HIV Vaccine. Sunee Sirivichayakul, Ph.D. Faculty of Medicine Chulalongkorn University. August 22, 2014

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1 HIV Vaccine Sunee Sirivichayakul, Ph.D. Faculty of Medicine Chulalongkorn University August 22, 2014

2 Immunity Natural immunity Active natural immunity e.g., infection Passive natural immunity e.g., trans-placental transfer of maternal antibody Acquired immunity Active acquired immunity e.g., vaccination Passive acquired immunity e.g., administration of antibody or immune T cells

3 Immunity: Active vs. passive acquired immunity Differences Active immunization Passive immunization Administration Vaccines, toxoids Antitoxin, -globulin Production of immunity Slowly Immediately Duration of immunity Long (from several months to years) Short (2 weeks to months) Usage Immunoprophylaxis Emergency prophylaxis and therapy

4 Downloaded from: StudentConsult (on 22 January :12 AM) 2005 Elsevier

5 Why do we need a vaccine? To teach the body s natural immune response to learn how to combat many disease-causing microorganisms, especially those lifethreatening microorganisms (to induce memory cells) When one encounters the same microorganisms later, the immune system will respond quicker and more efficient to get rid of them It is much cheaper to prevent than to treat a disease (In a 2005 study on the economic impact of routine childhood immunization in the US, researchers estimated that for every dollar spent, the vaccination program saved more than $5 in direct costs and approximately $11 in additional costs to society). If too many people in a community do not get vaccinations, diseases can reappear. In 1989, there was a measles outbreak in the US which resulted in > 55,000 cases of measles and 136 measlesassociated deaths.

6 Successful vaccine The success of vaccination in eradicating infectious disease is dependent on several properties of the microbes. Vaccines are effective if - the infectious agent does not establish latency - if it does not undergo much or any antigenic variation - if it does not interfere with the host immune response Vaccines are most effective against infections that are limited to human hosts and do not have animal reservoirs. Most vaccines in use today work by inducing humoral immunity (prevent infections by neutralizing and clearing microbes before they gain their foothold in the host) The best vaccines are those that stimulate the development of longlived plasma cells that produce high-affinity antibodies as well as memory B cells.

7 Effectiveness of Vaccines for Some Common Infectious Diseases Disease Maximum Number of Cases (year) Number of Cases in 2009 Percentage Change Diphtheria 206,939 (1921) Measles 894,134 (1941) Mumps 152,209 (1968) Pertussis 265,269 (1934) 13, Polio (paralytic) 21,269 (1952) Rubella 57,686 (1969) Tetanus 1,560 (1923) Haemophilus influenzae type B 20,000 (1984) Hepatitis B 26,611 (1985) 3,

8 Types of vaccines The type of vaccine is typically based on fundamental information about the microbe, such as how it infects cells and how the immune system responds to it, and the regions of the world where the vaccine would be used. Live, attenuated vaccines (closest to natural infection :- BCG, mumps, measles, rubella) Inactivated vaccines (Pertussis, typhoid) Subunit vaccines (recombinant subunit hepatitis B vaccine) Toxoid vaccines (Diphtheria and tetanus toxoid) Conjugate vaccines (Haemophilus influenzae type B (Hib) vaccine) DNA vaccines Recombinant vector vaccines

9 Why an HIV vaccine is needed? Global epidemic No cure once infected Treatment is life-long, expensive and not without side-effects Treatment is not universally accessible and affordable in many resource-limited settings Number of newly treated each year is less than number newly infected each year Treatment may fail

10 HIV/AIDS in Thailand MSM 10-30%

11 Three Decades of Antiretroviral Therapy and the Future Evidences and developments One/ two ARVs Improve survival Three ARVs (HAART) Durable undetectable VL Earlier HAART non-aids death Transmission New TB New strategies Long-acting ARV? Cure? 1981 late 1980 Mid Few ARVs More toxicity More class ARVs More potent PIs But high pill burden More new ARVs More tolerable More OD options More FDC options Single tablet regimens Monthly ARV? Cure? Availability and treatment options

12 HIV-1 vaccine concepts tested in humans in Asian countries (1) Vaccine concept Sponsor HIV-1 subtype or CRF V3 peptides UBI multiclade I (1994) Gp120 in MF59 Chiron B I (1995) Phase (year of initiation) AIDSVAX gp120 B/E in alum AIDSVAX gp120 B/E in alum ALVAC-HIV vcp gp140 or gp120 in polyphosphazene or alum or MF59 Genetech B/CRF01_AE I/II (1995/97) VaxGen B/CRF01_AE III (1998/2003) USAMRMC B/CRF01_AE I/II (2000) Nitayaphan S, et al. Expert Rev Vaccines 2012.

