Increased levels of anti-non-Gal IgG following pig-to-baboon bone marrow transplantation correlate with failure of engraftment.
Increased levels of anti-non-Gal IgG following pig-to-baboon bone marrow transplantation correlate with failure of engraftment.
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DOI：
10.1111/xen.12065
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发表时间：
2013-11
影响因子：
3.9
通讯作者：
Sachs DH
中科院分区：
文献类型：
--
作者：
Liang F;Wamala I;Scalea J;Tena A;Cormack T;Pratts S;Duran-Struuck R;Elias N;Hertl M;Huang CA;Sachs DH
缺乏α 1-3半乳糖基转移酶表达的遗传修饰的猪（GalT-KO猪）的开发通过避免由于预先存在的针对Gal表位的抗体而引起的超急性排斥，促进了猪器官和组织到灵长类动物中的异种移植。然而，在大多数灵长类动物的移植前血清中发现了不同水平的针对其他抗原的抗体（抗非Gal抗体）。我们以前发现，狒狒与高水平的移植前抗非半乳糖IgG，条件与非清髓性预处理方案，未能植入后，猪狒狒骨髓移植。移植前抗非Gal IgG水平低的两只狒狒，用相同的方案处理，在移植后28天显示出猪骨髓祖细胞，表明植入。这些狒狒也表现出对供体特异性低反应的证据。这一观察结果使我们研究了以下假设：选择具有低移植前抗非Gal IgG水平的狒狒受体可能改善GalT-KO猪至狒狒骨髓移植后的植入水平。移植前抗非Gal IgG水平较低的5只狒狒接受了来自GalT-KO猪的骨髓细胞移植（1-5 × 10^9/kg受体体重）。接受非清髓性预处理方案，包括低剂量全身照射（150 cGy）、胸腺照射（700 cGy）、抗胸腺细胞球蛋白（ATG）和他克莫司。此外，两只狒狒接受了利妥昔单抗和硼替佐米（万珂）治疗以及使用GalT-KO猪肝的皮质外免疫吸附。通过猪特异性PCR对第28天骨髓抽吸物的集落形成单位（CFU）进行骨髓植入评估。使用流式细胞术（FACS）通过血清与GalT-KO PBMC的结合来评估抗非Gal抗体水平。使用猪和狒狒特异性抗体通过FACS测量外周巨嵌合体，并通过混合淋巴细胞反应（MLR）评估狒狒抗猪细胞反应。如前所述，5只狒狒中有2只在移植后4周表现出可检测的骨髓植入。植入与抗-非-Gal IgG水平增加的缺乏以及对猪的细胞低反应性相关。具有类似低水平的预先存在的抗-非-Gal IgG的三只随后的狒狒显示没有植入，并且在移植后抗-非-Gal IgG抗体水平增加。外周巨嵌合体仅见于移植后几天，无论抗体的发展。选择具有低水平的移植前抗非Gal IgG的狒狒受体不能确保骨髓植入。移植失败与移植后抗非Gal IgG水平增加相关。这些结果表明，抗-非-Gal-IgG可能参与早期骨髓排斥反应，并且对抗抗-非-GalIgG发展的成功策略可能允许更好的植入。由于无论抗体产生如何，植入仅为低且短暂的，因此还可能需要解决先天免疫或种属相容性机制，以实现长期植入。
The development of genetically modified pigs which lack the expression of alpha 1–3 galactosyl transferase, (GalT-KO pigs) has facilitated the xenogeneic transplantation of porcine organs and tissues into primates by avoiding hyperacute rejection due to pre-existing antibodies against the Gal epitope. However, antibodies against other antigens (anti-non-Gal antibodies), are found at varying levels in the pre-transplant sera of most primates. We have previously found that baboons with high levels of pre-transplant anti-non-Gal IgG, conditioned with a non-myeloablative conditioning regimen, failed to engraft following pig-to-baboon bone marrow transplantation. Two baboons with low levels of pre-transplant anti-non-Gal IgG, conditioned with the same regimen, showed porcine bone marrow progenitors at 28 days following transplantation, suggesting engraftment. These baboons also showed evidence of donor-specific hypo-responsiveness. This observation led us to investigate the hypothesis that selecting for baboon recipients with low pre-transplant anti-non-Gal IgG levels might improve engraftment levels following GalT-KO pig-to-baboon bone marrow transplantation. Five baboons, with low pre-transplant anti-non-Gal IgG levels, received transplantation of bone marrow cells (1–5 × 10^9/kg of recipient weight) from GalT-KO pigs. They received a non-myeloablative conditioning regimen consisting of low-dose total body irradiation (150cGy), thymic irradiation (700cGy), anti-thymocyte globulin (ATG) and tacrolimus. In addition, two baboons received Rituximab and Bortezomib (Velcade) treatment as well as extra-corporeal immunoadsorption using GalT-KO pig livers. Bone marrow engraftment was assessed by porcine-specific PCR on colony forming units (CFU) of day 28 bone marrow aspirates. Anti-non-Gal antibody levels were assessed by serum binding towards GalT-KO PBMC using flow cytometry (FACS). Peripheral macro-chimerism was measured by FACS using pig and baboon-specific antibodies and baboon anti-pig cellular responses were assessed by mixed lymphocyte reactions (MLR). As previously reported, two of five baboons demonstrated detectable bone marrow engraftment at four weeks after transplantation. Engraftment was associated with lack of an increase in anti–non-Gal IgG levels as well as cellular hypo-responsiveness towards pig. Three subsequent baboons with similarly low levels of pre-existing anti-non-Gal IgG showed no engraftment and an increase in anti-non-Gal IgG antibody levels following transplantation. Peripheral macrochimerism was only seen for a few days following transplantation regardless of antibody development. Selecting for baboon recipients with low levels of pre-transplant anti-non-Gal IgG did not ensure bone marrow engraftment. Failure to engraft was associated with an increase in anti-non-Gal IgG levels following transplantation. These results suggest that anti-non-Gal-IgG is likely involved in early bone marrow rejection and that successful strategies for combating anti-non-Gal IgG development may allow better engraftment. Since engraftment was only low and transient regardless of antibody development, innate immune, or species compatibility mechanisms will likely also need to be addressed in order to achieve long term engraftment.
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影响因子：
82.9
作者：
Yamada, K;Yazawa, K;Sachs, DH
通讯作者：
Sachs, DH
DOI：
10.1073/pnas.0307819101
发表时间：
2004-05-11
影响因子：
11.1
作者：
Kolber-Simonds, D;Lai, LX;Hawley, RJ
通讯作者：
Hawley, RJ
影响因子：
6.2
作者：
Wong, Banny S.;Yamada, Kazuhiko;Sachs, David H.
通讯作者：
Sachs, David H.
影响因子：
6.2
作者：
Sablinski, T;Emery, DW;Sachs, DH
通讯作者：
Sachs, DH
由非致死性准备方案诱导的混合嵌合和永久特异性移植耐受。
影响因子：
15.3
作者：
Sharabi, Y;Sachs, D H
通讯作者：
Sachs, D H