B lymphocyte memory: role of stromal cell complement and FcgammaRIIB receptors - PubMed (original) (raw)

B lymphocyte memory: role of stromal cell complement and FcgammaRIIB receptors

Robert A Barrington et al. J Exp Med. 2002.

Abstract

To dissect the influence of CD21/CD35 and FcgammaRIIB in antigen retention and humoral memory, we used an adoptive transfer model in which antigen-primed B and T lymphocytes were given to sublethally irradiated wild-type mice or mice deficient in CD21/CD35 (Cr2(-/-)) or FcgammaRIIB receptors (FcgammaRIIB(-/-)). Cr2(-/-) chimeras showed impaired memory as characterized by a decrease in antibody titer, reduced frequency of antibody secreting cells, an absence of affinity maturation, and significantly reduced recall response. The impaired memory in Cr2(-/-) chimeras corresponded with the reduced frequency of antigen-specific memory B cells. Interestingly, FcgammaRIIB(-/-) chimeras showed a differential phenotype with impaired splenic but normal bone marrow responses. These data suggest that CD21/CD35 on stroma, including follicular dendritic cells, is critical to the maintenance of long-term B lymphocyte memory.

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Figures

Figure 1.

Figure 1.

Frequency of NP-specific ASCs in chimeric mice 16 wk after receiving memory B lymphocytes. Recipient BM (a and c) and spleen (b and d) before (a and b) or 1 wk after antigen challenge intravenously with 50 μg NP5-KLH (c and d) were analyzed by ELISPOT for IgG-secreting plasma cells specific for NP. Data are represented as means ± SEM (a and b shows three independent experiments and c and d shows four independent experiments). n = number of mice in each group. *, statistically significant differences upon comparisons to WT chimeras (P < 0.035 BM and P < 0.02 spleen).

Figure 2.

Figure 2.

Frequency of donor-derived, NP-specific memory B lymphocytes in chimeric mice 16 wk after adoptive transfer. (a) Representative scheme to identify antigen-specific, donor-derived memory B lymphocytes by FACS®. Frequency of NP-binding CD45.1+Igλ+ B lymphocytes (CD19+) was determined from mouse BM (b and c) and spleen (d and e) before (b and d) or 1 wk after antigen challenge intravenously with 50 μg NP-KLH (c and e). Cells were also confirmed as CD3−, IgM−, CD138−, and CD11b−. Data are represented as means ± SEM of three independent experiments. n = number of mice in each group. *, statistically significant differences upon comparisons to WT chimeras (P < 0.01 BM and spleen prechallenge and P < 0.04 after challenge).

Figure 3.

Figure 3.

Changes in relative serum affinity with time after antigen exposure. Sera collected from chimeric mice were titrated in ELISA for the amount of high affinity (NP5-BSA) and the amount of total (NP15-BSA) IgG. The ratio of NP5:NP15 titers is expressed as mean ratios ± SEM and are compiled from four independent experiments. ▪, WT chimeras (n = 34); ♦, Cr2−/− chimeras (n = 31); •, FcγRIIB−/− chimeras (n = 32); *, statistically significant differences. Statistical comparisons were made to the 2–3-wk time point for each group (P < 0.01).

Figure 4.

Figure 4.

Memory responses after secondary transfer. Splenic B lymphocytes from original chimeric mice were adoptively transferred intravenously with KLH-primed T cells and NP-KLH into sublethally irradiated WT (a and c) and Cr2−/− (b and d) recipients. 3 wk after secondary transfer into WT (a) and Cr2−/− (b) mice, NP-binding serum IgG was measured by ELISA. Each symbol represents a single mouse. Horizontal bars represent mean antibody titers. (c and d) ELISPOTs were performed on recipient splenocytes to determine ASC frequency. Data are represented as means ± SEM from a single experiment. *, statistically significant differences. Statistical comparisons to WT chimeras were made (P < 0.01).

Figure 5.

Figure 5.

Dependence on CD21/CD35+ stroma for localizing antigen in GCs. 16 wk after adoptive transfer of memory B lymphocytes, WT (a), FcgRIIB−/− (b), and Cr2−/− (c) chimeric mice were injected with biotinylated NP-BSA. (d) WT chimeras not receiving biotinylated antigen. 5-μM splenic cryosections were stained with anti-IgM (blue) and PNA (green) to delineate B lymphocyte zones and GCs, respectively. Arrows point to detectable antigen, as visualized using SA-TRITC (red). Colocalized IgM, PNA, and antigen are detected in white. Staining patterns shown are representative of multiple sections (>5) from five mice per chimeric group.

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