Expression of hCTLA4-Ig mediated by adeno-associated virus in porcine islets and their biological activity.
- Author:
Zhao-Hui MO
1
;
Wei WANG
;
Tao LIU
;
Qiu-Hua ZENG
;
Xiao-Bing WU
;
Yan-Hong XIE
Author Information
1. Cell Transplantation & Gene Therapy Institute of the Third Xiangya Hospital of Central South University, Department of Endocrinology, Changsha 410013, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Animals, Newborn;
Antigens, CD;
biosynthesis;
genetics;
Antigens, Differentiation;
biosynthesis;
genetics;
CTLA-4 Antigen;
Cells, Cultured;
Dependovirus;
genetics;
Enzyme-Linked Immunosorbent Assay;
Gene Expression;
Humans;
Immunoglobulin Fc Fragments;
biosynthesis;
genetics;
Immunohistochemistry;
Interferon-gamma;
analysis;
Interleukin-2;
analysis;
Islets of Langerhans;
cytology;
immunology;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Reverse Transcriptase Polymerase Chain Reaction;
Swine;
Transfection;
Tumor Necrosis Factor-alpha;
analysis
- From:
Journal of Central South University(Medical Sciences)
2007;32(1):36-40
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To evaluate the expression of hCTLA4-Ig and their biological function in newborn porcine islets (NPIs) transfected with AAV-hCTLA4-Ig.
METHODS:Cultured NPIs were transfected with AAV-hCTLA4-Ig. The expression of CTLA4-Ig in these NPIs was assayed by RT-PCR and immunocytochemistry. The levels of IL-2, IFN-gamma, and TNF-alpha in the culture medium were assayed by ELISA after these cells the co-cultured with human. The response of glucose-stimulated insulin secretion was observed in the transgene group and the control group.
RESULTS:The expressions of CTLA4-Ig mRNA and protein were detected in the transgene group. The levels of cytokines were obviously lower in the transgene group than those in the control group (P<0.01). There was no significant difference in the response of glucose-stimulated insulin release between the transgene group and the control group (P>0.05).
CONCLUSION:AAV mediated hCTLA4-Ig expression in NPIs could inhibit T lymphocyte to produce cytokines, while the endocrine functions of the NPIs were not significantly affected.
