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Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera.
- Source :
-
Microbial pathogenesis [Microb Pathog] 2013 Jan; Vol. 54, pp. 60-6. Date of Electronic Publication: 2012 Oct 05. - Publication Year :
- 2013
-
Abstract
- Toxoplasmosis is an infection caused by the parasite Toxoplasma gondii. Chronically-infected individuals with a compromised immune system are at risk for reactivation of the disease. In-vivo induced antigen technology (IVIAT) is a promising method for the identification of antigens expressed in-vivo. The aim of the present study was to apply IVIAT to identify antigens which are expressed in-vivo during T. gondii infection using sera from individuals with chronic toxoplasmosis. Forty serum samples were pooled, pre-adsorped against three different preparations of antigens, from each in-vitro grown T. gondii and Escherichia coli XLBlue MRF', and then used to screen a T. gondii cDNA expression library. Sequencing of DNA inserts from positive clones showed eight open reading frames with high homology to T. gondii genes. Expression analysis using quantitative real-time PCR showed that SAG1-related sequence 3 (SRS3) and two hypothetical genes were up-regulated in-vivo relative to their expression levels in-vitro. These three proteins also showed high sensitivity and specificity when tested with individual serum samples. Five other proteins namely M16 domain peptidase, microneme protein, elongation factor 1-alpha, pre-mRNA-splicing factor and small nuclear ribonucleoprotein F had lower RNA expression in-vivo as compared to in-vitro. SRS3 and the two hypothetical proteins warrant further investigation into their roles in the pathogenesis of toxoplasmosis.<br /> (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
Antigens, Protozoan genetics
Chronic Disease
Escherichia coli
Gene Expression
Gene Expression Profiling
Gene Library
Humans
Mice
Open Reading Frames
Real-Time Polymerase Chain Reaction
Sequence Analysis, DNA
Toxoplasma genetics
Antibodies, Protozoan immunology
Antigens, Protozoan immunology
Toxoplasma immunology
Toxoplasmosis immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1096-1208
- Volume :
- 54
- Database :
- MEDLINE
- Journal :
- Microbial pathogenesis
- Publication Type :
- Academic Journal
- Accession number :
- 23044055
- Full Text :
- https://doi.org/10.1016/j.micpath.2012.09.006