Annotation : A 500 bp fragment encoding an isoform of cysteine proteinase (CP) from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24. Immunization of BALB/c mice with rLacys24 plus CFA adjuvant resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice (Fedeli et al., 2010).
Related Vaccine(s): L. amazonensis CP Protein Vaccine

2. Gp46

Gene Name : Gp46
Sequence Strain (Species/Organism) : Leishmania amazonensis
VO ID : VO_0011292
NCBI Nucleotide GI : 159320
NCBI Protein GI : 159321
Protein Accession : AAA29234.1
Other Database IDs : CDD:178695
CDD:275380
Taxonomy ID : 5659
Gene Strand (Orientation) : ?
Protein Name : surface membrane glycoprotein GP46/M-2
Protein pI : 8.21
Protein Weight : 46877.99
Protein Length : 560
Protein Note : hypothetical protein; Provisional

DNA Sequence : Show Sequence

>gi|159320|gb|M38368.1|LEIGP46M2 L.amazonensis surface membrane glycoprotein GP46/M-2 gene, complete cds
ATGGCGCAGTGCGTGCGTCGGCTGGTGCTGGCGGCGCCCCTCGCCGCTGTGGTGGCGCTGCTGCTGTGCA
CGAGCAGTGCACCGGTGGCGCGTGCTGCGGGGACGAGCGACTTCACTGGTGCGCAGCAGAAGAACACGCT
GACGGTGCTGCAGGCGTTTGCGCGTGCGATCCCTGCGCTTGGGGACACGTGGACGGGCAGCGACTTCTGC
TCGTGGGAGCACATCATCTGCTACTCCTCCGGCGTCGGCGTGTGGATGCACAATGTGGATTATACCGGCA
CGCTGCCGGAGATGCCTGCGAGCGTCGACTACAAGGACGTCATGATCTTGGCACTGGACTTCGGCGCAAT
GGGCCAGGGGCTGAGCGGGACGCTGCCCCCCTCATGGAGCTCGATGAAGCACCTGATCGTTCTGGATCTG
GAGGGCACTAAGGTGTCCGGCACGCTGCCGCCCGAGTGGAGTGAGATGACGAGTGCTGAGGCCCTGCAGC
TGGAGAACTGCGGTCTGTCCGGGAGTCTGCCCACCTCGTGGTCTTCGATGCCGAAGCTGCGTATCGTCTC
ACTGAGCGGCAACCACTTCTGCGGGTGCGTGCCCGACTCGTGGAGGGAGAAGGACCGCCTCGATGTGACC
ATCGAGGAATGGCACATGGGCGAGGACTGCAAGCTTGCTAACGCCTGCCGCCCGACTGCTGCTCCGGGAA
CGACCACGACTAACCCGCCCACCACCACCGGCACCCCAGCAGCCTCCTCTACTCCTTCTCCAGGGTCGGG
GTGCGAGGTGGATGGGTGTGAGGTGTGCGAGGGGGACTCCGCTGCGCGGTGCGCCAGGTGCCGTGAGGGC
TACTCCCTGACGGACGAGAAGACGTGCCTGGGCGAACCACGATGGCGGCGTGGCGGCGGCGTCGAGCGGA
CGGCTGGCTGCCGCTGCTGTGTGGGCGGCTGTGCTGTTGAGCGTGGGGCTGGTGGCGTGAGGGTGCGGCG
GGCCCCTCTTCTCTGTGGTGCCCCTGGTGCCTGCCCTCGCCCCCGGCACGGCGTCGTCGCTGCCCTCTCA
CCCCCACCAGCCGACGGGGAGACCGACAGCCACACGCGCACGCGCACACGCCGTCGTGCATCGCGTGTGC
TTTCCGCCGTTGTGGCGCCTGCGCGGATGCACGGGCATGCGGAGGCGTGCATGCGTGTGCGCGTGCCGGC
TCTTGTGTGTCTCTCCGTGTGGCCAGCAGTCGGCACCCGCCGCCGATCGAATGTGCGCGCGGCGGCGGTG
TGTCGCCTTGGACAGCGGAGATGCGGCGCCCGCCCCTCGCCGTGTGCCTCGGTCTGCGTGTCGTGGCCGC
GCGAGCGACGTACGGAGTGCGCGTGTCCGGCCCTCTTCGACGGGGCTCGCTTGCGGTGCTGTGCTCTCGT
GGTCTGTGCCGGTGCTGCCCCGGCGGGGTGA

