The Increase of the Magnitude of Spontaneous Viral Blips in Some Participants of Phase II Clinical Trial of Therapeutic Optimized HIV DNA Vaccine Candidate.

We developed a candidate DNA vaccine called "DNA-4"consisting of 4 plasmid DNAs encoding Nef, Gag, Pol(rt), and gp140 HIV-1 proteins. The vaccine was found to be safe and immunogenic in a phase I clinical trial. Here we present the results of a phase II clinical trial of "DNA-4". This was a multicenter, double-blind, placebo-controlled clinical trial of safety, and dose selection of "DNA-4" in HIV-1 infected people receiving antiretroviral therapy (ART). Fifty-four patients were randomized into 3 groups (17 patients-group DNA-4 0.25 mg, 17 patients-group DNA-4 0.5 mg, 20 patients-the placebo group). All patients were immunized 4 times on days 0, 7, 11, and 15 followed by a 24-week follow-up period. "DNA-4" was found to be safe and well-tolerated at doses of 0.25 mg and 0.5 mg. We found that the amplitudes of the spontaneous viral load increases in three patients immunized with the candidate DNA vaccine were much higher than that in placebo group-2800, 180,000 and 709 copies/mL, suggesting a possible influence of therapeutic DNA vaccination on viral reservoirs in some patients on ART. We hypothesize that this influence was associated with the reactivation of proviral genomes.

Characterization of the Transmitted Virus in an Ongoing HIV-1 Epidemic Driven by Injecting Drug Use.

Understanding features of the HIV-1 transmission process has the potential to inform biological interventions for prevention. We have examined the transmitted virus in a cohort of people who inject drugs and who are at risk of HIV-1 infection through blood contamination when injecting in a group. This study focused on seven newly infected participants in St. Petersburg, Russia, who were in acute or early infection. We used end-point dilution polymerase chain reaction to amplify single viral genomes to assess the complexity of the transmitted virus. We also used deep sequencing to further assess the complexity of the virus. We interpret the results as indicating that a single viral variant was transmitted in each case, consistent with a model where the exposure to virus during transmission was limited. We also looked at phenotypic properties of the viral Env protein in isolates from acute and chronic infection. Although differences were noted, there was no consistent pattern that distinguished the transmitted variants. Similarly, despite the reduced genetic heterogeneity of the more recent subtype A HIV-1 epidemic in St. Petersburg, we did not see reduced variance in the neutralization properties compared to isolates from the more mature subtype C HIV-1 epidemic. Finally, in looking at members of injecting groups related to the acute HIV-1 infection/early subjects, we found examples of sequence linkage consistent with ongoing and rapid spread of HIV-1 in these groups. These studies emphasize the dynamic nature of this epidemic and reinforce the idea that improved prevention methods are needed.

The high frequency of HIV type 1-specific cellular immune responses in seronegative individuals with parenteral and/or heterosexual HIV type 1 exposure.

Some individuals remain HIV seronegative despite repeated unprotected exposure to the virus. Recent observations led to a concept that acquired immunity plays a role in protection or at least in altered susceptibility to HIV-1 infection in highly exposed seronegative (ESN) individuals. Our aim was to study HIV-specific cellular immune responses induced in parenterally and/or heterosexually ESN individuals. Nine seronegative injection drug users (IDUs), 10 seronegative individuals, and nine of their HIV-positive sexual and/or IDU partners from the cohort of IDUs were included in the study. The discordant couples had unprotected sex, and some of seronegative partners also had parenteral exposure. Cell-mediated responses were measured in peripheral blood mononuclear cells (PBMCs) by ex vivo interferon (IFN)-γ-ELISpot and ICS combining IFN-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-2 after stimulation with four consensus peptide pools (Nef, Gag, RT, Env, subtype A-EE). Thirteen out of 19 (68%) seronegative study subjects had strong Nef peptide pool-specific ELISpot responses, three (16%) subjects responded against the Gag peptide pool, and one subject had an RT peptide pool response. Nef peptide pool responses in ESN were as high as in seropositive subjects. The multiple HIV-specific cytokine production in both CD4(+) and CD8(+) T cells was shown for several ESN subjects. The functional profiles of the immune responses were different between seronegative and HIV-positive study groups. Whether the observed cellular responses have any protective role against HIV needs to be further investigated.