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Assessment of the lungs, bone marrow (BM) and spleens of saline- or DC10-treated asthmatic mice as reservoirs for OVA-specific IgG1-secreting B cells.

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posted on 02.01.2018, 18:57 by Yanna Ma, Wojciech Dawicki, Xiaobei Zhang, John R. Gordon

(A) Asthmatic BALB/c mice were given saline or DC10 treatments as indicated in Fig 1A; all mice were exposed to a recall allergen challenge 2 dy prior to sacrifice. Healthy, non-asthmatic mice were used as a negative control. At the indicated times (dy 2, 9, 16, 30, 58) we generated single cell suspensions from their lungs, BM and spleen, and assayed the numbers of OVA-specific IgG1-secreting B cells in 5 h ELISPOT assays, as noted in the Materials and Methods section. The data depicts the numbers of IgG1-secreting B cells per 1x106 input cells, presented as the mean (±SEM) of 4 wells/sample. (B) Plasma was obtained from the animals in panel A and assayed for OVA-specific IgG1 antibody by ELISA with duplicate wells per sample, represented as the mean (±SEM). The data are representative of three experiments (for each, n = 4 or 5 for experimental mice, and 2 for normal control mice). The statistical analyses were performed using one-way ANOVA assays with Tukey’s post-hoc testing. **, ***, **** and NS signify p<0.01, 0.001, or 0.0001, and p>0.05 respectively.