Protein Sequence : Show Sequence

>AAA29234.1 surface membrane glycoprotein GP46/M-2 [Leishmania amazonensis]
MAQCVRRLVLAAPLAAVVALLLCTSSAPVARAAGTSDFTGAQQKNTLTVLQAFARAIPALGDTWTGSDFC
SWEHIICYSSGVGVWMHNVDYTGTLPEMPASVDYKDVMILALDFGAMGQGLSGTLPPSWSSMKHLIVLDL
EGTKVSGTLPPEWSEMTSAEALQLENCGLSGSLPTSWSSMPKLRIVSLSGNHFCGCVPDSWREKDRLDVT
IEEWHMGEDCKLANACRPTAAPGTTTTNPPTTTGTPAASSTPSPGSGCEVDGCEVCEGDSAARCARCREG
YSLTDEKTCLGEPRWRRGGGVERTAGCRCCVGGCAVERGAGGVRVRRAPLLCGAPGACPRPRHGVVAALS
PPPADGETDSHTRTRTRRRASRVLSAVVAPARMHGHAEACMRVRVPALVCLSVWPAVGTRRRSNVRAAAV
CRLGQRRCGARPSPCASVCVSWPRERRTECACPALFDGARLRCCALVVCAGAAPAG

Molecule Role : Protective antigen
Molecule Role Annotation : cDNAs encoding L. m. amazonensis gp46 antigen were subcloned into the VR1012 plasmid, and susceptible BALB/c mice were immunised with two i.m. injections of 100 microg of plasmid DNA. Mice were challenged by the injection of 4 x 10^6 L. m. mexicana parasites in the foot pad to evaluate protection. Measurement of lesion size indicated that mice immunised with VR012-GP46 were partially protected against infection (Dumonteil et al., 2000).
Related Vaccine(s): L. amazonensis DNA vaccine encoding GP46

3. hypothetical protein [Leishmania braziliensis]

Gene Name : hypothetical protein [Leishmania braziliensis]
Sequence Strain (Species/Organism) : Leishmania amazonensis
NCBI Gene ID : 5418250
NCBI Protein GI : AKK31275
Other Database IDs : CDD:294530
Taxonomy ID : 5660
Protein Name : hypothetical protein
Protein pI : 5.06
Protein Weight : 17415.5
Protein Length : 237
Protein Note : Putative cyclase; cl00814

Protein Sequence : Show Sequence

>AKK31275.1 hypothetical protein [Leishmania braziliensis]
MRLVDLTLPLYDGMPVYDGDPPVRVTKVCTREKDGWEVRELRMSTHSGTHVDAPVHMHEGGRNLDEVPLT
QFCGPAVVVRIAAASFPQNKGLLFYEAVPADCVPRIVAANALFVGGPLEEEAERLLLSRGIITYTELVNV
EELIGESFTFYGLPLRIRGGDGSPVRAVAVIDDK

Molecule Role : Protective antigen
Molecule Role Annotation : (Duarte et al., 2017)

4. M2

Gene Name : M2
Sequence Strain (Species/Organism) : Leishmania amazonensis
VO ID : VO_0011291
NCBI Nucleotide GI : 2828562
NCBI Protein GI : 2828563
Protein Accession : AAC08302.1
Other Database IDs : CDD:240315
CDD:153108
Taxonomy ID : 5659
Gene Strand (Orientation) : ?
Protein Name : ribonucleotide reductase M2 subunit
Protein pI : 5.07
Protein Weight : 38155.95
Protein Length : 424
Protein Note : hydroxyurea-resistant strain derived from LV-78;
cloned from extrachromosomal amplified circular DNA

