Article Figures & Data
Figures
- FIGURE 1.
Immunohistology of the ST of patients with RA and ReA. Frozen tissue sections (6 μm) were labeled with mAbs specific for FDC (A, D, and E), plasma cells (B and F), CD4 (C and G), and CD20 (H). A double labeling with Abs specific for Ki-67 (brown color) and plasma cells (red color) is shown (B). A, B, and E–H, Consecutive sections from the ST of patient RA-PS: infiltrate VI is the right hand cluster seen in A and B, infiltrate I is shown in E–H. Infiltrate II from the ST of patient ReA-TS is shown in C and D.
- FIGURE 2.
Stepwise accumulation of somatic mutations. V gene diversification is shown for a heavy chain (VH3–8) (A) and a light chain (DPL23) (B) rearrangement. Numbered circles indicate isolated sequences, empty circles indicate hypothetical intermediates. Numbers besides the arrows refer to the number of nucleotide exchanges that distinguish one sequence from another. Data were obtained with two independent PCR reactions (indicated by open and shaded circles).
- FIGURE 3.
Diversity of V gene sequences isolated from infiltrate I of patient RA-PS. The diversity of B cells isolated from the network of FDC (A and D), the T cell-rich zone (B and E), and the ring of plasma cells (C and F) is shown. A–C, The repertoire of productively rearranged VH genes is shown. D–F, The number of nucleotide differences to the presumptive germline genes is shown. The somatic diversity is given for heavy and light chain sequences. For the two rearrangements where intraclonal diversity was found, the number of nucleotide exchanges are shown by open bars. For VH genes, the nomenclature of Matsuda et al. is given (26).
- FIGURE 4.
Diversity of V gene sequences isolated from infiltrate II of patient ReA-TS. The diversity of B cells isolated from the network of FDC (A and C) and the ring of plasma cells (B and D) is shown. A and B, The repertoire of productively rearranged VH genes is shown. C and D, The number of nucleotide differences to the presumptive germline genes is shown. Results from two consecutive sections are summarized. 4-Or corresponds to the VH-4 gene DP69, which is an orphan gene located on chromosome 15 (4-OR15-8; IMGT database, M.-P. Lefranc). For further details, see Fig. 3.
- FIGURE 5.
Intraclonal diversity of plasma cell V genes. Heavy and light chain sequences are compared with the most homologous germline gene. Only nucleotide differences are shown, CDR are indicated, and codons are numbered according to Kabat (41). Sequences were isolated from plasma cells of two patients with RA (infiltrate I from patient RA-PS and infiltrate V from patient RA-EK) and one patient with ReA (infiltrate II from patient ReA-TS). Genealogical relationship indicating the clonal V gene diversification is shown. For further details, see Fig. 2. E, The results of a single-cell analysis is shown.
- FIGURE 6.
A comparison of the VH gene repertoire expressed in CD20+ B (A) and plasma cells (B). Results from four different patients with RA and ReA are summarized. A total of 62 different rearrangements from CD20+ B cells and a total of 41 from plasma cells were isolated.
Tables
Patient Infiltratea Section CD20+ B Cells Plasma Cells Heavy chain Light chain Heavy chain Light chain κ λ κ λ Group of cells RA PS I 56 27/33 6/9 5/8 9/25 5/9 2/2 VII 56 1/1 14/35 1/5 59 10/23 61 2/2 2/4 VI 56 4/20 59 4/15 5/14 RA EK IV 158 1/14 160 9/15 5/7 1/6 1/5 V 164 11/20 7/14 6/19 AR III 87 4/10 5/16 ReA TS II 6 15/23 11/15 6/10 10/17 8/11 5/7 7 2/3 7/17 2/12 7/11 Single cellsb RA EK VIII 65 3 5 1 a For infiltrates I to VII, the number of different rearrangements/total sequenced V genes is given.
b For the single-cell analysis, the number of isolated V genes with a different rearrangement is given.
- Table II.
V gene repertoire in CD20+ B cells isolated from the network of FDC (patient PS infiltrate VI on section 59)
Gene Family Rearrangement No. of Isolated Sequences: PCR No. of Somatic Mutations Length of CDR3 1st 2nd Heavy chain VH1 1-2/JH4b 1 1 –a VH3 3-11/JH4b 4 3 4b 16 3-43/JH4b 5 6 12 VH4 4-30-2/JH4b 1 2 12 Light λ-chain Vλ1 DPL5/JL1 1 1 13 Vλ2 LV2046/JL2.3 2 1 10 Vλ3 DPL23/JL1 5 3 1b 9 DPL23/JL2.3 1 7 11 IgLV3S6/JL2.3 2 9 8 a Out of frame rearrangement.
b Number of common mutations.
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