case_images: PMC7552884_01
case_images: PMC7552884_02
case_images: PMC7552884_03
case_images: (A) NGS identified two heterozygous missense variants in trans in RAG1. Relative recombination activity level of the two alleles [16% (ref 16) and 40% (novel variant, measured in this study)] predict an overall relative recombinase activity of 28% that is characteristic of CID-G/AI phenotype (*based on gnomAD 12/2019). (B) CD3+ T cells analyzed by spectratyping using 28 probes to measure CDR3 length variability in 23 TCRVbeta families and sub-families. Families 2 to 18 are selected for representation. The following families demonstrated an oligoclonal distribution, i.e., <5 peaks: 3-1, 5, 6-4, 10-1, 10, 13, 14, and 18; family 16 had no peak. Each is identified with an * in the upper right corner. (C) Detection of Valpha7.2 segment in CD3+ T lymphocytes by flow cytometry shows that the patient had near absence of CD3+T cells expressing the TCR Valpha 7.2 chain [encoded by a distal V TCR locus] compared to healthy control and family members (measured in collaboration with the NIH). (D) Frequency of individual IGHJ gene usage of memory B cell receptor repertoire shows decrease in usage of the distal IGHJ5 and IGHJ6 gene elements in patient compared to age matched healthy donor.
case_images: fimmu-11-01954-g0001.jpg
case_images: PMC7552884_04_fimmu-11-01954-g0001.jpg
case_images: F1
case_images: Considering one variant had 16% predicted activity and the variant on the other allele 40% predicted activity, the calculated average recombinase activity is 28% (Figure 1A). In comparison to healthy controls, there was also a dramatic decline in usage of the distal IGHJ5 and IGHJ6 gene elements (BCR), suggesting an abnormal V (D) J recombination consistent with partial RAG deficiency (Figure 1D). TCRValpha7.2+ lymphocytes compared to controls (0.27% vs. 1.85-4.49%) (Figure 1C). The TCR/BCR repertoire studies, recombinase activity assay, and specific flow cytometric testing for distal Valpha7.2 element, all support a V (D) J recombination defect with partial RAG deficiency as the underlying cause of disease.
case_images: PMC7552884_04
case_images: (A) NGS identified two heterozygous missense variants in trans in RAG1. Relative recombination activity level of the two alleles [16% (ref 16) and 40% (novel variant, measured in this study)] predict an overall relative recombinase activity of 28% that is characteristic of CID-G/AI phenotype (*based on gnomAD 12/2019). (B) CD3+ T cells analyzed by spectratyping using 28 probes to measure CDR3 length variability in 23 TCRVbeta families and sub-families. Families 2 to 18 are selected for representation. The following families demonstrated an oligoclonal distribution, i.e., <5 peaks: 3-1, 5, 6-4, 10-1, 10, 13, 14, and 18; family 16 had no peak. Each is identified with an * in the upper right corner. (C) Detection of Valpha7.2 segment in CD3+ T lymphocytes by flow cytometry shows that the patient had near absence of CD3+T cells expressing the TCR Valpha 7.2 chain [encoded by a distal V TCR locus] compared to healthy control and family members (measured in collaboration with the NIH). (D) Frequency of individual IGHJ gene usage of memory B cell receptor repertoire shows decrease in usage of the distal IGHJ5 and IGHJ6 gene elements in patient compared to age matched healthy donor.
case_images: fimmu-11-01954-g0001.jpg
case_images: PMC7552884_04_fimmu-11-01954-g0001.jpg
case_images: F1
case_images: Considering one variant had 16% predicted activity and the variant on the other allele 40% predicted activity, the calculated average recombinase activity is 28% (Figure 1A). In comparison to healthy controls, there was also a dramatic decline in usage of the distal IGHJ5 and IGHJ6 gene elements (BCR), suggesting an abnormal V (D) J recombination consistent with partial RAG deficiency (Figure 1D). TCRValpha7.2+ lymphocytes compared to controls (0.27% vs. 1.85-4.49%) (Figure 1C). The TCR/BCR repertoire studies, recombinase activity assay, and specific flow cytometric testing for distal Valpha7.2 element, all support a V (D) J recombination defect with partial RAG deficiency as the underlying cause of disease.
case_images: PMC7552884_05
case_images: PMC7552884_06
case_images: PMC7552884_07