DNA Sequence : Show Sequence

>gi|2828562|gb|AF005247.1| Leishmania mexicana amazonensis extrachromosomal element ribonucleotide reductase M2 subunit gene, complete cds
ATGAAACCGGTTGCGGCTGGCGCCGAGGTGCTGCCGGCGGACAAGGTTGCCCAGGGGACGAACGCCGAGG
AGGAGCCGCTGCAGCAGGAGAACCCGTTCCGCTACGTCCTCTTCCCAATCCAGTACCATGACATCTGGCG
GAAGTACAAGGAGCAGGAGAGCTGCATCTGGACGGTGGAGGAGATCGACCTGGGCAACGACATGAAGGAC
TGGGCAACGCTGAACGACGGTGAGCGGCACTTCATCAAGCATGTGCTTGCCTTCTTCGCCGGCAGCGACG
GCATAGTCATCGAGAATCTTGCGCAGCGCTTCATGAGCGACGTGAAGGTGCCAGAGGCGCGCGCGTTCTA
CGGGTTCCAGCTGATGATGGAGAACATCCACTCGGAGACGTACTCTGTGCTGCTGGACACGTACATCACG
GACAGCGAGGAGAAGCTGCGGCTGCTGCACGCGATACAGACGATCCCGTGCATCCAGAAGAAGGCGGAGT
GGCCCGTGCGGTGGATCGGGAGCAGCGCGAGCTTTCAGCAGCGGCTGATCGGCTTTGCCGCGGTGGAGGG
CATTTTCTTTTCCGGGTCGTTCTGCGCGCTGTTCTGGCTGAAGAAGCGCGGTCTGATGCCAGGGCTGACG
TTCAGCAACGAGCTCATCTCGCGCGACGAGGGTCTACACACGGACTTCGCATGCCTGCTGTACAACTCGC
ACATCAAGCACAAGCTGCCGCGCGAGCGCGTGCTGGAGATCATCGTGGATGCGGTGAACATCGAGCGTGA
GTTCATCTGCGACGCGCTGCCGGTGCGGCTGATTGGCATGAACGCGGAGCTGATGGCGCAGTACATCGAG
TTCGTCGCGGACCGGCTGCTGGTGTCGCTCGGCGAGGAGAAGCACTACCGCTCGACGCAGCCGTTCGACT
TCATGGAGATGATCTCGCTGCAGGGCAAAACGAACTTCTTCGAGAAGAGGGTGGGGGAGTACCAGAAGGC
CGGCGTGATGAGCACGGAGGGCACGTCGAAAAAGTTCTCCCTCTCGGAGGACTTCTAA

Protein Sequence : Show Sequence

>AAC08302.1 ribonucleotide reductase M2 subunit [Leishmania amazonensis]
MKPVAAGAEVLPADKVAQGTNAEEEPLQQENPFRYVLFPIQYHDIWRKYKEQESCIWTVEEIDLGNDMKD
WATLNDGERHFIKHVLAFFAGSDGIVIENLAQRFMSDVKVPEARAFYGFQLMMENIHSETYSVLLDTYIT
DSEEKLRLLHAIQTIPCIQKKAEWPVRWIGSSASFQQRLIGFAAVEGIFFSGSFCALFWLKKRGLMPGLT
FSNELISRDEGLHTDFACLLYNSHIKHKLPRERVLEIIVDAVNIEREFICDALPVRLIGMNAELMAQYIE
FVADRLLVSLGEEKHYRSTQPFDFMEMISLQGKTNFFEKRVGEYQKAGVMSTEGTSKKFSLSEDF

Molecule Role : Protective antigen
Molecule Role Annotation : Immunization of CBA mice with the M-2 glycoprotein of L. amazonensis and C. parvum adjuvant resulted in complete protection against a challenge infection of 10^4 and 10^6 late log-phase promastigotes of L. amazonensis. In the BALB/c strain, complete protection was observed in some of the immunized animals (28 to 50%); in the rest of the mice the onset of infection was significantly delayed. Protective immunity for C57BL/6 mice was observed only at the low infecting dose (10(4) L. amazonensis organisms) (Champsi and McMahon-Pratt, 1988).
Related Vaccine(s): L. amazonensis M2 protein vaccine

5. P4

Gene Name : P4
Sequence Strain (Species/Organism) : Leishmania amazonensis
VO ID : VO_0011288
NCBI Nucleotide GI : 29165286
NCBI Protein GI : 29165287
Protein Accession : AAO65599.1
Other Database IDs : CDD:280435
CDD:211382
Taxonomy ID : 5659
Gene Strand (Orientation) : ?
Protein Name : P4 nuclease
Protein pI : 7.27
Protein Weight : 32555.78
Protein Length : 371
Protein Note : S1/P1 Nuclease; pfam02265

