Xu Lü-hong, Fang Jian-pei, Weng Wen-jun, Xu Hong-gui, Ye Qi-xiang
Department of Pediatrics, Zhongshan University, Guangzhou, China.
Zhonghua Xue Ye Xue Za Zhi. 2011 Nov;32(11):734-8.
To establish a murine model of sensitization, and investigate the effect and mechanism of sensitization on allogeneic donor bone marrow cells (BMCs).
Sensitized BALB/c mice were established by transfusions of allogeneic splenocytes. The donor reactive antibodies were detected by binding and complement-dependent cytotoxicity assays. After irradiation, 1 × 10(7) BMCs of C57BL/6 donor mice were injected into non-sensitized or sensitized BALB/c recipient mice. The distribution pattern of donor BMCs in peripheral blood, spleen and bone marrow of recipient mice were analyzed at different time points (2 h, 12 h and 48 h) post transplantation. Hematopoietic recovery post transplantation was assessed, and survival was monitored. Moreover, sera and splenocytes derived from non-sensitized or sensitized recipients were incubated with allogeneic BMCs in vitro, and the cytotoxic indexes were calculated in the immune experiments.
The binding and complement-dependent cytotoxicity assays showed that a high level of donor reactive antibodies was presented in sensitized sera. Compared with the non-sensitized recipients, the homing assay showed significantly decreased distributions of allogeneic donor BMCs in peripheral blood, spleen and femur of sensitized recipients. Non-sensitized recipients survived long term after irradiation, while all the sensitized recipients died within 12-15 days. Fourteen days post transplantation, the white blood cells and BMCs of non-sensitized recipients were (3240 ± 300) × 10(6)/L and (396 ± 27) × 10(6)/femur, respectively; while the white blood cells and BMCs of sensitized recipients were (320 ± 80) × 10(6)/L and (6 ± 2) × 10(6)/femur, respectively; the differences were statistically significant between this two groups (P < 0.05). Seven days post transplantation, the percentage of donor cells in bone marrow of non-sensitized and sensitized recipients was (48.07 ± 4.70)% and (0.77 ± 0.11)%, respectively, and the differences were statistically significant (P < 0.05). Furthermore, the white blood cells and BMCs following transplantation decreased along with time in sensitized recipients. The immune experiments of complement-dependent cytotoxicity reaction, cytotoxic T lymphocytes reaction and antibody-dependent cellular cytotoxicity showed the cytotoxic indexes were higher in sensitized group than the non-sensitized group.
A sensitized model was established by transfusions of allogeneic spleen cells. Allogeneic donor BMCs were rejected in sensitized recipients, and its mechanism might be through immune impairment pathways.
建立致敏小鼠模型，研究致敏对异基因供体骨髓细胞（BMCs）的影响及机制。
通过输注异基因脾细胞建立致敏BALB/c小鼠。采用结合及补体依赖细胞毒性试验检测供体反应性抗体。照射后，将1×10(7)个C57BL/6供体小鼠的BMCs注入未致敏或致敏的BALB/c受体小鼠。在移植后不同时间点（2小时、12小时和48小时）分析受体小鼠外周血、脾脏和骨髓中供体BMCs的分布模式。评估移植后的造血恢复情况，并监测生存情况。此外，将未致敏或致敏受体的血清和脾细胞与异基因BMCs在体外孵育，并在免疫实验中计算细胞毒性指数。
结合及补体依赖细胞毒性试验显示，致敏血清中存在高水平的供体反应性抗体。与未致敏受体相比，归巢试验显示致敏受体外周血、脾脏和股骨中异基因供体BMCs的分布显著减少。未致敏受体在照射后长期存活，而所有致敏受体在12 - 15天内死亡。移植后14天，未致敏受体的白细胞和BMCs分别为(3240 ± 300)×10(6)/L和(396 ± 27)×10(6)/股骨；而致敏受体的白细胞和BMCs分别为(320 ± 80)×10(6)/L和(6 ± 2)×10(6)/股骨；两组差异有统计学意义（P < 0.05）。移植后7天，未致敏和致敏受体骨髓中供体细胞的百分比分别为(48.07 ± 4.70)%和(0.77 ± 0.11)%，差异有统计学意义（P < 0.05）。此外，致敏受体移植后的白细胞和BMCs随时间下降。补体依赖细胞毒性反应免疫实验、细胞毒性T淋巴细胞反应和抗体依赖细胞毒性实验显示，致敏组的细胞毒性指数高于未致敏组。
通过输注异基因脾细胞建立了致敏模型。致敏受体中异基因供体BMCs被排斥，其机制可能是通过免疫损伤途径。
