Phylogenetic Tree Analysis of Protease/Reverse Transcriptase Sequences for CDC 310340 and CDC 310342 Viral Stocks.

<p>HIV-1 sequences were obtained using the Siemens' TRUGENE HIV-1 Genotype Test, analyzed using MegAlign version 9.0.4 (DNASTAR, Inc) and subtyped against the HIV Sequence Database using BLAST (<a href="http://www.hiv.lanl.gov/content/sequence/BASIC_BLAST.html" target="_blank">www.hiv.lanl.gov/content/sequence/BASIC_BLAST.html</a>). Both sequences were found to align with HIV-1 Group M, subtype CRF02_AG. The homology between the two sequences was 98% with 0.3% divergence. Both sequences are designated in the tree with a •. The dotted line indicates a negative branch length, which is a result of averaging.</p

Performance of two HIV-2 primers/probe sets in real-time RT-qPCR assays.

<p>Two HIV-2 primers/probe sets were assessed for amplification of serial dilutions of an HIV-2 isolate, NIH-Z, under standard ampification conditions that were not optimized for each primers/probe set. Primers/probe sets, designated as PD  =  Primer Design LTD HIV-2 PCR Kit and SM  =  Delarue et al <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096554#pone.0096554-Delarue1" target="_blank">[12]</a>, showed linear amplification profiles that were parallel. PD  =  •. SM  =  ▴.</p

Performance of HIV-1/2 viral stocks in real-time RT-qPCR and HIV-1 p24 antigen tests.

<p>Individual viral stocks identified as HIV-2 were obtained from the NIAID AIDS Reagent and Reference Repository and SeraCare, Inc. The HIV-1 subtype and HIV-2 group identification is based upon data sheets provided by NIAID and from publications. An HIV-1 Subtype B isolate from the United States (91US_4) was used as an HIV-1 control. Viral RNA was extracted and tested in HIV-2 real-time RT-qPCR assays. Viral isolates selected for testing in the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 v2.0 quantitative RNA assay and the Perkin-Elmer p24 Antigen test included the two viral isolates that were not amplified in the HIV-2 real-time RT-qPCR assays, three HIV-2 viral isolates that amplified well and the HIV-1 91US_4 isolate. Although the HIV-2 RT-qPCR assays were not optimized, the HIV-2 amplifications were robust for the majority of the isolates tested as reflected in the HIV-2 RNA concentrations reported based upon relative values extrapolated from the NIH-Z standard for each primers/probe set. The CDC 310340 and CDC 310342 viral stocks were not amplified in HIV-2 RT-qPCR assays but were quantified and reactive in HIV-1 specific tests. PD  =  Primer Design LTD HIV-2 PCR Kit primers/probe. SM  =  Delarue et al <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096554#pone.0096554-Delarue1" target="_blank">[12]</a> primers/probe set. TND  =  Target Not Detected. UNK is unknown group. Viral isolate was not tested (−).</p

Frequencies and incidence rates of HIV among United States Air Force personnel in service at any time from 1996 through 2011^*.

<p>*All personnel with available demographic records were included in this analysis.</p><p>**Other race included Asians or Pacific Islanders, American Indians or Alaskan natives, and individuals who self-reported their race as ‘Other’ on their personnel records.</p><p>Frequencies and incidence rates of HIV among United States Air Force personnel in service at any time from 1996 through 2011^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0126700#t001fn001" target="_blank">*</a>}.</p

Mobility-related factors among HIV-infected and matched HIV-uninfected United States Air Force active duty personnel, 1996–2011.

<p>Mobility-related factors among HIV-infected and matched HIV-uninfected United States Air Force active duty personnel, 1996–2011.</p

Socio-demographic factors of HIV-infected and matched HIV-uninfected United States Air Force active duty personnel, 1996–2011.

<p>*Other marital status included individuals who reported that they were neither married nor single on their personnel records.</p><p>**The occupation category, engineer, included non-combat engineer; ‘Other’ occupations included infantry/ combat engineer/ Special Forces/ artillery/ armor/ motor transport/ administration and other categories.^A service member’s residence at the time of entrance to the US military.</p><p>Socio-demographic factors of HIV-infected and matched HIV-uninfected United States Air Force active duty personnel, 1996–2011.</p

Incidence rates of new HIV diagnoses (and 95% confidence intervals) among active duty Air Force personnel, 1996–2011.

<p>Incidence rates of new HIV diagnoses (and 95% confidence intervals) among active duty Air Force personnel, 1996–2011.</p

DNA Vaccine Delivered by a Needle-Free Injection Device Improves Potency of Priming for Antibody and CD8+ T-Cell Responses after rAd5 Boost in a Randomized Clinical Trial

<div><p>Background</p><p>DNA vaccine immunogenicity has been limited by inefficient delivery. Needle-free delivery of DNA using a CO₂-powered Biojector® device was compared to delivery by needle and syringe and evaluated for safety and immunogenicity.</p><p>Methods</p><p>Forty adults, 18–50 years, were randomly assigned to intramuscular (IM) vaccinations with DNA vaccine, VRC-HIVDNA016-00-VP, (weeks 0, 4, 8) by Biojector® 2000™ or needle and syringe (N/S) and boosted IM at week 24 with VRC-HIVADV014-00-VP (rAd5) with N/S at 10¹⁰ or 10¹¹ particle units (PU). Equal numbers per assigned schedule had low (≤500) or high (>500) reciprocal titers of preexisting Ad5 neutralizing antibody.</p><p>Results</p><p>120 DNA and 39 rAd5 injections were given; 36 subjects completed follow-up research sample collections. IFN-γ ELISpot response rates were 17/19 (89%) for Biojector® and 13/17 (76%) for N/S delivery at Week 28 (4 weeks post rAd5 boost). The magnitude of ELISpot response was about 3-fold higher in Biojector® compared to N/S groups. Similar effects on response rates and magnitude were observed for CD8+, but not CD4+ T-cell responses by ICS. Env-specific antibody responses were about 10-fold higher in Biojector-primed subjects.</p><p>Conclusions</p><p>DNA vaccination by Biojector® was well-tolerated and compared to needle injection, primed for greater IFN-γ ELISpot, CD8+ T-cell, and antibody responses after rAd5 boosting.</p><p>Trial Registration</p><p>ClinicalTrials.gov <a href="http://clinicaltrials.gov/ct2/show/NCT00109629" target="_blank">NCT00109629</a></p></div

Worst Severity of Systemic Reactogenicity Following DNA and rAd5 Vaccinations.

<p>Worst Severity of Systemic Reactogenicity Following DNA and rAd5 Vaccinations.</p

Schematic diagram of study design and completion of vaccination schedule.

<p>The CONSORT diagram indicates the number of subjects screened to complete enrollment of 40 subjects in a factorial study design. Subjects were stratified by pre-existing Ad5 neutralizing antibody reciprocal titer of ≤500 or >500 then randomized to receive DNA priming by Biojector® or needle and syringe (N/S). In addition, 50% of each subgroup was boosted with rAd5 at 10¹⁰ PU and the other 50% received 10¹¹ PU.</p