DNA Sequence : Show Sequence

>gi|29165286|gb|AY219923.1| Leishmania amazonensis P4 nuclease mRNA, complete cds
ATGCCTGCGCCTGTCAGTCTCCGTCTGCCTCTCACAGCGCTGTGCCTCCTGGTGCTCAGCTCTGCGCTGT
GCGTGACTGAGGTGCTCGGATGGGGCTGCGTGGGCCACATGCTCCTCGCCGAGATCGCGCGGCGCCAGCT
GGACTTAGAGAATGAGGAAAAGATCGAGCTGATGGCGGCAGTATTTTCGGGCAGCGGTCCGTTCCCGATG
TCTCCGAGCATGGTGCAAGCGGCATGCTGGGCGGATGACGTGAAGCTTTGGCGTCAGTACGCCATGAGCA
CGTGGCACTTCTACGCCATGCCATATAACCCCGGAAACATCAATATCACCGATCCGGTCAATACAGTGAA
CGCCGTCACTGTGTGCCTTGATATGGTGACCTCGCTTAAGAACTCGAAAGCGCCGCTGTACTTGTTGAAC
TTTGCTTGGGTGAACCTGGTGCACATCTTTGGCGATCTGCATCAGCCCCTGCACACGATATCTAGGTACA
CTACCGCCTACCCGCACGGAGATCAAGGAGGCAACGCGATTTCGGTTCGTGTAGGCGGAAAAAAGGTGAA
GCTGCATGCCCTCTGGGATAATATTTGCAGTGCTACGCCACCACGCTACCAGCGGCCCCTCTCCCACACC
GACCTCTTCGCCCTCTCTGCGACAGCGGACGGGCTGGTGGAAACTTACACGTTTTCAGAGGCGTTGGAGA
CACTCGTGGACGTGATGGCGATTCATGAGGAGAGCTACATGTTTGCGGTAAACACCAGCTACCCCGGCGT
CACACCAGGTGGGACTCTCAGTCGTGCCTACCTTGATAAATGCAAGCGTGTCGCTGAGGCTCGACTGACG
CTTGGCGGCTACCGCCTCGGTTACCTTCTGAATCAGCTGCTGTCGGGCATCACTGTGGACAAGGCCGCAC
TGGAGGCGCACCGCGCTGCGCGTCCGAAGTGGAGCGCGTAA

Protein Sequence : Show Sequence

>AAO65599.1 P4 nuclease [Leishmania amazonensis]
MPAPVSLRLPLTALCLLVLSSALCVTEVLGWGCVGHMLLAEIARRQLDLENEEKIELMAAVFSGSGPFPM
SPSMVQAACWADDVKLWRQYAMSTWHFYAMPYNPGNINITDPVNTVNAVTVCLDMVTSLKNSKAPLYLLN
FAWVNLVHIFGDLHQPLHTISRYTTAYPHGDQGGNAISVRVGGKKVKLHALWDNICSATPPRYQRPLSHT
DLFALSATADGLVETYTFSEALETLVDVMAIHEESYMFAVNTSYPGVTPGGTLSRAYLDKCKRVAEARLT
LGGYRLGYLLNQLLSGITVDKAALEAHRAARPKWSA

Molecule Role : Protective antigen
Molecule Role Annotation : Susceptible BALB/c mice were immunized with the DNA encoding P4 alone, P4/IL-12, or P4/HSP70 prior to challenge with L. amazonensis promastigotes. Mice immunized with P4 alone had a delayed onset of lesion development and significantly smaller lesions than the infection control group at 11 weeks postinfection (P < 0.05). P4/IL-12 immunized animals showed no sign of lesion development in three independent experiments (P < 0.01). Our results indicate that different vaccine combinations, including DNA encoding P4, HSP70, or IL-12, can provide significant protection against both Old World and New World cutaneous leishmaniasis (Campbell et al., 2003).

