Marcheix B, Yacoub-Youssef H, Calise D, Thiers J-C, Benoist H, Blaes N, Ségui B, Thomsen M, Dambrin C
Inserm U466, CHU Rangueil, Toulouse, France.
Transplant Proc. 2005 Jul-Aug;37(6):2888-9. doi: 10.1016/j.transproceed.2005.05.031.
We developed an original experimental model to study chronic vascular rejection (CVR) consisting of a graft of human mesenteric artery followed by human immune reconstitution into CB.17 SCID/Beige mice. Human immune reconstitution achieved after human PBMC injection has often been variable and incomplete. The aim of this work was to develop an alternative method to achieve a complete, functional human immune reconstitution.
After institutional authorizations, spleen cells were recovered from cadaveric organ donors. Single intraperitoneal injections of various doses of spleen cells were made into 70 CB.17 SCID/Beige mice. Reconstitution of the human immune system was monitored by flow cytometry (circulating human cells) and ELISA (human IgG). Colonization of murine lymphoid organs by human cells was studied by immunohistochemistry and flow cytometry. Evaluation of the immune function consisted of examination of CVR lesions in human arterial grafts. The animals were humanely killed at day 28.
After injection of 30 to 40 x 10(6) spleen cells, the mice showed significant human CD3(+), CD19(+), and CD56(+) populations in peripheral blood. The mean human cells levels were, respectively, 8.2% +/- 5.4%, 2.9% +/- 1.2%, and 5.3% +/- 5.1%. Murine spleen and mesenteric lymph nodes were colonized by human T and B cells, while the murine thymus was only colonized by human T cells. Human IgG was detected in murine serum (65.9 +/- 63.3 mg/L) and typical CVR lesions were observed within the allogeneic grafts.
Intraperitoneal injection of 30 to 40 x 10(6) human spleen cells into CB.17 SCID/Beige mice induces complete and functional human immune reconstitution allowing the study of CVR under human allogeneic conditions.
我们开发了一种原创性实验模型来研究慢性血管排斥反应（CVR），该模型包括将人肠系膜动脉移植到CB.17 SCID/米色小鼠体内，随后进行人免疫重建。在注射人外周血单核细胞（PBMC）后实现的人免疫重建通常是可变且不完全的。这项工作的目的是开发一种替代方法来实现完全的、功能性的人免疫重建。
获得机构授权后，从尸体器官供体中回收脾细胞。将不同剂量的脾细胞单次腹腔注射到70只CB.17 SCID/米色小鼠体内。通过流式细胞术（循环人细胞）和酶联免疫吸附测定（ELISA，人IgG）监测人免疫系统的重建。通过免疫组织化学和流式细胞术研究人细胞在小鼠淋巴器官中的定植情况。免疫功能评估包括检查人动脉移植物中的CVR病变。在第28天对动物实施安乐死。
注射30至40×10⁶个脾细胞后，小鼠外周血中出现显著的人CD3⁺、CD19⁺和CD56⁺细胞群。人细胞的平均水平分别为8.2%±5.4%、2.9%±1.2%和5.3%±5.1%。小鼠脾脏和肠系膜淋巴结被人T细胞和B细胞定植，而小鼠胸腺仅被人T细胞定植。在小鼠血清中检测到人IgG（65.9±63.3mg/L），并且在同种异体移植物中观察到典型的CVR病变。
向CB.17 SCID/米色小鼠腹腔注射30至40×10⁶个人脾细胞可诱导完全的、功能性的人免疫重建，从而能够在人同种异体条件下研究CVR。
