1. Anti-Chlamydia Vaccine: From Bench to Bedside
Ashlesh Murthy, M.B.B.S., Ph.D
Research Assistant Professor
University of Texas at San Antonio

3. Chlamydia Obligate intracellular pathogen
C. trachomatis, C. pneumoniae, C. psittaci, C. pecorum, C. muridarum
C. trachomatis multiple serovars: A-K, L1-3

4. Chlamydia trachomatis Infections
Trachoma and Inclusion Conjunctivitis
Serovars - A, B, Ba, C
Urogenital Infections
Serovars - D, Da, E, F, G, Ga, H, I, J, K
LymphoGranuloma Venereum
Serovars - L1, L2, L3

5. Global Prevalence of Chlamydial STD

6. Chlamydial STD
Adapted from

7. Chlamydial STD-Ascending Infection
Eskilsden and Gupta et al., In Revision

8. Chlamydial STD-Ascending Infection
PID
No Symptoms
Dye alone
Dye+Chlamydia
Vaginitis
Cervicitis
Endometritis
Salpingitis
Antimicrobials

9. Chlamydial STD-Reproductive Damage
Pelvic Inflammatory Disease
Ectopic pregnancy
Infertility
Normal

10. Chlamydial STD- Statistics
90 million cases
50-70% of patients initially asymptomatic: not treated
40% of untreated females – pelvic inflammatory disease
20% of PID cases- infertility
Adapted from STD statistics

11. Anti-Chlamydial Vaccine

12. Goals for Chlamydial Vaccine
No Symptoms (75%)
Immunity
Transmission
Symptoms (25%)
Re-Infection
Sequelae
Infection
Prevent infection (Sterilizing immunity)- ideal
Reduce transmission / Duration of shedding- practical
Prevent pathological sequelae
Protection against multiple serovars

13. Chlamydial Vaccine Trials of 1960s
Formalin-killed whole chlamydial organisms
Reduction in incidence for ~ 1 year
Comparable/exaggerated ocular pathology in vaccinees
Focus on sub-unit vaccines

14. Chlamydial Developmental Cycle

15. Vaccines Against Chlamydial STD
Major outer membrane protein (MOMP)
Chlamydia-purified MOMP
Recombinant MOMP
MOMP DNA
MOMP synthetic peptides
MOMP plus outer membrane protein 2 (omp2)
Chlamydia-purified MOMP refolded to native configuration
Cons: MOMP - serovar-specific
Currently no licensed vaccine against C. trachomatis
Need for identification of new vaccine candidates

16. Chlamydial Developmental Cycle

17. Chlamydial Protease-Like Activity Factor (CPAF)
Secreted into host cytosol
Nucleus
Inclusion
CPAF
Overlay

18. CPAF-Human Antibody Responses
Sharma et al., Infect Immun. 2006

19. CPAF-Highly Conserved
Dong et al., Infect Immun. 2005

20. CPAF- A Potential Vaccine Candidate

21. Mouse Model of Chlamydial Infection
C. muridarum
106
Vaginal chlamydial shedding
Oviduct dilatation 30 80
Days after challenge

22. Protocol For Vaccination Studies
Intranasal
Day –1: IL-12 (0.5 mg)
Day 0:rCPAF(15mg)+ IL-12 (0.5 mg)
Day +1: IL-12 (0.5 mg)
Recombinant CPAF from C. trachomatis L2
cloned into E. coli
Day 14: rCPAF (15 mg)+IL-12 (0.5 mg)
Day 28: rCPAF (15 mg)+ IL-12 (0.5 mg)
30 day Rest
Vaginal challenge
Estimation of bacterial shedding
Examination of disease pathology
Day 60: 5X104 IFU C. muridarum

23. Early Responses to Immunization

24. CPAF Vaccine : Robust IFN-g Response
Murthy et al., Infect Immun. 2007

25. CPAF Vaccine: Systemic Antibodies
Murthy et al., Infect Immun. 2007

26. CPAF Vaccine: Mucosal Antibodies
Murthy et al., Infect Immun. 2007

27. CPAF Vaccine: Chlamydial Clearance
Murthy et al., Infect Immun. 2007

28. CPAF+IL-12: Prevents Hydrosalpinx
Murthy et al., Infect Immun. 2007

29. CPAF Vaccine: Preserves Fertility
Murthy et al., In Review, 2010

30. Role of Human HLA-DR
Murthy et al., Infect Immun. 2006

31. Minimal Role for Antibody
Murthy et al., FEMS Immunol. Med. Microbiol. 2009

32. Role of CD4+ T-cells CD4+ T cell adoptive transfer
Murphey et al., Cell. Immunol. 2006

33. Role of IFN-g
Murthy et al., J. Immunol. 2008

34. Summary CPAF vaccine administered with Th1 adjuvant:
Enhances clearance of infection
Protects against severe pathology
Induces robust cellular IFN-g response
Induces systemic and mucosal antibody
Protection is dependent upon CPAF-specific
cellular IFN-g responses but not antibody

35. Ongoing Collaborative Effort
US Patent Application No. 12/243,769: Inventors- Arulanandam, Murthy, Zhong

36. Bench to Bedside Anti-Chlamydia vaccine
Identification of pathogenic mechanisms
Vaccine antigen discovery
Validation of protection and pathogenesis
correlates in clinical samples
Routes, Delivery systems
Adjuvants, Formulations
Non-Human Primates
Clinical Trials
Anti-Chlamydia vaccine

37. Acknowledgements Bernard Arulanandam, Ph.D., M.B.A.
Weidang Li, M.D., Ph.D.
Bharat Chaganty, MS
Sangamithra Kamalakaran, B.Tech
Kishan Evani, MS
Rishein Gupta, Ph.D.
Yu Cong, M.D.
Madhulika Jupelli, Ph.D.
Cathi Murphey, M.S.
Guangming Zhong, M.D., Ph.D., UTHSCSA
M. Neal Guentzel, Ph.D., STCEID, UTSA
Aruna Mittal, Ph.D, IOP, India
1R03AI088342