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.

Add to citation manager EndNote|Ris|BibTeX

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

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

References

[1] BUGELSKI PJ, ACHUTHANANDAM R, CAPOCASALE RJ, et al.Monoclonal antibody-induced cytokine-release syndrome[J]. Expert Review of Clinical Immunology, 2009, 5(5): 499-521.
[2] TUDESQ JJ, YAKOUB-AGHA M, BAY JO, et al.Management of cytokine release syndrome and macrophage activation syndrome following CAR-T cell therapy: Guidelines from the SFGM-TC[J]. Bull Cancer, 2023, 110(2s): S116-s122.
[3] XIAO X, HUANG S, CHEN S, et al.Mechanisms of cytokine release syndrome and neurotoxicity of CAR T-cell therapy and associated prevention and management strategies[J]. J Exp Clin Cancer Res, 2021, 40(1): 367.
[4] WANG Y, DONG H, DONG T, et al. Treatment of cytokine release syndrome-induced vascular endothelial injury using mesenchymal stem cells[J/OL]. Mol Cell Biochem, 2023. https://pubmed.ncbi.nlm.nih.gov/37392343/.
[5] LAZOVIC B, DMITROVIC R, SIMONOVIC I, et al.Cytokine release syndrome (CRS) in severe COVID-19 patients: two controversial and interesting case reports and literature review[J]. Tanaffos, 2023, 22(1): 167-171.
[6] QUE Y, HU C, WAN K, et al.Cytokine release syndrome in COVID-19: a major mechanism of morbidity and mortality[J]. Int Rev Immunol, 2022, 41(2): 217-230.
[7] ETEMADIFAR M, SALARI M, SAERI M, et al.Rituximab induced cytokine release syndrome in an MS patient: a case report[J]. Clin Case Rep, 2021, 9(7): e04407.
[8] OJEMOLON PE, KALIDINDI S, AHLBORN TA, et al.Cytokine release syndrome following blinatumomab therapy[J]. Cureus, 2022, 14(1): e21583.
[9] HEGAZY AN, KRÖNKE J, ANGERMAIR S, et al. Anti-SARS-CoV2 antibody-mediated cytokine release syndrome in a patient with acute promyelocytic leukemia[J]. BMC Infect Dis, 2022, 22(1): 537.
[10] Center for Drug Evaluation, NMPA. Technical guidelines for drug immunogenicity research[EB/OL]. (2021-03-29)[2023-12-10]. https://www.cde.org.cn/main/news/viewInfoCommon/a0908879d6c54c7318f0881611b51122.
[11] Center for Drug Evaluation, NMPA. Technical Guidelines for Non-clinical Studies of Drug Immunotoxicity (Draft for Comment) [EB/OL]. (2022-09-30)[2024-4-10]. https://www.cde.org.cn/main/news/viewInfoCommon/a2f5f52dcdcd6cccae58e034c937 e986.
[12] YANG J, JIAO J, DRAHEIM KM, et al.Simultaneous evaluation of treatment efficacy and toxicity for bispecific T-cell engager therapeutics in a humanized mouse model[J]. FASEB J, 2023, 37(6): e22995.
[13] GRIMALDI C, IBRAGHIMOV A, KIESSLING A, et al.Current nonclinical approaches for immune assessments of immuno-oncology biotherapeutics[J]. Drug Discov Today, 2023, 28(2): 103440.
[14] RYAN JM, WASSER JS, ADLER AJ, et al.Enhancing the safety of antibody-based immunomodulatory cancer therapy without compromising therapeutic benefit: Can we have our cake and eat it too?[J]. Expert Opin Biol Ther, 2016, 16(5): 655-674.
[15] LIN Z, LYU JJ, QU Z, et al.Preliminary analysis of cytokine-release syndrome induced by monoclonal antibodies[J]. Chinese Journal of New Drugs(中国新药杂志), 2012, 21(7): 746-750, 756.
[16] FINCO D, GRIMALDI C, FORT M, et al.Cytokine release assays: current practices and future directions[J]. Cytokine, 2014, 66(2): 143-155.
[17] GRIMALDI C, FINCO D, FORT MM, et al.Cytokine release: a workshop proceedings on the state-of-the-science, current challenges and future directions[J]. Cytokine, 2016, 85: 101-108.
[18] VESSILLIER S, EASTWOOD D, FOX B, et al.Cytokine release assays for the prediction of therapeutic mAb safety in first-in man trials--Whole blood cytokine release assays are poorly predictive for TGN1412 cytokine storm[J]. J Immunol Methods, 2015, 424: 43-52.
[19] STEBBINGS R, EASTWOOD D, POOLE S, et al.After TGN1412: recent developments in cytokine release assays[J]. Journal of Immunotoxicology, 2013, 10(1): 75-82.
[20] CHOPRA RK, NAGEL JE, CHREST FJ, et al.Regulation of interleukin 2 and interleukin 2 receptor gene expression in human T cells: I. Effect of Ca²+-ionophore on phorbol myristate acetate co-stimulated cells[J]. Clin Exp Immunol, 1987, 69(2): 433-440.
[21] GAO R, LI X, GAO H, et al.Protein phosphatase 2A catalytic subunit β suppresses PMA/ionomycin-induced T-cell activation by negatively regulating PI3K/Akt signaling[J]. FEBS J, 2022, 289(15): 4518-4535.
[22] OLSEN I, SOLLID LM.Pitfalls in determining the cytokine profile of human T cells[J]. J Immunol Methods, 2013, 390(1-2): 106-112.
[23] TANAKA T, NARAZAKI M, KISHIMOTO T.Interleukin (IL-6) immunotherapy[J]. Cold Spring Harb Perspect Biol, 2018, 10(8): a028456.
[24] TEACHEY DT, RHEINGOLD SR, MAUDE SL, et al.Cytokine release syndrome after blinatumomab treatment related to abnormal macrophage activation and ameliorated with cytokine-directed therapy[J]. Blood, 2013, 121(26): 5154-5157.
[25] NORELLI M, CAMISA B, BARBIERA G, et al.Monocyte-derived IL-1 and IL-6 are differentially required for cytokine-release syndrome and neurotoxicity due to CAR T cells[J]. Nat Med, 2018, 24(6): 739-748.
[26] TANAKA T, NARAZAKI M, KISHIMOTO T.Immunotherapeutic implications of IL-6 blockade for cytokine storm[J]. Immunotherapy, 2016, 8(8): 959-970.
[27] JORGOVANOVIC D, SONG M, WANG L, et al.Roles of IFN-γ in tumor progression and regression: a review[J]. Biomark Res, 2020, 8: 49.
[28] BAILEY SR, VATSA S, LARSON RC, et al.Blockade or deletion of IFNγ reduces macrophage activation without compromising CAR T-cell function in hematologic malignancies[J]. Blood Cancer Discov, 2022, 3(2): 136-153.
[29] KURSUNEL MA, ESENDAGLI G.The untold story of IFN-γ in cancer biology[J]. Cytokine Growth Factor Rev, 2016, 31: 73-81.
[30] ALAM M, CHOUDHURY R, LAMERS RJ.An adaptable in vitro cytokine release assay (CRA): susceptibility to cytokine storm in COVID-19 as a model[J]. Curr Res Immunol, 2022, 3: 239-243.
[31] HUANG Y, HOU TT, QIN C, et al. Discussion on non-clinical safety evaluation of chimeric antigen receptor T cell products and main concerns[J]. Chinese Journal of Pharmacovigilance(中国药物警戒), 2202, 19(8): 813-816.