Optimization of methods for peripheral blood mononuclear cells isolation and expansion of human gamma delta T cells

Mohd Wajid Ali Khan

doi:10.6026//97320630017460

Abstract

Human Vγ9/Vδ2 T cells (γδ T cells) are immune surveillance cells both in innate and adaptive immunity and are a possible target for anticancer therapies, which can induce immune responses in a variety of cancers. Small non-peptide antigens such as zoledronate can do activation and expansion of T cells in vitro. It is evident that for adoptive cancer therapies, large numbers of functional cells are needed into cancer patients. Hence, optimization of methods needs to be carried out for the efficient expansion of these T cells. Standardization of peripheral blood mononuclear cells (PBMCs) isolation was devised. Cytokines (interleukin 2 (IL-2) and interleukin 15 (IL-15)) and zoledronate were also standardized for different concentrations. It was found that an increased number of PBMCs were recovered when washing was done at 1100 revolution per minute (rpm). Significantly high expansion fold was (2524 ± 787 expansion fold) achieved when stimulation of PBMCs was done with 1 μM of zoledronate and both cytokines IL-2 and IL-15 supported the expansion and survival of cells ISSN 0973-2063 (online) 0973-8894 (print) Bioinformation 17(3): 460-469 (2021) ©Biomedical Informatics (2021) 461 at the concentrations of 100 IU/ml and 10 ng/ml respectively. 14-day cultures showed highly pure (91.6 ± 5.1%) and live (96.5 ± 2.5%) expanded γδ T cells. This study aimed to standardize an easy to manipulate technique for the expansion of γδ T cells, giving a higher yield.

Highlights

Inflammation, including fatty liver diseases such as alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD), is an integral component of both all-acute and chronic liver disorder
It is of interest to report the molecular docking analysis of stachydrine and sakuranetin with IL-6 and TNF-α in the context of inflammation
Molecular docking means that the drug is correctly absorbed and The binding force, the number of hydrogen bond interactions and binds with the receptor

Summary

Background

Inflammation, including fatty liver diseases such as alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD), is an integral component of both all-acute and chronic liver disorder. It has been convincingly shown that immune mediators, especially pro-inflammatory cytokines, have been able to regulate many of the main characteristics of liver diseases, including acute liver failure, acute phase response, steatosis, cholestasis, hypergammaglobulinemia, and production of fibrosis [1,2]. It points out that classic signalling regulates interleukin-6 regenerative or anti-inflammatory actions, whereas interleukin-6 pro-inflammatory responses are more mediated by trans-signaling. This is relevant because the neutralising anti-interleukin-6 receptor monoclonal antibody tocilizumab has recently been licenced for therapeutic blockade of interleukin-6 in the therapy of inflammatory diseases [4]. Other chronic autoimmune disorders, such as systemic lupus erythematosus (SLE) [8], have a therapeutic benefit by blocking pro-inflammatory cytokines [8]. It is of interest to report the molecular docking analysis of stachydrine and sakuranetin with IL-6 and TNF-α in the context of inflammation

Materials and Methods

IL-6 2 TNF-α

Conclusion