6. WD

Gene Name : WD
Sequence Strain (Species/Organism) : Leishmania amazonensis
VO ID : VO_0011290
NCBI Nucleotide GI : 38455440
NCBI Protein GI : 38455441
Protein Accession : AAR20840.1
Other Database IDs : CDD:225201
CDD:238121
CDD:293791
Taxonomy ID : 5659
Gene Strand (Orientation) : ?
Protein Name : antigenic WD protein
Protein pI : 6.52
Protein Weight : 70626.83
Protein Length : 743
Protein Note : WD40 repeat [General function prediction only]; COG2319

DNA Sequence : Show Sequence

>gi|38455440|gb|AY457171.1| Leishmania amazonensis antigenic WD protein mRNA, complete cds
CCCTCCCACTACCACGCAGCTTGTCATATCATACTGTGTTTCATTGTATGCATCCGGCGTGAGCAATTCT
TTGTGTCCTCAACCGCTCTTTCCTTTCGTCTTTCATCGCTGTGCTTCCTCTCCCCTCAGCGCCCCCCCCT
CTCCCTTCTCGCACACGCACGCACAGTGGCACAGATACATACAGAGGCACTCCTCTTCTCCCTACACTAT
CTTGCAGTGCCCTCTTCTTCCCCCGCCTCTTACTCTGCCCGTCCCTCTCCGACTCTCTTTCACGTTTCGC
AACACCAGTATCAACCATGGCCAAGGAGAAACTCGGCTTCAAGATCCAGGGGAAGACCGTCTCCTACGAG
GGTTACCCCGAGCGTCCGCCAAGGGACCTGATGGACATCTTGGTCAAAATGGGTGTCGGCGAAGCTGCCG
TGGTGACCACCTTTCTAGAAAATATTGAGAACCACCGGGAGTTCACCGACGAGCAGGTGATGGTGATGTA
CAACGAAGCAAAAATGACGCGCGAGAAGAAGGCTCGTGCGAAGGCCGGCAACCAGGGTGACTCAGCCGCT
GCCGCCGCCGCTCCATCGGGCGCCGCCGATGAGGCGGAGAAGCCGCAGAAGATCCTTCTTTCCTACCGCG
GCAAGCAGCTTTCCTACGTAGGTTTGCCCAGCGGCAAGCGCAACGCCGTGCTGCAACTCCTCTCCAAGCA
AGAGGAGCTCACCCGGGAGCTTGCAGTGGAGACGTTCCGCACGAACCTGCCCAGTGATCCGTCCACGGAG
GAGGAGATTTGGATCTTGGTGCAGGCAGTCAAAGATCGCAAGAGTAGGAAGAGCCAAGCGAAGGGTCAGG
ATGGCGAAAGGTCCGGCGGTAACGGTGCGACTGCGTCGCTTGGGGCAGGCAGCAGCGCCATGGACAGCAT
AACCGGTATGTCCGAGAGGGAGCGCGACGACCTGAACAACTTCATTCTTCAGGTGCTGCAGCAGCCAGCG
CCGGATGTGGCCGCCCTCTTACATGAGAATCCGAAACCGCGCAAAGACACGGAAGACAATGCGGAGCTGG
CACTGACAGCGCGCATGTCGGGTAAGATTCCGGTTTACCTGCAGGAAGAGAACACCAGCAGCACGAAGCT
CATCTACGCCACGCCCCACATGAGCTTCGAAGATTTTTCTACCATTGTCGAGAAGAAGTGTGGCCGCAAG
TTGTCGCTGTCCTTCTACGACGGCGACGACCTCATCATGATGGATGATGACGACGTGCTGGCCATGTTTC
TCGAAATGACAGTGCAGAACGACCCAAAGAAGGTGCGCCTCATTTGCTCCAACCCTGGAGTCCCAAGGAA
GGCGGCGGATGACCGGATGATGGAAAACCGTCAAAATACCTTCAGCGGTCTTCCGCAGGTCTCAACGACC
AAGGCATTGGCGTACTCGAACGGCGAGCTCAACGTCGCGGAGGCTCGCACCTACTTTGGCCACTCTCTTG
CCGTCTACTGCTGCTGCTTCTCGCCAAGGGGCGACATGTTCGTTACGGCAAGTCGCGACCGCACCGTCCG
CCTGTGGAACTTGCGCACCGGTGTCTCCACAGTCATGAAAGGCGGGCACAACGGCTTTGTGCTGTCTTGC
GACTACTCGCCAAAGGGCAACCGCGTCGCTTCCTCTTCTGATGACCGCACTATCAAACTGTGGAACACGT
CGTCATGCAACAAGGTTGCCACACTTAAAGGGCACGAGGACAAGGTATACTGCGTCAAGTACAACTCTAG
CGGAGATCTCCTCGTGTCCGCCTCTTGCGACACCACAGTGCGCGTGTGGAACGCCGAAAGTCAGGCGAAG
CTGGTGACGCTGAGGGGCCACACCCTCGCCGTTTTTTCCTGCGCCTTCAGCAACAGCGATAATGGAAAGT
TCGTCGTCTCCGGCAGCGATGACCGCGTTATAAAGCTGTGGGACTGGGGCGCCGGCAGAGAGATCCTGTC
GCTGGTGGGTCACATCGGCACCGTGTGGACGGTTGTCTTCTCCCACAACGACAGGTACGTTCTCAGTGGC
AGCATGGACTATGAGCTCATTCTCTGGGATTCAATGACTGGCGCTCGCCTGCGCTCGATGGACGGGCACA
AGACATCGGTGCACCACGCCATCTTCTCCGAGGACGACGCATACATCTTCTCCTGCGCCCGCGACTGGAG
CGTCATGGTGTGGCGCACCGAAGATGGCGAGCATGTAGAGACCATCATCGGTCACCTCAGCACTGTCTAC
CACATGGACATCAGCGAGGACAAGTTGCTCACCTGCTCTCTTGATGACAAGATCAAGCTGTGGAATATCA
GGCGCCACTGAAGTTGAGCACCAGCGGAAAATGTGGTAGTGCGAACACTGGCGGGAGGGGCACGTGCCAG
CCGCAACCGAGAAAGAGGTGTGGGCGGGAAGGCACGTGATAGGTGATACAGACACTGTACCCGTACACCA
TTCCTCTTTTCAGCAGCAGTCCTGCACACAGAAAGACACTCGGATTTTTCATTGACAAATCAGGTGGGTC
TGAAGACGATTTCAAGAAGAAGCGAGCGTTCGTATGTACCGCCCTCCTCCCTCCACCGATACTGCAACAC
ACACACACACACATTTTACGGTCTTGTCTACTTTTCTTCGTTTCGCTCTCGTCAGTTCCACCGAGTCCTG
GCTGCGTTTGGGTGCCACGCGTGTGAATGTGTGAGCGCCACCGTTGGTCGACCCTTTTCGCTTTCCTGCA
TGCTTTTTTTCTTTTCGTATCTCTACGTCCTGCTCACAGCGCATCGACCTCTGCCAGAGGATATCATTTG
GCAGTGTCGTTTCTTTTGCTCTTTTTCTCTCTTTGCCACCGTTCTTGCGGGCCAACGGCACCGTGCTCCC
CGCTTCTCTGTCTCGCGACCTCGACATTCATCTCTTCCATGCTTCTCCCTCTCCCCCTCCGTCTCCCCAG
CTTTTTGTCGTTTGTGCATTCTCCTCTTTTCTTAAGTGATGCTTCAAAGGAAACGAAAAAAAAAAAAAAA
AA

