Identification of circulating antigens, including an immunoglobulin binding protein, from Toxoplasma gondii tissue cyst and tachyzoites in murine toxoplasmosis

Villavedra, M; Rampoldi, C; Carol, Hernán; Baz, Adriana; Battistoni, Julio; Nieto, Alberto

doi:10.1016/s0020-7519(00)00152-1

Cited by 10 publications

(8 citation statements)

References 26 publications

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“…Additionally, IgY antibodies recognized a broad range of proteins, with distinct isoelectric points determined by 2D-immunoblot. Previous studies have shown that serum samples from naturally infected humans react to a broad spectrum of T. gondii proteins [29] and experimentally infected mice produce specific antibodies against high and low molecular weight antigens of tachyzoites [30]. Consistently, our results demonstrated that IgY anti-STAg presented an antigen recognition profile similar to mouse serum, although some antigens were more immunogenic for hens than mice, probably due to the phylogenetic distance among these animals.…”

Section: Discussionsupporting

confidence: 86%

Production, Characterization and Applications for Toxoplasma gondii-Specific Polyclonal Chicken Egg Yolk Immunoglobulins

et al. 2012

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Background Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model.Methodology/Principal FindingsIn this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg) and purified egg yolk antibodies (IgY) by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers.Conclusions/SignificanceDue to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii.

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Section: Discussionsupporting

confidence: 86%

Production, Characterization and Applications for Toxoplasma gondii-Specific Polyclonal Chicken Egg Yolk Immunoglobulins

et al. 2012

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“…Bradyzoites form cysts in different tissues and persist throughout life. Both parasitic stages express specific antigens (23). During the chronic phase of the disease, bradyzoite antigens are able to maintain an efficient immune response (8).…”

Section: Discussionmentioning

confidence: 99%

Diagnosis of Congenital Toxoplasmosis by Using a Whole-Blood Gamma Interferon Release Assay

et al. 2010

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Congenital toxoplasmosis in newborns is generally subclinical, but infected infants are at risk of developing ocular lesions. Diagnosis at birth relies mainly on serological tests. Cell-mediated immunity plays the major role in resistance to infection but is not routinely investigated for diagnostic purposes. Here, we describe a simple test based on the gamma interferon (IFN-␥) response after stimulation of whole blood by crude parasitic antigens. One milliliter of heparinized blood was centrifuged; plasma was kept for routine serological tests, and pellets were resuspended in culture medium. After 24 h of culture in the presence of crude Toxoplasma gondii antigen, the cells were centrifuged and the supernatant was assayed for IFN-␥. For 62 infants under 1 year of age born to mothers who were infected during pregnancy, the sensitivity and specificity of the test were 94% (with positive results for 16 of 17 infected infants) and 98% (with negative results for 44 of 45 uninfected infants), respectively. The false-negative result was for a treated baby who gave positive results after the withdrawal of treatment. The false positive was obtained for a 3-month-old baby. For a cohort of 124 congenitally infected patients between 1 and 30 years of age, the sensitivity of the assay was 100%. We present a simple test based on IFN-␥ secretion to assess cell-mediated immunity in toxoplasmosis. As only 1 ml of blood is required to investigate humoral and cellular immunity, our assay is well adapted for the study of congenital toxoplasmosis in infants. Using purified antigens or recombinant peptides may improve the test performance.

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“…Currently, T. gondii tachyzoites are produced by serial passages in the peritoneal cavity of mice or cotton rats [1,3,35,43]. Although convenient and reliable, this technique is ethically undesirable.…”

Section: Discussionmentioning

confidence: 99%

Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells

et al. 2002

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-Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium.Neospora caninum / Toxoplasma gondii / cell culture / serum-free medium Résumé -Utilisation d'un milieu sans sérum pour la production in vitro de tachyzoites de Neospora caninum et Toxoplasma gondii sur cellules Vero. Neospora caninum et Toxoplasma gondii sont deux sporozoaires présentant un intérêt en médecine humaine et vétérinaire. La culture in vitro utilisant, entre autres, les cellules Vero comme support de la multiplication des deux parasites, est généralement employée en vue de l'obtention de tachyzoïtes. Au cours de cette étude, une évaluation quantitative et 159 Vet. Res. 33 (2002) 159-168

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Identification of circulating antigens, including an immunoglobulin binding protein, from Toxoplasma gondii tissue cyst and tachyzoites in murine toxoplasmosis

Cited by 10 publications

References 26 publications

Production, Characterization and Applications for Toxoplasma gondii-Specific Polyclonal Chicken Egg Yolk Immunoglobulins

Production, Characterization and Applications for Toxoplasma gondii-Specific Polyclonal Chicken Egg Yolk Immunoglobulins

Diagnosis of Congenital Toxoplasmosis by Using a Whole-Blood Gamma Interferon Release Assay

Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells

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