Human peripheral blood leukocyte-reconstituted severe combined immunodeficient mouse: analysis of the human immune response against porcine islet transplantation.
Human peripheral blood leukocyte-reconstituted severe combined immunodeficient mouse: analysis of the human immune response against porcine islet transplantation.
复制标题
人外周血白细胞重建严重联合免疫缺陷小鼠：针对猪胰岛移植的人体免疫反应分析。
DOI：
10.1097/00007890-199703270-00005
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发表时间：
1997
期刊：
影响因子：
6.2
通讯作者：
Mohanakumar,T
中科院分区：
文献类型：
--
作者：
Shiroki,R;Naziruddin,B;Shishido,S;Duffy,BF;Howard,T;Mohanakumar,T
人外周血白细胞（PBL）重建的严重联合免疫缺陷（SCID）小鼠（Hu-PBL-SCID）被用作研究人类异种移植排斥反应的模型。使用包括用抗人CD 3抗体致敏的细胞加强注射的方案用人PBL重建SCID小鼠。该方案增强了嵌合体在SCID小鼠中的建立，并且当与单独接受未致敏PBL的SCID小鼠相比时，导致检测到更高水平的人IG。用荧光细胞分析法在Hu-PBL-SCID小鼠血清中检测到识别猪胰岛（PI）的人异种反应性天然抗体（XNA），包括IgM和IgG两种亚型。用GS-IB 4凝集素预处理猪细胞抑制XNA结合，证明来自Hu-PBL-SCID的XNA的特异性。Western印迹分析显示，来自正常人血清和Hu-PBL-SCID血清的XNA识别PI上类似的异种抗原，表明Hu-PBL-SCID包含代表正常人血清的XNA谱系。组织切片的免疫荧光染色显示，人IgG和IgM两者在体内结合到植入Hu-PBL-SCID的肾囊下方的PI。此外，在异种移植的PI上检测到小鼠补体（C3）。异种移植胰岛的功能通过使用放射免疫测定法测量猪胰岛素浓度来监测。移植后14天内，Hu-PBL-SCID和PI异种移植的普通SCID血清中的猪胰岛素浓度相似，此后Hu-PBL-SCID中的胰岛素水平降低，从而表明排斥。因此，PI移植到Hu-PBL-SCID中应该是研究针对异种胰岛的细胞以及获得性体液免疫应答的有用模型。
Human peripheral blood leukocyte (PBL)-reconstituted severe combined immunodeficient (SCID) mice (Hu-PBL-SCID) were used as a model to study xenograft rejection in humans. SCID mice were reconstituted with human PBL using a protocol that included a booster injection with anti-human CD3 antibody-primed cells. This protocol enhanced chimera establishment in SCID mice and resulted in the detection of higher levels of human Ig when compared with SCID mice receiving unprimed PBL alone. Human xenoreactive natural antibodies (XNA), both IgM and IgG subtypes, which recognized porcine islets (PI), were detected in sera of Hu-PBL-SCID by cytofluorometric analysis. Pretreatment of porcine cells with GS-IB4 lectin inhibited the XNA binding, demonstrating the specificity of the XNA from Hu-PBL-SCID. Western blot analysis showed that XNA from normal human serum and Hu-PBL-SCID serum recognized similar xenoantigens on PI, indicating that Hu-PBL-SCID contained a XNA repertoire representative of normal human serum. Immunofluorescent staining of the tissue sections revealed that both human IgG and IgM bound in vivo to the PI engrafted beneath the kidney capsule of Hu-PBL-SCID. In addition, mouse complement (C3) was detected on xenografted PI. The function of xenografted islets were monitored by measuring porcine insulin concentration using a radioimmunoassay. Porcine insulin concentration in the sera of both Hu-PBL-SCID and plain SCID xenografted with PI was similar for up to 14 days after transplantation, after which the insulin levels in Hu-PBL-SCID decreased, thereby indicating rejection. Therefore, PI transplanted into the Hu-PBL-SCID should be a useful model for the study of cellular as well as acquired humoral immune response against xenoislets.