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Fc receptors on the surface of Toxoplasma gondii trophozoites: a confounding factor in testing for anti-Toxoplasma antibodies by indirect immunofluorescence.

Negative sera often produce false-positive polar patterns of fluorescence in indirect immunofluorescence tests for serum anti-Toxoplasma gondii antibodies, representing a confounding factor in the diagnosis of toxoplasmosis. In this work, we studied whether T. gondii trophozoites, the antigenic material used in the immunofluorescence tests, expressed surface Fc receptors that could cause the binding of normal immunoglobulins, thus producing false-positive results. We report here that T. gondii trophozoites indeed have Fc receptors on their surface. This was shown by the direct binding of purified human Fc to the parasite, evidenced by the subsequent binding of fluorescein-labeled Fab specific for human Fc. In addition, pretreatment of the parasite with excess purified Fc to saturate the surface Fc-binding sites abrogated the formation of polar fluorescence. The trophozoites appeared to express Fc receptors with different degrees of affinity for the specific ligand, since a diffuse fluorescence pattern was observed following incubation with 1 mg of Fc per ml (10 micrograms per well), whereas with 0.2 mg (0.2 micrograms per well), a majority of parasites showed polar fluorescence. This observation suggests that the Fc receptors accumulated at the polar cap are those with higher affinity. The present findings raise intriguing questions regarding the possible biological role(s) of the Fc receptors on T. gondii but, more immediately, indicated that pretreatment of the antigenic material with Fc, a commercially available reagent, constitutes a practical, simple way to avoid false-positive immunofluorescence test results due to the binding of nonspecific immunoglobulin to the parasite.

Animals↗

[Detection of toxoplasma-specific IgA and IgM antibodies in serum samples of adults with acquired toxoplasma infection].

In a study of persons with and without toxoplasmosis, IgA-ISAGA (immunoglobulin A-immunosorbent agglutination assay) was found to be specific. However, serum samples from subjects recently infected wtih toxoplasma were less reliable than the DNR test (IgM-ISAGA by the original method, 1981) and the IgM-IIFT. On the other hand, some serum samples from persons infected for up to a year or longer were IgA-positive. The IgA-ISAGA should therefore not be included in serodiagnosis in pregnancy as a substitute for IgM-ISAGA or IgM-IIFT, but, if at all, in addition to them.

Adult↗

A family of glycolipids from Toxoplasma gondii. Identification of candidate glycolipid precursor(s) for Toxoplasma gondii glycosylphosphatidylinositol membrane anchors.

Four major glycolipids were extracted from Toxoplasma gondii tachyzoites which were metabolically labeled with tritiated glucosamine, mannose, palmitic and myristic acid, ethanolamine, and inositol. Judging from their sensitivity to a set of enzymatic and chemical tests, these glycolipids share the following properties with the glycolipid moiety of the glycosylphosphatidylinositol anchor (GPI anchor) of the major surface protein, P30, of T. gondii: 1) a nonacetylated glucosamine-inositol phosphate linkage; 2) sensitivity toward phosphatidylinositol-specific phospholipase C and nitrous acid; 3) identity of HF-dephosphorylated GPI glycan backbone between three glycolipids and the HF-dephosphorylated core glycan of the GPI anchor of the major surface protein P30; 4) the presence of a linear core glycan structure blocked by an ethanolamine phosphate residue(s). Taken together with the nature of radiolabeled precursors incorporated into these glycolipids, the data indicate that these GPIs are involved in the biosynthesis of the GPI-membrane anchors of T. gondii.

Animals↗

Subcutaneous and intestinal vaccination with tachyzoites of Toxoplasma gondii and acquisition of immunity to peroral and congenital toxoplasma challenge.

Mice were immunized s.c. or intraintestinally with two injections of a temperature-sensitive mutant of Toxoplasma gondii (ts4). Nonpersistence of the vaccine strain was documented by subinoculation of tissues of a subgroup of mice 3 mo or more after the second immunization. Mice were immune to other-wise lethal parenteral challenges with tachyzoites of the M7741 strain or to peroral challenge with bradyzoites of the Me49 strain of T. gondii. Although two s.c. or intraintestinal immunizations did not completely protect against development of T. gondii in the brains of mice, fewer cysts developed in the s.c. immunized mice than in control mice (2 +/- 3 cysts/0.01 ml in immunized mice compared with 75 +/- 48 cysts/0.01 ml in controls (p less than 0.002)). Reduction in cyst number after intraintestinal immunization was more variable, but also statistically significant (p less than 0.02). Female mice were first immunized, then mated, and then challenged perorally. Neonates of the s.c. immunized mice were not protected. Neonates of intraintestinally immunized mice were protected in part (36% of 115) against congenital infection compared with controls (7% of 107).

Administration, Oral↗

[The effect of toxoplasma antibodies after reinfection with T. gondii. II. Communication: investigations over the incidence of toxoplasma in peripheral blood after primary and secondary infection (author's transl)].

After reinfection, parasites could be detected through transfusion of tail blood to healthy mice between 24 and 100 to 352 hours. A time point beyond this was not examined. Parasitaemia could be frequently established between 68 and 76 hours post reinfection. Infection immunity in mice does not protect against reinfection by Toxoplasma. It occurs after oral infection inspite of the presence of humoral antibodies. The parasites spread by blood circulation and increase the titer by one or two titer steps in the SABIN FELDMAN test.

Animals↗

[Studies on the therapeutics of experimental toxoplasmosis. I. Effect of acetylspiramycin and spiramycin on the intracardiac Toxoplasma multiplication in mice infected with Toxoplasma gondii (author's transl)].

