Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity

We adapted a previously described Agrobacterium-mediated transient expression system to test the expression level of three constructs carrying the surface antigen 1 (SAG1) of Toxoplasma gondii. Two constructs were based in a Potato virus X (PVX) amplicon. In one of them, the PVX movement protein gen...

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Autores principales:	Clemente, M., Curilovic, R., Sassone, A., Zelada, A., Angel, S.O., Mentaberry, A.N.
Formato:	JOUR
Materias:	Plant transient expression PVX amplicon Toxoplasma gondii Bacteria Diseases Immunology Plants (botany) Tobacco Viruses Subgenomic promoters Antigens coat protein immunoglobulin G2a membrane antigen plant extract signal peptide surface antigen 1 Toxoplasma gondii vaccine unclassified drug virus vector amplicon animal experiment animal model animal tissue antigen expression antigen specificity apoplast blood sampling body burden brain cyst controlled study female functional genomics gene construct gene expression regulation gene expression system gene fusion genetic strain heterozygote human humoral immunity immunization immunogenicity lethality motion mouse nonhuman plant leaf plant tissue plant virus potato virus x promoter region provocation test review Rhizobium solubility tobacco vaccination viral gene delivery system Western blotting Animals Antibodies, Protozoan Antigens, Protozoan Brain Cysts Female Gene Expression Immunoglobulin G Mice Mice, Inbred C3H Plant Leaves Potexvirus Protozoan Proteins Toxoplasma Toxoplasmosis Genes Plants Agrobacterium Nicotiana tabacum Potato virus X
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_10736085_v30_n1_p41_Clemente
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_10736085_v30_n1_p41_Clemente2023-10-03T16:02:54Z Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity Clemente, M. Curilovic, R. Sassone, A. Zelada, A. Angel, S.O. Mentaberry, A.N. Plant transient expression PVX amplicon Toxoplasma gondii Bacteria Diseases Immunology Plants (botany) Tobacco Viruses Plant transient expression PVX amplicon Subgenomic promoters Toxoplasma gondii Antigens coat protein immunoglobulin G2a membrane antigen plant extract signal peptide surface antigen 1 Toxoplasma gondii vaccine unclassified drug virus vector amplicon animal experiment animal model animal tissue antigen expression antigen specificity apoplast blood sampling body burden brain cyst controlled study female functional genomics gene construct gene expression regulation gene expression system gene fusion genetic strain heterozygote human humoral immunity immunization immunogenicity lethality motion mouse nonhuman plant leaf plant tissue plant virus potato virus x promoter region provocation test review Rhizobium solubility tobacco Toxoplasma gondii vaccination viral gene delivery system Western blotting Animals Antibodies, Protozoan Antigens, Protozoan Brain Cysts Female Gene Expression Immunoglobulin G Mice Mice, Inbred C3H Plant Leaves Potexvirus Protozoan Proteins Rhizobium Tobacco Toxoplasma Toxoplasmosis Bacteria Diseases Genes Plants Tobacco Viruses Agrobacterium Nicotiana tabacum Potato virus X Toxoplasma gondii We adapted a previously described Agrobacterium-mediated transient expression system to test the expression level of three constructs carrying the surface antigen 1 (SAG1) of Toxoplasma gondii. Two constructs were based in a Potato virus X (PVX) amplicon. In one of them, the PVX movement protein genes were replaced by the sag1 gene. In the other, the sag1 gene was placed under the control of an additional coat protein subgenomic promoter. In the third construct, the sag1 gene was fused to an apoplastic peptide signal under the CaMV 35S promoter. Western blot analysis of leaf extracts infiltrated with each construct revealed a protein of 35 kDa. SAG1 accumulation in leaves ranged from 0.1 to 0.06% of total soluble protein (equivalent to 10 μg and 6 μg of SAG1 per gram of fresh leaf tissue, respectively). Three of five human seropositive samples reacted with tobacco-expressed SAG1 in Western blot analysis. The C3H mice were immunized with SAG-expressing leaf extracts and perorally challenged with a nonlethal dose of the T. gondii Me49 strain. Mice vaccinated with SAG1 showed significantly lower brain cyst burdens compared to those from the control group. Immunization with SAG1-expressing leaves elicited a specific humoral response with predominant participation of type IgG2a. In conclusion, a functional SAG1 version could be transiently expressed in tobacco leaves. © 2005 Humana Press Inc. All rights of any nature whatsoever reserved. Fil:Clemente, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Curilovic, R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Sassone, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Zelada, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Angel, S.O. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Mentaberry, A.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_10736085_v30_n1_p41_Clemente