Protein Sequence : Show Sequence

>AAR20840.1 antigenic WD protein [Leishmania amazonensis]
MAKEKLGFKIQGKTVSYEGYPERPPRDLMDILVKMGVGEAAVVTTFLENIENHREFTDEQVMVMYNEAKM
TREKKARAKAGNQGDSAAAAAAPSGAADEAEKPQKILLSYRGKQLSYVGLPSGKRNAVLQLLSKQEELTR
ELAVETFRTNLPSDPSTEEEIWILVQAVKDRKSRKSQAKGQDGERSGGNGATASLGAGSSAMDSITGMSE
RERDDLNNFILQVLQQPAPDVAALLHENPKPRKDTEDNAELALTARMSGKIPVYLQEENTSSTKLIYATP
HMSFEDFSTIVEKKCGRKLSLSFYDGDDLIMMDDDDVLAMFLEMTVQNDPKKVRLICSNPGVPRKAADDR
MMENRQNTFSGLPQVSTTKALAYSNGELNVAEARTYFGHSLAVYCCCFSPRGDMFVTASRDRTVRLWNLR
TGVSTVMKGGHNGFVLSCDYSPKGNRVASSSDDRTIKLWNTSSCNKVATLKGHEDKVYCVKYNSSGDLLV
SASCDTTVRVWNAESQAKLVTLRGHTLAVFSCAFSNSDNGKFVVSGSDDRVIKLWDWGAGREILSLVGHI
GTVWTVVFSHNDRYVLSGSMDYELILWDSMTGARLRSMDGHKTSVHHAIFSEDDAYIFSCARDWSVMVWR
TEDGEHVETIIGHLSTVYHMDISEDKLLTCSLDDKIKLWNIRRH

