Immune responses induced by immunization with HIV-1 DNA followed by HIV-modified vaccinia virus Ankara with or without recombinant GP140 in healthy Tanzanian individuals

Detta är en avhandling från Stockholm : Karolinska Institutet, Dept of Microbiology, Tumor and Cell Biology

Sammanfattning: A vaccine against HIV is widely considered the most effective and sustainable way of preventing new infections. We previously conducted a phase I/II clinical trial using multi-clade, multigene HIV-DNA priming and boosting with the recombinant modified vaccinia Ankara (HIV-MVA) virus among healthy Tanzanian volunteers (HIVIS03 trial). The HIV-DNA vaccine contained seven plasmids expressing HIV-1 gp160 subtypes A, B, C, Rev B, Gag A, B and RTmut B and the recombinant HIV-MVA vaccine expressed CRF01_AE HIV-1 envelope (Env) subtype E and Gag-Pol subtype A. Sixty HIV-uninfected volunteers were randomized into three groups of 20 to receive HIV-DNA or placebo intradermally (id) 1 mg or 3.8 mg intramuscularly (im) at 0, 1 and 3 months with a needle free device and were boosted with HIV-MVA 108-pfu or placebo im at 9 and 21 months. The vaccine regimen was safe and induced strong and potent immune responses. In this thesis we further explored the immune responses elicited by the HIV-DNA and HIV-MVA vaccine regimen and the effect of boosting with a third HIV-MVA or an envelope protein. In study I, we evaluated the HIV vaccine-induced antibody responses in sera collected from 29 HIVIS03 vaccinees at baseline and four weeks after the second HIV-MVA. High titers of neutralizing antibodies (NAbs) (median 357) were detected using an infectious molecular clone (IMC)/peripheral blood mononuclear cell (PBMC) assay. The NAbs were significantly (but not completely) removed upon depletion of natural killer (NK) cells from PBMC (p=0.0039), indicating a role for Fc–receptor mediated antibody function. ADCC-mediating antibodies were demonstrated in the majority (97%) of the vaccinees against CRF01_AE and/or subtype B. The magnitude of ADCC-mediating antibodies against CM235 CRF01_AE IMC-infected cells correlated with NAbs against CM235 in the IMC/PBMC assay. In studies II and III we explored the duration of immune responses in individuals primed with HIV-DNA and boosted with HIV-MVA in the HIVIS03 trial and the effect of a late third HIV-MVA. Twenty volunteers who had previously received three HIV-DNA and two HIV-MVA immunizations in the HIVIS03 trial were given a third HIV-MVA, three years after the second HIV-MVA boost (HIVIS06). A high proportion of vaccinees showed durable binding antibodies, 90% to HIV-1 subtype C gp140 (median titer 200) and 85% to subtype B gp160 (median titer 100) three years after the second HIV-MVA. The majority of vaccinees had detectable ADCC–mediating antibodies, 70% against CRF01_AE virus–infected cells (median titer 239) and 84% against CRF01_AE gp120–coated cells (median titer 499). Furthermore, 74% of vaccinees still had IFN- γ ELISpot responses, 63% to Gag and 42% to Env. After the third HIV-MVA, there was an increase in Env-binding antibodies and ADCC-mediating antibodies relative to the response seen at the 3-years time point. The frequency of IFN- γ ELISpot responses increased to 95% against Gag or Env, and 90% to both Gag and Env, p = 0.064 and p = 0.002, respectively, after the third HIV-MVA. All 19 (100%) evaluable vaccinees had IgG antibodies to V1V2 CRF01_AE A244 after the second HIV-MVA with a median titer of 3200. A high proportion (75%) of the vaccinees still had V1V2 IgG antibodies to CRF01_AE A244 three years after the second HIV-MVA, which increased to 95% after the third HIV-MVA. The magnitude of response before and after the third MVA increased significantly from a median titer of 400 to 1600, p<0.0001, but not to the same level as after the second HIV-MVA (p=0.025). Surface plasmon resonance/Biacore analysis data supported the ELISA findings. V1V2- specific IgG1 antibody responses were more frequently detected than V1V2-specific IgG3 antibodies. AntiV1V2 IgG1 responses decreased after three years but could be boosted by the third HIV-MVA in the majority of the vaccinees. In study IV, we evaluated the safety and impact of boosting with subtype C CN54rgp140 Env protein adjuvanted in glucopyranosyl lipid A-aqueous formulation (GLA-AF) in volunteers previously given three HIV-DNA, followed by two HIV-MVA in the TaMoVac 01 trial. Forty volunteers (35 vaccinees and five placebo recipients) were given two CN54rgp140/ GLA-AF immunizations 30–71 weeks after the second HIV-MVA. The vaccine was safe and well tolerated, except for one incident of asymptomatic hypoglycemia. After the second HIV-MVA vaccination, 34 (97%) of the vaccinees developed Env-specific binding antibodies, whereas 79% and 84% exhibited IFN- γ ELISpot responses to Gag and Env, respectively. Binding antibodies to subtype C, B and CRF01_AE Env were significantly boosted by the CN54rgp140/GLA-AF immunizations, while functional antibodies were not significantly boosted. In contrast, T-cell proliferative responses to subtype B MN antigen and IFN- γ ELISpot responses to Env peptides were significantly enhanced. In conclusion, the HIV-DNA prime/HIV-MVA boost regimen elicited potent antibody and cellular immune responses with remarkable durability, and a third HIV-MVA significantly boosted both antibody and cellular immune responses. Binding antibody responses and Env-specific cell-mediated immune responses but not functional antibody responses, increased after boosting with two CN54rgp140/GLA-AF immunizations following priming with HIV-DNA and HIV-MVA.

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