异基因造血干细胞移植(allo-HSCT)后,供者来源的自然杀伤细胞(NK)可在保留移植物抗白血病(GVL)效应的同时发挥控制移植物抗宿主病(GVHD)作用。NK细胞表面的1磷酸鞘氨醇受体5(S1PR5)通过与胞外的第一信使1磷酸鞘氨醇(S1P)相互作用,调节N...异基因造血干细胞移植(allo-HSCT)后,供者来源的自然杀伤细胞(NK)可在保留移植物抗白血病(GVL)效应的同时发挥控制移植物抗宿主病(GVHD)作用。NK细胞表面的1磷酸鞘氨醇受体5(S1PR5)通过与胞外的第一信使1磷酸鞘氨醇(S1P)相互作用,调节NK细胞的迁移和在外周血、脾、淋巴结等器官中的分布。本研究旨在探讨allo-HSCT后外周血NK细胞S1PR5表达变化及对GVHD的影响。分离纯化17例供者及相应受者移植后1个月外周血NK细胞,通过荧光探针实时定量PCR技术检测S1PR5 mRNA的表达情况,并结合患者临床资料和随访结果进行分析。结果表明,供者与allo-HSCT后患者外周血中NK细胞S1PR5表达变化(0.235±0.191 vs0.330±0.261,P>0.05)不具有统计学意义;急性GVHD(aGVHD)患者组移植后外周血NK细胞S1PR5表达水平较无aGVHD的患者组明显下降(0.973±0.834 vs 6.166±5.32,P<0.05);移植后患者与相应供者相比,外周血NK细胞S1PR5表达水平下降超过10%时易发生aGVHD;NK细胞S1PR5表达变化与慢性GVHD发生率无明显相关性(3.401±2.324 vs 2.762±1.972,P>0.05)。结论:NK细胞S1PR5表达水平下调与aGVHD发生有关,可能机制是由于S1PR5的低表达影响了NK细胞在体内的分布,从而影响到调控aGVHD的作用。展开更多
Graft-versus-host disease （GVHD） is the most common complication after hematopoietic stem cell transplantation. To clarify the role of Toll-like receptor 4 （TLR4）, which is a major receptor for bacterial lipopolys...Graft-versus-host disease （GVHD） is the most common complication after hematopoietic stem cell transplantation. To clarify the role of Toll-like receptor 4 （TLR4）, which is a major receptor for bacterial lipopolysaccharides （LPS）, in the development of acute GVHD, we used a TLR4-knockout （TLR4-/-） mouse GVHD model and analyzed the underlying immunological mechanisms. When TLR4-/- mice were used as bone marrow and splenocyte cell graft donors or recipients, GVHD symptom occurrence and mortality were delayed compared to wild-type （TLR4＋/＋） mice. In addition, histopathological analyses revealed that in TLR4-/-→BALB/c chimeras, liver and small intestine tissue damage was reduced with minimal lymphocytic infiltration. In contrast to TLR4＋/＋, TLR4-/- mice dendritic cells did not express CD80, CD86, CD40, MHC-II or IL-12 during LPS induction and remained in an immature state. Furthermore, the ability of TLR4-/- mice spleen dendritic cells to promote allogeneic T-cell proliferation and, in particular, T-helper cell 1 （Th 1） development was obviously attenuated compared with TLR4＋/＋ mice dendritic cells, and the levels of interferon-T （IFN-γ） and IL-IO, Th2-cell specific cytokines, were significantly higher in the serum of TLR4-/-→BALB/c than in TLR4＋/＋→BALB/c chimeric mice. Overall, our data revealed that TLR4 may play a role in the pathogenesis of GVHD and that targeted TLR4 gene therapy might provide a new treatment approach to reduce the risk of GVHD.展开更多
The successful of transplantation is determined by the shared human leukocyte antigens(HLAs) and ABO blood group antigens between donor and recipient. In recent years, killer cell receptor [i.e., killer cell immunoglo...The successful of transplantation is determined by the shared human leukocyte antigens(HLAs) and ABO blood group antigens between donor and recipient. In recent years, killer cell receptor [i.e., killer cell immunoglobulinlike receptor(KIR)] and major histocompatibility complex(MHC) class I chain-related gene molecule(i.e., MICA) were also reported as important determinants of transplant compatibility. At present, several different genotyping techniques(e.g., sequence specific primer and sequence based typing) can be used to characterize blood group, HLA, MICA and KIR and loci. These molecular techniques have several advantages because they do not depend on the availability of anti-sera, cellular expression and have greater specificity and accuracy compared with the antibody-antigen based typing. Nonetheless, these molecular techniques have limited capability to capture increasing number of markers which have been demonstrated to determine donor and recipient compatibility. It is now possible to genotype multiple markers and to the extent of a complete sequencing of the human genome using next generation sequencer(NGS). This high throughput genotyping platform has been tested for HLA, and it is expected that NGS will be used to simultaneously genotype a large number of clinically relevant transplantation genes in near future. This is not far from reality due to the bioinformatics support given by the immunogenetics community and the rigorous improvement in NGS methodology. In addition, new developments in immune tolerance based therapy, donor recruitment strategies and bioengineering are expected to provide significant advances in the field of transplantation medicine.展开更多
