中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2013年
10期
625-629
,共5页
徐三荣%李伟%周庆%韩博
徐三榮%李偉%週慶%韓博
서삼영%리위%주경%한박
骨髓移植%PTD-mFoxp3融合蛋白%嵌合体%移植物抗宿主病
骨髓移植%PTD-mFoxp3融閤蛋白%嵌閤體%移植物抗宿主病
골수이식%PTD-mFoxp3융합단백%감합체%이식물항숙주병
Bone marrow transplantation%PTD-mFoxp3 fusion protein%Chimerism%Graft-versus-host disease
目的 研究PTD-mFoxp3融合蛋白减轻同种异基因小鼠骨髓移植后急性移植物抗宿主病(aGVHD)的作用及机理.方法 以雌性C57BL/6小鼠(H-2Kb)和BALB/c小鼠(H-2Kd)作为骨髓移植的供、受鼠,建立异基因小鼠骨髓移植模型.术前受鼠接受全身照射预处理,并于4～6 h内输注供鼠全骨髓细胞.将受鼠分为3组,PTD组通过尾静脉间断多次输注PTD-mFoxp3融合蛋白,mFoxp3组输注等剂量mFoxp3蛋白,对照组输入等体积生理盐水,以未经任何处理的正常BALB/c小鼠作为阴性对照.移植后每天观察各组aGVHD的发生情况;移植后1、2、4、8和12周,采集受鼠外周血,用流式细胞术测定受鼠的嵌合率,取受鼠肝脏和小肠组织进行病理检测;移植后第4和第12周通过混合淋巴细胞反应检测各组受鼠脾细胞经供鼠淋巴细胞刺激后的增殖反应.结果 移植后第12周,PTD组仍保持较高的嵌合体水平,嵌合体率为(38.16±3.09)％,明显高于mFoxp3组的(20.12±4.75)％和对照组的(15.72±2.36)％(P＜0.05);混合淋巴细胞反应示.移植后4周和12周时,PTD组淋巴细胞对供者来源的淋巴细胞增殖率明显降低,较mFoxp3组和对照组显著降低(P＜0.05),PTD组肝脏和小肠组织淋巴细胞浸润明显轻于mFoxp3组和对照组.结论 异基因骨髓移植时输注PTD-mFoxp3融合蛋白可有效建立稳定的嵌合体模型,并减轻移植后的aGVHD.
目的 研究PTD-mFoxp3融閤蛋白減輕同種異基因小鼠骨髓移植後急性移植物抗宿主病(aGVHD)的作用及機理.方法 以雌性C57BL/6小鼠(H-2Kb)和BALB/c小鼠(H-2Kd)作為骨髓移植的供、受鼠,建立異基因小鼠骨髓移植模型.術前受鼠接受全身照射預處理,併于4～6 h內輸註供鼠全骨髓細胞.將受鼠分為3組,PTD組通過尾靜脈間斷多次輸註PTD-mFoxp3融閤蛋白,mFoxp3組輸註等劑量mFoxp3蛋白,對照組輸入等體積生理鹽水,以未經任何處理的正常BALB/c小鼠作為陰性對照.移植後每天觀察各組aGVHD的髮生情況;移植後1、2、4、8和12週,採集受鼠外週血,用流式細胞術測定受鼠的嵌閤率,取受鼠肝髒和小腸組織進行病理檢測;移植後第4和第12週通過混閤淋巴細胞反應檢測各組受鼠脾細胞經供鼠淋巴細胞刺激後的增殖反應.結果 移植後第12週,PTD組仍保持較高的嵌閤體水平,嵌閤體率為(38.16±3.09)％,明顯高于mFoxp3組的(20.12±4.75)％和對照組的(15.72±2.36)％(P＜0.05);混閤淋巴細胞反應示.移植後4週和12週時,PTD組淋巴細胞對供者來源的淋巴細胞增殖率明顯降低,較mFoxp3組和對照組顯著降低(P＜0.05),PTD組肝髒和小腸組織淋巴細胞浸潤明顯輕于mFoxp3組和對照組.結論 異基因骨髓移植時輸註PTD-mFoxp3融閤蛋白可有效建立穩定的嵌閤體模型,併減輕移植後的aGVHD.
목적 연구PTD-mFoxp3융합단백감경동충이기인소서골수이식후급성이식물항숙주병(aGVHD)적작용급궤리.방법 이자성C57BL/6소서(H-2Kb)화BALB/c소서(H-2Kd)작위골수이식적공、수서,건립이기인소서골수이식모형.술전수서접수전신조사예처리,병우4～6 h내수주공서전골수세포.장수서분위3조,PTD조통과미정맥간단다차수주PTD-mFoxp3융합단백,mFoxp3조수주등제량mFoxp3단백,대조조수입등체적생리염수,이미경임하처리적정상BALB/c소서작위음성대조.이식후매천관찰각조aGVHD적발생정황;이식후1、2、4、8화12주,채집수서외주혈,용류식세포술측정수서적감합솔,취수서간장화소장조직진행병리검측;이식후제4화제12주통과혼합림파세포반응검측각조수서비세포경공서림파세포자격후적증식반응.결과 이식후제12주,PTD조잉보지교고적감합체수평,감합체솔위(38.16±3.09)％,명현고우mFoxp3조적(20.12±4.75)％화대조조적(15.72±2.36)％(P＜0.05);혼합림파세포반응시.이식후4주화12주시,PTD조림파세포대공자래원적림파세포증식솔명현강저,교mFoxp3조화대조조현저강저(P＜0.05),PTD조간장화소장조직림파세포침윤명현경우mFoxp3조화대조조.결론 이기인골수이식시수주PTD-mFoxp3융합단백가유효건립은정적감합체모형,병감경이식후적aGVHD.
Objective To establish a stable transplantation tolerance model by combined treatment with PTD-mFoxp3 fusion protein and allogeneic bone marrow transplantation and study its application to reduce the incidence of graft-versus-host disease (GVHD).Method BALB/c mice as recipients were randomly divided into four groups,accepted medical linear accelerator ray 3.0-Gy total body irradiation (TBI) before bone marrow transplantation (BMT),and injected with donor C57BL/6 mice bone marrow cells 3 × 107withinn 4-6 h.The BALB/c mice in group A were given PTD-mFoxp3 fusion protein through tail vein intermittently (day-2,0,1,3,5,7,9,11,13),those in group B given the same dose mFoxp3 protein,those in group C given normal saline,and those in blank control group given no treatment.The symptoms of GVHD were observed after BMT.Chimerisms were determined on the week 1,2,4,8 and12 post-BMT by flow cytometry.Liver and intestinal tissues were collected for pathological examination.Recipient immune response was assessed on the week 4 and 12 by mixed lymphocyte reactions (MLR) after BMT.Results The chimerism level in group A was high [(38.16 ± 3.09) ％] in the first 12 weeks after BMT,and that in group B and group C was reduced [(20.12 ± 4.75) ％ and (15.72 ± 2.36) ％ respectively,P＜0.05].MLR revealed that shown lymphocyte donor-derived lymphocyte proliferation rate at 4th and 12th week in group A was significantly lower than in other groups (P＜0.05).Pathological examination showed infiltration of lymphocytes in the liver and intestine was milder in group A than in other groups.Conclusion PTD-mFoxp3 fusion protein combined with allogeneic bone marrow transplantation can effectively establish a stable transplantation chimeric model and reduce the incidence of GVHD.