13 HIV-1 vaccine concepts tested in humans in Asian countries (2) Vaccine concept Sponsor HIV-1 subtype or CRF ALVAC-HIV vcp gp120 in alum (RV144) Phase (year of initiation) USAMRMC B/CRF01_AE III (2003) (mostly heterosexual) Adenovirus type 5 Merck B I/II (2003) DNA + fowlpox NCHECR CRF01_AE I (2004) MVA USAMRMC CRF01_AE II (2012) Nitayaphan S, et al. Expert Rev Vaccines 2012.

14 HIV-1 vaccine concepts planned to be tested in Thailand Vaccine concept Sponsor HIV-1 subtype or CRF ALVAC-HIV vcp gp120 in alum late boosts of RV144 vaccinees USAMRMC B/CRF01_AE II (2012) AIDSVAX gp120 B/E in alum USAMRMC B/CRF01_AE II (2012) Phase (year of initiation) ALVAC-HIV vcp AIDSVAX gp120 B/E in alum DNA biojector vs. electroporation + MVA USAMRMC B/CRF01_AE II (2012) USAMRMC Env A, C, D, and gag consensus, CRF01_AE II (2012) Adenovirus type 26 + MVA USAMRMC Mosaic I (2013) Nitayaphan S, et al. Expert Rev Vaccines 2012.

15 Phases of HIV vaccine development Pre-clinical trials: - mice, rabbits and rhesus macaques - have to move ultimately in human trials because no animals have the same immune systems as humans Clinical trials phase I: - small number (20-100) of healthy, uninfected participants at low risk - to test safety (usually by comparing with a control placebo) - can provide initial data on the dose and administration schedule that achieve the optimal immune response - usually last months

16 Phases of HIV vaccine development (2) Phase II trials : - involves hundreds of participants with varying degrees of risk to better characterize the safety and the immune response - can last 2 or more years Phase III trials : - involves several thousand high-risk volunteers to further assess if the vaccine works in preventing infection - can last 3-5 years An experimental vaccine must successfully complete at least three stages of testing in people before it can be licensed.

17 Results of all efficacy trials of HIV vaccine worldwide Vax003 (USA) gp120 (B/B ) + alum Vax004 (Thailand) gp120 (B/E) + alum STEP (USA + Australia), Phambili (South Africa) Ad5-HIV-1 trivalent vaccine (Gag, Pol, Nef) enhanced rate of infection in uncircumcised Ad5-seropositive male volunteers RV144 (Thailand) Canarypox (ALVAC)-Gag Pol Env E + gp120b/e + alum HVTN505 (USA) DNA-Gag, Pol, Env priming + Ad5-Gag Pol Env (A,B,C) boost

18 RV144 Vaccine efficacy of 31.2 % An analysis of the correlates of risk showed that - IgG-binding Abs to scaffolded V1V2 recombinant protein correlated inversely with infection rate - envelope binding plasma IgA correlated directly with infection rate (may diminish the effects of potentially protective Abs) - Neither low levels of V1V2 Abs nor high levels of envspecific IgA Abs were associated with higher rates of infection than we found in the placebo group.

19 HIV nomenclature HIV-1 M (Major) Group : A1, A2, B, C, D, F1, F2, G, H, J, K, and at least 55 CRFs (circulating recombinant forms) i.e., CRF_A/E, CRF_A/G, CRF_A/B HIV-1 N (not-m, not-o) Group (identified in a few individuals in Cameroon, West Africa) HIV-1 O Group (outlier), most commonly found in West Africa HIV-1 P Group (A human isolate closely related to SIVgor was isolated) HIV-2 (closely related to SIVs isolated from sooty mangabeys : A, B, C, F and G)

20 M Group Subtypes A1, A2, B, C, D, F1, F2, G, H, J and K Subtypes and sub-subtypes (a distinctive lineage that is very closely related to a particular subtype, and is not genetically distinct enough to justify calling a new subtype) The sequences within any one subtype or sub-subtype are more similar to each other than to sequences from other subtypes throughout their genomes. These subtypes represent different lineages of HIV, and have some geographical associations.

21 There is nothing for Asian epidemic (CRF01_AE and Asian subtype B!!!

22

23

24 Why mosaic? Extreme diversity of HIV-1 Coverage of viral diversity using mosaics was greatly increased as compared to natural-sequence vaccine candidates, for both variable and conserved proteins (Fisher W, et al; Nat Med 2007; 13 : ) Possibility of matching with 9-amino acid potential epitopes in global gag sequences Mosaic protein Natural protein 9-amino acid matched 74 % 37 % 8/9-amino acid matched 87% 67 %

25 Mosaics can provide excellent coverage of both CRF01 and Asian B GAG with a three protein cocktail Bette Korber, Thomas Leitner. et al LANL, USA

26 Origin of replication Initiate transcription Improve RNA stability Antibiotics resistance gene Directing expression in mammalian cells Basic structure of vector for DNA vaccine

