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CXCR4/IgG-expressing plasma cells are associated with human gastrointestinal tissue inflammation  Clarisa M. Buckner, PhD, Susan Moir, PhD, Lela Kardava,

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Presentation on theme: "CXCR4/IgG-expressing plasma cells are associated with human gastrointestinal tissue inflammation  Clarisa M. Buckner, PhD, Susan Moir, PhD, Lela Kardava,"— Presentation transcript:

1 CXCR4/IgG-expressing plasma cells are associated with human gastrointestinal tissue inflammation 
Clarisa M. Buckner, PhD, Susan Moir, PhD, Lela Kardava, PhD, Jason Ho, PhD, Brian H. Santich, BA, Leo Jin Young Kim, BA, Emily K. Funk, BA, Amy K. Nelson, BSN, Britanny Winckler, BA, Cheryl L. Chairez, BSN, Narda L. Theobald-Whiting, BS, Sandra Anaya-O’Brien, MSN, Meghna Alimchandani, MD, Martha M. Quezado, MD, Michael D. Yao, MD, Joseph A. Kovacs, MD, Tae-Wook Chun, PhD, Anthony S. Fauci, MD, Harry L. Malech, MD, Suk See De Ravin, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 133, Issue 6, Pages e5 (June 2014) DOI: /j.jaci Copyright © Terms and Conditions

2 Fig 1 Identification of gut-derived PCs by using flow cytometry. A, PCs of a representative HD were defined as CD19+CD3−CD20−CD27++ within the lymphocyte gate. Each plot title identifies the gated cell population, and numbers indicate cell percentages in each quadrant or gate. FSC, Forward scatter; SSC, side scatter. B, The immunoglobulin isotype profile was determined with 2 separate stains: one for IgA and IgG and the other for IgM and IgD. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

3 Fig 2 Characterization of dual IgA/IgG-expressing PCs in the gut. A, Microscopy of sorted PCs stained for IgA (Alexa Fluor 488, green) and IgG (Alexa Fluor 594, red). Arrows show dual positivity. B, Expression of immunoglobulin light chains for single IgG- or IgA-expressing and dual IgG/IgA-expressing PCs. Numbers indicate cell percentages in each quadrant. The dotted box identifies cells sorted for transcriptional analysis (see Fig E1). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

4 Fig 3 Immunoglobulin isotype distribution among gut-derived and peripheral blood PCs/plasmablasts. A and B, Immunoglobulin isotype mean distribution (Fig 3, A) and median comparisons of gut-derived PCs (Fig 3, B) from the groups described in Table I. Light gray triangles, Patients with CGD with no gastrointestinal symptoms. C and D, Analyses were performed on peripheral blood PCs/plasmablasts from participants in Table I. Horizontal bars represent medians. NS, Not significant. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

5 Fig 4 Expression of CCR10 on gut-derived PCs. A, Representative plots of CCR10 expression by IgA and IgG are shown by group. Numbers indicate cell percentages in each quadrant. B and C, Histograms from plots in Fig 4, A (Fig 4, B), and group comparisons of CCR10 expression (Fig 4, C) were obtained by gating on single IgG- or IgA-expressing and dual IgG/IgA-expressing PCs. Horizontal bars represent medians and asterisks and light gray triangles identify IgA-deficient subjects and patients with CGD with no gastrointestinal symptoms from Table I, respectively. NS, Not significant. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

6 Fig 5 A-C, Expression of CXCR4 on gut-derived PCs. Analyses for CXCR4 were identical to those presented for CCR10 in Fig 4. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

7 Fig 6 Correlation and association between CXCR4, IgG, and combined scores. Comparisons were performed between fraction of gut-derived PCs expressing only IgG and the intensity (MFI) of CXCR4 expression on PCs expressing only IgG (A) and the percentage of CXCR4+ PCs expressing only IgG and the combined pathology and gastrointestinal symptom scores described in Table I (B). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

8 Fig E1 Properties of surface IgA+/IgG− gut-derived PCs. A, Flow cytometry with and without permeabilization of cells was performed to evaluate the expression of CD32, IgG, and IgA on (Surface) and within (Intracellular) gut-derived PCs. B, RT-PCR was performed to evaluate the expression of IgG and IgA in gut-derived PCs that were sorted by IgA. CD19+ B cells from the peripheral blood and sorted by IgA+/IgG− were used as positive controls, and amplification of RNaseP was included as a housekeeping gene. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

9 Fig E2 A, Graphs are identical to those of Fig 3, B, with the exception that the CGD group does not include patients without colitis. Only those P values that changed are shown. B, Graphs are identical to those of Fig 5, B, with the exception that the CGD group does not include patients without colitis. Only P values that changed are shown. NS, Not significant. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

10 Fig E3 Comparison of homing receptor expression on gut-derived PCs by immunoglobulin isotype. Data are from Figs 4, B, and 5, B, with all groups combined. Intensities of CCR10 (A) and CXCR4 (B) were compared between PCs expressing only IgA, dual IgA/G-expressing PCs, and PCs expressing only IgG. Horizontal bars represent medians. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © Terms and Conditions

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