TY - JOUR
T1 - Quantitative analysis of interleukin-2-induced proliferation in the presence of inhibitors using a mathematical model
AU - Svirshchevskaya, E. V.
AU - Sidorov, I. A.
AU - Viskova, N. Y.
AU - Dozmorov, I. M.
N1 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1993/2/26
Y1 - 1993/2/26
N2 - 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 × 103 to 4 × 104 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.
AB - 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 × 103 to 4 × 104 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.
KW - Anti-IL-2 receptor antibody
KW - CTLL-2
KW - Enzyme kinetics equation
KW - IL-2
KW - Serum IL-2 inhibitor
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U2 - 10.1016/0022-1759(93)90137-V
DO - 10.1016/0022-1759(93)90137-V
M3 - Article
C2 - 8445249
AN - SCOPUS:0027512950
SN - 0022-1759
VL - 159
SP - 17
EP - 27
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -