CD4+CXCR5+IL-4+ TFH cells are abundant around GCs in IgG4-RD and sometimes physically contact AID-expressing B cells. CD4+CXCR5+IL-4+ TFH cells are abundant.

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CD4+CXCR5+IL-4+ TFH cells are abundant around GCs in IgG4-RD and sometimes physically contact AID-expressing B cells. CD4+CXCR5+IL-4+ TFH cells are abundant around GCs in IgG4-RD and sometimes physically contact AID-expressing B cells. (A) CD4+CXCR5+IL-4+ T cells were enriched around TLOs in IgG4-RD SMGs. Immunofluorescence staining of CD4 (red), IL-4 (magenta), CXCR5 (green), and DAPI (blue) in IgG4-RD SMGs (patient G3). Quantification of CD4+CXCR5+IL-4+ TFH cells and CD4+CXCR5+ TFH cells comparing those outside and within GCs from each of five different areas in TLOs of 17 patients with IgG4-RD (G1–G17). The P-value is based on the Mann–Whitney U test. (B) IgG4+B cells were enriched outside GC, especially some these cells express AID. Immunofluorescence staining of CD4 (red), CD19 (blue), and Bcl6 (green) in IgG4-RD SMGs (patient G3). Immunofluorescence staining of AID (red), Bcl6 (magenta), IgG4 (green), and DAPI (blue) in IgG4-RD SMGs (patient G3). The numbers of IgG4+ cells (per square micrometer) outside and within GCs were quantified from each of five different areas in TLOs of 17 patients with IgG4-RD (G1–G17). The P-value is based on the Mann–Whitney U test. (C) AID-expressing B cells outside GCs in IgG4-RD lymph nodes were abundant. Immunofluorescence staining of Bcl6 (red), AID (green), and ICOS (orange) in a normal tonsil and IgG4-RD lymph node. (D) Immunofluorescence staining of ICOS (green), IL-4 (red), and AID (magenta) in IgG4-RD SMGs. (E) Immunofluorescence staining of IgG4 (green), ICOS (magenta), IL-4 (red), and DAPI (blue) in TLOs with an IgG4-RD patient (G3). Magenta arrows indicate ICOS+IL-4+ TFH cells. Green arrows indicate IgG4+ B cells. A number of IL-4–expressing TFH cells and IgG4+ B cells formed close and extensive intercellular plasma membrane contacts. The distance was measured between the edge of each IgG4+ B cell nucleus to the edge of the closest IL-4–expressing TFH-cell nucleus, and an internuclear distance of less than 0.4 μm was indicative of a T-B conjugate. (F) Immunofluorescence staining of ICOS (green), IgG4 (magenta), BATF (red), and DAPI (blue) in the lymph nodes of an IgG4-RD patient. Green arrows indicate ICOS+BATF+ TFH cells. Magenta arrows indicate IgG4+ B cells. A number of BATF+ICOS+ TFH cells and IgG4+ B cells formed close and extensive intercellular plasma membrane contacts. The distance was measured between the edge of each IgG4+ B-cell nucleus to the edge of the closest BATF+ICOS+ TFH-cell nucleus. All internuclear distances in this figure were <0.2 μm. LZ, light zone; DZ, dark zone. Takashi Maehara et al. LSA 2018;1:e201800050 © 2018 Maehara et al.