PMC6831519_01
(A) Sequence alignment of the patient's BAM file on IGV (Integrative Genomics Viewer) and the two heterozygous mutation found by IL7R DNA sequencing. (B) Patient's genealogical tree.
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F1
Torrent), revealed two novel compound heterozygous mutations in IL-7Ralpha gene confirmed by Sanger sequencing (Figure 1A). The C82 was involved in disulfide bond formation with C74 (Supplementary Figure 1A) and the S54 was involved in moderate electrostatic bond formation with Q45 (hydrogen bond distance 2.7A, Supplementary Figure 1B), both destabilizing the IL7R antiparallel beta-sheets.
Phenotypic and pSTAT5 signaling in patient's relatives and in patient derived lymphocytes. (A) IL-7Ralpha (CD 127) surface expression were assessed by means of multiparameter flow cytometry in T cell subset. (B) The phosphorylation status of STAT5 (Y694) was determined by means of Phosflow analysis after IL-7 treatment in T cell subsets.
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F2
T lymphocytes and the cutaneous histopathology a diagnosis of SCID with maternal T-cell engraftment was suspected. The variable number of tandem repeat analysis performed on both peripheral blood and the skin biopsy revealed that all T lymphocytes (CD4+ and CD8+) in the patient were of maternal origin while CD19+ and CD16+CD56+ cells were autologous (Supplementary Figure 2). IL-7Ralpha expression was reduced in total CD3+ T cells with a bimodal pattern explained by normal expression on CD3+CD4+cells (data not showed) and marked reduction on CD3+CD4- T cells (Figure 2). In line with these results, STAT5 phosphorylation in response to IL-7 stimulation (50 ng/mL) was reduced in total CD3+, normal in CD3+CD4+cells and almost absent in CD3+CD4- T lymphocytes compared to her parents (Figure 2).