Marmor M D, Benatar T, Ratcliffe M J
Department of Microbiology and Immunology, McGill University, Montreal, Canada.
J Exp Med. 1993 Mar 1;177(3):647-56. doi: 10.1084/jem.177.3.647.
Exposure of normal juvenile chicken bone marrow cells to the replication defective avian reticuloendotheliosis virus strain T (REV-T) (chicken syncytial virus [CSV]) in vitro resulted in the generation of transformed cell lines containing T cells. The transformed T cells derived from bone marrow included cells expressing either alpha/beta or gamma/delta T cell receptors (TCRs) in proportions roughly equivalent to the proportions of TCR-alpha/beta and TCR-gamma/delta T cells found in the normal bone marrow in vivo. Essentially all TCR-alpha/beta-expressing transformed bone marrow-derived T cells expressed CD8, whereas few, if any, expressed CD4. In contrast, among TCR-gamma/delta T cells, both CD8+ and CD8- cells were derived, all of which were CD4-. Exposure of ex vivo spleen cells to REV-T(CSV) yielded transformed polyclonal cell lines containing > 99% B cells. However, REV-T(CSV) infection of mitogen-activated spleen cells in vitro resulted in transformed populations containing predominantly T cells. This may be explained at least in part by in vitro activation resulting in dramatically increased levels of T cell REV-T(CSV) receptor expression. In contrast to REV-T(CSV)-transformed lines derived from normal bone marrow, transformed lines derived from activated spleen cells contained substantial numbers of CD4+ cells, all of which expressed TCR-alpha/beta. While transformed T cells derived from bone marrow were stable for extended periods of in vitro culture and were cloned from single cells, transformed T cells from activated spleen were not stable and could not be cloned. We have therefore dissociated the initial transformation of T cells with REV-T(CSV) from the requirements for long-term growth. These results provide the first demonstration of efficient in vitro transformation of chicken T lineage cells by REV-T(CSV). Since productive infection with REV-T(CSV) is not sufficient to promote long-term growth of transformed cells, these results further suggest that immortalization depends not only upon expression of the v-rel oncogene but also on intracellular factor(s) whose expression varies according to the state of T cell physiology and/or activation.
将正常幼年鸡骨髓细胞在体外暴露于复制缺陷型禽网状内皮组织增生症病毒T株(REV-T)(鸡合胞体病毒[CSV]),导致产生了含有T细胞的转化细胞系。源自骨髓的转化T细胞包括表达α/β或γ/δ T细胞受体(TCR)的细胞,其比例大致等同于体内正常骨髓中TCR-α/β和TCR-γ/δ T细胞的比例。基本上所有表达TCR-α/β的转化骨髓源T细胞都表达CD8,而表达CD4的细胞极少,甚至没有。相比之下,在TCR-γ/δ T细胞中,既有CD8 +细胞也有CD8 -细胞,所有这些细胞都是CD4 -。将离体脾细胞暴露于REV-T(CSV)产生了含有> 99% B细胞的转化多克隆细胞系。然而,体外将有丝分裂原激活的脾细胞用REV-T(CSV)感染,导致产生的转化群体主要是T细胞。这至少可以部分解释为体外激活导致T细胞REV-T(CSV)受体表达水平大幅增加。与源自正常骨髓的REV-T(CSV)转化系不同,源自激活脾细胞的转化系含有大量CD4 +细胞,所有这些细胞都表达TCR-α/β。虽然源自骨髓的转化T细胞在体外长时间培养时是稳定的,并且可以从单细胞克隆,但来自激活脾细胞的转化T细胞不稳定,无法克隆。因此,我们将T细胞与REV-T(CSV)的初始转化与长期生长的需求分离开来。这些结果首次证明了REV-T(CSV)能在体外有效转化鸡T系细胞。由于用REV-T(CSV)进行有效感染不足以促进转化细胞的长期生长,这些结果进一步表明,永生化不仅取决于v-rel癌基因的表达,还取决于细胞内因子,其表达根据T细胞生理状态和/或激活状态而变化。