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n D3 deficient mice were analyzed, P 0.0001, error bars are SEM. (E) Loss of cyclin D3 caused a 2 fold decrease in GC size. Each column in the graph corresponds to an individual mouse; each point is an individual GC, P 0.01. P values are based on two tailed Student t tests.
Full figure and legend (56K)
Defective antibody response in Ccnd3 miceIn order to determine whether the structural defect in GC formation also translates to a functional perturbation in the T cell dependent antibody response, CSR and affinity maturation were evaluated in vivo. Basal serum levels of most Ig subtypes were comparable between WT and Ccnd3 animals, with the latter showing a modest but statistically significant elevation of IgG1 and a decrease in IgG2b concentrations (Figure 4A). Next, animals were immunized with the T dependent antigen 4 hydroxy 3 nitrophenyl acetyl (NP) KLH and boosted 21 days and again 42 days later to elicit secondary and tertiary antibody responses 24 (experimental scheme summarized in Supplementary information, Figure S2). Ccnd3 animals produced significantly lower serum anti NP titers of all subclasses tested except IgM and IgA (Figure 4B). We studied the IgG1 response in more detail, as this is the main anti NP subclass elicited in vivo. An impairment in total NP specific IgG1 primary titers was observed in Ccnd3 mice (Figure 4C) (P = 0.0264), which persisted after the first NP KLH boost (P = 0.0010) but was ameliorated by the tertiary immunization (P = 0.32). The abnormalities in GC development are likely to be the main cause of these defects, although in vitro CSR assays in isolated B cells suggested that a minor contribution may also come from suboptimal switching to IgG1 (Supplementary information, Figure S3). Ccnd3 mice exhibited statistically significant impairments in affinity maturation as measured either by anti NP3 titers (Figure 4D) or by anti NP3 titers normalized to the total anti NP response (Figure 4E) (P 0.0001). Interestingly, the secondary immunization markedly raised the anti NP affinity of Ccnd3 sera such that Cheap UGG Boots the genotype difference essentially disappeared (P = 0.16) and this trend persisted through the tertiary response (P = 0.53) (Figure 4E). Collectively, these results indicate that cyclin D3 plays an important role in affinity maturation in vivo. However, the initial impairments in both the affinity and magnitude of the T dependent antibRequisites Of The Home Vegetable Garden
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Full figure and legend (56K)
Defective antibody response in Ccnd3 miceIn order to determine whether the structural defect in GC formation also translates to a functional perturbation in the T cell dependent antibody response, CSR and affinity maturation were evaluated in vivo. Basal serum levels of most Ig subtypes were comparable between WT and Ccnd3 animals, with the latter showing a modest but statistically significant elevation of IgG1 and a decrease in IgG2b concentrations (Figure 4A). Next, animals were immunized with the T dependent antigen 4 hydroxy 3 nitrophenyl acetyl (NP) KLH and boosted 21 days and again 42 days later to elicit secondary and tertiary antibody responses 24 (experimental scheme summarized in Supplementary information, Figure S2). Ccnd3 animals produced significantly lower serum anti NP titers of all subclasses tested except IgM and IgA (Figure 4B). We studied the IgG1 response in more detail, as this is the main anti NP subclass elicited in vivo. An impairment in total NP specific IgG1 primary titers was observed in Ccnd3 mice (Figure 4C) (P = 0.0264), which persisted after the first NP KLH boost (P = 0.0010) but was ameliorated by the tertiary immunization (P = 0.32). The abnormalities in GC development are likely to be the main cause of these defects, although in vitro CSR assays in isolated B cells suggested that a minor contribution may also come from suboptimal switching to IgG1 (Supplementary information, Figure S3). Ccnd3 mice exhibited statistically significant impairments in affinity maturation as measured either by anti NP3 titers (Figure 4D) or by anti NP3 titers normalized to the total anti NP response (Figure 4E) (P 0.0001). Interestingly, the secondary immunization markedly raised the anti NP affinity of Ccnd3 sera such that Cheap UGG Boots the genotype difference essentially disappeared (P = 0.16) and this trend persisted through the tertiary response (P = 0.53) (Figure 4E). Collectively, these results indicate that cyclin D3 plays an important role in affinity maturation in vivo. However, the initial impairments in both the affinity and magnitude of the T dependent antibRequisites Of The Home Vegetable Garden
ARTICLE REPRINTING IS PERMITTED
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