The patients were asked to gargle for 30 s with 20 ml of 0.9% sodium chloride. EBV IgG antibody titers to EA and VCA was determined in plasma by conventional Selleckchem CHIR-99021 immunofluoroscence applied to antigen positive cells. IgG

and IgM titers were determined against EBNA 1 with peptide (p107) based ELISA. The patients gargled with 10 mL of RPMI medium for 1 min. The throat wash was centrifuged at 2000 rpm (approximately 600 × g) for 10 min, and then the supernatant was frozen at −70 °C until testing. Half mL of the sample was lysed in 0.5 mL of PCR-lysate buffer [18]. EBV DNA analysis and statistics were performed as previously reported by Friis et al. [18]. This method is as sensitive and gives similar results as quantitative PCR (qPCR) [2]. In addition it provides results in all samples, while qPCR may fail more often due to inhibition and quenching. One hundred μL of plasma were lysed in 100 μL PCR-lysate buffer. Plasma samples were tested for positive

respectively negative Onalespib supplier reaction using the same PCR condition as for blood. Non-parametric Mann Whitney or Kruskal Wallis tests were applied, using StatView II (Abacus Concepts Inc.). Multivariate analysis was also performed using Simca-P 8.0 (Umetrics AB) but did not add anything to our interpretation based on univariate analysis. HIV-1 infected patients included in the rgp160 vaccine trials showed higher median EBV-DNA load, 2.4 copies per 1000 B cells (n = 42)

compared to non-vaccinated HIV-carriers, 0.49 per 1000 B cells (n = 18; p < 0.01, Fig. 1A). Although the patients were recruited from two slightly different vaccination trials (see Materials and Methods), we found no statistical difference in EBV-DNA load between the two groups. A considerable individual variation was observed. much There was no significant statistical difference as regards age, sex, and antiretroviral treatment when comparing immunised and non-immunised patients ( Table 1). However, in the rgp160 study group higher CD4+ cell counts were detected, which is most likely a result of the selection criteria for the vaccine trial. The immunised group had a median value of 270 × 106 cells/L (n = 42) as compared to a median of 120 × 106 cells/L (n = 18) in the HIV-1 positive patients not included in the vaccine trial. We observed no significant correlation between the CD4+ cell counts and the EBV load, although there was a tendency to inverted correlation between these variables that patients with a high EBV load had low CD4+ cell counts, and patients with a low EBV load had a high CD4+ cell count. The highest EBV values were exclusively found in the immunised group, while low values could be seen both in immunised and non-immunised patients. In the non-immunised HIV-1 carriers, the asymptomatic patients had a median EBV load of 0.