Donor MHC gene to mitigate rejection of transplantation in recipient mice  

作　　者：LI Tong YAN Jun TAN Jia-li LU Yue-ping HOU Sheng-cai LI Shen-tao XU Qing TONG Xue-hong DING Jie ZHANG Zhi-tai LI Hui 

机构地区：[1]Department of Thoracic Surgery Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China [2]Department of Cardiae Surgery Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China [3]Experimental Centre Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China [4]Department of Orthodontics, School of Stomatology, FourthMilitary Medical University, Xi＇an, Shaanxi 710032, China [5]Department of Molecular Biology, Capital Medical University, Beijing 100069, China [6]Department of Thoracic Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China

出　　处：《Chinese Medical Journal》2011年第24期4279-4285,共7页中华医学杂志（英文版）

摘　　要：Background Donor organ rejection continues to be a significant problem for patients receiving transplants. We therefore tested whether transferring a donor＇s major histocompatibility complex （MHC） gene to the recipient would mitigate the rejection of transplanted hearts in mice. Methods H-2K^kgene from donor mice was amplified using nested polymerase chain reaction （PCR） and ligated into a mammalian expression vector, which was then transfected into thymus ground mass cells collected from the recipients. Clones stably expressing the transgene were then injected into the recipients＇ thymus visualized using ultrasound. Control mice were administered cells previously transfected with empty vector. Following heart transplantation, cardiac activity was monitored electrocardiographically. Recipient thymus cells were tested for MHC antigenicity using flow cytometry and spleen cells were subjected to mixed lymphocyte culture tests. Finally, the transplanted hearts were sectioned, stained and examined under light microscopy. Results Southern analysis following nested PCR revealed clear expression of H-2K^ gene. Following transplantation, electrocardiosignals were detectable highly significantly longer in recipients administered thymal cells expressing donor H-2K^ than in those receiving control cells. Flow cytometric analysis using an anti-H-2K^ antibody confirmed its expression in H-2K^ treated recipients but not in control mice. Mixed lymphocyte cultures containing H-2K^ treated cells showed significantly less proliferation than those containing control cells. Hearts from control mice showed substantially greater lymphocyte infiltration than those from H-2K^ treated mice and large areas of necrosis. Conclusion Rejection of transplanted hearts can be mitigated substantially by introducing the donor＇s MHC into the recipient.Background Donor organ rejection continues to be a significant problem for patients receiving transplants. We therefore tested whether transferring a donor＇s major histocompatibility complex （MHC） gene to the recipient would mitigate the rejection of transplanted hearts in mice. Methods H-2K^kgene from donor mice was amplified using nested polymerase chain reaction （PCR） and ligated into a mammalian expression vector, which was then transfected into thymus ground mass cells collected from the recipients. Clones stably expressing the transgene were then injected into the recipients＇ thymus visualized using ultrasound. Control mice were administered cells previously transfected with empty vector. Following heart transplantation, cardiac activity was monitored electrocardiographically. Recipient thymus cells were tested for MHC antigenicity using flow cytometry and spleen cells were subjected to mixed lymphocyte culture tests. Finally, the transplanted hearts were sectioned, stained and examined under light microscopy. Results Southern analysis following nested PCR revealed clear expression of H-2K^ gene. Following transplantation, electrocardiosignals were detectable highly significantly longer in recipients administered thymal cells expressing donor H-2K^ than in those receiving control cells. Flow cytometric analysis using an anti-H-2K^ antibody confirmed its expression in H-2K^ treated recipients but not in control mice. Mixed lymphocyte cultures containing H-2K^ treated cells showed significantly less proliferation than those containing control cells. Hearts from control mice showed substantially greater lymphocyte infiltration than those from H-2K^ treated mice and large areas of necrosis. Conclusion Rejection of transplanted hearts can be mitigated substantially by introducing the donor＇s MHC into the recipient.

关 键 词：major histocompatibility complex TRANSPLANTATION gene transference TOLERANCE 

分 类 号：R657.3[医药卫生—外科学]