Subcellular localization and quantitative analysis of <i>Toxoplasma gondii</i> target antigens of specific immunoglobulins G, M, A, and E

Kumolosasi, Endang; Bonhomme, A.; Lepan, H.; Marx, C; Foudrinier, F.; Pinon, J. M.; Beorchia, A.; Pluot, M

doi:10.1002/jemt.1070290309

Cited by 5 publications

(4 citation statements)

References 22 publications

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“…Here, we employed a tachyzoite lysate enriched in membrane fractions, including the apical complex. The latter is associated with active motility during parasite invasion and is a strong immunogen for IgM (32). Interestingly, the presently generated IgM SIA based on this antigen showed 100% agreement with the Vidas IgM test employing the tachyzoite lysate.…”

Section: Discussionmentioning

confidence: 76%

Microsphere-Based IgM and IgG Avidity Assays for Human Parvovirus B19, Human Cytomegalovirus, and Toxoplasma gondii

Wang

Hedman

Nurmi

et al. 2020

mSphere

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Human parvovirus B19 (here B19), human cytomegalovirus (HCMV), and Toxoplasma gondii infections during pregnancy can lead to severe complications. While traditional diagnosis of infections is mostly confined to one pathogen at a time, a multiplex array is a feasible alternative to improve diagnostic management and cost-efficiency. In the present study, for these three pathogens, we developed microsphere-based suspension immunoassays (SIAs) in multiplex and monoplex formats for the detection of antimicrobial IgM antibodies as well as corresponding chaotrope-based IgG avidity SIAs. We determined the diagnostic performances of the SIAs versus in-house and commercial reference assays using a panel of 318 serum samples from well-characterized clinical cohorts. All the newly developed assays exhibited excellent performance compared to the corresponding high-quality reference methods. The positive and negative percent agreements of the IgM SIAs in comparison with reference methods were 95 to 100% and 98 to 100%, and those of the IgG avidity SIAs were 92 to 100% and 95 to 100%, respectively. Kappa efficiency values between the SIAs and the corresponding reference assays were 0.91 to 1. Furthermore, with another panel comprising 391 clinical samples from individuals with primary infection by B19, HCMV, or T. gondii, the IgM SIAs were highly sensitive for the detection of acute infections, and the IgG avidity SIAs were highly specific for the separation of primary infections from past immunity. Altogether, the strategy of IgM multiplex screening followed by IgG avidity reflex testing can provide high-throughput and accurate means for the detection and stage determination of B19, HCMV, and T. gondii infections. IMPORTANCE Human parvovirus B19, human cytomegalovirus, and Toxoplasma gondii are ubiquitous pathogens. Their infections are often asymptomatic or mild in the general population yet may be transmitted from mother to fetus during pregnancy. Maternal infections by these pathogens can cause severe complications to the fetus or congenital abnormalities. As a rule, the risk of maternal transmission is critically related to the infection time; hence, it is important to determine when a pregnant woman has acquired the infection. In this study, we developed new diagnostic approaches for the timing of infections by three pathogens. All the new assays appeared to be highly sensitive and specific, providing powerful tools for medical diagnosis.

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Section: Discussionmentioning

confidence: 76%

Microsphere-Based IgM and IgG Avidity Assays for Human Parvovirus B19, Human Cytomegalovirus, and Toxoplasma gondii

Wang

Hedman

Nurmi

et al. 2020

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“…GRA proteins are secreted into the tachyzoite parasitophorous vacuole after invasion of the host cell and are also found in the parasitophorous vacuole membrane and in the bradyzoite cyst wall. Dense granule antigens are recognized by an important fraction of the specific G, M, and A immunoglobulins in the serum of patients suffering from acute toxoplasmosis (31).…”

Section: Resultsmentioning

confidence: 99%

Selection and Identification of Dense Granule Antigen GRA3 by Toxoplasma gondii Whole Genome Phage Display

