The role of the bound peptide in alloreactive T-cell recognition is controversial, ranging from pep-tide-independent to peptide-specific recognition of alloreactive T-cells. The aim of this study is to find the eviden...The role of the bound peptide in alloreactive T-cell recognition is controversial, ranging from pep-tide-independent to peptide-specific recognition of alloreactive T-cells. The aim of this study is to find the evidence that there exist peptide/MHC complex (pMHC)-specific CTLs among alloreactive T cells generated with long-term mixed lymphocytes culture (LTMLC). A single pMHC was manipulated by loading the TAP-defective, HLA-A2 expressing T2 cells with a viral peptide (LMP2A426-434) or a self-peptide (Tyr369-377). The PBLs samples from 4 HLA-A2 positive (HLA-A2+ve) and 4 HLA-A2 negative (HLA-A2-ve) donors were included in this study. The HLA-A2+ve PBL co-cultured with the LMP2A426-434 pulsed T2 (T2/LMP) stands for the nominal T-cell response to a viral antigen, and the HLA-A2-ve PBLs co-cultured with the Tyr369-377 pulsed T2 (T2/Tyr) for alloreactive T-cell response to an allogeneic antigen. The specificity of the expanded CTLs after the LTMLC was detected by their specific cytotoxicity and binding ability to specific pMHC-tetramer. An HLA-A2 restricted, HIV peptide (Gag77-85)was included for control. The cultural bulk of HLA-A2+ve PBLs with the T2/LMP showed an elevated specific cytotoxicity against the T2/LMP compared to that against the T2/HIV (26.52%±3.72% vs 7.01%±0.87%, P<0.001), and an increased frequency of binding to LMP-tetramer compared to that binding to HIV-tetramer (0.98%±0.33% vs 0.05%±0.01%, P=0.0014). The cultural bulk of HLA-A2-ve PBLs with the T2/Tyr showed a more active cytotoxicity against the T2/Tyr than that against T2/HIV (28.07%±2.58% vs 6.87%±0.01 %, P<0.001), and a higher frequency of binding to the Tyr-tetramer than that binding to the HIV-tetramer (0.88%±0.3% vs 0.06%±0.03%, P=0.0018). Our results indicate that the LTMLC is able to expand the viral antigen-specific CTLs as well as allogeneic antigen-specific CTLs. A relatively large proportion of alloreactive CTLs should be pMHC-specific, i.e., the specificity of the alloreactive lines depends on both the bound peptide and the展开更多
目的观察参附汤联合同种异体NK细胞(alloreactive naturall killer cells,alloNK)对移植小鼠术后外周血象、骨髓有核细胞计数(nucleated cell counts,NCC)、骨髓造血组织容量(hematopoietic volume of bone marrow,HV)及骨髓病理的影响...目的观察参附汤联合同种异体NK细胞(alloreactive naturall killer cells,alloNK)对移植小鼠术后外周血象、骨髓有核细胞计数(nucleated cell counts,NCC)、骨髓造血组织容量(hematopoietic volume of bone marrow,HV)及骨髓病理的影响。方法BALB/c小鼠经过13.0 Gy剂量的全身照射(total body irradiation,TBI),4 h内经尾静脉注入供鼠骨髓单个核细胞(mononuclear cells,MNCs)8×10^(6)/kg,制作移植小鼠模型,然后随机分为模型组、alloNK组、参附汤组、参附+alloNK组4组,分别给予生理盐水、单用alloNK、单用参附汤和参附汤联合alloNK灌胃,观察治疗前及治疗后第7、14、30天,小鼠外周血象、骨髓NCC、HV及组织病理的变化。结果移植前及移植后第7天各组小鼠外周血象、骨髓NCC、HV无显著性差异。移植后第14天,模型组小鼠白细胞仍处于粒缺期,而单纯alloNK组和参附组小鼠的白细胞均有所上升,参附联合alloNK组小鼠上升幅度高于其他组。移植后第30天,模型组、alloNK组及参附组小鼠白细胞、骨髓NCC、HV均较前回升,但仍低于移植前水平,而参附联合alloNK组小鼠的白细胞、骨髓NCC、HV恢复至移植前水平,与其他组相比差异显著。结论参附汤可保护骨髓免受放射损伤、促进造血功能恢复、缩短粒细胞缺乏时间,而联合alloNK细胞促进造血功能恢复的作用更明显。展开更多
