Chlamydophila pneumoniae is a species of Chlamydophila bacteria[1][2][3] that infects humans and is a major cause of pneumonia. C. pneumoniae has a complex life cycle and must infect another cell in order to reproduce and thus is classified as an obligate intracellular pathogen. This atypical bacterium commonly causes pharyngitis, bronchitis and atypical pneumonia mainly in elderly and debilitated patients but in healthy adults also. C. pneumoniae infection has been implicated in several chronic lung diseases by serology and direct antigen detection. Acute lower respiratory tract infection caused by C. pneumoniae seems often to precede attacks of asthma in both children and adults but is also involved in some exacerbations of chronic bronchitis. More importantly it seems to be strongly associated with chronic obstructive lung disease irrespective of exacerbation status. Moreover, persistently elevated C. pneumoniae antibody titers have been observed in sarcoidosis and lung cancer (Wiki: Chlamydophila pneumoniae).
4. Microbial Pathogenesis
Chlamydophila pneumoniae is a small bacterium (0.2 to 1 micrometer) that undergoes several transformations during its life cycle. It exists as an elementary body (EB) in between hosts. The EB is not biologically active but is resistant to environmental stresses and can survive outside of a host for a limited time. The EB travels from an infected person to the lungs of a non-infected person in small droplets and is responsible for infection. Once in the lungs, the EB is taken up by cells in a pouch called an endosome by a process called phagocytosis. However, the EB is not destroyed by fusion with lysosomes as is typical for phagocytosed material. Instead, it transforms into a reticulate body and begins to replicate within the endosome. The reticulate bodies must utilize some of the host's cellular machinery to complete its replication. The reticulate bodies then convert back to elementary bodies and are released back into the lung, often after causing the death of the host cell. The EBs are thereafter able to infect new cells, either in the same organism or in a new host. Thus, the life cycle of C. pneumoniae is divided between the elementary body, which is able to infect new hosts but can not replicate, and the reticulate body ,which replicates but is not able to cause new infection (Wiki: Chlamydophila pneumoniae).
5. Host Ranges and Animal Models
In addition to infecting humans, C. pneumoniae also infects and causes disease in Koalas, emerald tree boa (Corallus caninus), iguanas, chameleons, frogs, and turtles (Wiki: Chlamydophila pneumoniae).
Molecule Role Annotation :
Results of this study showed that intranasal immunization of BALB/c mice with heat-aggregated CopN protein and an Escherichia coli heat-labile toxin (LT) induced a strong immune response. The immunization induced statistically significant protection against intranasal C. pneumoniae challenge, the level of which correlated with the magnitude of CopN-specific lymphocyte proliferation (Tammiruusu et al., 2007).
Molecule Role Annotation :
Four of the six in vitro neutralizing antigens (Pmp2, Pmp10, OmpH-like and enolase) could inhibit Cpn dissemination in a hamster model. The results show that these Cpn proteins are immunoaccessible in infectious EBs, and recommend further investigation on their value as vaccine components.(Finco et al., 2005)
Molecule Role Annotation :
M-ID vaccination with fabD generated a response that resulted in moderately, but significantly reduced total C. pneumoniae lung loads as compared to control mice vaccinated with a plasmid expressing a non-Chlamydia ORF (p ≤ 0.019). This resulted in the ability of fabD to mediate a moderate, but statistically significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
Molecule Role Annotation :
Immunization of mice with a single construct containing multiple epitopes derived from ApoB100, hHSP60 and Cpn was more effective in reducing early atherosclerotic lesions.(Lu et al., 2012)
Immunization with pmomp or phsp60 showed 1.2–1.5 log reduction in the mean lung bacterial counts after the challenge.(Penttilä et al., 2000)
>NP_224529.1 low calcium response protein E [Chlamydia pneumoniae CWL029]
MAASGGTGGLGGTQGVNLAAVEAAAAKADAAEVVASQEGSEMNMIQQSQDLTNPAAATRTKKKEEKFQTL
ESRKKGEAGKAEKKSESTEEKPDTDLADKYASGNSEISGQELRGLRDAIGDDASPEDILALVQEKIKDPA
LQSTALDYLVQTTPPSQGKLKEALIQARNTHTEQFGRTAIGAKNILFASQEYADQLNVSPSGLRSLYLEV
TGDTHTCDQLLSMLQDRYTYQDMAIVSSFLMKGMATELKRQGPYVPSAQLQVLMTETRNLQAVLTSYDYF
ESRVPILLDSLKAEGIQTPSDLNFVKVAESYHKIINDKFPTASKVEREVRNLIGDDVDSVTGVLNLFFSA
LRQTSSRLFSSADKRQQLGAMIANALDAVNINNEDYPKASDFPKPYPWS
Molecule Role :
Protective antigen
Molecule Role Annotation :
The immunogenicity and protective effect of recombinant LcrE protein combined either with Freund's or Alum adjuvant were investigated in mice. The immunization with both protocols resulted in a significant reduction of the number of viable C. pneumoniae in the lungs after challenge. Results confirm that LcrE induces protective immunity in mice (Faludi et al., 2009).
