Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells

Abstract Background Producing an appropriate number of engineered cells is considered as one of the influential factors in the successful treatments with chimeric antigen receptor (CAR) T cells. To this aim, the transduction rate of the viral vectors can play a significant role. In addition, improvi...

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Autores principales:	Alireza Rajabzadeh, Amir Ali Hamidieh, Fatemeh Rahbarizadeh
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Chimeric antigen receptor Immunotherapy Polybrene Retronectin T cell Cytology QH573-671
Acceso en línea:	https://doaj.org/article/15094b31441a4be193eacbf51293d0ca
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spelling	oai:doaj.org-article:15094b31441a4be193eacbf51293d0ca2021-11-28T12:23:26ZSpinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells10.1186/s12860-021-00397-z2661-8850https://doaj.org/article/15094b31441a4be193eacbf51293d0ca2021-11-01T00:00:00Zhttps://doi.org/10.1186/s12860-021-00397-zhttps://doaj.org/toc/2661-8850Abstract Background Producing an appropriate number of engineered cells is considered as one of the influential factors in the successful treatments with chimeric antigen receptor (CAR) T cells. To this aim, the transduction rate of the viral vectors can play a significant role. In addition, improving transduction rates can affect the success rate of this treatment due to hard-transduced T lymphocytes. Results In this study, activated T cells were transduced using different transduction methods such as spinoculation, retronectin, polybrene, spinoculation + retronectin, and spinoculation + polybrene after selecting the most efficient transfection method to produce recombinant viral particles containing MUC1 CAR. PEI and lipofectamine with the amount of 73.72 and 72.53%, respectively, showed the highest transfection rates with respect to calcium phosphate (14.13%) for producing lentiviral particles. However, the cytotoxicity of transfection methods was not significantly different. Based on the results, spinoculation + retronectin leads to the highest transduction rates of T cells (63.19 ± 4.45%) relative to spinoculation + polybrene (34.6 ± 4.44%), polybrene (10.23 ± 0.79%), retronectin (10.37 ± 1.85%), and spinoculation (21.11 ± 1.55%). Further, the polybrene (40.02%) and spinoculation + polybrene (48.83% ± 4.83) increased cytotoxicity significantly compared to other groups. Conclusion Improving transduction conditions such as using spinoculation with retronectin can ameliorate the production of CAR-T cells by increasing the rate of transduction, as well as the success rate of treatment.Alireza RajabzadehAmir Ali HamidiehFatemeh RahbarizadehBMCarticleChimeric antigen receptorImmunotherapyPolybreneRetronectinT cellCytologyQH573-671ENBMC Molecular and Cell Biology, Vol 22, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Chimeric antigen receptor Immunotherapy Polybrene Retronectin T cell Cytology QH573-671
spellingShingle	Chimeric antigen receptor Immunotherapy Polybrene Retronectin T cell Cytology QH573-671 Alireza Rajabzadeh Amir Ali Hamidieh Fatemeh Rahbarizadeh Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
description	Abstract Background Producing an appropriate number of engineered cells is considered as one of the influential factors in the successful treatments with chimeric antigen receptor (CAR) T cells. To this aim, the transduction rate of the viral vectors can play a significant role. In addition, improving transduction rates can affect the success rate of this treatment due to hard-transduced T lymphocytes. Results In this study, activated T cells were transduced using different transduction methods such as spinoculation, retronectin, polybrene, spinoculation + retronectin, and spinoculation + polybrene after selecting the most efficient transfection method to produce recombinant viral particles containing MUC1 CAR. PEI and lipofectamine with the amount of 73.72 and 72.53%, respectively, showed the highest transfection rates with respect to calcium phosphate (14.13%) for producing lentiviral particles. However, the cytotoxicity of transfection methods was not significantly different. Based on the results, spinoculation + retronectin leads to the highest transduction rates of T cells (63.19 ± 4.45%) relative to spinoculation + polybrene (34.6 ± 4.44%), polybrene (10.23 ± 0.79%), retronectin (10.37 ± 1.85%), and spinoculation (21.11 ± 1.55%). Further, the polybrene (40.02%) and spinoculation + polybrene (48.83% ± 4.83) increased cytotoxicity significantly compared to other groups. Conclusion Improving transduction conditions such as using spinoculation with retronectin can ameliorate the production of CAR-T cells by increasing the rate of transduction, as well as the success rate of treatment.
format	article
author	Alireza Rajabzadeh Amir Ali Hamidieh Fatemeh Rahbarizadeh
author_facet	Alireza Rajabzadeh Amir Ali Hamidieh Fatemeh Rahbarizadeh
author_sort	Alireza Rajabzadeh
title	Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
title_short	Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
title_full	Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
title_fullStr	Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
title_full_unstemmed	Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells
title_sort	spinoculation and retronectin highly enhance the gene transduction efficiency of mucin-1-specific chimeric antigen receptor (car) in human primary t cells
publisher	BMC
publishDate	2021
url	https://doaj.org/article/15094b31441a4be193eacbf51293d0ca
work_keys_str_mv	AT alirezarajabzadeh spinoculationandretronectinhighlyenhancethegenetransductionefficiencyofmucin1specificchimericantigenreceptorcarinhumanprimarytcells AT amiralihamidieh spinoculationandretronectinhighlyenhancethegenetransductionefficiencyofmucin1specificchimericantigenreceptorcarinhumanprimarytcells AT fatemehrahbarizadeh spinoculationandretronectinhighlyenhancethegenetransductionefficiencyofmucin1specificchimericantigenreceptorcarinhumanprimarytcells
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Spinoculation and retronectin highly enhance the gene transduction efficiency of Mucin-1-specific chimeric antigen receptor (CAR) in human primary T cells

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