基金the National Natural Science Foundation of China (Grant Nos.30271201 and 30490241), and the "973" Project of the Ministry of Science and Technology of China (Grant No. 2001CB510008)
文摘The role of the bound peptide in alloreactive T-cell recognition is controversial, ranging from pep-tide-independent to peptide-specific recognition of alloreactive T-cells. The aim of this study is to find the evidence that there exist peptide/MHC complex (pMHC)-specific CTLs among alloreactive T cells generated with long-term mixed lymphocytes culture (LTMLC). A single pMHC was manipulated by loading the TAP-defective, HLA-A2 expressing T2 cells with a viral peptide (LMP2A426-434) or a self-peptide (Tyr369-377). The PBLs samples from 4 HLA-A2 positive (HLA-A2+ve) and 4 HLA-A2 negative (HLA-A2-ve) donors were included in this study. The HLA-A2+ve PBL co-cultured with the LMP2A426-434 pulsed T2 (T2/LMP) stands for the nominal T-cell response to a viral antigen, and the HLA-A2-ve PBLs co-cultured with the Tyr369-377 pulsed T2 (T2/Tyr) for alloreactive T-cell response to an allogeneic antigen. The specificity of the expanded CTLs after the LTMLC was detected by their specific cytotoxicity and binding ability to specific pMHC-tetramer. An HLA-A2 restricted, HIV peptide (Gag77-85)was included for control. The cultural bulk of HLA-A2+ve PBLs with the T2/LMP showed an elevated specific cytotoxicity against the T2/LMP compared to that against the T2/HIV (26.52%±3.72% vs 7.01%±0.87%, P<0.001), and an increased frequency of binding to LMP-tetramer compared to that binding to HIV-tetramer (0.98%±0.33% vs 0.05%±0.01%, P=0.0014). The cultural bulk of HLA-A2-ve PBLs with the T2/Tyr showed a more active cytotoxicity against the T2/Tyr than that against T2/HIV (28.07%±2.58% vs 6.87%±0.01 %, P<0.001), and a higher frequency of binding to the Tyr-tetramer than that binding to the HIV-tetramer (0.88%±0.3% vs 0.06%±0.03%, P=0.0018). Our results indicate that the LTMLC is able to expand the viral antigen-specific CTLs as well as allogeneic antigen-specific CTLs. A relatively large proportion of alloreactive CTLs should be pMHC-specific, i.e., the specificity of the alloreactive lines depends on both the bound peptide and the
文摘目的观察参附汤联合同种异体NK细胞(alloreactive naturall killer cells,alloNK)对移植小鼠术后外周血象、骨髓有核细胞计数(nucleated cell counts,NCC)、骨髓造血组织容量(hematopoietic volume of bone marrow,HV)及骨髓病理的影响。方法BALB/c小鼠经过13.0 Gy剂量的全身照射(total body irradiation,TBI),4 h内经尾静脉注入供鼠骨髓单个核细胞(mononuclear cells,MNCs)8×10^(6)/kg,制作移植小鼠模型,然后随机分为模型组、alloNK组、参附汤组、参附+alloNK组4组,分别给予生理盐水、单用alloNK、单用参附汤和参附汤联合alloNK灌胃,观察治疗前及治疗后第7、14、30天,小鼠外周血象、骨髓NCC、HV及组织病理的变化。结果移植前及移植后第7天各组小鼠外周血象、骨髓NCC、HV无显著性差异。移植后第14天,模型组小鼠白细胞仍处于粒缺期,而单纯alloNK组和参附组小鼠的白细胞均有所上升,参附联合alloNK组小鼠上升幅度高于其他组。移植后第30天,模型组、alloNK组及参附组小鼠白细胞、骨髓NCC、HV均较前回升,但仍低于移植前水平,而参附联合alloNK组小鼠的白细胞、骨髓NCC、HV恢复至移植前水平,与其他组相比差异显著。结论参附汤可保护骨髓免受放射损伤、促进造血功能恢复、缩短粒细胞缺乏时间,而联合alloNK细胞促进造血功能恢复的作用更明显。