DNA immunization given as a priming and followed by a protein booster significantly reduced the number of viable bacteria in the lungs after challenge with C. pneumoniae. These results confirm that immunization with pΔRCLcrE can be an effective part of a vaccination schedule against C. pneumoniae. (Faludi and Szab�, 2011)
>WP_010883333.1 porin [Chlamydia pneumoniae]
MKKLLKSALLSAAFAGSVGSLQALPVGNPSDPSLLIDGTIWEGAAGDPCDPCATWCDAISLRAGFYGDYV
FDRILKVDAPKTFSMGAKPTGSAAANYTTAVDRPNPAYNKHLHDAEWFTNAGFIALNIWDRFDVFCTLGA
SNGYIRGNSTAFNLVGLFGVKGTTVNANELPNVSLSNGVVELYTDTSFSWSVGARGALWECGCATLGAEF
QYAQSKPKVEELNVICNVSQFSVNKPKGYKGVAFPLPTDAGVATATGTKSATINYHEWQVGASLSYRLNS
LVPYIGVQWSRATFDADNIRIAQPKLPTAVLNLTAWNPSLLGNATALSTTDSFSDFMQIVSCQINKFKSR
KACGVTVGATLVDADKWSLTAEARLINERAAHVSGQFRF
Molecule Role :
Protective antigen
Molecule Role Annotation :
The use of MOMP in C. pneumoniae as a possible vaccine target and the role of MOMP-derived peptides as vaccine candidates for immune-therapy in chronic inflammation.(Atanu et al., 2013)
Molecule Role Annotation :
Four of the six in vitro neutralizing antigens (Pmp2, Pmp10, OmpH-like and enolase) could inhibit Cpn dissemination in a hamster model. The results show that these Cpn proteins are immunoaccessible in infectious EBs, and recommend further investigation on their value as vaccine components.(Finco et al., 2005)
Protein Note :
PknD; responsible for phosphorylation of proteins on serine and threonine residues; similar to eukaryotic Ser/Thr kinases; in Chlamydia trachomatis itseems to interact with Pkn1, another serine/threonine-protein kinase
Molecule Role Annotation :
IM-ID vaccination with CPn0095 (pknD) generated a response that resulted in moderately, but significantly reduced total C. pneumoniae lung loads as compared to control mice vaccinated with a plasmid expressing a non-Chlamydia ORF (p ≤ 0.019). This resulted in the ability of CPN0095 to mediate a moderate, but statistically significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
Molecule Role Annotation :
Four of the six in vitro neutralizing antigens (Pmp2, Pmp10, OmpH-like and enolase) could inhibit Cpn dissemination in a hamster model. The results show that these Cpn proteins are immunoaccessible in infectious EBs, and recommend further investigation on their value as vaccine components.(Finco et al., 2005)
Molecule Role Annotation :
Four of the six in vitro neutralizing antigens (Pmp2, Pmp10, OmpH-like and enolase) could inhibit Cpn dissemination in a hamster model. The results show that these Cpn proteins are immunoaccessible in infectious EBs, and recommend further investigation on their value as vaccine components.(Finco et al., 2005)
>NP_224586.1 single-stranded DNA-binding protein [Chlamydia pneumoniae CWL029]
MMFGHFAGYLGADPEERMTSKGKRVITLRLGVKTRVGMKDETVWCKCNIWHNRYDKMLPYLKKGSGVIVA
GDISVESYMSKDGSPQSSLVISVDSLKFSPFGRNEGSRSPSLEDNHQQVGYESVSVGFEGEALDAEAIKD
KDMYAGYGQEQQYVCEDVPF
Molecule Role :
Protective antigen
Molecule Role Annotation :
Mice vaccinated with candidate gene ssb showed significant reduction of spleen chlamydial loads as compared to naïve, non-protected control mice (p ≤ 0.048). This resulted in the ability of ssb to mediate a modest, but significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
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).
