Frequencies of ASC ranged from 52 to 1065 sfu/106 MNC for total IgG and from 115 to 906 sfu/106 MNC for total IgA in all tissues other than bone marrow, where frequencies for both isotypes exceeded 2500/106 MNC ( Table 4). In most instances, only low frequencies of anti-gp140 ASC were detected; notably however, IgG anti-gp140-specific ASC represented 6% and 16% of total IgG secreting cells recovered from the interior iliac lymph nodes of animals E53 and E56 respectively; the animals that failed to seroconvert after intravaginal immunisation but responded following intramuscular immunisation ( Table find more 4). This is the first demonstration that intravaginally delivered buy Sirolimus soluble

recombinant HIV-1 gp140 is immunogenic in primates in the absence of a conventional mucosal adjuvant. Although intravaginal immunisation alone was less efficient in macaques compared to in rabbits at inducing serum and mucosally-detected antibodies, where a single cycle of immunisation was sufficient to induce responsiveness [21], women in a parallel clinical trial, were also essentially unresponsive following a single cycle of immunisation (Lewis et al., personal communication). This may reflect inefficiency in accessing sites of inductive immunity,

despite the strategy of repeated inoculation designed to increase the likelihood of exposure to immunogen during the menstrual cycle. However, there may only be a narrow window for induction of immune function during any one cycle as shown by studies in women [19]. It was therefore

encouraging that two macaques responded with serum and mucosally-detected responses after multiple cycles of intravaginal administration and that a third macaque was primed for a response as revealed by subsequent intramuscular immunisation. This “silent priming” of a memory B-cell response may be similar to that observed in macaques following mucosal exposure to “subinfectious” doses of SIV where antigen-primed B cells were detected in vitro by amplified Parvulin ELISpot in the absence of seroconversion [32]. In the present study, for logistical reasons it was not possible to sample lymphoid tissues over time; intriguingly however, high frequencies of gp140-specific IgG ASC were detected post mortem in the interior iliac lymph nodes from the two intravaginally immunised macaques that seroconverted only after intramuscular immunisation. These lymph nodes, which drain both the female genital and the rectal tracts, have been associated with protective immunity in SIV challenge studies [33]. In the present study we were unable to recover sufficient cells directly from cervico-vaginal tissues for ELISpot analysis and therefore assessment of the frequency of mucosal ASC was not possible.