Resistance of porcine Sertoli cells to xenoreactive antibodies mediated complement lysis
10.3760/cma.j.issn.0254-5101.2009.02.010
- VernacularTitle:猪睾丸Sertoli细胞抵御人血清中天然抗体介导的补体杀伤作用
- Author:
Zhuzeng YIN
;
Lu WANG
;
Feng HU
;
Ying XIANG
;
Lin XIE
;
Jing XU
;
Junhua LI
;
Gang CHEN
;
Shi CHEN
- Publication Type:Journal Article
- Keywords:
Sertoli cells;
α-Gal;
Complement
- From:
Chinese Journal of Microbiology and Immunology
2009;29(2):137-141
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether porcine Sertoli cells eould resist xenoreactive antibodies mediated complement lysis. Methods Sertoli cells were isolated from testes of 10 to 15 day-old landrace pigs. Α-Gal expression on Sertoli cells was measured by FACS and cytoimmunofluorescence. The binding of human se-rum IgG and IgM with Sertoli cells was assayed by FACS. After the incubation of the cultured Sertoli cells with 20% human B serum in vitro, the cellular lysis and cytotoxicity assay were detected with CytoTox-ONETM homogeneous membrane integrity assay and MTT method, and then activation of the complement cascade was examined by immunohistochemistry and immunofluorescence. The SV40-porcine endothelium cells line (SV40-PED) was served as control cells. Results α-Gal expression was found on Sertoli cells by FACS and cytoimmunofluorescence. After incubation with 20% human serum, cellular lysis ratio of the SV40-PED was 53. 13% ± 14.53%, while lysis ratio of the Sertoli cells was significantly lower (24.38% ±0.50%, P<0.01), the viability of Sertoli cells and SV40-PED were 98.73% ± 18.84% and 52.43% ± 8.08%, respectively. With immunohistochemistry and immunofluorescence, C3c and C4d were found binding on both the Sertoli cells and SV40-PED cells, however, C5b-9 was only detected on SV40-PED cells. Conclusion In vitro, compared with the SV40-PED, Sertoli cells could resist xenoreactive antibodies of human serum mediated complement lysis by preventing the C5b-9 formation.
