A.K.I. Kuroki, Masayuki Iyoda, Takanori Shibata, Tetsuzo Sugisaki

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Th2 cytokines increase and stimulate B cells to produce IgG4 in idiopathic membranous nephropathy A.K.I. Kuroki, Masayuki Iyoda, Takanori Shibata, Tetsuzo Sugisaki Kidney International Volume 68, Issue 1, Pages 302-310 (July 2005) DOI: 10.1111/j.1523-1755.2005.00415.x Copyright © 2005 International Society of Nephrology Terms and Conditions

Figure 1 Peripheral lymphocyte subpopulations in patients with idiopathic membranous nephropathy (MN) and normal controls (NC). The numbers of lymphocytes were calculated based on the results of flow cytometry. (A) Numbers of the peripheral lymphocytes. (B) Percentages of CD3+ T cells and CD19+ B cells. (C) Numbers of CD3+ T cells and CD19+ B cells. (D) Percentages of CD4+ T cells and CD8+ T cells. (E) Numbers of CD4+ T cells and CD8+ T cells. (F) CD4+/8+ ratio. Each column and bar represents the mean and 1 SD. The numbers of lymphocytes, both T and B cells were identical in the two groups. The idiopathic membranous nephropathy patients showed an increased percentage of CD4+ T cells, and a decreased number of CD8+ T cells, resulting in a high CD4+/8+ ratio. Kidney International 2005 68, 302-310DOI: (10.1111/j.1523-1755.2005.00415.x) Copyright © 2005 International Society of Nephrology Terms and Conditions

Figure 2 Cytokine mRNA levels in peripheral blood mononuclear cells (PBMCs). Cytokine mRNA levels were quantified using real-time reverse transcription-polymerase chain reaction (RT-PCR) and results were expressed in relative copy numbers normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA level. (A) Interferon (IFN)-γ. (B) Interleukin (IL)-4. (C) IL-10. (D) IL-13. Each column and bar represents the mean and 1 SD. The levels of Th2 cytokines IL-10 and IL-13 increased in the idiopathic membranous nephropathy (MN) group. NC is normal controls. Kidney International 2005 68, 302-310DOI: (10.1111/j.1523-1755.2005.00415.x) Copyright © 2005 International Society of Nephrology Terms and Conditions

Figure 3 IgG subclass production by B cells. B cells were cultured in the absence (A) or presence of cytokines. (B) Interferon (IFN)-γ. (C) Interleukin (IL)-4. (D) IL-10. (E) IL-13. After 7 days of incubation, the levels of IgG subclasses in the supernatant were quantified. For the concentrations, the scales on the left side are for total IgG, IgG1, and IgG2, and the scales on the right side are for IgG3 and IgG4. Percentages are expressed as the mean and 1 SD. Open circles and bars represent normal controls, and closed circles and bars represent idiopathic membranous nephropathy. Although some B cells from idiopathic membranous nephropathy patients augmented the production of IgG4 in response to IL-4, IL-10, or IL-13, there was no significant difference in the concentration and percentage of each IgG subclass between the two groups. Kidney International 2005 68, 302-310DOI: (10.1111/j.1523-1755.2005.00415.x) Copyright © 2005 International Society of Nephrology Terms and Conditions

Figure 4 To assess the effect of cytokines on the production of IgG subclasses in individual samples, the increase ratio was calculated. (A) Interferon (INF)-γ. (B) Interleukin (IL)-4. (C) IL-10. (D). IL-13. Columns and bars indicate the mean and 1 SD of increase ratios for each group. The mean increase ratio of IgG4 in the presence of IL-4 in the idiopathic membranous nephropathy group was significantly higher than that in the normal control group. Kidney International 2005 68, 302-310DOI: (10.1111/j.1523-1755.2005.00415.x) Copyright © 2005 International Society of Nephrology Terms and Conditions