Additional file 1 of Both T and B cells are indispensable for the development of a PBMC transfer-induced humanized mouse model for SSc
Creators: name: Shu, Yaqing
name: Yue, Xiaoyang
name: Wax, Jacqueline
name: Kasper, Brigitte
name: Yin, Junping
name: Wang, Xiaoqing
name: Zhang, Liang
name: Ahmadi, Marjan
name: Heidecke, Harald
name: Müller, Antje
name: Lamprecht, Peter
name: Yu, Xinhua
name: Riemekasten, Gabriela
name: Petersen, Frank
Publisher: name: figshare
Year: 2022
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Supplementary Table 1. Demographic and clinical features of SSc patients recruited for the in vitro T or B cell-depletion. Supplementary Table 2. Overview on medications given to all individual SSc patients with SSc treated with or without immunosuppressive drugs in the present and previous study [5]. Supplementary Figure 1. Efficiency of depletion of human T or B cells from PBMC. Levels of CD3+ T cells (A), CD4+ T cells (B), CD8+ T cells (C), and CD20+ B cells (D) were determined by FACS analysis and presented as % of total human leukocytes in PBMC. Statistical significance of comparison was determined using the paired t test. Supplementary Figure 2. Levels of human leukocytes in peripheral blood of recipient mice. Mice were transferred with whole PBMC, T-cell depleted, or B-cell depleted PBMC, and peripheral blood was taken at the 6th week (A, B) and 12th week (C, D) after the transfer. Subsequently, human CD45+ leukocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells and CD20+ B cells were identified by flow cytometry. Levels of human leukocytes are presented as percentage of total leukocytes including murine and human leukocytes in murine blood. Comparison on levels of human leukocytes indicated between mice transferred with whole PBMC (n = 7) and mice transferred with T-cell depleted PBMC (n = 7) (A) and between mice transferred with whole PBMC (n = 10) and mice transferred with B-cell depleted PBMC (n = 10) (B) by week by the 6th week after the transfer. Comparison on levels of human leukocytes indicated between mice transferred with whole PBMC (n = 4) and mice transferred with T-cell depleted PBMC (n = 4) (C) and between mice transferred with whole PBMC (n = 16) and mice transferred with B-cell depleted PBMC (n = 6) (D) by week by the 12th week after the transfer. P values reflect comparisons between mice transferred with whole PBMC and mice transferred with T- or B-cell depleted PBMC. Statistical significance was determined using the Wilcoxon matched pairs test. Supplementary Figure 3. Presence of ANA in sera of PBMC-transferred mice. Antinuclear antibody (ANA) pattern of murine sera were detected using (HEp-2) cell-based immunofluorescence staining (EUROPattern Suite, Euroimmun, Germany). Two out of 7 mice which received whole PBMC from SSc patients scored positive for ANA, and their ANA patterns were consistent with those of two corresponding SSc patients. By contrast, mice which received T cell-depleted or B cell-depleted PBMC isolated from the 2 SSc patients were ANA negative. Representative micrographs of the ANA test for mice received whole PBMC, T cell-depleted PBMC (PBMC T-/-) or B cell-depleted PBMC (PBMC B-/-) are shown. Supplementary Figure 4. Matrix of Spearman’s correlation coefficients between immunological and histopathological variables. Severity of inflammation in the lung, kidney and liver as well as levels of circulating human CD45+ leukocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells and CD20+ B cells at 6th week after the cell transfer, and levels of total IgG, anti-AT1R IgG and anti-ETAR IgG in sera obtained after the sacrifice of mice were used for the analysis. Samples were tested for normal distribution by using Shapiro-Wilk normality test. Since most variables were not normally distributed, Spearman correlation was applied for the analysis. A color-coded correlation scale is provided on the right of the plot. Blue and ellipses represent negative and positive correlations, respectively, and darker color tones representd larger correlation coefficient magnitudes. Statistically significant differences are indicated by asterisks (*p&lt;0.05, **p&lt;0.01 and ***p&lt;0.001).