Hosseinzadeh H, Goldschneider I
Department of Pathology, School of Medicine, University of Connecticut Health Center, Farmington 06030.
J Immunol. 1993 Mar 1;150(5):1670-9.
Studies of the functional properties and developmental potentials of immediate post-thymic cells have been hampered by the lack of reliable markers with which to distinguish recent thymic emigrants (RTE) from the bulk of peripheral T cells. In the present study, the intrathymic FITC-labeling technique was used in concert with three-color flow-cytometric analysis to identify, phenotypically characterize, and study the short term fate of RTE in normal rats. The results indicated that between 3 and 4% of total T cells in lymph node and spleen of 5- to 8-wk-old rats had been released from the thymus within the preceding 24 h. Unlike the bulk of the peripheral T cells, which had a Thy1-, CD45RC+, and/or RT6+ phenotype, these RTE were Thy1+, CD45RC-, and RT6-. Furthermore, two discrete subsets of RTE were defined: a major subset (approximately 95%) of CD4+ or 8+ (single positive), TCR-alpha beta hi T cells that resembled medullary thymocytes; and a minor subset (approximately 5%) of CD4+ and 8+ (double positive), TCR-alpha beta low T cells that resembled cortical thymocytes. The following evidence suggested that most if not all Thy1+ T cells in young adult rats are RTE and their immediate descendants: 1) thymectomy (but not sham thymectomy) selectively depleted Thy1+ T cells from lymph node within 3 to 7 days, even in adrenalectomized rats; 2) most FITC-labeled RTE differentiated into Thy1-, CD45RC+, RT6+ T cells within 7 days of release from the thymus; 3) transitional phenotypes of Thy1+ T cells, including Thy1low, CD45RC+, and RT6+, were observed in normal, as well as in intrathymic, FITC-injected rats; 4) most T cells in neonatal rats were Thy1+ and RT6-, whereas their descendants were Thy1- and RT6+. These experiments demonstrate that most RTE in normal rats are phenotypically (and presumably developmentally) immature at the time of release from the thymus, and progressively acquire the phenotypic attributes of more mature T cells post-thymically. These unique phenotypic attributes should expedite the isolation of RTE and their immediate descendants for definitive studies of their developmental and functional properties.
由于缺乏可靠的标志物来区分新近胸腺迁出细胞(RTE)与外周T细胞主体,对胸腺后即刻细胞的功能特性和发育潜能的研究受到了阻碍。在本研究中,胸腺内异硫氰酸荧光素(FITC)标记技术与三色流式细胞术分析协同使用,以鉴定、表型特征化并研究正常大鼠中RTE的短期命运。结果表明,在5至8周龄大鼠的淋巴结和脾脏中,3%至4%的总T细胞在之前24小时内从胸腺释放。与具有Thy1-、CD45RC+和/或RT6+表型的外周T细胞主体不同,这些RTE是Thy1+、CD45RC-和RT6-。此外,定义了两个离散的RTE亚群:一个主要亚群(约95%)为CD4+或8+(单阳性)、TCR-αβ高表达的T细胞,类似于髓质胸腺细胞;一个次要亚群(约5%)为CD4+和8+(双阳性)、TCR-αβ低表达的T细胞,类似于皮质胸腺细胞。以下证据表明,在年轻成年大鼠中,大多数(如果不是全部)Thy1+ T细胞是RTE及其直接后代:1)胸腺切除术(而非假胸腺切除术)在3至7天内选择性地从淋巴结中耗尽Thy1+ T细胞,即使在肾上腺切除大鼠中也是如此;2)大多数FITC标记的RTE在从胸腺释放后7天内分化为Thy1-、CD45RC+、RT6+ T细胞;3)在正常以及胸腺内注射FITC的大鼠中观察到Thy1+ T细胞的过渡表型,包括Thy1低表达、CD45RC+和RT6+;4)新生大鼠中的大多数T细胞是Thy1+和RT6-,而它们的后代是Thy1-和RT6+。这些实验表明,正常大鼠中的大多数RTE在从胸腺释放时在表型上(可能在发育上)不成熟,并在胸腺后逐渐获得更成熟T细胞的表型特征。这些独特的表型特征应有助于加速RTE及其直接后代的分离,以便对其发育和功能特性进行确定性研究。