Robben

Hertveldt

Bosmans

et al. 2002

Journal of Biological Chemistry

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Toxoplasma gondii is a ubiquitous, unicellular, eukaryotic parasite with a complex intracellular life cycle capable of invading and chronically infecting a wide variety of vertebrate host species, including man. Although normally opportunistic in healthy adults, it is a lethal pathogen in immunocompromised humans, particularly in AIDS patients. We present the application of a genomic phage display as a tool for the direct identification of antigens with potential value in diagnosis and/or as subunit vaccine components. Using a polycosmid cloning strategy, we constructed a large phagemid display library (>10 9 independent clones) of mixed short genomic restriction fragments (< 500 bp) of T. gondii genomic DNA (80 Mbp genome size) fused to gene III of the filamentous phage M13. Biopanning of the library with monoclonal Toxoplasma antibodies resulted in the isolation and identification of an epitope of GRA3, an antigen located in the dense granules of T. gondii tachyzoites. The reactivity of the phage displaying the GRA3 epitope with the monoclonal antibody was confirmed by an enzyme-linked immunosorbent assay. These results demonstrate the accessibility of midsized eukaryotic genomes to display technology and the feasibility to screen these whole genome display libraries with antibodies for isolating novel antigenic determinants.

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“…The conditions for sample preparation were determined so as to preserve the ultrastructure, to identify the labeled organelles, and to retain immunoreactivity (Kumolosasi et al 1994). Each IgA fraction was tested at increasing concentrations up to an optimal immunolabeling response, i.e., when the ratio of total signal to nonspecific signal was at its maximum.…”

Section: Quantitative Immunolabeling Analysis Of Iga Fractionsmentioning

confidence: 99%

“…As a reference, ten isolated markers were used and the mean area of a marker was determined. In the case of coalescent particles, the count was estimated from the ratio of the area occupied by these particles and the area occupied by the reference marker (Boulanger et al 1991;Kumolosasi et al 1994). The quantitation was expressed as the number of gold particles counted per square micrometer of area.…”

Section: Quantitative Immunolabeling Analysis Of Iga Fractionsmentioning

confidence: 99%

Kinetics study of the localization and quantitation of target antigens of immunoglobulin a antibodies in acquired and congenital toxoplasmosis

et al. 1996

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The cellular distribution (localization and quantitation) of the target parasite's antigens in the tachyzoite along the IgA kinetics was determined in the course of acquired toxoplasmosis and congenital toxoplasmosis. In the case of acquired toxoplasmosis, throughout the IgA kinetics a correlation was noted between the membrane and submembrane immunolabeling and the results of the immunocapture and enzyme-linked immunosorbent assay IgA (ELISA-A) tests. The rhoptries' immunolabeling remained higher. The immunolabeling evolution and the results of the immunology tests were not closely related to the treatment (Rovamycin). From the congenital toxoplasmosis cases it was observed that membrane immunolabeling correlated with the results of the serology tests and with the treatment (Fansidar). The rhoptry antigens were recognized throughout the IgA kinetics; even when the serology tests became negative, immunolabeling persisted. Rhoptries appeared as secretory organelles of antigens recognized during acute, chronic, and congenital stages of Toxoplasma infection.

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Subcellular localization and quantitative analysis of Toxoplasma gondii target antigens of specific immunoglobulins G, M, A, and E

Cited by 5 publications

References 22 publications

Microsphere-Based IgM and IgG Avidity Assays for Human Parvovirus B19, Human Cytomegalovirus, and Toxoplasma gondii

Microsphere-Based IgM and IgG Avidity Assays for Human Parvovirus B19, Human Cytomegalovirus, and Toxoplasma gondii

Selection and Identification of Dense Granule Antigen GRA3 by Toxoplasma gondii Whole Genome Phage Display

Kinetics study of the localization and quantitation of target antigens of immunoglobulin a antibodies in acquired and congenital toxoplasmosis

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