Medical notes were reviewed retrospectively and the following data collected: baseline demographics [age, sex, year of HIV diagnosis, antiretroviral
treatment (ART) status, viral load (VL) and CD4 T-cell count] and pre- and post-vaccination H1N1 antibody levels, where available. Patients Selleckchem Cyclopamine had only one post vaccination H1N1 antibody titre measured at one of the three time-points (months 3, 6 or 9). This audit was approved by the South Eastern Sydney and Illawarra Area Health Service Research Ethics Committee. Patients who had consented to vaccination received a single 0.5-ml intramuscular injection of the Panvax® monovalent nonadjuvant split virion H1N1 vaccine, equivalent to 15 μg of haemagglutinin, in the deltoid muscle. The haemagglutination inhibition (HI) assay method used was based on established techniques , with modifications. In the HI assay, the pdf H1N1 reference strain A/California/7/2009 was used as the assay antigen (obtained from the WHO Collaborating Centre for Reference and Research on Influenza, Melbourne, Victoria). Patient sera were
treated with a receptor-destroying enzyme (RDE) (Denka Seiken, Tokyo, Japan) at a ratio of four parts RDE to one part serum and incubated overnight at 37°C. Five parts of 1.6% sodium citrate were then added, and the treated serum incubated at 56°C for 30 min. After titration of the treated sera in phosphate-buffered saline (PBS) Hydroxychloroquine concentration containing 0.8% bovine serum albumin, the antigen was added. Following a 1-h incubation period, fresh human group O red blood cells from a single donor were added and incubated Oligomycin A clinical trial for a further 3 h. Positive and negative control sera were included in each testing run. Two independent
operators read the plate to determine the HI titre and no discordance was assumed. The endpoint titre was taken as the highest dilution of serum completely inhibiting agglutination. An antigen titration was performed in duplicate with each run to confirm the presence of four units of haemagglutinin. The data were analysed using the Statistical Package for Social Sciences (spss) software version 10 (SPSS, Chicago, IL). Descriptive statistics were used to describe the study population. The significance of differences between pre- and post-vaccination HI H1N1 antibody geometric mean titres (GMTs) was assessed using the paired t-test. Spearman’s rank correlation (rho) was used to determine the association between age and pre-vaccination HI titre. Geometric mean antibody levels at baseline and months 3, 6 and 9 were compared using the Kruskal–Wallis test. The Mann–Whitney U-test with Bonferroni correction (adjusted P-value of 0.0083) was used for post hoc comparison. The P-value for all other statistical analyses was set at 0.05. The seroprotection rate (SPR; i.e. the percentage of vaccine recipients with HI titre ≥ 40 after vaccination) and seroconversion rate (SCR; i.e.