2b). All subjects responded against all antigens, except one who only had FHA- and PRN-specific responses. Between days 28 and 150–180 after vaccination the numbers of antigen-specific BVD 523 memory B cells had declined. Some subjects

were back to background levels, whereas others had maintained higher levels of antigen-specific memory B cells compared to day 0. One subject had maintained the level of FHA-specific memory B cells between days 28 and 150–180. No vaccine-responders were seen in the culture-negative group ( Fig. 2b) or against the Libraries control antigen TTd (data not shown). For an in-depth evaluation of the memory B-cell response two panels were included in the flow cytometric analysis. Panel I identified different memory B-cell subpopulations (activated, resting and tissue-like) and panel II identified IgG-switched memory B cells. Detection and analysis were performed for 12 subjects (4 culture positives, 4 culture negatives and 4 placebos). Not all subjects had samples available for all time points. No differences were found between the culture positives, culture negatives or placebo when antibody isotype-switch was evaluated

(IgD+/− and IgG+/−), data not shown. However, there was an increase in the culture-positive group at days 7 and 14 of the activated memory B cells, as well as the tissue-like memory B cells (fig. 3). This was not seen in the naïve and resting memory B-cell subpopulations, nor did the FcLR4 staining differ between the groups (data not shown). The number of responding subjects was insufficient crotamiton for a thorough correlation analysis. Therefore, a more general comparison of the B-cell responses detected was made. The Ixazomib order serological response (as detected by ELISA, reported in detail in Ref. [16]), the plasma blast response and the memory B-cell response were compared in all seven culture-positive subjects (Fig. 4). As expected, the cellular response had declined in blood at day 150–180, whereas the serological response was maintained. There were minor exceptions where subjects differed between their cellular and humoral responses, but in general the subjects

responded similarly in the antigen-specific responses detected by both ELISpot and ELISA. The novel, live attenuated pertussis vaccine candidate, BPZE1, was tested for the first time in man and showed to be safe and able to induce serological responses [16]. In this study, we evaluated the B-cell responses evoked by BPZE1 during the same trial. In total 48 subjects were recruited to the study. Out of the 36 subjects that received the vaccine 7 were colonized by BPZE1 and mounted a response against the vaccine-related antigens. Since it was a first-in-man study, the dosages used in this study were based on studies in mice [19]. An optimization of the doses may perhaps lead to a better vaccine take. The results obtained in this study are considered exploratory due to the novelty of the vaccine.