Induction of immunity against experimental Chlamydia pneumoniae infection

7 1. REVIEW OF THE LITERATURE . 8 1.1. Vaccines against intracellular microbes. 8 1.2. Chlamydiae, common human pathogens . 8 1.2. 1. Chlamydiae. 8 1.2. 2. Acute infection caused by C. pneumoniae . 11 1.2. 3. Chronic infection caused by C. pneumoniae . 12 1.3. Vaccination against C. pneumoniae . 13 1.3. 1. History of Chlamydial vaccine . 13 1.3. 2. The correlates of protective immunity to C. pneumoniae. 16 1.4. Vaccines inducing cellular immunity . 21 1.4.1. General requirements . 21 1.4.2. DNA vaccines . 21 1.4.3. Viral vectors . 23 1.4.4. Heterologous prime-boost regimens. 25 1.5. Antigenic proteins of C. pneumoniae  possible vaccine candidates. 26 1.5. 1. Outer membrane proteins. 27 1.5. 2. Inclusion membrane proteins (Incs) . 29 1.5. 3. Type III secretion system (TTSS) . 29 1.5. 4. Heat shock proteins (Hsps). 30 2. AIMS OF THE STUDY.32 3. MATERIALS AND METHODS.33 4. RESULTS AND DISCUSSION.37 4.1. Selection of ORFs . 37 4.2. Production of C. pneumoniae proteins in B. subtilis (I, IV) . 38 4.3. Utilization of C. pneumoniae recombinant proteins in immunological assays (I, IV) . 39 4.3. 1. Assays for CMI . 39 4.3. 2. Use of recombinant C. pneumoniae proteins in characterization of murine systemic and mucosal antibody response after infection. 39 4.4. DNA immunization against C. pneumoniae in mice (II) . 42 4.4.1. In vitro protein expression. 43 4.4.2. Immune response to immunization . 44 4.4.3. Response to challenge . 45 4.5. DNA priming and rSFV boosting (III) . 45 4.5. 1. In vitro protein expression. 47 4.5. 2. Immune response to immunization . 47 4.5. 3. Response to challenge . 48 4.6. Why only partial protection was achieved? 50 5. CONCLUDING REMARKS AND FUTURE PROSPECTS . 53 6. ACKNOWLEDGEMENTS.54 7. REFERENCES.55 Induction of immunity against experimental Chlamydia pneumoniae infection 4 List of original publications This thesis is based on the following original articles referred to in the text by their Roman numerals. The articles in this thesis have been reproduced with the permission of the copyright holders. In addition, some unpublished data are presented. I Airaksinen U, Penttilä T, Wahlström E, Jackson L, Vuola JM, Puolakkainen M and Sarvas M (2003). Production of Chlamydia pneumoniae proteins in Bacillus subtilis and their use in characterizing immune responses in the experimental infection model. Clin and Diag Lab Immunol 10:367-375. II Penttilä T, Vuola JM, Puurula V, Anttila M, Sarvas M, Rautonen N, Mäkelä PH and Puolakkainen M (2000). Immunity to Chlamydia pnemoniae induced by vaccination with DNA vectors expressing the cytoplasmic protein (Hsp60) or outer membrane proteins (MOMP and Omp2). Vaccine 19:1256-1265. III Penttilä T*, Tammiruusu A*, Liljeström P, Sarvas M, Mäkelä PH, Vuola JM and Puolakkainen M (2004). DNA immunization followed by a viral vector booster in a Chlamydia pneumoniae mouse model. Vaccine 22:3386-3394. * equal contribution IV Penttilä T, Wahlström E, Vuola JM, Sarvas M and Puolakkainen M. Systemic and local antigen-specific antibody response in murine Chlamydia pneumoniae infection (submitted). Induction of immunity against experimental Chlamydia pneumoniae infection 5