Article Figures & Data
Figures
- FIGURE 1.
Anti-TNF treated RA patients have reduced fractions of PB memory B cells. A, Representative phenotypic analysis of B cells in human PB of healthy control (left), RA patient on MTX (middle), or RA patient on anti-TNF (right). PBMCs were stained with Abs to CD19, CD27, and IgD and analyzed by multicolor flow cytometry. Data are presented as dot plots on CD19-gated B cells. B cell subsets are defined as indicated (N, naïve; pre-M, unswitched memory; post-M, switched memory), with the numbers in each dot plot representing the percentage of B cells in each subset. B, Percentage of memory B cells. Mean values ± SD are indicated for each group.
- FIGURE 2.
TNF blockade increases PB transitional and naive B cells while decreasing memory B cell numbers. A, Absolute numbers of naive (N) and memory (M) B cells were calculated from the fractions defined in Fig. 1, the percentage of lymphocytes, and the percentage of CD19 cells identified on flow cytometry. The increase in naive and the decrease in memory B cell numbers was reflected in the highly significant increase in the ratio of absolute naive to memory B cells with anti-TNF. B, Percentage of transitional B cells was determined by staining PBMCs with Abs to CD19, CD24, and CD38 as described in Materials and Methods. Mean values are indicated by the horizontal bars.
- FIGURE 3.
Anti-TNF treatment inhibits GC reactions and FDC networks in peripheral lymphoid tissue. A, Immunohistochemistry of tonsillar tissue in a normal control, a RA patient on MTX, and a RA patient on anti-TNF. FDCs are labeled with the CNA.42 FDC-specific Ab in the top panels. Images represent serial sections. B, Primary follicles are indicated by diffuse IgD staining (∗) (also distinguished by the absence of polarized dark/light zones and the paucity of T cells; data not shown for the latter) and secondary follicles (arrowhead points to GC) by polarization with peripheral IgD staining (follicular naive B cells), dark zone staining for Ki67, and light zone staining with anti-CD23 (arrowhead) (CD23 is also expressed at a low level on follicular B cells in the mantle). C, Phenotypic analysis of B cells in human tonsil.
Tables
Healthy RA: MTX RA: Anti-TNF HAQ (0–3) 0.24 (0.22) 0.3 (0.47) DAS (0–9.3) 3.69 (1.0) 3.3 (1.3) RF+ (%) 77% 79% GC (Bm3/4) 33 ± 12 27 ± 18 13 ± 9b FDC (%) 12.2 ± 4.8 11.9 ± 5.1 7.5 ± 1.6c FDC/IgD 0.63 ± 0.29 0.71 ± 0.10 0.42 ± 0.14d GC (%) 8.4 ± 3.6 9 ± 5.2 2.3 ± 1.6e 1°:2° 2.8:13.2 1:14 4.7:7.3 a The first three rows from the top summarize select patient characteristics (HAQ, health assessment questionnaire; DAS, disease activity score) with mean ± SD or positive percentage (RF, rheumatoid factor) in RA on MTX (n = 17) and RA on anti-TNF (n = 28) recruited for PB analysis. The next four rows characterize tonsil tissue from healthy subjects (n = 25 for flow cytometry analysis of GC cells), RA on MTX (n = 2), and RA on anti-TNF (n = 4). FDC network area is quantitated as a percentage of the total tissue area (FDC (%)) or relative to the IgD+ areas (FDC/IgD) and is based on morphometric analysis of immunohistochemistry stains. The fraction of the total tissue area occupied by GC FDC networks (GC (%)) and the mean primary number (1°) to secondary follicle number (2°) are shown. The statistical significance of the differences is indicated as follows:
b p = 0.003 compared to healthy controls;
c p = 0.036 compared to healthy controls;
d p < 0.0001 compared to RA MTX; and
e p < 0.0001 compared to RA MTX and healthy controls.
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