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

Mohd Wajid Ali Khan; Ahmed Al Otaibi; Subuhi Sherwani; Eida Mohammed Alshammari; Salma Ahmed Al-Zahrani; Wahid Ali Khan; Abdulmohsen Khalaf Dhahi Alsukaibi; Sultan Alouffi; Shahper Nazeer Khan

doi:10.6026/97320630017460

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

Bioinformation. 2021 Mar 31;17(3):460-469. doi: 10.6026/97320630017460. eCollection 2021.

Affiliations

¹ Department of Chemistry, College of Sciences, University of Ha'il, Ha'il-2440, Saudi Arabia.
² Molecular Diagnostic and Personalised Therapeutics Unit, University of Ha'il, Ha'il-2440, Saudi Arabia.
³ Department of Biology, College of Sciences, University of Ha'il, Ha'il-2440, Saudi Arabia.
⁴ Department of Clinical Biochemistry,College of Medicine, King Khalid University, Abha-62529, Saudi Arabia.
⁵ Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, University of Ha'il, Ha'il-2440, Saudi Arabia.
⁶ Interdisciplinary Nanotechnology Centre, Aligarh Muslim University, Aligarh-202002, U.P,India.

Abstract

Human Vg9/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 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.

Keywords: PBMCs; cancer therapeutics; cytokines; expanded; zoledronate; γδT cell.