[Expression of hCTLA4-Ig mediated by adeno-associated virus in porcine islets and their biological activity]

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2007 Feb;32(1):36-40.
[Article in 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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Antigens, CD / biosynthesis*
  • Antigens, CD / genetics
  • Antigens, Differentiation / biosynthesis*
  • Antigens, Differentiation / genetics
  • CTLA-4 Antigen
  • Cells, Cultured
  • Dependovirus / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression
  • Humans
  • Immunoglobulin Fc Fragments / biosynthesis*
  • Immunoglobulin Fc Fragments / genetics
  • Immunohistochemistry
  • Interferon-gamma / analysis
  • Interleukin-2 / analysis
  • Islets of Langerhans / cytology
  • Islets of Langerhans / immunology
  • Islets of Langerhans / metabolism*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Swine
  • Transfection
  • Tumor Necrosis Factor-alpha / analysis

Substances

  • Antigens, CD
  • Antigens, Differentiation
  • CTLA-4 Antigen
  • CTLA4 protein, human
  • Immunoglobulin Fc Fragments
  • Interleukin-2
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma