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. 2023 Mar 18;8:43. doi: 10.1038/s41541-023-00638-6

Fig. 5. SpFN + ALFQ-induced significantly higher frequencies of plasmablasts in the spleen and higher frequencies of S-2P protein-specific long-lived plasma cells (LLPCs) in the bone marrow.

Fig. 5

Spleens from naive, or vaccinated SpFN + ALFQ or SpFN + AH mice (n = 5 in each group) were collected at week 6 (day 42, Fig. 5d). ELISpot and flow cytometry analysis were performed. a Percentage of plasmablasts (CD3-CD45R+CD138+) in all the three groups. b ELISpot images of the triplicate wells showing S-2P protein-specific- IgG (blue spots) and IgA (red spots) for each group. c Number of S-2P protein-specific IgG and IgA spot-forming units per million splenocytes are presented as a bar graph (Mean ± SEM). Each dot in the bar graph represents the average of triplicate wells for each mouse in each group. The responses were considered positive when the spot count exceeded the Mean ± 3 SD of the negative control wells. d A separate group of mice (n = 6 in each group) were vaccinated three times with SpFN + ALFQ or SpFN + AH. Bone marrows were collected at week 21 and the percentage of e plasma cells (CD3-CD45R-CD138+) and f intracellular S-2P protein-specific plasma cells were analyzed. The data are represented as bar graphs (Mean ± SD) and each dot represents the data from an individual spleen or bone marrow. The right panels show the representative contour or dot plots for each subset. The flow panel and the gating strategy for the respective subsets are shown in Supplementary Table 5 and Supplementary Fig. 6, respectively. The statistical differences between the two vaccinated groups were evaluated by Mann–Whitney U-test with P ≤ 0.05 considered as significant.