Knudsen E, Seierstad T, Vaage J T, Naper C, Benestad H B, Rolstad B, Maghazachi A A
Department of Physiology, Institute of Basic Medical Sciences, University of Oslo, Norway.
Int Immunol. 1997 Jul;9(7):1043-51. doi: 10.1093/intimm/9.7.1043.
We have successfully cloned nine NKR-P1+ TCR alpha beta + cells from PVG rat spleens, utilizing murine macrophage inflammatory protein-1 alpha (MIP-1 alpha) and IL-2. These clones are either double negative (DN, CD4-CD8-), which included clones 3.31, 3.71, 4.19, 4.59 and 4.65, or single positive (SP, CD4+CD8-), which included clones 1.64, 3.8, 3.76 and 3.78. No CD8+ clone was recovered. All nine clones are restricted in terms of their expression of the V beta antigens, since they express V beta 8.2 but not V beta 8.5, V beta 10 or V beta 16. These clones are agranular and they fall to generate NK or LAK activity upon incubation with IL-2, IL-12 or their combination. On the basis of their production of intracellular cytokines they can be divided into three categories: (I) SP clones (1.64, 3.8, 3.76 and 3.78) do not produce IL-2 or IL-4, but produce IFN-gamma and IL-12, and they vary in their production of IL-1, RANTES or tumor necrosis factor (TNF)-alpha; (II) DN clones 4.59 and 4.65 produce IL-1 alpha and IFN-gamma only, and fall to produce other cytokines; and (III) DN clones 3.31, 3.71 and 4.19 produce IL-1 alpha, IL-1 beta, IL-2, IL-12, IFN-gamma, RANTES and TNF-alpha. From all the clones examined only DN clones 3.31 and to a lesser degree 4.19 produce IL-4. In vivo tissue localization of clones 3.8, 3.31 and 4.59 shows that these cells distribute into the liver and bone marrow 24 h post i.v. administration. Their accumulation in the liver and bone marrow along with their ability to secrete various cytokines suggest that these cells may influence the generation, differentiation or apoptosis of immune or hematopoietic cells.
我们利用小鼠巨噬细胞炎性蛋白-1α（MIP-1α）和白细胞介素-2（IL-2），成功从PVG大鼠脾脏中克隆出9个NKR-P1⁺TCRαβ⁺细胞。这些克隆细胞要么是双阴性（DN，CD4⁻CD8⁻），包括克隆3.31、3.71、4.19、4.59和4.65，要么是单阳性（SP，CD4⁺CD8⁻），包括克隆1.64、3.8、3.76和3.78。未获得CD8⁺克隆。所有9个克隆细胞在Vβ抗原表达方面都具有限制性，因为它们表达Vβ8.2，但不表达Vβ8.5、Vβ10或Vβ16。这些克隆细胞无颗粒，并且在与IL-2、IL-12或它们的组合孵育后不会产生自然杀伤（NK）或淋巴因子激活的杀伤（LAK）活性。根据它们细胞内细胞因子的产生情况，可将它们分为三类：（I）SP克隆（1.64、3.8、3.76和3.78）不产生IL-2或IL-4，但产生干扰素-γ（IFN-γ）和IL-12，并且它们在IL-1、调节激活正常T细胞表达和分泌因子（RANTES）或肿瘤坏死因子（TNF）-α的产生上存在差异；（II）DN克隆4.59和4.65仅产生IL-1α和IFN-γ，不产生其他细胞因子；（III）DN克隆3.31、3.71和4.19产生IL-1α、IL-1β、IL-2、IL-12、IFN-γ、RANTES和TNF-α。在所检测的所有克隆中，只有DN克隆3.31以及程度稍低的4.19产生IL-4。克隆3.8、3.31和4.59在体内组织中的定位显示，静脉注射后24小时，这些细胞分布到肝脏和骨髓中。它们在肝脏和骨髓中的聚集以及它们分泌各种细胞因子的能力表明，这些细胞可能影响免疫细胞或造血细胞的生成、分化或凋亡。
