Van Hoffen E, Gmelig-Meyling F H, Bosboom-Kalsbeek K C, Hu H, De Jonge N, Tilanus M G, Lahpor J R, De Weger R A
Department of Pathology, University Hospital, Utrecht, The Netherlands.
J Heart Lung Transplant. 1997 Feb;16(2):216-21.
After heart transplantation, expression of various cytokines can be detected in endomyocardial biopsy specimens both in the presence and in the absence of rejection. In this study we have analyzed the contribution of donor-specific CD8+ cytotoxic T cells to intragraft cytokine expression.
T cells were propagated from endomyocardial biopsy specimens in medium containing interleukin-2 and interleukin-4. From the T-cell lines obtained, T-cell clones were generated by limiting dilution. A number of 15 CD8+ donor-specific cytotoxic T-cell clones were generated from a single T-cell line and were analyzed for their cytokine messenger RNA expression. The cytokine profile of the clones was studied on the mRNA level by reverse transcriptase polymerase chain reaction.
Almost all clones expressed mRNA for interleukin-1 beta, interleukin-4, and interleukin-10, and 75% of the clones expressed mRNA for interleukin-1 alpha, interleukin-9, and tumor necrosis factor-beta. In about half of the clones expression of interleukin-2, interleukin-6, interleukin-8, and interferon-gamma was detected. Tumor necrosis factor-alpha was only detected in one of the clones. The cytokine profiles exhibited a considerable heterogeneity. The 15 clones had previously been analyzed for their T-cell receptor V beta-gene family expression and T-cell receptor V-D-J region sequence. Homology in the V-D-J regions of the clones indicated that the 15 clones were derived from a limited number of progenitor cells. Interestingly, clones derived from one progenitor expressed a different mRNA cytokine profile.
This study shows that donor-specific cytotoxic T cells can contribute to the spectrum of locally produced cytokines. The cytokine expression of these cytotoxic T cells seems not to be limited to a distinct profile.
心脏移植后，无论有无排斥反应，均可在心肌内膜活检标本中检测到多种细胞因子的表达。在本研究中，我们分析了供体特异性CD8 + 细胞毒性T细胞对移植内细胞因子表达的作用。
从心肌内膜活检标本中，在含有白细胞介素-2和白细胞介素-4的培养基中培养T细胞。从获得的T细胞系中，通过有限稀释产生T细胞克隆。从单个T细胞系中产生了15个CD8 + 供体特异性细胞毒性T细胞克隆，并分析了它们的细胞因子信使RNA表达。通过逆转录聚合酶链反应在mRNA水平上研究克隆的细胞因子谱。
几乎所有克隆均表达白细胞介素-1β、白细胞介素-4和白细胞介素-10的mRNA，75%的克隆表达白细胞介素-1α、白细胞介素-9和肿瘤坏死因子-β的mRNA。在约一半的克隆中检测到白细胞介素-2、白细胞介素-6、白细胞介素-8和干扰素-γ的表达。仅在其中一个克隆中检测到肿瘤坏死因子-α。细胞因子谱表现出相当大的异质性。之前已对这15个克隆的T细胞受体Vβ基因家族表达和T细胞受体V-D-J区域序列进行了分析。克隆的V-D-J区域的同源性表明这15个克隆源自有限数量的祖细胞。有趣的是，源自一个祖细胞的克隆表达不同的mRNA细胞因子谱。
本研究表明，供体特异性细胞毒性T细胞可促成局部产生的细胞因子谱。这些细胞毒性T细胞的细胞因子表达似乎不限于特定的谱。
