Exploratory study of cytokine release syndrome in vitro

doi:10.19803/j.1672-8629.20240149

Abstract

Abstract: Objective To establish an in vitro evaluation method of cytokine release syndrome(CRS), and explore the cell density, positive drug types and concentrations of the test system. Methods Peripheral blood mononuclear cells (PBMC) were stimulated with CD3 antibody (OKT3) (0.1~5μg), OKT3 (0.1~1μg) + CD28 antibody (anti-CD28) (1 and 2 μg·mL^-1), phorbol-12-myristate-13-acetate (PMA) (10~100 ng·mL^-1), and PMA (10~100 ng·mL^-1) + ionomycin (ION) (1 and 5 μg·mL^-1) for 48 h, respectively. The proliferation of PBMC was assessed by the CCK-8 method to determine the optimal cell density and drug concentration. There were seven groups in the experiment: Control group, OKT3 0.1μg+anti-CD28 1 μg·mL^-1group, OKT3 0.5μg+anti-CD28 1μg·mL^-1 group, OKT3 1μg+anti-CD28 1μg·mL^-1 group, PMA 10 ng·mL^-1+ION 1μg·mL^-1 group, PMA 25 ng·mL^-1+ION 1μg·mL^-1 group and PMA 50 ng·mL^-1+ION 1μg·mL^-1 group. The cell supernatant was collected after 48 h of exposure to the respective treatments and cytokine IL-6 and IFN-γ was detected by ELISA. Results Compared with the control group, both OKT3 and PMA induced significant proliferation of medium density and high density PBMC, and OKT3 and PMA induced cell proliferation at all doses. The combination of drugs produced a stronger cell activation effect than the positive drug alone. Compared with the control group, the release of IL-6 and IFN-γ from PBMC was significantly increased under OKT3 (0.1~1μg) +anti-CD28 (1μg·mL^-1) treatment. Under PMA (10~50 ng·mL^-1) +ION (1μg·mL^-1), the secretion of IFN-γ by PBMC was significantly increased compared with the control group, and under PMA (25-50 ng·mL^-1) +ION (1μg·mL^-1), the secretion of IL-6 by PBMC was significantly increased compared with the control group. Conclusion The study determines the cell density, positive drug types and concentrations, and drug combination of CRS risk assessment methods for PBMC in vitro.

Key words: cytokine release syndrome, orthoclone, CD28 antibody, phorbol-12-myristate -13-acetate, ionomycin, peripheral blood mononuclear cell, cell density

CLC Number:

R965.3

FU Yingshuang, LI Shuangxing, JIANG Hua, QU Zhe, HUO Guitao, YANG Yanwei, ZHANG Di, HUANG Ying, LI Bo, LIN Zhi. Exploratory study of cytokine release syndrome in vitro[J]. Chinese Journal of Pharmacovigilance, 2024, 21(6): 625-631.

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URL: https://www.zgywjj.com/EN/10.19803/j.1672-8629.20240149

https://www.zgywjj.com/EN/Y2024/V21/I6/625

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