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Plant transient expression PVX amplicon Toxoplasma gondii Bacteria Diseases Immunology Plants (botany) Tobacco Viruses Plant transient expression PVX amplicon Subgenomic promoters Toxoplasma gondii Antigens coat protein immunoglobulin G2a membrane antigen plant extract signal peptide surface antigen 1 Toxoplasma gondii vaccine unclassified drug virus vector amplicon animal experiment animal model animal tissue antigen expression antigen specificity apoplast blood sampling body burden brain cyst controlled study female functional genomics gene construct gene expression regulation gene expression system gene fusion genetic strain heterozygote human humoral immunity immunization immunogenicity lethality motion mouse nonhuman plant leaf plant tissue plant virus potato virus x promoter region provocation test review Rhizobium solubility tobacco Toxoplasma gondii vaccination viral gene delivery system Western blotting Animals Antibodies, Protozoan Antigens, Protozoan Brain Cysts Female Gene Expression Immunoglobulin G Mice Mice, Inbred C3H Plant Leaves Potexvirus Protozoan Proteins Rhizobium Tobacco Toxoplasma Toxoplasmosis Bacteria Diseases Genes Plants Tobacco Viruses Agrobacterium Nicotiana tabacum Potato virus X Toxoplasma gondii
spellingShingle	Plant transient expression PVX amplicon Toxoplasma gondii Bacteria Diseases Immunology Plants (botany) Tobacco Viruses Plant transient expression PVX amplicon Subgenomic promoters Toxoplasma gondii Antigens coat protein immunoglobulin G2a membrane antigen plant extract signal peptide surface antigen 1 Toxoplasma gondii vaccine unclassified drug virus vector amplicon animal experiment animal model animal tissue antigen expression antigen specificity apoplast blood sampling body burden brain cyst controlled study female functional genomics gene construct gene expression regulation gene expression system gene fusion genetic strain heterozygote human humoral immunity immunization immunogenicity lethality motion mouse nonhuman plant leaf plant tissue plant virus potato virus x promoter region provocation test review Rhizobium solubility tobacco Toxoplasma gondii vaccination viral gene delivery system Western blotting Animals Antibodies, Protozoan Antigens, Protozoan Brain Cysts Female Gene Expression Immunoglobulin G Mice Mice, Inbred C3H Plant Leaves Potexvirus Protozoan Proteins Rhizobium Tobacco Toxoplasma Toxoplasmosis Bacteria Diseases Genes Plants Tobacco Viruses Agrobacterium Nicotiana tabacum Potato virus X Toxoplasma gondii Clemente, M. Curilovic, R. Sassone, A. Zelada, A. Angel, S.O. Mentaberry, A.N. Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
topic_facet	Plant transient expression PVX amplicon Toxoplasma gondii Bacteria Diseases Immunology Plants (botany) Tobacco Viruses Plant transient expression PVX amplicon Subgenomic promoters Toxoplasma gondii Antigens coat protein immunoglobulin G2a membrane antigen plant extract signal peptide surface antigen 1 Toxoplasma gondii vaccine unclassified drug virus vector amplicon animal experiment animal model animal tissue antigen expression antigen specificity apoplast blood sampling body burden brain cyst controlled study female functional genomics gene construct gene expression regulation gene expression system gene fusion genetic strain heterozygote human humoral immunity immunization immunogenicity lethality motion mouse nonhuman plant leaf plant tissue plant virus potato virus x promoter region provocation test review Rhizobium solubility tobacco Toxoplasma gondii vaccination viral gene delivery system Western blotting Animals Antibodies, Protozoan Antigens, Protozoan Brain Cysts Female Gene Expression Immunoglobulin G Mice Mice, Inbred C3H Plant Leaves Potexvirus Protozoan Proteins Rhizobium Tobacco Toxoplasma Toxoplasmosis Bacteria Diseases Genes Plants Tobacco Viruses Agrobacterium Nicotiana tabacum Potato virus X Toxoplasma gondii
description	We adapted a previously described Agrobacterium-mediated transient expression system to test the expression level of three constructs carrying the surface antigen 1 (SAG1) of Toxoplasma gondii. Two constructs were based in a Potato virus X (PVX) amplicon. In one of them, the PVX movement protein genes were replaced by the sag1 gene. In the other, the sag1 gene was placed under the control of an additional coat protein subgenomic promoter. In the third construct, the sag1 gene was fused to an apoplastic peptide signal under the CaMV 35S promoter. Western blot analysis of leaf extracts infiltrated with each construct revealed a protein of 35 kDa. SAG1 accumulation in leaves ranged from 0.1 to 0.06% of total soluble protein (equivalent to 10 μg and 6 μg of SAG1 per gram of fresh leaf tissue, respectively). Three of five human seropositive samples reacted with tobacco-expressed SAG1 in Western blot analysis. The C3H mice were immunized with SAG-expressing leaf extracts and perorally challenged with a nonlethal dose of the T. gondii Me49 strain. Mice vaccinated with SAG1 showed significantly lower brain cyst burdens compared to those from the control group. Immunization with SAG1-expressing leaves elicited a specific humoral response with predominant participation of type IgG2a. In conclusion, a functional SAG1 version could be transiently expressed in tobacco leaves. © 2005 Humana Press Inc. All rights of any nature whatsoever reserved.
format	JOUR
author	Clemente, M. Curilovic, R. Sassone, A. Zelada, A. Angel, S.O. Mentaberry, A.N.
author_facet	Clemente, M. Curilovic, R. Sassone, A. Zelada, A. Angel, S.O. Mentaberry, A.N.
author_sort	Clemente, M.
title	Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
title_short	Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
title_full	Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
title_fullStr	Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
title_full_unstemmed	Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
title_sort	production of the main surface antigen of toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity
url	http://hdl.handle.net/20.500.12110/paper_10736085_v30_n1_p41_Clemente
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Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity

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