T-cell immune tolerance of HLA haploidentical donor induced by CTLA4Ig-modified bone marrow stromal cells

doi:10.3969/j.issn.2095-4344.2015.01.003

Abstract

Abstract:

BACKGROUND: CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention.
OBJECTIVE: To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cells mediated by adenovirus to induce T-cell tolerance of haploidentical donors.
METHODS: The bone marrow stromal cells isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cells were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cells (2×10⁴, 4×10⁴ and 8×10⁴) were respectively co-cultured with 10⁵ T cells from the peripheral blood of HLA haploidentical donors and 10⁵ peripheral blood mononuclear cells from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cells (1×10⁵/well) were co-cultured with CTLA4Ig-modified bone marrow stromal cell layers
constructed in 6-well plates. The number of bone marrow mononuclear cells and colony-forming unit-granulocyte macrophages were calculated after 5-day culture.
RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×10⁴, 4×10⁴, and 8×10⁴ CTLA4Ig-modified bone marrow stromal cells on proliferation of T cells were higher than that of untransfected cells. The levels of interluekin-2 in the corresponding cell groups were significantly lower than that in the untransfected cells (P < 0.05). At 5 days of culture, there was no significant difference in the number of bone marrow mononuclear cells and colony-forming unit-granulocyte macrophages between the transfected and untransfected cell groups (P > 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cells mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors in vitro.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Graft vs Host Disease, Hematopoietic Stem Cell Transplantation, Haploidy, Immune Tolerance

CLC Number:

R394.2

Wang Ji-gang, Zhou Fan, Liu Yan-qin, Bai Ying, Liu Jing-hua, Zhang Hai-ting, Li Min-yan. T-cell immune tolerance of HLA haploidentical donor induced by CTLA4Ig-modified bone marrow stromal cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(1): 12-17.

Figures/Tables 2

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