Description: C. pneumoniae CopN (gene lcrE; position 0324 of C. pneumoniae CWL029), was produced in a Bacillus subtilis protein expression system as a soluble protein. Recombinant CopN protein was dissolved in PBS at a concentration of 1 mg/ml and heated to 100 °C for 10 min after which the visible precipitation of protein was discernible. C. pneumoniae preparation was boiled for 10 min in a water bath at a concentration of 2.5 × 10^7 IFU/ml in SPG. E. coli heat-labile toxin, LT (kindly provided by Prof. G. Dougan, Imperial Collage, London, UK) was added to heat-aggregated protein suspension to a final concentration of 12.5 μg/ml (Tammiruusu et al., 2007).
Vaccination Protocol:
Mice were immunized intranasally with 40 μg of heat-aggregated CopN/ 40 μl dose or 106 heat-treated C. pneumoniae inclusion forming unit (IFU) (approximately 1 μg of protein)/40 μl dose. Mice immunized intranasally with disrupted HL cells (Mock) or PBS were used as control. Fourteen days after the first immunization, the mice were boosted once with the same dose of antigen. All immunizations were performed under methoxyflurane anaesthesia (Metofane, Pitman-Moore, Mundelein, IL, USA) (Tammiruusu et al., 2007).
Challenge Protocol:
At 14 days after the second immunization, the mice were challenged intranasally with 10^5 IFU of C. pneumoniae in 40 μl of SPG under Metofane anaesthesia. At certain time points after infection, three to six mice were sacrificed, lungs were mechanically homogenized in SPG and dilutions of lung supernatant were cultured on HL cell monolayers (Tammiruusu et al., 2007).
Efficacy:
Intranasal immunization of BALB/c mice with heat-aggregated CopN protein and an Escherichia coli heat-labile toxin (LT) induced a strong immune response. The immunization induced statistically significant protection against intranasal C. pneumoniae challenge, the level of which correlated with the magnitude of CopN-specific lymphocyte proliferation (Tammiruusu et al., 2007).
Host Gene Response of
Ifng (Interferon gamma)
Gene Response:
The results of this study showed that intranasal immunization of BALB/c mice with heat-aggregated CopN protein and an Escherichia coli heat-labile toxin (LT) induced a strong immune response, detected as IFN-gamma production. The response was significant as compared to PBS-vaccinated mice in the lungs, spleen, and mediastinal lymph nodes 14 days after challenge (Tammiruusu et al., 2007).
Description:
The genome sequence of C. pneumoniae isolate CDC/CWL-029 (ATCC strain VR-1310) was extracted from Genbank (AE001363, 1,230,230 bp). The 1052 annotated genes of C. pneumoniae were imported into a gene-splitting and primer prediction program; primer pairs to amplify 1263 ORFs of 1.5 kb or less were exported. A 1.5 kb maximum ORF length was chosen to ensure sufficient PCR quality and yields, and this generated a few additional fragments (Li et al., 2006).
Vaccination Protocol:
For intranasal inoculation, mice received a light isoflurane inhalation anesthesia. Vaccine protection control mice were inoculated with a low dose of 5 × 10^6 C. pneumoniae elementary bodies in 30 μl SPG buffer (Li et al., 2006).
Challenge Protocol:
High-dose challenge infection was performed 4 weeks after the last gene gun genetic vaccination or low dose inoculation of live C. pneumoniae, and 6 weeks after the last intramuscular-intradermal genetic vaccination, by intranasal inoculation of 1 × 10^8 C. pneumoniae elementary bodies in 30 μl SPG buffer. Mice were sacrificed by CO2 inhalation 2 h, 3 days, 10 days, or 15 days after inoculation, and lungs and spleen were weighed, snap frozen in liquid nitrogen, and stored at −80 °C until further processing (Li et al., 2006).
Efficacy:
M-ID vaccination with fabD generated a response that resulted in moderately, but significantly reduced total C. pneumoniae lung loads as compared to control mice vaccinated with a plasmid expressing a non-Chlamydia ORF (p ≤ 0.019). This resulted in the ability of fabD to mediate a moderate, but statistically significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
C. pneumoniae serine/threonine-protein kinase, PknD
e. Gene Engineering of
PknD
Type:
DNA vaccine construction
Description:
The genome sequence of C. pneumoniae isolate CDC/CWL-029 (ATCC strain VR-1310) was extracted from Genbank (AE001363, 1,230,230 bp). The 1052 annotated genes of C. pneumoniae were imported into a gene-splitting and primer prediction program; primer pairs to amplify 1263 ORFs of 1.5 kb or less were exported. A 1.5 kb maximum ORF length was chosen to ensure sufficient PCR quality and yields, and this generated a few additional fragments (Li et al., 2006).
