小鼠非清髓性半相合骨髓移植造血的重建

目的：探讨在非清髓性预处理条件下，纯化的供鼠NK细胞对MHC半相合小鼠骨髓移植（BMT）造血重建的影响。 方法：以CB6F₁^H-2d/b（BALB/c^H-2d×C57BL／6^H-2b）小鼠为受者，C57BL／6^H-2b小鼠为供者。60只受者小鼠根据不同照射剂量及不同NK细胞输注量随机分为10组，每组6只，经9.0、7.0、6.0 Gy 3个剂量⁶⁰Co照射。经9.0 Gy照射后分为单纯照射组（Ⅰ）、半相合BMT组（Ⅱ）。经7.0 Gy照射后分为单纯照射组（Ⅲ）、半相合BMT组（Ⅳ）、NK细胞移植1组（Ⅴ）和NK细胞移植2组（Ⅵ）。经6.0 Gy照射后分为单纯照射组（Ⅶ）、半相合BMT组（Ⅷ）、NK细胞移植1组（Ⅸ）和NK细胞移植2组（X）。在照射后4 h移植供鼠骨髓细胞，其中NK细胞移植1组（Ⅴ、Ⅸ）每只受鼠移植前输注纯化供鼠NK细胞1×106，NK细胞移植2组（Ⅵ、X）每只受鼠移植前输注供鼠NK细胞5×10⁵。以血常规、体重、生存期等变化为指标，观察60 d。 结果：①生存期：Ⅰ组为（5.83±0.98）d，其他组小鼠生存期均大于60 d。②移植后第10天时白细胞及血小板计数：Ⅴ、Ⅵ组WBC和PLT分别为（1.05±0.30）×10⁹?L^-1、（53.50±12.11）×10⁹?L^-1及（0.95±0.26）×10⁹?L^-1、（47.17±11.82）×10⁹?L^-1，明显高于Ⅲ、Ⅳ组（P<0.01）；Ⅸ、X组WBC和PLT分别为（0.98±0.34）×10⁹?L^-1、（50.67±12.58）×10⁹?L^-1及（0.88±0.29）×10⁹?L^-1、（45.33±14.36）×10⁹?L^-1，明显高于Ⅶ、Ⅷ组（P<0.05）。 结论：MHC半相合BMT小鼠移植前输注纯化的供鼠NK细胞促进其造血重建。

Hematopoietic reconstitution of nonmyeloablative MHC haplotype-mismatched BMT in mice

Objective To study the relationship between donor-derived NK cells and hematopoietic reconstitution after nonmyeloablative MHC haplotype-mismatched BMT in mice. Methods The murine model of MHC haplotype-mismatched BMT was established by using (BALB/c H-2d ×C57BL/6 H-2b ) CB6F 1 H-2d/b mouse as the recipient, and C57BL/6 H-2b mouse as the donor. 60 recipient mice were divided into 10 groups after irradiation (TBI 60 Co 9.0,7.0, and 6.0 Gy):9.0 Gy-irradiation groups including simple-irradiation group (Ⅰ), MHC haplotype-mismatched BMT group(Ⅱ); 7.0 Gy-irradiation groups including simple-irradiation group (Ⅲ), MHC haplotype-mismatched BMT group (Ⅳ), the first of NK cells-transplantation group (Ⅴ), the second of NK cells-transplantation group (Ⅵ), 6.0 Gy-irradiation groups including simple-irradiation group (Ⅶ), MHC haplotype-mismatched BMT group (Ⅷ), the first of NK cells-transplantation group (Ⅸ), the second of NK cells-transplantation group (Ⅹ). The recipients were infused into the bone marrow cells of donors when they were irradiated after 4 hours. The recipients, the first of NK cells-transplantation groups, were infused into NK cells of 1×10 6/one; The recipients, the second of NK cells-transplantation groups, were infused into NK cells of 5×10 5/one. The effects were assessed by the count of WBC and PLT, survival time, body weight in the recipients. Results ①Life span: survival time was (5.83±0.98) days in group Ⅰ, beyond 60 days in other groups. ②Leukocyte and platelet counts after 10 days: leukocyte and platelet counts were (1.05±0.30)×10 9· L -1 , (53.50±12.11) ×10 9·L -1 and (0.95±0.26)×10 9·L -1 ,(47.17±11.82)×10 9·L -1 in group Ⅴ and group Ⅵ, which were much higher than those in group Ⅲ and group Ⅳ (P<0.01); In the cases of group Ⅸ and group Ⅹ, leukocyte and platelet counts were (0.98±0.34)×10 9·L -1 ,(50.67±12.58)×10 9·L -1 ;(0.88±0.29)× 10 9·L -1 , (45.33±14.36)×10 9·L -1 , which were much higher than those in group Ⅶ and group Ⅷ (P<0.05). Conclusion Donor-derived NK cells can promote hematopoietic reconstitution after nonmyeloablative MHC haplotype-mismatched BMT in mice.