Immune Netw.  2013 Dec;13(6):240-248. 10.4110/in.2013.13.6.240.

Increased Lymphocyte Infiltration in Rheumatoid Arthritis Is Correlated with an Increase in LTi-like Cells in Synovial Fluid

• Koo J
• Kim S
• Jung WJ
• Lee YE
• Song GG
• Kim KS
• Kim MY

• KMID: 2150784
• DOI: http://doi.org/10.4110/in.2013.13.6.240

Abstract

In this study, we compared the immune cell populations in rheumatoid arthritis (RA) synovial fluid, which shows lymphoid tissue-like structure, with those in tonsils, which are normal secondary lymphoid tissues. Firstly, we found that CD4-CD11b+ macrophages were the major population in RA synovial fluid and that B cells were the major population in tonsils. In addition, synovial fluid from patients with osteoarthritis, which is a degenerative joint disease, contained CD4+CD11b+ monocytes as the major immune cell population. Secondly, we categorized three groups based on the proportion of macrophages found in RA synovial fluid: (1) the macrophage-high group, which contained more than 80% macrophages; (2) the macrophage-intermediate group, which contained between 40% and 80% macrophages; and (3) the macrophage-low group, which contained less than 40% macrophages. In the macrophage-low group, more lymphoid tissue inducer (LTi)-like cells were detected, and the expression of OX40L and TRANCE in these cells was higher than that in the other groups. In addition, in this group, the suppressive function of regulatory T cells was downregulated. Finally, CXCL13 expression was higher in RA synovial fluid than in tonsils, but CCL21 expression was comparable in synovial fluid from all groups and in tonsils. These data demonstrate that increased lymphocyte infiltration in RA synovial fluid is correlated with an increase in LTi-like cells and the elevation of the chemokine expression.

Keyword

Lymphoid tissue inducer; Rheumatoid arthritis; Tonsil

MeSH Terms

Arthritis, Rheumatoid*
B-Lymphocytes
Humans
Joint Diseases
Lymphocytes*
Lymphoid Tissue
Macrophages
Monocytes
Osteoarthritis
Palatine Tonsil
Synovial Fluid*
T-Lymphocytes, Regulatory

Figure

• 

  Figure 1 Flow cytometric analysis of cell populations in RA, OA, and tonsil showing by dot plots (A) and contour plots (B). X-axis shows forward-scattered light (FSC) and Y-axis shows side-scattered light (SSC). The cell populations were costained with anti-CD11b and anti-CD4 and analyzed in 'a' area of A and B (C), 'b' area of A and B (D), and 'c' area of A and B (E). The results are representative of each RA (n=17), OA (n=3), and tonsil (n=14) group.

• 

  Figure 2 Comparison of percentage of cell populations including B cells, NK cells, NKT cells, CD4 T cells, CD8 T cells, CD4+CD11b+ monocytes, and CD4-CD11b+ macrophages in RA (n=17), OA (n=3), and tonsil (n=14). Error bar shows the standard deviation.

• 

  Figure 3 Comparison of percentage of cell populations in RA patients. Synovial fluids which contain over 80% of CD4-CD11b+ macrophages were grouped as macrophage high group (high, n=4), and synovial fluids which contain between 40% and 80% of macrophages were grouped as macrophage intermediate group (int, n=10). Synovial fluids which contain less than 40% of macrophages were grouped as macrophage low group (low, n=3).

• 

  Figure 4 Identification and comparison of LTi/LTi-like cells in RA and tonsil. (A) Flow cytometric analysis of LTi-like cells. According to FSC and SSC, lymphocytes were selected and CD3-negative and CD117-positive population were gated. The expression of CD127 and RORC in CD117+CD3-CD56- cells was analyzed. Shaded histograms indicate isotype staining. (B) Correlation between the portion of macrophages and LTi-like cells in synovial fluid (SF). Percentage and absolute cell numbers of LTi-like cells in macrophage-high, intermediate (int), and low groups of RA synovial fluid were compared with tonsils. The statistical evaluation of the data was performed with Student's t-test and one-way analysis of variance. *p<0.05, and **p<0.005 compared to macrophage-high group. (C) Expression of OX40L and TRANCE in LTi-like cells in macrophage-high, intermediate, and low groups of RA synovial fluid and tonsil. Shaded histograms indicate isotype staining.

• 

  Figure 5 Flow cytometric analyses of OX40 expression on activated CD4 T cells and GITR and CD25 expression on Tregs in macrophage-high, intermediate (int), and low groups of RA synovial fluid and tonsil. Shaded histograms indicate isotype staining. The results are representative of each macrophage high (n=4), intermediate (n=10), and low (n=3) group.

• 

  Figure 6 Comparison of CXCL13 and CCL21 expression in macrophage-high, intermediate (int), and low groups of RA synovial fluid and tonsil. Error bar shows the standard deviation.

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