Thorp K M, Verschueren H, De Baetselier P, Southern C, Matthews N
Yamanouchi Research Institute, Littlemore Hospital, Oxford, UK.
Immunology. 1996 Mar;87(3):434-8. doi: 10.1046/j.1365-2567.1996.486567.x.
Lymphocyte migration into inflammatory sites involves a change from a spherical, non-motile phenotype to an irregular, constantly shape-changing, motile phenotype. We have previously shown that lymphocytes are maintained in the non-motile state by the constitutive activity of protein kinase C (PKC). In this paper we have attempted to identify the PKC isotype which regulates these morphological changes by three different approaches. (a) Motile and non-motile T-cell lines were compared for expression of the alpha, beta I, beta II, gamma, delta, epsilon, eta, zeta and theta isotypes by Western blotting. There was no obvious correlation of isotype expression with motility. (b) Two different PKC inhibitors, one specific for classical isotypes, Go6976 and the other GF109203X, which inhibits both classical and non-classical isotypes were compared for induction of motility in non-motile lymphocytes. Only GF109203X induced motility implying that a non-classical isotype is involved. (c) Non-motile lymphocytes were chronically treated with the PKC activator bryostatin and the time courses of induction of motility and downregulation of PKC isotypes were compared. Induction of motility correlated better with downregulation of epsilon, eta and theta than with alpha or beta. It is concluded that the data fit best with the involvement of a non-classical PKC isotype in regulating lymphocyte motility although no association with a particular isotype was found.
淋巴细胞迁移至炎症部位涉及从球形、非运动表型转变为不规则、不断改变形状的运动表型。我们之前已经表明，蛋白激酶C（PKC）的组成性活性可使淋巴细胞维持在非运动状态。在本文中，我们尝试通过三种不同方法来确定调节这些形态变化的PKC同工型。（a）通过蛋白质印迹法比较运动性和非运动性T细胞系中α、βI、βII、γ、δ、ε、η、ζ和θ同工型的表达。同工型表达与运动性之间没有明显的相关性。（b）比较两种不同的PKC抑制剂，一种对经典同工型具有特异性的Go6976，另一种抑制经典和非经典同工型的GF109203X，观察它们对非运动淋巴细胞运动性的诱导作用。只有GF109203X能诱导运动性，这意味着涉及一种非经典同工型。（c）用PKC激活剂苔藓抑素长期处理非运动淋巴细胞，并比较运动性诱导和PKC同工型下调的时间进程。运动性的诱导与ε、η和θ同工型的下调相关性比与α或β同工型的下调更好。得出的结论是，尽管未发现与特定同工型相关，但数据最符合非经典PKC同工型参与调节淋巴细胞运动性的情况。
