Figure 1.
Comparison of constitutive allele-specific HLA class I expression on stem cells and PBMC.
Flow cytometry analysis of allele-specific HLA class I expression. Histograms showing high expression of HLA-A2 and -A3 and a strongly downregulated HLA-B7, -B8 and -B27 expression on: (A) Four different primary human satellite cell lines, (B) Two bone marrow-derived primary MSC lines, (C) PBMCs from three different individuals.
Table 1.
Cells and cell lines of this study, their genomic types and semi-quantitative cell-surface expression of HLA class I.
Figure 2.
Kinetics of HLA-A and -B upon IFNγ stimulation of MSC and human satellite cells.
Overlay histograms showing expression of HLA class I alleles before and after IFNγ (25 ng/mL) stimulation. Kinetics (0 h, 24 h, 48 h and 72 h) is shown for different alleles, different MSC lines, human satellite cells and PBMC.
Figure 3.
Kinetics of allele-specific HLA class I surface expression.
Quantitative measurements of constitutive and up-regulated allele-specific HLA class I protein expression pre- and post-IFNγ stimulation of DD8 cells measured by indirect flow cytometry.
Table 2.
Primer sequences and quantitative allele-specific HLA-class I mRNA expression data on DD8 cells.
Figure 4.
Analysis of allele-specific HLA class I expression performed by complement-dependent microcytotoxicity assay.
(A) Histogram showing percentages of dead cells from three independent experiments. (B) Images showing CDC results for HLA-A2, -A3 and -B7 and -B27 for IFNγ-stimulated and non-stimulated DD8 cells. Green: living cells, red: dead cells.
Figure 5.
Kinetics of HLA class I gene transcripts upon IFNγ stimulation and correlation between allele-specific surface protein expression and mRNA expression.
(A) Quantitative measurements of constitutive and up-regulated allele-specific HLA class I gene transcripts pre- and post-IFNγ stimulation by quantitative RT-PCR for DD8 cells. (B) Correlation between constitutive and up-regulated expression of HLA-A and -B allelic gene transcripts and the capacity of antibody binding to the surface HLA class I allelic proteins by indirect flow cytometry staining for HLA-A2, -B7 and -B27. HLA-B*07 showed the lowest constitutive mRNA expression while HLA-B27 showed the lowest constitutive protein expression.