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The Optimum Dose of Beta-Glucan for Stimulating Peripheral Blood Mononuclear Cells (PBMCs) to Produce Cytokines: In Vitro Study

Meira Erawati

Abstract

Background: Beta-glucan has been frequently used in laboratory trials as an immunomodulator in both in vivo and in vitro studies, but the ef­fective dose for measuring its performance has not been established. Like other immunomodula­tors, researchers must establish the right dose of beta-glucan in their laboratory experiments for the purpose of testing substances as immuno­modulators without achieving false-positive or –negative results. This study aimed to determine the optimum dose of beta-glucan to induce cyto­kine production by peripheral blood mononu­clear cells (PBMCs) in vitro.

Subjects and Method: This was a laboratory experimental study. This study measur­ed the production of two cytokines, including in­terferon gamma (IFN-γ) and interleukin 12 (IL-12), from the isolated PBMCs of healthy subjects. The doses of beta-glucan used as immunomodu­lator in­clu­ded 1, 5, 10, 20, and 50 μg/ml. Beta-glucan was add­ed to the PBMC culture medium, and the PBMCs were cultured for 6 days. On the sixth day, the supernatant was harvested and the cytokine production was analyzed using sandwich enzyme-linked immunosorbent assay (ELISA). Cytokines were also analyzed using the human IFN-γ ELISA kit and the human IL-12 ELISA kit, and data analysis was performed by one-way ANOVA.

Results: IFN-γ levels were found to be increased in the group treated with 5 μg/ml beta-glucan. The highest IFN-γ levels (70.0 pg/ml) were observed in the group treated with 10 μg/ml beta-glucan. The production of IL-12 increased sharply in the group treated with5 μg/ml beta-glucan but decreased in the group treated with 10 μg/ml beta-glucan. The mean cytokine levels of the beta-glucan group were found to be significantly different from those of the control group (p=0.001). One-way ANOVA revealed that the highest IL-12 production (77.2 pg/ml) occurred at a dose of 5 μg/ml beta-glucan. This average value was significantly different from the average production of IL-12 in the control group (p=0.001).

Conclusion: The optimum dose of beta-glucan for stimulating PBMCs to produce IFN-γ in vitro was 10 μg/ml, while for the production of IL-12, the dose was 5 μg/ml. Both cytokines can be mea­sured within 6 days of cell culture.

Keywords: beta-glucan, immunomodulator, cy­to­kines

Correspondence: Meira Erawati. Faculty of Medicine, Diponegoro University. Jl. Prof. Soedarto, S.H, Tembalang, Semarang, Indonesia. Email: meira­e­rawati­@­gmail.com. Mobile: +62-81915339685

Indonesian Journal of Medicine (2020), 5(2): 170-177
https://doi.org/10.26911/theijmed.2020.05.02.12

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