异种造血嵌合体预防急性GVHD的作用机制
发布时间：2019-03-08 07:53
【摘要】：目的 异种移植被认为是有潜力的解决器官供源不足的方法,但异种移植面临的免疫排斥比同种异体移植更复杂,移植物抗宿主病(GVHD)更为严重。我们应用大鼠-小鼠造血嵌和体模型,进一步研究反向骨髓移植对aGVHD的预防机制。 方法 1、异种骨髓嵌合体模型的建立:术前5天SD大鼠(8～10周龄,雌性,SPF级)开始饮含红霉素(250mg/L)和庆大霉素(320mg/L)抗生素溶液净化肠道。无菌条件下取BALB/C小鼠(8～10周龄,雌性,SPF级)的骨髓细胞,制成单细胞悬液,SD大鼠接受7.5Gy(照射率0.5Gy/min)全身照射(TBI)后4小时内经尾静脉输入上述BALB/c小鼠骨髓细胞2×10~8/只,48小时后腹腔注射环磷酰胺50mg/kg(此预处理方案为非清髓性),将该嵌合了BALB/c小鼠骨髓细胞的供体大鼠饲养30天后,检测外周血嵌合率,并做皮片移植,结果证实小鼠源性骨髓已在大鼠体内植活,并诱导出对BALB/c小鼠的特异性耐受。再将该嵌合体SD大鼠的骨髓细胞移植到目标受体(BALB/C小鼠)。 2、实验分组:A,B,C组以BALB/C(8～10周龄,雌性,SPF级)小鼠为受鼠,D组以无关第3者B6小鼠为受鼠,肠道净化后接受9Gy~(60)Coγ射线致死性TBI:A组经尾静脉输注正常SD大鼠的骨髓细胞4×10~7/只;B组经尾静脉输注嵌合体SD大鼠的骨髓细胞4×10~7/只;C组经尾静脉输注生理盐水;D组经尾静脉输注嵌合体SD大鼠的骨髓细胞4×10~7/只。观察各组小鼠aGVHD的临床表现及生存时间,濒死小鼠做病理分析。移植后15天测定TNF-α、INF-γ与IL-4的产量,移植后
[Abstract]:Aim xenotransplantation is considered to be a potential method to solve the shortage of organ supply and source, but the immune rejection of xenotransplantation is more complicated than that of allotransplantation, and (GVHD) is more serious. We used the rat-mouse hemopoietic chimerism model to further study the preventive mechanism of reverse bone marrow transplantation (BMT) on aGVHD. Methods 1. Establishment of xenogeneic bone marrow chimera model: SD rats (8 weeks, 10 weeks old, female, 5 days before operation); SPF grade) began to drink erythromycin (250mg/L) and gentamicin (320mg/L) antibiotic solution to purify the intestinal tract. Bone marrow cells of BALB/C mice (8-10-week-old, female, SPF grade) were taken under sterile conditions to make single-cell suspension. The bone marrow cells of the above-mentioned BALB/c mice were injected into the tail vein of SD rats within 4 hours after the whole body irradiation with 7.5Gy (irradiation rate 0.5Gy/min) 2 脳 10 ~ 8 / mouse. 48 hours later, cyclophosphamide 50mg/kg (a non-myeloablative regimen) was injected intraperitoneally. After 30 days of feeding the donor rat chimeric with BALB/c mouse bone marrow cells, the peripheral blood chimerism rate was detected and skin graft was performed. The results showed that mouse-derived bone marrow had been implanted in rats and induced specific tolerance to BALB/c mice. The bone marrow cells of the chimeric SD rats were transplanted to the target receptor (BALB/C mice). 2. The experimental group: a, B, C: BALB/C (8-week-old, female, SPF grade) mice as recipient mice, group D: B-6 mice with no third group as recipient mice, group C (8-10 weeks old, female, SPF grade) mice as recipient mice. After intestinal purification, 4 脳 10 ~ 7 bone marrow cells of normal SD rats were infused through tail vein in 9 Gy ~ (60) Co 纬-ray lethal TBI:A group. Bone marrow cells of chimeric SD rats were infused into group B (4 脳 10 ~ 7), group C (normal saline), and group D (4 脳 10 ~ 7) of chimeric SD rats. The clinical manifestation and survival time of aGVHD in each group were observed, and pathological analysis was made in dying mice. The production of TNF- 伪, INF- 纬 and IL-4 was measured 15 days after transplantation.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
本文编号：2436591
[Abstract]:Aim xenotransplantation is considered to be a potential method to solve the shortage of organ supply and source, but the immune rejection of xenotransplantation is more complicated than that of allotransplantation, and (GVHD) is more serious. We used the rat-mouse hemopoietic chimerism model to further study the preventive mechanism of reverse bone marrow transplantation (BMT) on aGVHD. Methods 1. Establishment of xenogeneic bone marrow chimera model: SD rats (8 weeks, 10 weeks old, female, 5 days before operation); SPF grade) began to drink erythromycin (250mg/L) and gentamicin (320mg/L) antibiotic solution to purify the intestinal tract. Bone marrow cells of BALB/C mice (8-10-week-old, female, SPF grade) were taken under sterile conditions to make single-cell suspension. The bone marrow cells of the above-mentioned BALB/c mice were injected into the tail vein of SD rats within 4 hours after the whole body irradiation with 7.5Gy (irradiation rate 0.5Gy/min) 2 脳 10 ~ 8 / mouse. 48 hours later, cyclophosphamide 50mg/kg (a non-myeloablative regimen) was injected intraperitoneally. After 30 days of feeding the donor rat chimeric with BALB/c mouse bone marrow cells, the peripheral blood chimerism rate was detected and skin graft was performed. The results showed that mouse-derived bone marrow had been implanted in rats and induced specific tolerance to BALB/c mice. The bone marrow cells of the chimeric SD rats were transplanted to the target receptor (BALB/C mice). 2. The experimental group: a, B, C: BALB/C (8-week-old, female, SPF grade) mice as recipient mice, group D: B-6 mice with no third group as recipient mice, group C (8-10 weeks old, female, SPF grade) mice as recipient mice. After intestinal purification, 4 脳 10 ~ 7 bone marrow cells of normal SD rats were infused through tail vein in 9 Gy ~ (60) Co 纬-ray lethal TBI:A group. Bone marrow cells of chimeric SD rats were infused into group B (4 脳 10 ~ 7), group C (normal saline), and group D (4 脳 10 ~ 7) of chimeric SD rats. The clinical manifestation and survival time of aGVHD in each group were observed, and pathological analysis was made in dying mice. The production of TNF- 伪, INF- 纬 and IL-4 was measured 15 days after transplantation.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
【参考文献】
相关期刊论文 前1条
1 唐湘凤,李春富,裴夫瑜;小鼠→大鼠异种移植耐受模型的建立[J];免疫学杂志;2003年06期
，本文编号：2436591
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