Vaccination Protocol:
For intranasal inoculation, mice received a light isoflurane inhalation anesthesia. Vaccine protection control mice were inoculated with a low dose of 5 × 10^6 C. pneumoniae elementary bodies in 30 μl SPG buffer (Li et al., 2006).
Challenge Protocol:
High-dose challenge infection was performed 4 weeks after the last gene gun genetic vaccination or low dose inoculation of live C. pneumoniae, and 6 weeks after the last intramuscular-intradermal genetic vaccination, by intranasal inoculation of 1 × 10^8 C. pneumoniae elementary bodies in 30 μl SPG buffer. Mice were sacrificed by CO2 inhalation 2 h, 3 days, 10 days, or 15 days after inoculation, and lungs and spleen were weighed, snap frozen in liquid nitrogen, and stored at −80 °C until further processing (Li et al., 2006).
Efficacy:
IM-ID vaccination with CPn0095 (pknD) generated a response that resulted in moderately, but significantly reduced total C. pneumoniae lung loads as compared to control mice vaccinated with a plasmid expressing a non-Chlamydia ORF (p ≤ 0.019). This resulted in the ability of CPN0095 to mediate a moderate, but statistically significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
C. pneumoniae single-stranded DNA-binding protein, ssb
e. Gene Engineering of
Ssb
Type:
DNA vaccine construction
Description:
The genome sequence of C. pneumoniae isolate CDC/CWL-029 (ATCC strain VR-1310) was extracted from Genbank (AE001363, 1,230,230 bp). The 1052 annotated genes of C. pneumoniae were imported into a gene-splitting and primer prediction program; primer pairs to amplify 1263 ORFs of 1.5 kb or less were exported. A 1.5 kb maximum ORF length was chosen to ensure sufficient PCR quality and yields, and this generated a few additional fragments (Li et al., 2006).
Vaccination Protocol:
For intranasal inoculation, mice received a light isoflurane inhalation anesthesia. Vaccine protection control mice were inoculated with a low dose of 5 × 10^6 C. pneumoniae elementary bodies in 30 μl SPG buffer (Li et al., 2006).
Challenge Protocol:
High-dose challenge infection was performed 4 weeks after the last gene gun genetic vaccination or low dose inoculation of live C. pneumoniae, and 6 weeks after the last intramuscular-intradermal genetic vaccination, by intranasal inoculation of 1 × 10^8 C. pneumoniae elementary bodies in 30 μl SPG buffer. Mice were sacrificed by CO2 inhalation 2 h, 3 days, 10 days, or 15 days after inoculation, and lungs and spleen were weighed, snap frozen in liquid nitrogen, and stored at −80 °C until further processing (Li et al., 2006).
Efficacy:
Mice vaccinated with candidate gene ssb showed significant reduction of spleen chlamydial loads as compared to naïve, non-protected control mice (p ≤ 0.048). This resulted in the ability of ssb to mediate a modest, but significant level of protection in an inbred A/J mouse respiratory challenge model (Li et al., 2006).
Efficacy:
Immunization with this vaccine resulted in signi®cant protection, as measured by a lower bacterial load and a less severe pathological outcome after infection with C. pneumoniae (Svanholm et al., 2000).
Description:
A 1218-kb DNA fragment containing the lcrE gene (GenBank ID 15618244, Locus tag CPn0324) was amplified by PCR, using C. pneumoniae (CWL029 ATCC) DNA as template. The PCR was performed in a GeneAmp II (Applied Biosystems, Foster City, CA, USA) thermocycler with Advantage GC cDNA polymerase (Clontech, Mountain View, CA, USA), and the amplification conditions were set as recommended by the manufacturer. The amplicon was digested with NdeI and BamHI and inserted into p6HisF-11d (icl) pET vector by digesting it with the same enzymes and replacing the icl gene (Faludi et al., 2009).
Vaccination Protocol:
The mice in groups of 25 were immunized subcutaneously into the tail base with the purified LcrE protein diluted in phosphate buffered saline (PBS) at a dose of 20 μg mixed with 25 μl Alum (Aluminum hydroxide Gel, Sigma) or 75 μl Freund's adjuvants (Chemicon International, Temecula, CA, USA; 1st inoculation with complete and 2nd and 3rd inoculations with incomplete Freund's adjuvant) in 0.15-ml volume 3 times at 3-week intervals (Faludi et al., 2009).
Challenge Protocol:
Two weeks after the last immunization, the immunized and non-immunized mice (absolute naive animals) were challenged with 4×10^5 inclusion forming unit (IFU) C. pneumoniae (CWL029, ATCC) in 25 μl PBS intranasally under pentobarbital sodium anesthesia (Faludi et al., 2009).
