Objective. To study thymus-dependent T cell development and T cell repertoire in human s ever com-bined immunodeficiencypatients after HLA-identical or haploid entical T cell-depleted allogeneic bone marrow transplant...Objective. To study thymus-dependent T cell development and T cell repertoire in human s ever com-bined immunodeficiencypatients after HLA-identical or haploid entical T cell-depleted allogeneic bone marrow transplantation.Methods .Blood samples were obtained from15SCID patients before transplantation and a t varying intervals thereafter.Quantitative competitive PCR assay and immunosco pe analysis of the T cell receptorVarepertoire were performed.Results. Before and within the first100days after transplantation,patients’ periphera l blood mononuclear cellpresented an oligoclonal or polyclonal skewed T cell repertoire,low T cell re-ceptor excision circlesvalues and pred ominance of CD45RO + T cell.In contrast,the presence of high numbers of CD45RA + T cells in bone marrowcirculation reconstituted SCID patients(>10 0days post-transplantation)correlated with active T cell production by the th ymus as revealed by high TREC values,and a polyclonal T cell repertoire demonst rated by a Gaussian distribution of Va-specific peaks.Conclusions.Within one year after BMT ,a normal T cell repertoire develops in SCID patients as a resu lt of thymic output.The T cell receptor diversity is highly and positively corr elated in these patients with TREC levels.展开更多
Analysis of complementarity determining region 3 （CDR3） length of T lymphocyte receptors （TCRs） by immunoscope spectratyping technique has been used successfully to investigate the diversity of TCR in autoimmune d...Analysis of complementarity determining region 3 （CDR3） length of T lymphocyte receptors （TCRs） by immunoscope spectratyping technique has been used successfully to investigate the diversity of TCR in autoimmune diseases and infection diseases. In this study, we investigated the patterns of CDR3 length distribution for all 32 TCR AV gene families in human peripheral blood lymphocytes of four normal volunteers by the immunoscope spectratyping technique. It was found that PCR products exhibited an obscure band on 1.5% agarose gel electrophoresis. Each TCR AV family exhibited more than 8 bands on 6% sequencing gel electrophoresis. The CDR3 spectratyping of all TCR AV families showed a standard Gaussian distribution with different CDR3 length, and the expression frequency of CDR3 was similar among the gene families. Most of CDR3 in TCR AV family recombine in frame. However, some of the CDR3 showed out-of frame gene rearrangement. Additionally, we found that in some of TCR AV families there were 18 amino acid discrepancies between the longest CDR3 and shortest CDR3. These results may be helpful to further study the recombination mechanism of human TCR genes, the TCR CDR3 gene repertoire, and the repertoire drift in health people and disease state. Cellular ＆ Molecular Immunology.展开更多
文摘Objective. To study thymus-dependent T cell development and T cell repertoire in human s ever com-bined immunodeficiencypatients after HLA-identical or haploid entical T cell-depleted allogeneic bone marrow transplantation.Methods .Blood samples were obtained from15SCID patients before transplantation and a t varying intervals thereafter.Quantitative competitive PCR assay and immunosco pe analysis of the T cell receptorVarepertoire were performed.Results. Before and within the first100days after transplantation,patients’ periphera l blood mononuclear cellpresented an oligoclonal or polyclonal skewed T cell repertoire,low T cell re-ceptor excision circlesvalues and pred ominance of CD45RO + T cell.In contrast,the presence of high numbers of CD45RA + T cells in bone marrowcirculation reconstituted SCID patients(>10 0days post-transplantation)correlated with active T cell production by the th ymus as revealed by high TREC values,and a polyclonal T cell repertoire demonst rated by a Gaussian distribution of Va-specific peaks.Conclusions.Within one year after BMT ,a normal T cell repertoire develops in SCID patients as a resu lt of thymic output.The T cell receptor diversity is highly and positively corr elated in these patients with TREC levels.
文摘Analysis of complementarity determining region 3 （CDR3） length of T lymphocyte receptors （TCRs） by immunoscope spectratyping technique has been used successfully to investigate the diversity of TCR in autoimmune diseases and infection diseases. In this study, we investigated the patterns of CDR3 length distribution for all 32 TCR AV gene families in human peripheral blood lymphocytes of four normal volunteers by the immunoscope spectratyping technique. It was found that PCR products exhibited an obscure band on 1.5% agarose gel electrophoresis. Each TCR AV family exhibited more than 8 bands on 6% sequencing gel electrophoresis. The CDR3 spectratyping of all TCR AV families showed a standard Gaussian distribution with different CDR3 length, and the expression frequency of CDR3 was similar among the gene families. Most of CDR3 in TCR AV family recombine in frame. However, some of the CDR3 showed out-of frame gene rearrangement. Additionally, we found that in some of TCR AV families there were 18 amino acid discrepancies between the longest CDR3 and shortest CDR3. These results may be helpful to further study the recombination mechanism of human TCR genes, the TCR CDR3 gene repertoire, and the repertoire drift in health people and disease state. Cellular ＆ Molecular Immunology.
