Yacoub-Youssef Houda, Marcheix Bertrand, Calise Denis, Thiers Jean-Claude, Therville Nicole, Benoist Hervé, Blaes Nelly, Ségui Bruno, Dambrin Camille, Thomsen Mogens
INSERM U466, University Hospital of Toulouse, France.
Transpl Immunol. 2005 Dec;15(2):157-64. doi: 10.1016/j.trim.2005.07.002. Epub 2005 Aug 8.
Models of severe combined immuno-deficient (SCID) mice reconstituted with a competent human immune system represent a valuable tool for the study of human immune responses in vivo. Reconstitution with human cells can be achieved using large numbers of peripheral blood lymphocytes, but levels of engraftment are poor and graft versus host disease (GVHD) frequently occurs. SCID/beige mice are at the same time deficient for adaptive and innate immunity and the objective of this study was to develop a safe and efficient way to achieve human lymphocyte engraftment in these mice using human spleen cells. After institutional authorisations and informed consent of relatives, a piece of spleen was obtained from cadaveric organ donors and the splenocytes were isolated and cryopreserved for later use. Single intraperitoneal injections of 5-100 x10(6) splenocytes were performed into SCID/beige mice. Reconstitution of a human immune system was monitored weekly by the presence of human cells and IgG in peripheral blood. The mice were sacrificed 4 weeks after the injection and the engraftment in lymphoid organs was studied. A reproducible reconstitution was obtained with intraperitoneal injection of 30-40 x10(6) spleen cells. Human T, B and NK cells as well as human IgG were present in peripheral blood. In lymphoid tissues, the same lymphocytic subpopulations were detected and in addition some antigen presenting cells. The reconstitution was functional because graft rejection was observed after transplantation of human allogeneic tissues. When less than 30 x10(6) cells were injected, the reconstitution was variable. When more than 40 x10(6) cells were injected, GVHD occurred with increasing frequency. In conclusion, we show that intraperitoneal injection of 30-40 x10(6) human splenocytes into SCID/beige mice induces a quick and functional engraftment of human T, B and NK cells with no risk of GVHD. This model may be used to study human transplantation immunobiology in vivo.
用有功能的人类免疫系统重建的严重联合免疫缺陷（SCID）小鼠模型，是研究体内人类免疫反应的一种有价值的工具。使用大量外周血淋巴细胞可实现用人细胞进行重建，但植入水平较差且经常发生移植物抗宿主病（GVHD）。SCID/米色小鼠同时缺乏适应性免疫和先天性免疫，本研究的目的是开发一种安全有效的方法，利用人脾细胞在这些小鼠中实现人淋巴细胞植入。在获得机构授权并征得亲属的知情同意后，从尸体器官供体获取一块脾脏，分离脾细胞并冷冻保存以备后用。将5 - 100×10⁶个脾细胞单次腹腔注射到SCID/米色小鼠体内。通过外周血中人类细胞和IgG的存在情况每周监测人类免疫系统的重建。注射后4周处死小鼠，研究淋巴器官中的植入情况。腹腔注射30 - 40×10⁶个脾细胞可实现可重复的重建。外周血中存在人类T、B和NK细胞以及人类IgG。在淋巴组织中，检测到相同的淋巴细胞亚群，此外还有一些抗原呈递细胞。这种重建是有功能的，因为在移植人类同种异体组织后观察到了移植物排斥反应。当注射少于30×10⁶个细胞时，重建情况不稳定。当注射超过40×10⁶个细胞时，GVHD的发生频率增加。总之，我们表明向SCID/米色小鼠腹腔注射30 - 40×10⁶个人脾细胞可诱导人T、B和NK细胞快速且有功能的植入，且无GVHD风险。该模型可用于体内研究人类移植免疫生物学。
