Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant

Context: The co-delivery of adjuvant and antigen has shown to be more effective for targeting the immune response than antigen alone. Therefore, designing an efficient bicistronic system is more assuring for production of both elements in the same tobacco cells as a plant model system. Objective: Co...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sara Mohammadzadeh, Hamideh Ofoghi, Mina Ebrahimi-Rad, Parastoo Ehsani
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2019
Materias:	vaccine plant-produced gm-csf hbsag co-expression Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/da7de49ad7d94246bcd95b2d19674618
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:da7de49ad7d94246bcd95b2d19674618
record_format	dspace
spelling	oai:doaj.org-article:da7de49ad7d94246bcd95b2d196746182021-11-17T14:21:56ZConstruction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant1388-02091744-511610.1080/13880209.2019.1662458https://doaj.org/article/da7de49ad7d94246bcd95b2d196746182019-01-01T00:00:00Zhttp://dx.doi.org/10.1080/13880209.2019.1662458https://doaj.org/toc/1388-0209https://doaj.org/toc/1744-5116Context: The co-delivery of adjuvant and antigen has shown to be more effective for targeting the immune response than antigen alone. Therefore, designing an efficient bicistronic system is more assuring for production of both elements in the same tobacco cells as a plant model system. Objective: Comparing the efficient transient co-expression of hepatitis B surface antigen (HBsAg) and mouse granulocyte macrophage colony stimulating factor (mGM-CSF) in tobacco leaves by designing either mono or bicistronic cassettes. Materials and methods: Four expression cassettes containing tobacco etch virus (TEV) leader sequence were constructed with and without above genes in different orders. The cassettes were transferred into tobacco, Nicotiana tabacum L. (Solanaceae), leaves by agroinfiltration technique. The expression levels were compared using ELISA and western blotting and bioactivity of cytokine was assessed by in vitro proliferation of mouse GM-CSF-responsive progenitor cells. Results: Agroinfiltrated leaves contained recombinant HBsAg protein at 20–50 ng/mg and mGM-CSF at 0.2–4 ng/mg in both nonglycosylated and glycosylated forms. The highest expression obtained in HBsAg and mGM-CSF monocistronic co-agroinfiltrated leaves. The expression of mGM-CSF was 1.1 and 0.2 ng/mg in two different orders of bicistronic cassettes. The growth frequency of GM progenitors was approximately 1/187 cells for standard rGM-CSF and 3.2 times less activity for the plant produced. Discussion and conclusions: The recombinant mGM-CSF was produced less in bicistronic cassette than other forms; however, co-presenting of both vaccine candidate and adjuvant is confirmed and could be promising for amelioration of plant expression system as a means for vaccine production.Sara MohammadzadehHamideh OfoghiMina Ebrahimi-RadParastoo EhsaniTaylor & Francis Grouparticlevaccineplant-producedgm-csfhbsagco-expressionTherapeutics. PharmacologyRM1-950ENPharmaceutical Biology, Vol 57, Iss 1, Pp 669-675 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	vaccine plant-produced gm-csf hbsag co-expression Therapeutics. Pharmacology RM1-950
spellingShingle	vaccine plant-produced gm-csf hbsag co-expression Therapeutics. Pharmacology RM1-950 Sara Mohammadzadeh Hamideh Ofoghi Mina Ebrahimi-Rad Parastoo Ehsani Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
description	Context: The co-delivery of adjuvant and antigen has shown to be more effective for targeting the immune response than antigen alone. Therefore, designing an efficient bicistronic system is more assuring for production of both elements in the same tobacco cells as a plant model system. Objective: Comparing the efficient transient co-expression of hepatitis B surface antigen (HBsAg) and mouse granulocyte macrophage colony stimulating factor (mGM-CSF) in tobacco leaves by designing either mono or bicistronic cassettes. Materials and methods: Four expression cassettes containing tobacco etch virus (TEV) leader sequence were constructed with and without above genes in different orders. The cassettes were transferred into tobacco, Nicotiana tabacum L. (Solanaceae), leaves by agroinfiltration technique. The expression levels were compared using ELISA and western blotting and bioactivity of cytokine was assessed by in vitro proliferation of mouse GM-CSF-responsive progenitor cells. Results: Agroinfiltrated leaves contained recombinant HBsAg protein at 20–50 ng/mg and mGM-CSF at 0.2–4 ng/mg in both nonglycosylated and glycosylated forms. The highest expression obtained in HBsAg and mGM-CSF monocistronic co-agroinfiltrated leaves. The expression of mGM-CSF was 1.1 and 0.2 ng/mg in two different orders of bicistronic cassettes. The growth frequency of GM progenitors was approximately 1/187 cells for standard rGM-CSF and 3.2 times less activity for the plant produced. Discussion and conclusions: The recombinant mGM-CSF was produced less in bicistronic cassette than other forms; however, co-presenting of both vaccine candidate and adjuvant is confirmed and could be promising for amelioration of plant expression system as a means for vaccine production.
format	article
author	Sara Mohammadzadeh Hamideh Ofoghi Mina Ebrahimi-Rad Parastoo Ehsani
author_facet	Sara Mohammadzadeh Hamideh Ofoghi Mina Ebrahimi-Rad Parastoo Ehsani
author_sort	Sara Mohammadzadeh
title	Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
title_short	Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
title_full	Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
title_fullStr	Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
title_full_unstemmed	Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
title_sort	construction of bicistronic cassette for co-expressing hepatitis b surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant
publisher	Taylor & Francis Group
publishDate	2019
url	https://doaj.org/article/da7de49ad7d94246bcd95b2d19674618
work_keys_str_mv	AT saramohammadzadeh constructionofbicistroniccassetteforcoexpressinghepatitisbsurfaceantigenandmousegranulocytemacrophagecolonystimulatingfactorasadjuvantintobaccoplant AT hamidehofoghi constructionofbicistroniccassetteforcoexpressinghepatitisbsurfaceantigenandmousegranulocytemacrophagecolonystimulatingfactorasadjuvantintobaccoplant AT minaebrahimirad constructionofbicistroniccassetteforcoexpressinghepatitisbsurfaceantigenandmousegranulocytemacrophagecolonystimulatingfactorasadjuvantintobaccoplant AT parastooehsani constructionofbicistroniccassetteforcoexpressinghepatitisbsurfaceantigenandmousegranulocytemacrophagecolonystimulatingfactorasadjuvantintobaccoplant
_version_	1718425514230677504

Construction of bicistronic cassette for co-expressing hepatitis B surface antigen and mouse granulocyte-macrophage colony stimulating factor as adjuvant in tobacco plant

Ejemplares similares