Journal: Cell reports

Article Title: Monocytes Acquire the Ability to Prime Tissue-Resident T Cells via IL-10-Mediated TGF-β Release

doi: 10.1016/j.celrep.2019.06.087

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: IL-10 Secretion Assay , Miltenyi , 130-090-435.

Techniques: Purification, Recombinant, Antibody Labeling, Enzyme-linked Immunosorbent Assay, Luminex, Cell Isolation, Activation Assay

Journal: Life Science Alliance

Article Title: The expansion in lymphoid organs of IL-4 + BATF + T follicular helper cells is linked to IgG4 class switching in vivo

doi: 10.26508/lsa.201800050

Figure Lengend Snippet: (A) Immunofluorescence staining of CD4 (red), ICOS (green), IL-4 (magenta), and DAPI (blue) in tissues from normal tonsils. The yellow broken line demarcates the area within GCs. The white broken line demarcates the area outside GC. (B) Immunofluorescence staining of CD4 (red), ICOS (green), IL-4 (magenta), and DAPI (blue) in tissues from normal tonsils. (C) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent antibody used to stain tissue from an IgG4-RD patient, normal tonsils, and normal mesenteric lymph nodes. (D) CD4 + ICOS + IL-4 + (red) and CD4 + ICOS + IL-4 − (green) cells were quantified in tissue from 12 tonsils. (E) Scatter plots depict the mean fluorescence intensity per cell quantified using TissueQuest software for each fluorescent antibody used to stain normal tonsils and mesenteric lymph nodes.

Article Snippet: The stimulated cells were then enriched for IL-4–secreting cells using the vendor’s protocol (IL-4 Secretion Assay Kit; Miltenyi Biotec, #130-054-101).

Techniques: Immunofluorescence, Staining, Fluorescence, Software

Journal: Life Science Alliance

Article Title: The expansion in lymphoid organs of IL-4 + BATF + T follicular helper cells is linked to IgG4 class switching in vivo

doi: 10.26508/lsa.201800050

Figure Lengend Snippet: (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 T FH and CD4 + Bcl-6 + IL-4–negative T FH 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 + T FH 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 + T FH 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 T FH and CD4 + IL-4 + CXCR5–negative non-T FH 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).

Article Snippet: The stimulated cells were then enriched for IL-4–secreting cells using the vendor’s protocol (IL-4 Secretion Assay Kit; Miltenyi Biotec, #130-054-101).

Techniques: Immunofluorescence, Staining, Fluorescence, Software, MANN-WHITNEY, Expressing

Journal: Life Science Alliance

Article Title: The expansion in lymphoid organs of IL-4 + BATF + T follicular helper cells is linked to IgG4 class switching in vivo

doi: 10.26508/lsa.201800050

Figure Lengend Snippet: (A) Gating strategy used to sort IL-4–secreting and nonsecreting tonsillar CD45RA + CXCR5 + T FH cells following anti-CD3/anti-CD28 stimulation. CD45RA + CXCR5 − cells and CD45RA − CXCR5 − IL-4 + cells were sorted as additional controls. (B) A heatmap of differentially expressed genes across all four conditions depicting the Z -scores for normalized expected read counts. (C) A correlation matrix of differentially expressed genes. (D) Expression pattern of individual ILs is depicted on a log scale. (E) Expression of CD molecules, cytokines, and transcription factors clustered into patterns using k -means. Z -scores of the expected read counts for each cluster are shown.

Article Snippet: The stimulated cells were then enriched for IL-4–secreting cells using the vendor’s protocol (IL-4 Secretion Assay Kit; Miltenyi Biotec, #130-054-101).

Techniques: Expressing

Journal: Life Science Alliance

Article Title: The expansion in lymphoid organs of IL-4 + BATF + T follicular helper cells is linked to IgG4 class switching in vivo

doi: 10.26508/lsa.201800050

Figure Lengend Snippet: (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 + T FH cells and CD4 + CXCR5 + T FH 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 + T FH cells. Green arrows indicate IgG4 + B cells. A number of IL-4–expressing T FH 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 T FH -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 + T FH cells. Magenta arrows indicate IgG4 + B cells. A number of BATF + ICOS + T FH 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 + T FH -cell nucleus. All internuclear distances in this figure were <0.2 μm. LZ, light zone; DZ, dark zone.

Article Snippet: The stimulated cells were then enriched for IL-4–secreting cells using the vendor’s protocol (IL-4 Secretion Assay Kit; Miltenyi Biotec, #130-054-101).

Techniques: Immunofluorescence, Staining, MANN-WHITNEY, Expressing, Membrane

Journal: Life Science Alliance

Article Title: The expansion in lymphoid organs of IL-4 + BATF + T follicular helper cells is linked to IgG4 class switching in vivo

doi: 10.26508/lsa.201800050

Figure Lengend Snippet: (A) Quantification of CD4 + CXCR5 + IL-4 + and CD4 + CXCR5 + T FH cells in 17 IgG4- RD SMGs, seven SS LSGs, 12 tonsils, two cervical lymph nodes, and three mesenteric lymph nodes. The P -value is based on the Mann–Whitney U test. (B) Correlations of the proportion of CD4 + CXCR5 + IL-4 + T FH cells and CD4 + BATF + IL-4 + T cells in SMGs from patients with IgG4-RD and their serum IgG, IgA, IgE, IgM, or IgG4 levels. The r and P -value were determined using Spearman’s rank correlations. (C) Correlations of the proportions of CD4 + CXCR5 + IL-4 + T FH cells in SMGs from patients with IgG4-RD and their ratio of IgG4 + /IgG + cells (n = 17). The r and P -value were determined using Spearman’s rank correlations. (D) The frequency of CD4 + CXCR5 + IL-4 + T FH cells in SMGs from patients with IgG4-RD (n = 17) correlated with the frequency of CD4 + BATF + IL-4 + T cells and serum IgG4 concentrations. The r and P -value were determined using Spearman’s rank correlations.

Article Snippet: The stimulated cells were then enriched for IL-4–secreting cells using the vendor’s protocol (IL-4 Secretion Assay Kit; Miltenyi Biotec, #130-054-101).

Techniques: MANN-WHITNEY