IL-4+BATF+ TFH cells are rare in normal SLOs but abundant in IgG4-RD.

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IL-4+BATF+ TFH cells are rare in normal SLOs but abundant in IgG4-RD. IL-4+BATF+ TFH cells are rare in normal SLOs but abundant in IgG4-RD. (A) Immunofluorescence staining of CD4 (red), Bcl-6 (green), IL-4 (magenta), and DAPI (blue) in tissues from normal tonsils. Quantification of CD4+Bcl-6+IL-4–positive TFH and CD4+Bcl-6+IL-4–negative TFH in 12 tonsils. (B) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent immunostain in IgG4-RD SMGs (G1) and normal tonsils. Quantification of CD4+CXCR5+IL-4+ and CD4+CXCR5+ TFH cells in 17 IgG4-RD SMGs, 12 tonsils, and 7 SS salivary glands. The P-value is based on the Mann–Whitney U test. (C) Almost all IL-4–expressing CD4+ T cells in IgG4-RD lymph nodes were CXCR5+ TFH cells. Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent immunostain in IgG4-RD lymph node (LN1). The quantification of CD4+IL-4+CXCR5–positive TFH and CD4+IL-4+CXCR5–negative non-TFH cells in lymph nodes from two patients with IgG4-RD (LN1 and LN2) is shown. (D) CD4+IL-4+BATF+ T cells were enriched in IgG4-RD SMGs. Immunofluorescence staining of CD4 (red), IL-4 (green), BATF (magenta), and DAPI (blue) in IgG4-RD SMGs (G12). White arrows indicate CD4+IL-4+BATF+ T cells. (E) Quantification of CD4+BATF+IL-4+ T cells and CD4+ T cells in 17 IgG4-RD SMGs and 12 normal tonsils. The P-value is based on the Mann–Whitney U test. (F) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent antibody used to stain normal tonsils and IgG4-RD lymph node (LN1). (G) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent immunostain in IgG4-RD SMGs (G3). (H) Immunofluorescence staining of ICOS (green), BATF (magenta), and IL-4 (Red) in IgG4-RD LNs (LN1). (I) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent immunostain in IgG4-RD LNs (LN1). The quantification of ICOS+BATF+IL-4–positive T and ICOS+BATF+IL-4–negative T cells in lymph nodes from six patients with IgG4-RD (LN1–LN6). (J) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent immunostain in IgG4-RD SMGs (G3). Takashi Maehara et al. LSA 2018;1:e201800050 © 2018 Maehara et al.