Protective role of expression of human complement regulatory protein hCD55 in islet xenotransplantation
10.3969/j.issn.1674-7445.2022.04.010
- VernacularTitle:表达人源补体调节蛋白hCD55对异种胰岛移植的保护作用
- Author:
Xin LI
1
;
Dengke PAN
;
Jia ZHOU
;
Jie YAN
;
Jun CHEN
;
Jiaxiang DU
;
Manlin GONG
Author Information
1. School of Medicine University of Electronic Science and Technology of China, Chengdu 610054, China
- Publication Type:Research Article
- Keywords:
Diabetes mellitus;
Porcine islet cell;
Transgene;
Human cluster of differentiation55(hCD55);
Complement;
Instant blood-mediated inflammatory reaction (IBMIR);
C3c;
C5b-9
- From:
Organ Transplantation
2022;13(4):475-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To validate whether the expression of human cluster of differentiation 55 (hCD55) protein in porcine islet cells could inhibit the activation of complement components in human serum. Methods Four adult pigs with WT (wild type), GTKO [α-1, 3-galactosyltransferase (GGTA1) knockout], GTKO/hCD55 and hCD55 genotypes were selected. Islet cells were isolated from WT, GTKO and GTKO/hCD55 pigs, and the purity and insulin secretion function were detected. The expression of hCD55 at the DNA, RNA and protein levels was analyzed by agarose gel electrophoresis, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry, respectively. Complement-dependent cytotoxicity assay and complement deposition assay were performed under the incubation conditions with fresh human serum. Results The purity of isolated porcine islet cells from three genotype pigs was > 75%, and the glycemic index was > 1. The expression of hCD55 messenger RNA(mRNA) and protein in GTKO/hCD55 porcine islet cells decreased the deposition of human complement component C3c and membrane-attacking complex C5b-9, and reduced the cytotoxicity. Conclusions The expression of hCD55 protein in porcine islet cells could inhibit the activation of human complement and reduce complement-mediated killing effect, indicating that hCD55 protein could exert complement protection effect on porcine islet cells. These findings provide theoretical basis for the application of hCD55 in islet xenotransplantation.
