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Knockdown of Core 1 Beta 1, 3-galactosyltransferase Prolongs Skin Allograft Survival with Induction of Galectin-1 Secretion and Suppression of CD8+T Cells

T synthase knockdown effects on galectin-1 and CD8+T cells

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Abstract

Core 1 beta 1,3-galactosyltransferase also known as T-antigen-synthase or T-synthase is a key enzyme for the synthesis of the common core 1 O-glycan structure (T-antigen). Although T-synthase is known to be important in human immune-related diseases, the effects of T-synthase and T-antigen on host immune responses remain poorly defined. In this study, a T-synthase-specific short hairpin RNA (shRNA) was transfected into murine colon carcinoma CT26 cells or mouse muscle tissues via intramuscular electroporation to assess the effects of T-synthase on T cells and cytokines. T-synthase knockdown significantly induced galectin-1 secretion both in vivo and in vitro and strongly enhanced Th2 cytokine (IL-10 and IL-4) production in vivo. Further, the increased production of galectin-1 induced by T-synthase knockdown promoted CD8+ T-cell apoptosis, which, when combined with the increased production of CD4+ T cell-derived Th2 cytokines prolonged the survival of skin allografts in mice. Our data suggest core 1 beta 1,3-galactosyltransferase-shRNA could serve not only as a useful tool in organ transplantation but also as a powerful tool for investigating O-glycans and glycoprotein synthesis and function.

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Abbreviations

shRNA:

Short hairpin RNA

T synthase or C1GalT:

Core 1 beta 1, 3-galactosyltransferase

CsA:

Cyclosporin A

T-antigen:

Thomsen-Friedenreich antigen

MTT:

3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

FCM:

Flow Cytometry

PNA:

Peanut agglutinin

PI:

Propidium iodide

CFSE:

Carboxyfluorescein succinimidyl ester

ECL:

Enhanced chemiluminescence

ELISA:

Antigen-capture enzyme-linked immunosorbent assay

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Acknowledgments

The plasmid pPCR-hC1GalT was kindly provided by Dr. Narimatsu H and Dr. Ju T.

This work was supported by grants from the National Grand Program on Key Infectious Disease, National Natural Science Foundation of China (30921001, 81000714 and 30800038), and National Outstanding Youth Foundation of China (81025008), the 973 Program of China (2009CB522507), 211 program (303-581045), the program for Innovative Research Team of University, the Science and Technology Program of Wuhan (#201150530141) and DFG Transregio 60.

Competing Interests

The authors have dedared that no competing interests exist.

Author information

Author notes

  1. Gang Yu

    Present address: The third hospital of Wuhan, Wuhan, 430071, China

Authors and Affiliations

  1. Department of Immunology and State Key Laboratory of Virology and Hubei province Key Laboratory of Allergy and Immune-related diseases, Wuhan University School of Medicine, Wuhan, 430071, People’s Republic of China

    Hai-Dan Chen, Xia Zhou, Gang Yu, Ying-Lan Zhao, Yushan Ren & Xiao-Lian Zhang

  2. College of Life Science, Department of Biotechnology, Jianghan University, Wuhan, 430056, People’s Republic of China

    Yi-Dan Zhou

  3. University of Michigan Medical Center, Ann Arbor, MI, 48109, USA

    Qiao Li

  4. Department of Immunology, Wuhan University School of Medicine, Donghu Road 165, Wuhan, 430071, People’s Republic of China

    Xiao-Lian Zhang

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  1. Hai-Dan Chen
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Correspondence to Xiao-Lian Zhang.

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Chen, HD., Zhou, X., Yu, G. et al. Knockdown of Core 1 Beta 1, 3-galactosyltransferase Prolongs Skin Allograft Survival with Induction of Galectin-1 Secretion and Suppression of CD8+T Cells. J Clin Immunol 32, 820–836 (2012). https://doi.org/10.1007/s10875-012-9653-8

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  • DOI: https://doi.org/10.1007/s10875-012-9653-8

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