Efficacy:
The immunogenicity and protective effect of recombinant LcrE protein combined either with Freund's or Alum adjuvant were investigated in mice. The immunization with both protocols resulted in a significant reduction of the number of viable C. pneumoniae in the lungs after challenge. Results confirm that LcrE induces protective immunity in mice (Faludi et al., 2009).
Host Gene Response of
Ifng (Interferon gamma)
Gene Response:
The presence of LcrE-specific IFN-gamma-producing cells in LcrE+Alum-immunized mice indicates Th1 type response. IFN-gamma responses were measured in spleen cells collected 2 weeks after last immunization and were significantly higher than mock-immunized mice (Faludi et al., 2009).
Gene Response:
LcrE-specific IgA level was higher in both the sera and the lungs after using Freund's adjuvant than non-immunized mice at the time of the challenge (Faludi et al., 2009).
1. Atanu et al., 2013: Atanu FO, Oviedo-Orta E, Watson KA. A novel transport mechanism for MOMP in Chlamydophila pneumoniae and its putative role in immune-therapy. PloS one. 2013; 8(4); e61139. [PubMed: 23637791].
2. Faludi and Szabó, 2011: Faludi I, Szabó ÁM. Vaccination with DNA vector expressing chlamydial low calcium response protein E (LcrE) against Chlamydophila pneumoniae infection. Acta microbiologica et immunologica Hungarica. 2011; 58(2); 123-134. [PubMed: 21715282].
3. Faludi et al., 2009: Faludi I, Burian K, Csanadi A, Miczak A, Lu X, Kakkar VV, Gonczol E, Endresz V. Adjuvant modulation of the immune response of mice against the LcrE protein of Chlamydophila pneumoniae. International journal of medical microbiology : IJMM. 2009; 299(7); 520-528. [PubMed: 19451031].
4. Finco et al., 2005: Finco O, Bonci A, Agnusdei M, Scarselli M, Petracca R, Norais N, Ferrari G, Garaguso I, Donati M, Sambri V, Cevenini R, Ratti G, Grandi G. Identification of new potential vaccine candidates against Chlamydia pneumoniae by multiple screenings. Vaccine. 2005; 23(9); 1178-1188. [PubMed: 15629361].
5. Li et al., 2006: Li D, Borovkov A, Vaglenov A, Wang C, Kim T, Gao D, Sykes KF, Kaltenboeck B. Mouse model of respiratory Chlamydia pneumoniae infection for a genomic screen of subunit vaccine candidates. Vaccine. 2006; 24(15); 2917-2927. [PubMed: 16434129].
6. Lu et al., 2012: Lu X, Xia M, Endresz V, Faludi I, Szabo A, Gonczol E, Mundkur L, Chen D, Kakkar V. Impact of multiple antigenic epitopes from ApoB100, hHSP60 and Chlamydophila pneumoniae on atherosclerotic lesion development in Apob(tm2Sgy)Ldlr(tm1Her)J mice. Atherosclerosis. 2012; 225(1); 56-68. [PubMed: 22959702].
7. Penttilä et al., 2000: Penttilä T, Vuola JM, Puurula V, Anttila M, Sarvas M, Rautonen N, Mäkelä PH, Puolakkainen M. Immunity to Chlamydia pneumoniae induced by vaccination with DNA vectors expressing a cytoplasmic protein (Hsp60) or outer membrane proteins (MOMP and Omp2). Vaccine. 2000; 19(9-10); 1256-1265. [PubMed: 11137265].
8. Svanholm et al., 2000: Svanholm C, Bandholtz L, Castaños-Velez E, Wigzell H, Rottenberg ME. Protective DNA immunization against Chlamydia pneumoniae. Scandinavian journal of immunology. 2000; 51(4); 345-353. [PubMed: 10736106].
9. Tammiruusu et al., 2007: Tammiruusu A, Penttilä T, Lahesmaa R, Sarvas M, Puolakkainen M, Vuola JM. Intranasal administration of chlamydial outer protein N (CopN) induces protection against pulmonary Chlamydia pneumoniae infection in a mouse model. Vaccine. 2007; 25(2); 283-290. [PubMed: 16949182].
10. Tammiruusu et al., 2007: Tammiruusu A, Penttilä T, Lahesmaa R, Sarvas M, Puolakkainen M, Vuola JM. Intranasal administration of chlamydial outer protein N (CopN) induces protection against pulmonary Chlamydia pneumoniae infection in a mouse model. Vaccine. 2007; 25(2); 283-290. [PubMed: 16949182].
