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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Immunol. 2011 Nov 30;188(1):404–416. doi: 10.4049/jimmunol.1102124

Figure 5. Number of DENV-specific memory B cells and PCs after 1° and 2° heterotypic DENV infection.

Figure 5

A. DENV-1-specific memory B cells detected by ELISPOT. AG129 mice were infected either with 105 PFU of DENV-2 D2S10 i.v. (1° infection) or with 105 PFU of DENV-1 98J s.c. (1° infection) then 6 to 8 weeks p.i., infected i.v. with 107 PFU of DENV-2 D2S10 (2° infection). A control group was mock-infected (Naïve or Post1°/Pre2°). After 3, 6, 9 and 14 days, splenocytes were harvested and polyclonally stimulated in vitro for 6 days to obtain ASCs. ASCs against DENV-1 were tested by ELISPOT using cellular Ag to detect DENV-1-specific memory B cells. The number of spots from control wells coated with Mock-Ag was subtracted from the number of spots counted in DENV-coated wells. Statistical analysis was performed using the Mann-Whitney test to compare the number of DENV-1-specific memory B cells at each time-point to the number of DENV-1-specific memory B cells from naïve or post1°/pre2° mock-infected mice. A statistically significant difference was found at day 9 and day 14 post-1° infection; p-values are shown below each graph. B. DENV-2-specific memory B cells detected by ELISPOT. Cells were prepared as in Figure 5A and were tested by ELISPOT using DENV-2 cellular Ag to detect DENV-2-specific memory B cells. Statistical analysis was performed using the Mann-Whitney U test to compare the number of DENV-2-specific memory B cells at each time-point to the number of DENV-2-specific memory B cells from naïve or post1°/pre2° mock-infected mice. A statistically significant difference was found at day 9 and day 14 post-1° infection and at day 6 and day 9 post-2° infection; p-values are shown below each graph. C. DENV-1-specific PCs detected by ELISPOT. Cells were prepared as in Figure 5A and tested ex vivo by ELISPOT using DENV-1 cellular Ag to detect DENV-1-specific PCs. Statistical analysis was performed using the Mann-Whitney U test to compare the number of DENV-1-specific PCs at each time-point to the number of DENV-1-specific PCs from naïve or post1°/pre2° mock-infected mice. A statistically significant difference was found at day 6, day 9 and day 14 post-1° infection and at day 6 and day 9 post-2° infection; p-values are shown below each graph. D. DENV-2-specific PCs detected by ELISPOT. Cells were prepared as in Figure 5A and were tested by ELISPOT ex vivo using DENV-2 cellular Ag to detect DENV-2-specific PCs. Statistical analysis was performed using the Mann-Whitney U test to compare the number of DENV-2-specific PCs at each time-point to the number of DENV-2-specific PCs from naïve or post1°/pre2° mock-infected mice. A statistically significant difference was found at day 6, day 9 and day 14 post-1° infection and at day 9 post-2° infection; p-values are shown below each graph.