Molecule Role : Protective antigen
Molecule Role Annotation : DNA vaccination in BALB/c mice with the LAWD (for Leishmania antigenic WD protein) and IL-12 genes significantly delayed lesion development from challenge with Leishmania amazonensis, which correlated with a dramatic reduction in parasite burdens (Campbell et al., 2004).
Related Vaccine(s): L. amazonensis DNA vaccine encoding WD protein

III. Vaccine Information

1. L. amazonensis CP Protein Vaccine

a. Vaccine Ontology ID:

VO_0004199

b. Type:

Subunit vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

mouse

e. Antigen

A 500 bp fragment encoding an isoform of cysteine proteinase (CP) from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24 (Fedeli et al., 2010).

f. Gene Engineering of CP cysteine proteinase

Type: Recombinant protein preparation
Description:
Detailed Gene Information: Click here.

g. Adjuvant:

VO ID: VO_0000139

h. Immunization Route

subcutaneous injection

i. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Female BALB/c mice (six per group) were immunized thrice with a 2 week interval with 25 μg of rLacys24 plus complete Freund’s adjuvant (CFA) by the subcutaneous route in the base of the tail. Control animals received PBS or only adjuvant (Fedeli et al., 2010).
Challenge Protocol: Two weeks after the last dose animals were challenged with 2.5 × 10^5 L. (L.) amazonensis amastigotes in PBS in the hind footpad (Fedeli et al., 2010).
Efficacy: Immunization of BALB/c mice with rLacys24 plus CFA adjuvant resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice (Fedeli et al., 2010).

2. L. amazonensis DNA vaccine encoding GP46

a. Vaccine Ontology ID:

VO_0011354

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

mouse

e. Antigen

L. amazonensis gp46

f. Gene Engineering of Gp46

Type: DNA vaccine construction
Description: cDNAs encoding L. m. amazonensis gp46 antigen was subcloned into the VR1012 plasmid (Dumonteil et al., 2000)
Detailed Gene Information: Click here.

g. Vector:

VR1012 plasmid

h. Immunization Route

Intramuscular injection (i.m.)

i. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Susceptible BALB/c mice were immunised with two i.m. injections of 100 microg of plasmid DNA (Dumonteil et al., 2000).
Challenge Protocol: Mice were challenged by the injection of 4 x 10^6 L. m. mexicana parasites in the foot pad to evaluate protection (Dumonteil et al., 2000).
Efficacy: Measurement of lesion size indicated that mice immunised with VR012-GP46 were partially protected against infection (Dumonteil et al., 2000).

3. L. amazonensis DNA vaccine encoding WD protein

a. Vaccine Ontology ID:

VO_0011349

b. Type:

DNA vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

mouse

e. Antigen

L. amazonensis WD

f. Gene Engineering of WD

Type: DNA vaccine construction
Description: The whole gene insert from the pBK-CMV clone containing the LAWD gene was subcloned into the pET32a plasmid (Invitrogen) for His tag protein expression. To ensure that the ORF would be read in frame, the plasmid was first digested with BamHI and then blunt-ended by a Klenow reaction (Invitrogen) (Campbell et al., 2004).
Detailed Gene Information: Click here.

g. Vector:

pcDNA3.1/Hygro(−) (Invitrogen)

h. Immunization Route

Subcutaneous injection

i. Mouse Response

Host Strain: BALB/c
Vaccination Protocol: Mice (five per group) were immunized in five locations with a total of 100 μg of DNA (50 μg of LAWD and 50 μg of IL-12) per mouse: four injections in both sides of the inner and outer thigh muscles of the hind legs (∼50 μl/site) and one subcutaneous injection in the left hind foot (∼5 μl/site) (Campbell et al., 2004).
Challenge Protocol: Mice were boosted twice at 3-week intervals and then challenged 3 weeks after the last immunization with 2 × 10^5 metacyclic promastigotes in the right hind foot (Campbell et al., 2004).
Efficacy: DNA vaccination in BALB/c mice with the LAWD (for Leishmania antigenic WD protein) and IL-12 genes significantly delayed lesion development from challenge with Leishmania amazonensis, which correlated with a dramatic reduction in parasite burdens (Campbell et al., 2004).