27 Immunization routes Intramuscular (IM) Intradermal (ID) Needle-free (NF) Electroporation

28 Needle Free Injector: New Device to improve immunogenicity Electroporation

29

30

31 BALB/c mouse

32 Anti-p24 WB Results: ID vs NFI ID 100 mg pdna Day 3 Day 7 after last boost Needle Free 50 µg pdna

33

34 Sources of HIV-1 subtypes for the gag, pol, nef mosaic DNA Immunogen Designs of the Chula DNA vaccine In collaboration with Bette Korber (Los Alamos, USA) Gag (3 mosaic cocktail) o CRF01AE: 156 sequences (114 Thai) o B: 72 Asian sequences (8 Thai) Pol (3 mosaic cocktail) o CRF01AE: 101 sequences (76 Thai) o B: 30 Asian sequences (8 Thai) Nef (4 mosaic cocktail) o CRF01AE: 150 sequences (121 Thai) o B: 210 Asian sequences (14 Thai) (sample collection starting 2005)

35 Mosaic envelope Envelope CRF01_AE (68 sequences from , 9 sequences from , 20 sequences from 2011) - 59 HIV subtype B - 3 mosaic : mosaic 1 = CRF01_AE : mosaic 2 = CRF01_AE + B : mosaic 3 = with some modification of mosaic 2 (by filling in some of the gaps)

36 Overall immunogenicity study Pilot immunogenicity study: DNA vaccine alone DNA priming / recombinant vaccinia boosting Mouse immunogenicity study Non-human primate (monkey) immunogenicity study

37 IFN- ELISpot results of HIV-1 gag, pol, nef AE/B Mosaic DNA vaccines (without boosting) in mice P<0.001 P< Methods: Immunization with a combination of 3 gag mosaic, 3 pol mosaic and 4 nef mosaic with a total amount of DNA 100 g (10 g each) on Week 0, 2, 4; mice sacrificed on Week 5 Interpretations: Unlike the gag mosaic, the pol and nef mosaic cocktail results showed an imbalance of specific T cell response as seen by a significant bias toward subtype B specific responses. Thus, approaches to improve AE-specific responses is needed.

38 Immunization Experiments Objective: To study immunogenicity of Asian mosaic HIV-1 AE/B env DNA vaccine in mice by IM electroporation DNA = 100 µg/dose Recombinant vaccinia = 5 x 10 6 pfu/dose (positive control and boosting) Immunization Ags : DNA alone (Week 0, 2, 4) : DNA prime/vaccinia boost (Week 6) Mice sacrificed : 1 week after last immunization

39 Immunogenicity study Cell-mediated immunity: ELISpot using truncated peptides from subtype B and AE Humoral immunity: Anti-gp120/160 titer by western blot Neutralizing antibody

40 Conclusion : Mosaic primed and boosted with env recombinant vaccinia AE induced the highest response when stimulated with AE pooled peptides as compared to boosted with env recombinant vaccinia B or stimulated with B pooled peptides.

41 IFN- ELISpot results of HIV-1 Env AE/B DNA Mosaic (3 cocktail) with/without recombinant vaccine boosting in mice * p<0.005 (vs all other groups) Conclusions: 1. AE/B DNA mosaic alone induced good env-specific IFN producing T cell responses 2. Boosted with HIV-1 AE rvaccinia, but not B rvac, significantly enhanced the responses

42 Pigtailed macaque (Macaca nemestrina)

43 Monkey (pigtailed macaques) experiment N = 12 DNA: gag and env (total of 2 mg per immunization) Intramuscular or Intradermal, both followed by electroporation on Months 0, 1, 2 Agent Dose (mg) Route of administration Frequency and duration pcmvkan 2 mg/dose 1 ID and 1 IM 3x (mo 0, 1 and 2) HIV-1 gag/env DNA vaccine HIV-1 gag/env DNA vaccine 2 mg/dose EP IM 3x (mo 0, 1 and 2) [2 injecton sites/vaccination] 2 mg/dose EP ID 3x (mo 0, 1 and 2) [3 injecton sites/vaccination] Blood collection : Month 0, and 2 weeks after the last immunization for CMI and HMI (binding and neutralizing Abs)

44 ID immunization could induce better responses to both HIV-1 AE or HIV-1 B gag peptide stimulation.

45 There is no different responses to either HIV-1 AE or HIV-1 B peptide stimulations in both IM and ID immunizations.

46 Monkey # Route Tattoo ELISpot (SFU/million PBMC) week 10 Neutralizing Ab Env E Gag E Env B Gag B TH023 1 IM (C) B <20 2 ID (C) B <20 3 IM B IM B (21 against MN) 5 IM B (22 against MN) 6 IM B111103A (30 against MN) 7 IM B <20 8 ID B ID B ID B <20 11 ID B ID B All unstimulated wells (neg) had 0 and anti-cd3-stimulated (pos cont.) wells had >1200 SFU/million PBMC Neutralizing Ab was kindly performed by Dr.Chitraporn Kannasutra and team at AFRIMS.

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