Kelso A, Gough N M
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9189-93. doi: 10.1073/pnas.85.23.9189.
Lymphokine gene expression was examined in a panel of 116 short-term murine T-lymphocyte clones derived by single-cell micromanipulation from allogeneic mixed leukocyte cultures. About 30% of clonable T cells, including both CD4+ CD8- and CD4- CD8+ cells, could be expanded for assay at an average of 22 days after cloning. By RNA blot-hybridization analysis, most clones (85-96%) expressed detectable granulocyte-macrophage colony-stimulating factor, interleukin 3, and gamma interferon mRNAs, and 11% expressed interleukin 4 mRNA. Although no differences were noted between CD4+ and CD8+ clones in the combinations of lymphokines produced, CD4+ clones on average transcribed and secreted higher levels. When the frequencies of coexpression of any pair of lymphokine mRNAs were determined, all were found to correspond to the values predicted for random assortment of the individual frequencies. For example, among 13 interleukin 4-positive clones, 11 also transcribed gamma interferon, giving the frequency of double-positive clones expected for random association (9.6% versus 10.8%). Therefore, expression of the four lymphokine genes segregated independently among the clones and did not allow the division of T cells into subsets with distinct patterns of lymphokine synthesis.
通过单细胞显微操作从同种异体混合淋巴细胞培养物中获得了116个短期小鼠T淋巴细胞克隆,对其淋巴因子基因表达进行了检测。约30%的可克隆T细胞,包括CD4+CD8-和CD4-CD8+细胞,在克隆后平均22天可扩增用于检测。通过RNA印迹杂交分析,大多数克隆(85%-96%)表达可检测到的粒细胞-巨噬细胞集落刺激因子、白细胞介素3和γ干扰素mRNA,11%表达白细胞介素4 mRNA。虽然在产生的淋巴因子组合方面,CD4+和CD8+克隆之间未观察到差异,但CD4+克隆平均转录和分泌水平更高。当确定任意一对淋巴因子mRNA共表达的频率时,发现所有频率均与个体频率随机排列预测的值相符。例如,在13个白细胞介素4阳性克隆中,11个也转录γ干扰素,给出了随机关联预期的双阳性克隆频率(9.6%对10.8%)。因此,四个淋巴因子基因的表达在克隆间独立分离,不允许将T细胞分为具有不同淋巴因子合成模式的亚群。