HIV persists in a reservoir of latently infected CD4+ T cells in individuals treated with highly active antiretroviral therapy (HAART). Here we identify central memory (TCM) and transitional memory (TTM) CD4+ T cells as the major cellular reservoirs for HIV and find that viral persistence is ensured by two different mechanisms. HIV primarily persists in TCM cells in subjects showing reconstitution of the CD4+ compartment upon HAART. This reservoir is maintained through T cell survival and low-level antigen-driven proliferation and is slowly depleted with time. In contrast, proviral DNA is preferentially detected in TTM cells from aviremic individuals with low CD4+ counts and higher amounts of interleukin-7–mediated homeostatic proliferation, a mechanism that ensures the persistence of these cells. Our results suggest that viral eradication might be achieved through the combined use of strategic interventions targeting viral replication and, as in cancer, drugs that interfere with the self renewal and persistence of proliferating memory T cells.

The Roche PGMY primer-based research prototype line blot assay (PGMY-LB) is a convenient tool in epidemiological studies for the detection and typing of human papillomavirus (HPV) DNA. This assay has been optimized and is being commercialized as the Linear Array HPV genotyping test (LA-HPV). We assessed the agreement between LA-HPV and PGMY-LB for detection and typing of 37 HPV genotypes in 528 anogenital samples ( Infection by human papillomavirus (HPV) causes squamous intraepithelial lesions and invasive cancer of the uterine cervix and anus (3). HPV testing relies on the detection and analysis of viral DNA. Epidemiological studies and vaccine clinical trials require reliable and reproducible identification and genotyping of genital HPV infections. Since only a fraction of the 40 HPV genotypes infecting the anogenital tract are associated with malignant lesions, the detection method has to identify types individually. Specific genotyping also provides information on mixed HPV infections (26). Type-specific PCR assays are impractical for epidemiological studies because of the multiplicity of relevant genotypes infecting the anogenital tract. Consensus PCR assays that target conserved regions of the HPV genome have been devised to amplify all relevant genital types in one reaction, with analysis of amplicons by direct sequencing, restriction fragment length polymorphism analysis, or type-specific hybridization.The most common PCR methods use the consensus primer set MY09/MY11/HMB01 (20, 25), GP5ϩ/GP6ϩ (9, 21), PGMY09/ PGMY11 (13,34), or SPF10 (30,34). Convenient assays for detection and typing of HPV have been developed for all of these primer sets. HPV amplicons generated by PGMY or MY primers can easily be detected and typed by a nonisotopic

Human papillomavirus DNA was detected in 32 (11.2%) of 287 individuals. Associated with oral human papillomavirus infection on univariate analyses were human immunodeficiency virus infection (odds ratio, 6.9; 95% confidence interval, 2.0-23.2), homosexuality (odds ratio, 3.7; 95% confidence interval, 1.5-9.4), unprotected oral sex (odds ratio, 5.5; 95% confidence interval, 1.6-18.4), syphilis (odds ratio, 2.5; 95% confidence interval, 1.1-6.3), gonorrhea (odds ratio, 4.2; 95% confidence interval, 1.9-9.1), Chlamydia trachomatis (odds ratio, 4.4; 95% confidence interval, 1.8-10.6), and genital herpes (odds ratio, 2.9; 95% confidence interval, 1.3-6.5). Human immunodeficiency virus infection and C. trachomatis were independently predictive of human papillomavirus infection in multivariate stepwise logistic regression.

A novel real-time PCR assay for detection of human papillomavirus type 52 (HPV-52) DNA (RT-52) was evaluated on 265 anogenital samples. RT-52 had a sensitivity of 98.4% and a specificity of 100% compared to conventional HPV-52 typing assays, including hybridization of PGMY products with an HPV-52-specific probe and PCR sequencing of HPV-52 E6.The detection of high-risk human papillomavirus (HPV) types in genital specimens has now been approved in several countries for the triage of women with a cytological diagnosis of atypical squamous cells of undetermined significance (ASCUS) and for the primary screening for cervical cancer for women aged 30 years and above as an adjunct to cytology (18). HPV DNA genotyping has proven useful in the study of the natural history and transmission of HPV infection (9,15). Genotyping assays will be instrumental in assessing the impact of HPV vaccination on the risk of acquisition and on the distribution of individual HPV types in a population. Moreover, HPV genotyping could identify HPVinfected women at greater risk for high-grade cervical intraepithelial neoplasia or cancer (2, 13, 15).The Linear Array HPV genotyping test (LA-HPV) from Roche Molecular Systems is approved by Health Canada for HPV genotyping. LA-HPV identifies 36 genotypes by hybridization on a linear array of PGMY-generated amplicons with 34 type-specific probes for , two probes for two subtypes of HPV-82, and one probe that cross-reacts with . A sample is thus considered positive for HPV-52 when it reacts with the HPV-52-cross-reactive probe but not with the HPV-33, -35, or -58 type-specific individual probes. However, LA-HPV cannot establish the presence of HPV-52 DNA in a sample reacting with the HPV-33/35/52/58-cross-reactive probe and with at least one of the HPV type-specific probe for HPV-33, -35, or -58. If these samples are considered positive for HPV-52, the prevalence of HPV-52 will be overestimated, and if they are considered negative, HPV-52 prevalence may be underestimated.HPV-52 is the seventh most frequently detected high-risk type in invasive cervical cancer worldwide (5) and causes 8.8% of low-grade and 2.5% of high-grade squamous intraepithelial lesions (4). It is thus imperative in epidemiological studies to be able to determine if HPV-52 is truly present in samples reacting with the HPV-52-cross-reactive probe and containing another type reacting with that probe. We present here the results of the validation of a novel real-time PCR assay for the rapid confirmation of HPV-52 infection in specimens positive with the HPV-52 cross-reacting probe in the LA-HPV test.Clinical specimens. Overall, 265 genital specimens collected from 95 women and 170 men participating in four studies described elsewhere (8,11,14,16) were selected on the basis of previous testing of PGMY-generated amplicons with the cross-reacting HPV-33/35/52/58 probe and with conventional HPV-52 typing methods (see below). Anal swabs were collected in Preservcyt (Cytyc Corporation, Boxborough, MA) from 170 men in the HIPVIRG cohort ...