Callewaert D M, Mahle N H, Genyea J, Wilusz J R, Chores J B, Baker S, Thomas R, Ruedisueli E
Nat Immun Cell Growth Regul. 1983;3(4):210-21.
A multistep kinetic model for natural cytotoxicity reactions in vitro is described that includes consideration of the effect of nonlytic target-binding lymphocytes on the experimentally determined kinetic parameters. The expression for the maximal velocity, Vmax, obtained using this model is essentially identical to that obtained using simpler models, whereas the expression for the apparent Michaelis constant, KappM, is considerably more complex. As a first step in the application of this model, experiments were designed to determine the relative contribution of some of the component terms to the value of KappM. Methods were developed that allow for experimental determination of the rate constants for both lytic programming and killer-cell-independent lysis (KCIL) steps in the cytolytic process. The results obtained support lytic programming as the rate-determining step in natural cytotoxicity reactions and demonstrate that terms related to nonlytic target-binding lymphocytes contribute significantly to experimentally determined values for KappM. In addition, the methods developed for the determination of the rates of lytic programming and KCIL should prove useful for various studies of the mechanism of cytotoxicity and the effects of drugs and disease on such phenomena.
本文描述了一种体外自然细胞毒性反应的多步动力学模型，该模型考虑了非裂解性靶标结合淋巴细胞对实验测定的动力学参数的影响。使用该模型获得的最大速度Vmax的表达式与使用更简单模型获得的表达式基本相同，而表观米氏常数KappM的表达式则要复杂得多。作为该模型应用的第一步，设计了实验来确定一些组成项对KappM值的相对贡献。开发了一些方法，可用于实验测定细胞溶解过程中裂解编程和非杀伤细胞依赖性裂解（KCIL）步骤的速率常数。获得的结果支持裂解编程是自然细胞毒性反应中的速率决定步骤，并表明与非裂解性靶标结合淋巴细胞相关的项对实验测定的KappM值有显著贡献。此外，开发的用于测定裂解编程速率和KCIL的方法对于细胞毒性机制以及药物和疾病对此类现象影响的各种研究应该是有用的。
