For qualitative analysis, blood IUPM values 0.3219 were considered positive and 0.3219 negative3. HIV RNA was assayed in oral swab, plasma, CVL and unspun saliva with Nuclisens(Organon Teknika Corporation, Raleigh, Andrographolide NC), while cervical swabs were assayed with Roche Amplicor HIV-1 Monitor test(Roche Rabbit Polyclonal to UTP14A Molecular Systems, Branchburg, NJ). adaptive mucosal defense factors examined, only IgA was associated with HIV RNA shedding. However, rather than being protective, there was a striking increase in probability of detectable HIV RNA shedding in women with highest total IgA. was assessed by wet mount and bacterial vaginosis by Amsel19. HIV-1 evaluations Semi-quantitative Andrographolide HIV cultures of blood and qualitative HIV cultures of genital(unspun CVL, swab) and oral(unspun saliva, swab) specimens were performed within 6hr by protocol20. Infectious virus was expressed as infectious units/106 cells(IUPM). For qualitative analysis, blood IUPM values 0.3219 were considered positive and 0.3219 negative3. HIV RNA was assayed in oral swab, plasma, CVL and unspun saliva with Nuclisens(Organon Teknika Corporation, Raleigh, NC), while cervical swabs were assayed with Roche Amplicor HIV-1 Monitor test(Roche Molecular Systems, Branchburg, NJ). Lower limit of detection(LLD) for plasma and cervical swabs was 400 copies/ml, and CVL, oral swabs and saliva had LLD of 80 copies/ml21. Immunologic evaluations Centrifuged saliva(1:3 in PBS), CVL and plasma were tested for SLPI and TSP-1 by ELISA(R&D Systems, Minneapolis, MN) or as described12. Total IgG and IgA and HIV-specific antibodies were determined by capture ELISA22. IgG and IgA anti-gp120 were measured against standard curves of polyclonal secretory IgA or serum(Moni-Trol ES, Baxter, Stone Mountain, GA) with high anti-HIV-1 Ig as positive control and mucosal secretions and sera of uninfected individuals as negative controls. The cutoff(nonspecific) was set at Andrographolide 100ng/ml. Study definitions For our analyses, HIV shedding was defined as presence of HIV RNA in oral swab or saliva and in genital swab or CVL using LLD above. We used DHHS/Kaiser Panel[DHHS/Kaiser 2004] guidelines to define HAART usage: (a) 2 NRTIs in combination with 1 PI or one NNRTI(88% of observations classified as HAART); (b) one NRTI in combination with 1 PI and 1 NNRTI(5%); (c) regimen containing ritonavir and saquinavir in combination with one NRTI and no NNRTIs(1%); and (d) abacavir or tenofovir containing regimen of 3 NRTIs in absence of PI and NNRTI(6%), except for three-NRTI regimens consisting of: abacavir+tenofovir+lamivudine or didanosine+tenofovir+lamivudine. Statistical analysis Frequency distributions were calculated for categorical variables and median(range) for continuous variables. Associations among detection of HIV RNA in three compartments(oral, genital, blood) were assessed using a log-linear model; models were analyzed in total sample and also stratified by ART. HIV RNA was considered detectable in mucosal compartments if either swab or fluid was above LLD; women with both tests not completed were treated as unknown. To compare median levels of innate and adaptive molecules between compartments, Wilcoxon signed rank tests were used. Using logistic regression to assess relationships between HIV RNA in oral and genital compartments with clinical and laboratory assessments, odds ratios(ORs) and 95% confidence limits were calculated. The relationship between each variable and presence of HIV RNA in Andrographolide oral cavity and genital tract was analyzed in multivariate logistic regression models adjusting for ART(none, monotherapy, combination therapy, HAART) and plasma HIV RNA( 400, 400-10,000, 10,000 copies/ml) or ART only where appropriate. Additional multivariate models adjusted for CD4+ T cells( 200, 200- 500, 500+ cells/mm3). In logistic regression models, SLPI, TSP, IgA and IgG were categorized by tertile. Relationships between total IgA and HIV RNA within each compartment were estimated by Spearman rank correlation coefficients. Two-sided hypotheses were used throughout, testing at 5% significance level. Results HIV RNA and infectious HIV in blood, oral cavity and genital tract Demographic and clinical characteristics of the WIHS cohort, including this subset (n=115)3, 14 are summarized in Supplemental Table 1. The majority were African American(57%) and Hispanic(20%) women who reported heterosexual Andrographolide contact as primary risk factor for transmission; only 24% reported injection drug use (IDU). Most had detectable plasma viral burden with approximately one third having HIV RNA levels 400, one third from 400-10,000.