4. L. amazonensis M2 protein vaccine

a. Vaccine Ontology ID:

VO_0011355

b. Type:

Subunit vaccine

c. Status:

Research

d. Host Species as Laboratory Animal Model:

mouse

e. Antigen

L. amazonensis M2

f. Gene Engineering of M2

Type: Recombinant protein preparation
Description: M-2, a 46-kDa promastigote-specific glycoprotein was isolated. The protein was further purified by removal of detergent with an anionexchange column(Champsi and McMahon-Pratt, 1988).
Detailed Gene Information: Click here.

g. Adjuvant:

VO ID: VO_0001259

h. Immunization Route

Intraperitoneal injection (i.p.)

i. Mouse Response

Host Strain: CBA
Vaccination Protocol: BALB/c, CBA, and C57BL/6 mice were immunized intraperitoneally with M-2 at a final concentration of 0.03 mg/ml. The amount of C. parvum used in immunizations 2 and 3 was reduced to 0.05 mg per immunization (Champsi and McMahon-Pratt, 1988).
Challenge Protocol: Animals were rested for 2 to 4 weeks after final immunization and challenged in the right hindfoot with late-log-phase promastigotes. Parasites used for infections were passaged a maximum of four times. Challenge doses of 10^3, 10^4, 10^5, and 10^6 were used (Champsi and McMahon-Pratt, 1988).
Efficacy: Immunization of CBA mice with the M-2 glycoprotein of L. amazonensis and C. parvum adjuvant resulted in complete protection against a challenge infection of 10^4 and 10^6 late log-phase promastigotes of L. amazonensis. In the BALB/c strain, complete protection was observed in some of the immunized animals (28 to 50%); in the rest of the mice the onset of infection was significantly delayed. Protective immunity for C57BL/6 mice was observed only at the low infecting dose (10(4) L. amazonensis organisms) (Champsi and McMahon-Pratt, 1988).

IV. References

1. Campbell et al., 2003: Campbell K, Diao H, Ji J, Soong L. DNA immunization with the gene encoding P4 nuclease of Leishmania amazonensis protects mice against cutaneous Leishmaniasis. Infection and immunity. 2003; 71(11); 6270-6278. [PubMed: 14573646].

2. Campbell et al., 2004: Campbell K, Popov V, Soong L. Identification and molecular characterization of a gene encoding a protective Leishmania amazonensis Trp-Asp (WD) protein. Infection and immunity. 2004; 72(4); 2194-2202. [PubMed: 15039343].

3. Champsi and McMahon-Pratt, 1988: Champsi J, McMahon-Pratt D. Membrane glycoprotein M-2 protects against Leishmania amazonensis infection. Infection and immunity. 1988; 56(12); 3272-3279. [PubMed: 3182080].

4. Duarte et al., 2017: Duarte MC, Lage DP, Martins VT, Costa LE, Carvalho AMRS, Ludolf F, Santos TTO, Vale DL, Roatt BM, Menezes-Souza D, Fernandes AP, Tavares CAP, Coelho EAF. A vaccine composed of a hypothetical protein and the eukaryotic initiation factor 5a from Leishmania braziliensis cross-protection against Leishmania amazonensis infection. Immunobiology. 2017; 222(2); 251-260. [PubMed: 27693018].

5. Dumonteil et al., 2000: Dumonteil E, Andrade-Narvarez F, Escobedo-Ortegon J, Ramirez-Sierra MJ, Valencia-Pacheco G, Flores-Serrano A, Canto-Lara S, Arjona-Torres A. Comparative study of DNA vaccines encoding various antigens against Leishmania mexicana. Developments in biologicals. 2000; 104; 135-141. [PubMed: 11713811].

6. Fedeli et al., 2010: Fedeli CE, Ferreira JH, Mussalem JS, Longo-Maugéri IM, Gentil LG, dos Santos MR, Katz S, Barbiéri CL. Partial protective responses induced by a recombinant cysteine proteinase from Leishmania (Leishmania) amazonensis in a murine model of cutaneous leishmaniasis. Experimental parasitology. 2010; 124(2); 153-158. [PubMed: 19735658].