Secretion of autoimmune antibodies in the human subcutaneous adipose tissue
Fig 8
Autoantibody production in the SAT.
A. Total IgG antibodies were detected by ELISA in supernatants of unstimulated and CpG-stimulated SVF cultures. B. Fat-specific IgG antibodies were detected by ELISA after enrichment in IgG antibodies. ELISA plates were coated with protein lysates from adipocytes from the same individuals. C. Detection of GC B and T cells in the SVF, and in the blood (from the same individuals) as control, was performed by flow cytometry. GC B cells were CD19+CD10+IgD- and also expressed intracellular Bcl-6. Grey line, filled histogram: blood; black line, unfilled histogram: SVF. GC T cells were CD3+CD4+PD-1+CXCR5+. D. Naïve B cells were sorted from the peripheral blood of 8 lean healthy individuals and stimulated in the presence of ACM for 5–8 days to detect mRNA expression of AID and BLIMP-1, respectively. Cells (106/ml of ACM) were stimulated with 5 μg/ml of CpG. Results show qPCR values (2-ΔΔCt) of RNA expression of AID (top) and BLIMP-1 (bottom). Mean comparisons between groups were performed by Student’s t test (two-tailed). **p<0.01.