Objective To induce the donor-specific tolerance by intrathymic inoculation of recipient T cells expressing donor MHC antigen.Methods Plasmid PXN (N2-B19-H-2K b) encoding donor(C57BL/6, H-2 b) MHC classⅠgene cDNA (H-2K b) was introduced into producer cells PA317 to get recombinant virus. T lymphocytes of recipient were infected by recombinant virus, and then the T cells expressing donor and recipient MHC antigen (H-2K db) were injected into recipient thymus to induce tolerance. Following operations were performed in which donor mice (C57BL/6,H-2 b) skin allografts were transplanted to recipient mice (Balb/c,H-2 d). Results Allogenic MHC gene had integrated into the chromosomal DNA of the T cells and was effectively transcripted in the transfected T cells. Flow cytometry assay showed that allogenic MHC molecules were expressed on the membrane of transfected T cells 72 h after transfection. In the control group, median survival time (MST) of skin allografts was 9 days. MST of skin allografts was 11 days in the received monolonal antibodies control group. MST of skin allografts was 35 days in the induced tolerance group while normally rejected 3rd-party CBA/J skin grafts (MST 11.5 days). Responder cells of tolerant recipients to C57BL/6 stimulators showed significantly less incorporation in one way mixed lymphocyte reaction but in normal fashion to third-party CBA/J stimulators and to ConA.Conclusion Donor-specific tolerance can be induced by intrathymic inoculation of transfected recipient T cells expressing allogeneic MHC products.

Objective To investigate the effects of interleukin 10(IL 10) gene transfer on secretion of inflammatory cytokines in rat synoviocytes (RSC) induced by lipopolysaccharide (LPS) in vitro. Methods Constructed retroviral vector pLX (IL 10) SN was introduced into packaging cell line PA317. PA317 was selected by G418 and then retrovirus produce cells titre was determined by NIH3T3 for identification of producer clones making high titre virus. Through the highest titre produce cells clone (6×10 5 CFU/ml)exogenous gene IL 10 was transferred into RSC. 72 hours after gene transfer, DNA and RNA were prepared from rat transfected RSC for the polymerase chain reaction (PCR) and the reverse transcription polymerase chain reaction (RT PCR) ;expression of IL 10 in transfected RSC was checked by ELISA. Effects of IL 10 gene transfer on secretion of inflammatory cytokines induced by LPS in rat RSC cells were detected by RT PCR and ELISA in vitro.Results Exogenous IL 10 gene was integrated into the chromosal DNA of the transfected RSC cells. RT PCR and ELISA showed that exogenous IL 10 gene was expressed in the transfected RSC 48 hours after transfection and the expression lasted for over 28 days. Exogenous IL 10 gene transfer can inhibit secretion of inflammatory cytokines, IL 1β and TNF α in the transfected RSC at mRNA and protein level induced by LPS. Conclusion Exogenous IL 10 gene transfer can inhibit secretion of inflammatory cytokines in RSC, it is suggested that exogenous IL 10 gene transferring into RSC can be used in the treatment of joint injury caused by immunological diseases.

目的：观察人白细胞介素１０ （ｈｕｍａｎ Ｉｎｔｅｒｌｅｕｋｉｎ １０ ，ｈ Ｉ Ｌ１０） 基因转导对脂多糖（ Ｌ Ｐ Ｓ） 诱导大鼠心肌细胞（ Ｃ Ｍ Ｃ） 的炎症细胞因子的产生及其基因表达的影响。方法：通过重组逆转录病毒载体ｐ Ｌ Ｘ（ Ｉ Ｌ１０） Ｓ Ｎ 将人 Ｉ Ｌ１０ 基因导入大鼠 Ｃ Ｍ Ｃ，应用 Ｐ Ｃ Ｒ， Ｒ Ｔ Ｐ Ｃ Ｒ 和 Ｅ Ｌ Ｉ Ｓ Ａ：①检测 Ｉ Ｌ１０ 基因的整合和表达；②观察 Ｉ Ｌ１０ 基因转导对 Ｌ Ｐ Ｓ诱导的 Ｃ Ｍ Ｃ 肿瘤坏死因子α（ Ｔ Ｎ Ｆα） 的ｍ Ｒ Ｎ Ａ 表达及白细胞介素１β（ Ｉ Ｌ１β） 和 Ｔ Ｎ Ｆα的蛋白质的产生的影响。结果：外源性 Ｉ Ｌ１０ 基因已整合到靶细胞染色体 Ｄ Ｎ Ａ 并有效地表达，并能抑制 Ｌ Ｐ Ｓ诱生 Ｃ Ｍ Ｃ 产生 Ｉ Ｌ１β， Ｔ Ｎ Ｆα。结论：逆转录病毒载体介导的人 Ｉ Ｌ１０ 基因转导能抑制 Ｃ Ｍ Ｃ 炎症效应中细胞因子的产生及其基因表达。

目的:通过胸腺内注射表达外源性主要组织相容性抗原复合物(MHC)抗原质粒PXN(N2-B19-H-2Kb) , 诱导异基因小鼠心肌移植耐受.方法:给BALB/C小鼠胸腺内注射质粒PXN(N2-B19-H-2Kb),将外源性的编码C57BL/6小鼠MHC I 类抗原的H-2KbcDNA转移到BALB/C小鼠胸腺,2周后行C57BL/6小鼠心肌移植.用聚合酶链反应(PCR),反转录聚合酶链反应(RT-PCR),单克隆抗体免疫萤光染色流式细胞仪检测BALB/C小鼠胸腺细胞DNA,mRNA和MHC蛋白质表达.结果: BALB/C小鼠胸腺细胞表面有外源性MHC分子表达,转染效率为5.1%,胸腺内注射质粒PXN(N2-B19-H-2Kb)能明显延长移植小鼠心肌的存活时间,平均17 d,对照为8 d (P＜0.01).结论:同种MHC基因转移至受体鼠胸腺可以诱导特异性的免疫耐受.

1991至1994年4月中山医科大学附一医院共完成101例肾移植.随访时间28.6±9.6个月.术前供受者淋巴细胞毒性试验阴性.术中及术后2天,每日用甲基泼尼松龙(MP)0.25～0.5 g,同时选用二联或三联免疫抑制方案.

Intrathymic (IT) inoculation of allogeneic major histocompatibility complex (MHC) soluble antigen extracted from C57BL/6 inbred mice splenocytes using 3mol/L KCl was performed to induce donor-specific tolerance to skin allografts in adult BALB/C mice. The recipients (BALB/C) received anti-T lymphocyte monoclonal antibodies (mAb) 8.4μg on 0 and 3rd day relative to IT injection of allogeneic MHC antigen without other immunosuppressant. The control group received anti-T lymphocyte mAb 8.4μg but IT injection of normal saline (NS). Median survival time (MST) of transplanted skin allografts was 83 days in tolerant recipients while normally rejected 3rd-party CBA/J skin grafts (MST 12 days). In the control group, skin grafts'MST was 11 days. Responder cells of tolerant recipients to C57BL/6 stimulators showed significantly less incorporation in oneway mixed lymphocyte reaction but in normal fashion to third-party CBA/J stimulators. The results suggested that unresponsiveness can be induced by IT inoculation of soluble allogeneic MHC antigen which was donor-specific.