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



Mohd Wajid Ali Khan1, 2 ,*, Ahmed Al Otaibi1, Subuhi Sherwani3, Eida Mohammed Alshammari1,
Salma Ahmed Al-Zahrani1, Wahid Ali Khan4, Abdulmohsen Khalaf Dhahi Alsukaibi1, Sultan
Alouffi2, 5 & Shahper Nazeer Khan6



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



Mohd Wajid Ali Khan - Email; wajidkhan11@gmail.com; mw.khan@uoh.edu.sa; * Corresponding author


Article Type

Research Article



Received February 28, 2021; Revised March 30, 2021; Accepted March 30, 2021, Published March 31, 2021



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 ア 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) ゥBiomedical Informatics (2021) 461 at the concentrations of 100 IU/ml and 10 ng/ml respectively. 14-day cultures showed highly pure (91.6 ア 5.1%) and live (96.5 ア 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.



γδ T cell, expanded, cytokines, zoledronate, cancer therapeutics, PBMCs



Ali Khan et al. Bioinformation 17(3): 460-470 (2021)


Edited by

P Kangueane






Biomedical Informatics



This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.