Article Figures & Data
Figures
- FIGURE 1.
GC formation in TCR β−/− mice. Spleens were analyzed on day 12 after immunization. A, The percentages of GC B cells were assessed by staining splenocytes with PNA, anti-B220, and anti-Fas Abs. B, The volumes of GCs of wild-type (▪) and TCR β−/− (▪) were determined by both flow cytometry and in situ staining as described in Materials and Methods. Data are representative of two similar experiments with four to six animals in each group (mean ± SE).
- FIGURE 2.
Phenotype of GC T cells in TCR β−/− mice. Consecutive spleen sections of a TCR β−/− mouse were costained with PNA-HRP and biotinylated Abs to CD4 (A), CD8 (B), αβ TCR (C), γδ TCR (D), CD3 (E), or CD90 (F), followed by streptavidin-alkaline phosphatase. GCs were stained red and T cell markers were stained blue. Original magnification, ×200.
- FIGURE 3.
Clonotypic repertoire shift of GC B cells in TCR β−/− mice. VH segments from individual GCs were sequenced and analyzed. Percentages of GCs containing VH186.2 gene (open segments), analog genes (gray segments), or both VH186.2 and analog genes (black segments) from wild-type (A) or mutant (B) mice are indicated around the periphery of each chart. Percentages of unique clones carrying VH186.2 (open segments) or analog (gray segments) genes are shown in charts C and D. Percentages of all VDJ sequences containing VH186.2 (open segments) or analog (gray segments) genes are shown in charts E and F. The total number of GCs, clones, or VDJ sequences analyzed in each group is indicated in the center of each chart.
- FIGURE 4.
Use of κL chain by NP-specific Abs in TCR β−/− mice. A, Adjacent splenic sections of a wild-type or mutant mouse were stained with PNA (red) for GCs and anti-λ1 or κL chain Ab (blue). Original magnification, ×200. B, NP-specific Abs 12 days after primary or secondary immunization were determined using biotin-conjugated anti-λ1 or κL chain Ab as secondary Ab. The ratios of NP-specific Abs using λ1 or κL chain produced in wild-type (□) or TCR β−/− (▪) mice are shown (mean ± SE). Data are representative of two similar experiments with four to six animals in each group.
Tables
- Table I.
Mutational characteristics in VH gene segments from GCs of TCR β−/− and wild-type mice
TCR β−/− Wild-Type VH186.2 segments Mutation frequencya Total 1.4 1.8 CDRs 2.9 4.6 FRs 0.9 1.0 R/S ratio CDRs 61/0 13/1 FRs 3/1 3/1 Position 33b 42 35 Position 50c 0 0 DFL16.1d 61 41 Tyr95e 51 74 Analogue segments Mutation frequency Total 0.5 NA CDRs 0.8 NA FRs 0.4 NA R/S ratio CDRs 4/1 NA FRs 1/1 NA DFL16.1 (%) 2 NA Tyr95 (%) 12 NA -
a Point mutations (percentage (%) of base pairs). R, replacement; S, silent.
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b Percentage (%) of sequences containing mutation (W → L) at codon 33.
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c Percentage (%) of sequences containing mutation (R → G) at codon 50.
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d Percentage (%) of rearrangements using DFL16.1 segments.
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e Percentage (%) of rearrangements encoding tyrosine at codon 95 in CDR3. NA, not applicable.
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