Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries

Beghetto, Elisa; Spadoni, Andrea; Buffolano, Wilma; Pezzo, Mariassunta Del; Minenkova, Olga; Pavoni, Emiliano; Pucci, Andrea; Cortese, Riccardo; Felici, Franco; Gargano, Nicola

doi:10.1016/s0020-7519(02)00256-4

Cited by 56 publications

(63 citation statements)

References 38 publications

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“…In a previous study, we identified and characterized a panel of immunodominant regions of T. gondii proteins (2,3). In this study, we investigated further the IgG avidities of specific Toxoplasma antibodies directed against four distinct antigenic regions.…”

Section: Discussionmentioning

confidence: 99%

“…Four T. gondii antigens, GRA3 (amino acids 35 to 133) (4), GRA7 (amino acids 24 to 103) (10), MIC3 (amino acids 233 to 307) (13), and SAG1 (amino acids 182 to 303) (6), were used. Recombinant antigens were expressed in bacterial cells as fusion proteins either with glutathione S-transferase (GST) or with Lambda protein D (3) and were purified by affinity chromatography as previously described (3). Briefly, recombinant Escherichia coli was induced with isopropyl-␤-D-thiogalactopyranoside (IPTG), centrifuged, and suspended in STE buffer (10 mM Tris-HCl [pH 8], 150 mM NaCl) containing 100 g of lysozyme per ml and protease inhibitor cocktail (Boehringer, Mannheim, Germany).…”

Section: Methodsmentioning

confidence: 99%

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Use of an Immunoglobulin G Avidity Assay Based on Recombinant Antigens for Diagnosis of PrimaryToxoplasma gondiiInfection during Pregnancy

Beghetto¹,

Buffolano²,

Spadoni³

et al. 2003

J Clin Microbiol

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The objective of this work was to develop an antibody-specific immunoglobulin G (IgG) avidity assay to discriminate between acute and latent phases of Toxoplasma gondii infection by using recombinant antigens. One hundred twenty-one serum samples from women who developed IgG antibodies against Toxoplasma during pregnancy were used. The IgG avidities of antibodies directed against epitopes carried by fragments of GRA3, GRA7, MIC3, and SAG1 antigens were measured by performing parallel enzyme immunoassays. The avidity index for Toxoplasma-specific antibodies against a homogeneous mixture of recombinant GRA3, GRA7, MIC3, and SAG1 antigens correlated closely with the IgG avidity of antibodies against lysed whole-cell T. gondii antigen. The avidity assay performed with the recombinant MIC3 antigen highlighted the presence of avidity low-antibodies IgG exclusively in sera collected within 2 months after primary infection. The presence of T. gondii-specific, low-avidity IgG antibodies against recombinant MIC3 antigen can be used to determine the point of infection with T. gondii within a 2-month time frame after infection.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Use of an Immunoglobulin G Avidity Assay Based on Recombinant Antigens for Diagnosis of PrimaryToxoplasma gondiiInfection during Pregnancy

Beghetto¹,

Buffolano²,

Spadoni³

et al. 2003

J Clin Microbiol

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“…Immunodominant regions encoded by GRA3 (and MIC3) genes on the human B-cell response against T. gondii infection is identified in a panel of recombinant phage clones carrying Bcell epitopes [35].…”

Section: Gra3mentioning

confidence: 99%

GRA Proteins of Toxoplasma gondii: Maintenance of Host-Parasite Interactions across the Parasitophorous Vacuolar Membrane

Nam

2009

Korean J Parasitol

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Abstract:The dense granule of Toxoplasma gondii is a secretory vesicular organelle of which the proteins participate in the modification of the parasitophorous vacuole (PV) and PV membrane for the maintenance of intracellular parasitism in almost all nucleated host cells. In this review, the archives on the research of GRA proteins are reviewed on the foci of finding GRA proteins, characterizing molecular aspects, usefulness in diagnostic antigen, and vaccine trials in addition to some functions in host-parasite interactions.

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“…Seven Toxoplasma gondii antigens were used: MIC2a, MIC2b, MIC4, M2AP, AMA 1 (5), MIC3, and SAG1 (3). Recombinant antigens were expressed in bacterial cells as fusion proteins with glutathione S-transferase (GST) and purified by affinity chromatography as previously described (3). Briefly, recombinant Escherichia coli cells were induced with IPTG (isopropylthiogalactopyranoside), centrifuged and suspended in STE buffer (10 mM TrisHCl [pH 8], 150 mM NaCl) containing 100 g/ml of lysozyme and protease inhibitor cocktail (Boehringer, Germany).…”

Section: Patientsmentioning

confidence: 99%

Use of Recombinant Antigens for Early Postnatal Diagnosis of Congenital Toxoplasmosis

Buffolano

Beghetto²,

Pezzo

et al. 2005

J Clin Microbiol

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The main objective of this work was to improve the early serologic diagnosis of toxoplasmosis in children at risk of congenital infection by using recombinant antigens. Serum samples from 104 infants born to mothers with primary Toxoplasma gondii infection acquired during pregnancy, of which 35 were congenitally infected and 22 had clinical silent toxoplasmosis at birth, were included. Immunoglobulin M (IgM), IgG, and IgG subtype antibodies against epitopes carried by fragments of T. gondii MIC2, MIC3, MIC4, M2AP, AMA1, and SAG1 gene products were measured by performing parallel enzyme immunoassays (Rec-ELISAs). Recombinant antigens preferentially reacted with IgG antibodies from infected infants compared to uninfected subjects (P < 0.0001), indicating that sera from infected children recognized a more diverse repertoire of antigens than sera transferred over the placenta from the mothers. Using two serial samples collected within 3 months of life, it was possible to demonstrate a neosynthesis of specific anti-MIC2 and anti-SAG1 immunoglobulin G, mainly of the IgG2 subtype, in 13 out of 20 infants with congenital toxoplasmosis. IgM antibodies in 97% of infected infants reacted with at least one of the recombinant antigens, confirming the diagnosis of congenital infection as soon as 2 months after birth (P < 0.0001). The use of recombinant antigens is effective in distinguishing T. gondii-infected from uninfected infants and shows that assays based on recombinant antigens improve the diagnosis of newborns with congenital toxoplasmosis.

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Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries

Cited by 56 publications

References 38 publications

Use of an Immunoglobulin G Avidity Assay Based on Recombinant Antigens for Diagnosis of PrimaryToxoplasma gondiiInfection during Pregnancy

Use of an Immunoglobulin G Avidity Assay Based on Recombinant Antigens for Diagnosis of PrimaryToxoplasma gondiiInfection during Pregnancy

GRA Proteins of Toxoplasma gondii: Maintenance of Host-Parasite Interactions across the Parasitophorous Vacuolar Membrane

Use of Recombinant Antigens for Early Postnatal Diagnosis of Congenital Toxoplasmosis

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