Lubin I, Segall H, Marcus H, David M, Kulova L, Steinitz M, Erlich P, Gan J, Reisner Y
Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Blood. 1994 Apr 15;83(8):2368-81.
Transplantation of bone marrow from SCID mice into lethally irradiated normal mice can potentially endow the normal recipients with characteristics typical of the immune-deficient SCID mouse. In the present study, we investigated whether intraperitoneal grafting of human peripheral blood lymphocytes (PBLs), which has been documented in the SCID mouse, can also be achieved in irradiated BALB/c mice radioprotected with SCID bone marrow. Evaluation of different radiation protocols suggested that, considering the quality of engraftment and rate of survival, optimal results were obtained with split dose total body irradiation (TBI; 4 Gy followed 3 days later by 10 Gy). Monitoring of mouse T cells in peripheral blood indicated an inverse correlation between the presence of such cells and the engraftment of human CD45+ cells in the peritoneum. Also, engraftment of human PBLs in nude BALB/c mice, conditioned with the same radiation protocol, was significantly higher than that achieved in their normal counterparts. Further improvement of human PBL engraftment was found when the mice were thymectomized 2 weeks before conditioning with split TBI. After transplantation of 80 x 10(6) human PBLs in such recipients, a marked engraftment of human T cells and B cells in the peritoneum cavity could be detected for at least 2 months, whereas significant amounts of human Ig could be detected for more than 3 months. Migration of human PBLs into internal organs such as spleen, liver, kidney, and lungs (and into thymus in nonthymectomized mice) was found within a few days of grafting and also persisted for 2 to 3 months. The majority of the engrafted lymphocytes were single-positive CD4+ and CD8+ T lymphocytes, about 50% of which were activated, as judged by their expression of HLA-DR. Staining with anti-CD25 antibody was lower compared with that found with anti-HLA-DR. CD20+ B cells were detected in all of the above-mentioned internal organs, but were mainly concentrated in the spleen. CD14+ monocytes could be detected only during the first week posttransplant of PBLs. Total human Ig in peripheral blood reached an average of 2.8 mg/mL 14 days posttransplant, and continued to be significant for several months. In vitro transformation by Epstein-Barr virus of human B cells from different tissues could be established 30 days after transplantation and led to outgrowth of two IgG+ cell lines, two IgM+ cell lines, and one IgA+ cell line producing 0.6 to 4.2 micrograms/mL human Ig in the supernatant.(ABSTRACT TRUNCATED AT 400 WORDS)
将严重联合免疫缺陷（SCID）小鼠的骨髓移植到经致死剂量照射的正常小鼠体内，有可能使正常受体具备免疫缺陷SCID小鼠的典型特征。在本研究中，我们探究了已在SCID小鼠中得到证实的人外周血淋巴细胞（PBL）腹腔移植，是否也能在接受SCID骨髓辐射防护的经照射BALB/c小鼠中实现。对不同辐射方案的评估表明，考虑到植入质量和存活率，分次剂量全身照射（TBI；4 Gy，3天后再给予10 Gy）能取得最佳效果。对外周血中小鼠T细胞的监测表明，此类细胞的存在与腹膜中人CD45+细胞的植入呈负相关。此外，用相同辐射方案预处理的裸BALB/c小鼠中人PBL的植入率，显著高于其正常同系小鼠。当小鼠在分次TBI预处理前2周进行胸腺切除时，人PBL的植入情况得到进一步改善。在此类受体中移植80×10⁶个人PBL后，可在腹腔中检测到明显的人T细胞和B细胞植入，至少持续2个月，而在3个多月的时间里都能检测到大量的人Ig。移植后几天内即可发现人PBL迁移至脾脏、肝脏、肾脏和肺等内部器官（在未进行胸腺切除的小鼠中还迁移至胸腺），且这种迁移也持续2至3个月。大多数植入的淋巴细胞为单阳性CD4⁺和CD8⁺ T淋巴细胞，根据其HLA-DR表达判断，其中约50%为活化状态。与抗HLA-DR染色相比，抗CD25抗体染色较低。在上述所有内部器官中均检测到CD20⁺ B细胞，但主要集中在脾脏。仅在PBL移植后的第一周能检测到CD14⁺单核细胞。移植后14天，外周血中人总Ig平均达到2.8 mg/mL，并在数月内持续保持较高水平。移植后30天，可建立来自不同组织的人B细胞经爱泼斯坦-巴尔病毒的体外转化，并导致两个IgG⁺细胞系、两个IgM⁺细胞系和一个IgA⁺细胞系生长，其培养上清液中产生0.6至4.2微克/毫升的人Ig。（摘要截选至400字）
