Chen C, Johnston T D, Jeon H, Gedaly R, McHugh P, Ranjan D
Department of Surgery, Transplant Section, University of Kentucky, College of Medicine, Lexington, Kentucky 40536, USA.
Transplant Proc. 2009 Jan-Feb;41(1):366-70. doi: 10.1016/j.transproceed.2008.10.085.
We have reported that cyclosporine (CsA) has direct effect to promote Epstein-Barr virus (EBV) transformation of human peripheral blood B lymphocytes. In this article, we have reported that CsA promoted EBV-infected, human B-cell transformation as assayed by three methods of colony number counting, cell number counting, and (3)H-thymidine incorporation. At first, we sought to correlate the three methods in EBV-infected human B-cell transformation, observing that they are convenient correlate with each other, and only vary in the degree when transformed cells are compared to the controls. Based on these pilot experiments, the three assay methods were then applied to CsA-treated and nontreated, EBV-infected human B cells to investigate whether CsA treatment promoted EBV-infected human B-cell transformation. We observed that CsA treatment increased colony formation above the control value of 28 +/- 4.5/well to 49 +/- 4.3 (colonies/well; n = 5; P < .05). CsA treatment increased the cell number from the control of 33,025 +/- 1900 to 50,925 +/- 4194 (cells/well; n = 5; P < .05). CsA treatment increased (3)H-thymidine incorporation from the control result of 12,481 +/- 1341 to 26,514 +/- 5464 (CPM/well; n = 5; P < .05). In conclusion, CsA promoted EBV-B-cell transformation in three correlated assay methods in vitro using a model of posttransplant lymphoproliferative disorder.
我们曾报道环孢素(CsA)具有直接促进爱泼斯坦-巴尔病毒(EBV)转化人外周血B淋巴细胞的作用。在本文中,我们报道了通过集落计数、细胞计数和³H-胸腺嘧啶核苷掺入这三种方法检测发现,CsA促进了EBV感染的人B细胞转化。首先,我们试图关联这三种用于检测EBV感染的人B细胞转化的方法,发现它们彼此关联方便,只是在将转化细胞与对照相比时程度有所不同。基于这些初步实验,然后将这三种检测方法应用于经CsA处理和未经处理的EBV感染的人B细胞,以研究CsA处理是否促进了EBV感染的人B细胞转化。我们观察到,CsA处理使集落形成从对照值28±4.5/孔增加到49±4.3(集落/孔;n = 5;P <.05)。CsA处理使细胞数量从对照的33,025±1900增加到50,925±4194(细胞/孔;n = 5;P <.05)。CsA处理使³H-胸腺嘧啶核苷掺入从对照结果12,481±1341增加到26,514±5464(CPM/孔;n = 5;P <.05)。总之,在体外利用移植后淋巴细胞增生性疾病模型,通过三种相关检测方法证实CsA促进了EBV-B细胞转化。