Svirshchevskaya E V, Sidorov I A, Viskova N Y, Dozmorov I M
Shemyakin Institute of Bioorganic Chemistry, Russian Acad. Sci., Moscow.
J Immunol Methods. 1993 Feb 26;159(1-2):17-27. doi: 10.1016/0022-1759(93)90137-v.
The proliferative response of CTLL-2 cells to human recombinant interleukin-2 (IL-2) can be modeled mathematically using enzyme kinetic equations. This approach has been used to analyze dose-response curves (IL-2 concentration vs. level of proliferation) measured by MTT and [3H]TdR assays. The values of functional dissociation constants, equivalent to IL-2 concentrations giving 50% of the maximal response, depended on the cell concentration and increased from 4 to 60 pM for the [3H]TdR assay and from 40 to 140 pM for the MTT assay when the cell concentration was increased from 2 x 10(3) to 4 x 10(4) cells/well. The types of inhibition and dissociation constants for various inhibitors of IL-2-dependent proliferation such as mAbs against IL-2 receptor (7D4 and AMT13) and normal mouse serum (NMS) were also analyzed. Both mAbs exhibited competitive mechanisms of inhibition whereas NMS inhibited IL-2-driven proliferation in a mixed manner. Two gel-filtration fractions of NMS with inhibitory activity manifested different types of inhibition: purely competitive type of inhibition in the case of a 10-15 kDa fraction and a mixed type of inhibition for a 100-150 kDa fraction. The proposed model can also be used for quantitative analysis of the influence of various factors (pH, temperature, cultivation condition) on the level of proliferation.
CTLL-2细胞对人重组白细胞介素-2（IL-2）的增殖反应可用酶动力学方程进行数学建模。该方法已用于分析通过MTT和[3H]TdR测定法测得的剂量反应曲线（IL-2浓度与增殖水平）。功能解离常数的值，相当于产生最大反应50%时的IL-2浓度，取决于细胞浓度，当细胞浓度从2×10³增加到4×10⁴个细胞/孔时，[3H]TdR测定法的值从4增加到60 pM，MTT测定法的值从40增加到140 pM。还分析了IL-2依赖性增殖的各种抑制剂的抑制类型和解离常数，如抗IL-2受体的单克隆抗体（7D4和AMT13）和正常小鼠血清（NMS）。两种单克隆抗体均表现出竞争性抑制机制，而NMS以混合方式抑制IL-2驱动的增殖。具有抑制活性的NMS的两个凝胶过滤级分表现出不同类型的抑制：10 - 15 kDa级分为纯竞争性抑制类型，100 - 150 kDa级分为混合抑制类型。所提出的模型还可用于定量分析各种因素（pH、温度、培养条件）对增殖水平的影响。