文摘异基因造血干细胞移植(allo-HSCT)后,供者来源的自然杀伤细胞(NK)可在保留移植物抗白血病(GVL)效应的同时发挥控制移植物抗宿主病(GVHD)作用。NK细胞表面的1磷酸鞘氨醇受体5(S1PR5)通过与胞外的第一信使1磷酸鞘氨醇(S1P)相互作用,调节NK细胞的迁移和在外周血、脾、淋巴结等器官中的分布。本研究旨在探讨allo-HSCT后外周血NK细胞S1PR5表达变化及对GVHD的影响。分离纯化17例供者及相应受者移植后1个月外周血NK细胞,通过荧光探针实时定量PCR技术检测S1PR5 mRNA的表达情况,并结合患者临床资料和随访结果进行分析。结果表明,供者与allo-HSCT后患者外周血中NK细胞S1PR5表达变化(0.235±0.191 vs0.330±0.261,P>0.05)不具有统计学意义;急性GVHD(aGVHD)患者组移植后外周血NK细胞S1PR5表达水平较无aGVHD的患者组明显下降(0.973±0.834 vs 6.166±5.32,P<0.05);移植后患者与相应供者相比,外周血NK细胞S1PR5表达水平下降超过10%时易发生aGVHD;NK细胞S1PR5表达变化与慢性GVHD发生率无明显相关性(3.401±2.324 vs 2.762±1.972,P>0.05)。结论:NK细胞S1PR5表达水平下调与aGVHD发生有关,可能机制是由于S1PR5的低表达影响了NK细胞在体内的分布,从而影响到调控aGVHD的作用。
基金We are grateful to Miao Chen, Qiangguo Gao and Yiqi Liu （Second Military Medical University, Shanghai, China） for technical support and offer special thanks to Professor Qing Yi （M.D. Anderson Cancer Center Houston, TX, USA） for helpful guidance in the experiments. We thank Shizuo Akira （Osaka University, Osaka, Japan） for originally providing key mouse strains. This work was supported by grants of the National Natural Science Foundation of China （no. 30772502 and 30973455）, Zhejiang Major Medical and the Health Science and Technology ＆ Ministry of Health of the Chinese Government （no. WKJ2009-2-022）. This work was also supported by the Major Research Plan of the Chinese National Natural Science Foundation （no. 91029740）, Zhejiang Province Science and Technology Department Foundation （no. 2009C03012-2） and Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents.
文摘Graft-versus-host disease （GVHD） is the most common complication after hematopoietic stem cell transplantation. To clarify the role of Toll-like receptor 4 （TLR4）, which is a major receptor for bacterial lipopolysaccharides （LPS）, in the development of acute GVHD, we used a TLR4-knockout （TLR4-/-） mouse GVHD model and analyzed the underlying immunological mechanisms. When TLR4-/- mice were used as bone marrow and splenocyte cell graft donors or recipients, GVHD symptom occurrence and mortality were delayed compared to wild-type （TLR4＋/＋） mice. In addition, histopathological analyses revealed that in TLR4-/-→BALB/c chimeras, liver and small intestine tissue damage was reduced with minimal lymphocytic infiltration. In contrast to TLR4＋/＋, TLR4-/- mice dendritic cells did not express CD80, CD86, CD40, MHC-II or IL-12 during LPS induction and remained in an immature state. Furthermore, the ability of TLR4-/- mice spleen dendritic cells to promote allogeneic T-cell proliferation and, in particular, T-helper cell 1 （Th 1） development was obviously attenuated compared with TLR4＋/＋ mice dendritic cells, and the levels of interferon-T （IFN-γ） and IL-IO, Th2-cell specific cytokines, were significantly higher in the serum of TLR4-/-→BALB/c than in TLR4＋/＋→BALB/c chimeric mice. Overall, our data revealed that TLR4 may play a role in the pathogenesis of GVHD and that targeted TLR4 gene therapy might provide a new treatment approach to reduce the risk of GVHD.
文摘The successful of transplantation is determined by the shared human leukocyte antigens(HLAs) and ABO blood group antigens between donor and recipient. In recent years, killer cell receptor [i.e., killer cell immunoglobulinlike receptor(KIR)] and major histocompatibility complex(MHC) class I chain-related gene molecule(i.e., MICA) were also reported as important determinants of transplant compatibility. At present, several different genotyping techniques(e.g., sequence specific primer and sequence based typing) can be used to characterize blood group, HLA, MICA and KIR and loci. These molecular techniques have several advantages because they do not depend on the availability of anti-sera, cellular expression and have greater specificity and accuracy compared with the antibody-antigen based typing. Nonetheless, these molecular techniques have limited capability to capture increasing number of markers which have been demonstrated to determine donor and recipient compatibility. It is now possible to genotype multiple markers and to the extent of a complete sequencing of the human genome using next generation sequencer(NGS). This high throughput genotyping platform has been tested for HLA, and it is expected that NGS will be used to simultaneously genotype a large number of clinically relevant transplantation genes in near future. This is not far from reality due to the bioinformatics support given by the immunogenetics community and the rigorous improvement in NGS methodology. In addition, new developments in immune tolerance based therapy, donor recruitment strategies and bioengineering are expected to provide significant advances in the field of transplantation medicine.