A total of 329 human serum samples were collected from the clinics of internal medicine of Obihiro city and its suburbs and were screened for Toxoplasma antibody level using the latex agglutination microtiter method (LA). Ninety-one (27.7%) were positive, 4 (1.2%) were equivocal and 234 (71.1%) were negative. Positive reactants were highest in the age group of 55-74 years old. A relatively high percentage on positive reactants was observed in patients with accompanying cardiac and cerebral disorders, i.e., 33.3% and 35.6%, respectively. ICR-JCL mice were infected with 10(2) tachyzoites of the S-273 strain of T. gondii intraperitoneally. Chronic cases were obtained from those mice which survived the first infection without any further challenge 1 month later. Therapy was carried out by administering acetylspiramycin or spiramycin per os at a dose of 8 mg/0.5 ml per mouse per day for 30 days. After the end of therapy, brain and heart emulsions were made, and passaged to clean mice and observed for 30 days. Antibody titers were also monitored during the course of treatment using the LA microtiter method. In the acute cases, antibody titers were observed to gradually increase from the end of treatment up to 10 weeks. Antibody titers of the chronic cases were maintained at high levels before the start of treatment and up to its termination. Most of the mice subinoculated with brain emulsions obtained from the acute cases, both in the control and treatment groups, died. However, all of the mice subinoculated with heart emulsions from both treatment groups, survived. In the chronic cases, all mice subinoculated with brain emulsions from all groups died. A few survived the heart passages from both treatment groups, however, they showed very high reactions to the LA microtiter method.

Adolescent↗

Neonatal serologic screening and early treatment for congenital Toxoplasma gondii infection. The New England Regional Toxoplasma Working Group.

BACKGROUND: Most infants with congenital Toxoplasma gondii infection have no symptoms at birth, but many will have retinal disease or neurologic abnormalities later in life. Early detection and treatment of congenital toxoplasmosis may reduce these sequelae. METHODS: In Massachusetts since January 1986, and in New Hampshire since July 1988, newborns have been screened for intrauterine infection with T. gondii by means of an IgM capture immunoassay of blood specimens routinely collected for screening for metabolic disorders. Congenital infection is confirmed by assays for specific IgG and IgM antibodies in serum from infants and their mothers. For this study, infants with serologic evidence of infection underwent extensive clinical evaluation and received one year of treatment. RESULTS: Through June 1992, 100 of 635,000 infants tested had positive screening tests. Congenital infection was confirmed in 52 infants, 50 of whom were identified only through neonatal screening and not through initial clinical examination. However, after the serologic results became available, more detailed examinations revealed abnormalities of either the central nervous system or the retina in 19 of 48 infants evaluated (40 percent). After treatment, only 1 of 46 children had a neurologic deficit (hemiplegia attributable to a cerebral lesion present at birth). Thirty-nine treated children had follow-up ophthalmologic examinations when one to six years old; four (10 percent) had eye lesions that may have developed postnatally (a macular lesion in one child and minor retinal scars in three). CONCLUSIONS: Routine neonatal screening for toxoplasmosis identifies congenital infections that are subclinical, and early treatment may reduce the severe long-term sequelae.

Antibodies, Protozoan↗

Toxoplasma gondii Hsp70 as a danger signal in toxoplasma gondii-infected mice.

Toxoplasma gondii Hsp70, T gondii Hsp30/bag1, and surface antigen 1 messenger RNAs were shown to be useful in analyzing stage conversion of T gondii between bradyzoites and tachyzoites. The high-level expression of T gondii Hsp70 was correlated with mortality in interferon-gamma knockout mice infected with T gondii. Tgondii Hsp70 inhibited the induction of nitric oxide release by peritoneal macrophages of T gondii-infected mice. These findings identify T gondii Hsp70 as a danger signal during lethal, acute T gondii infection.

Animals↗

High levels of IgM antibodies specific for Toxoplasma gondii in pregnancy 12 years after primary toxoplasma infection. Case report.

A case of high levels of specific IgM antibodies registered by the immunosorbent agglutination assay (ISAgA) in a pregnant woman with a history of toxoplasmosis is reported. The patient had acute lymphoglandular toxoplasmosis diagnosed serologically by increases in the specific antibody titer detected by the Sabin-Feldman test (SFT) and pathohistologically 12 years before pregnancy. In pregnancy, she had stable titers of specific antibodies registered by the SFT, enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody test (IFAT), indirect hemagglutination test (IHAT) and direct agglutination test. Specific IgM antibodies were detected by the ISAgA but not by the IgM-IFAT, IgM-ELISA and IgM-IHAT. She had a normal pregnancy and gave birth to a clinically healthy baby who had a negative ISAgA finding at 7 days of age. This case indicates that ISAgA is not necessarily a marker of recent infection and is therefore not reliable for the diagnosis of toxoplasmosis in pregnancy.

Adult↗

Evaluation of intrathecal synthesis of specific IgG antibodies against Toxoplasma gondii in the diagnosis assessment of presumptive toxoplasma encephalitis in AIDS patients.

The diagnosis of neurotoxoplasmosis in patients with acquired immunodeficiency syndrome is mainly based on tomographic or magnetic resonance findings and on the response to specific treatment. We studied 55 patients with AIDS and neurotoxoplasmosis according to these diagnostic criteria (group 1), 37 patients with AIDS and neurological involvement of other etiology (group 2), and 16 anti-HIV-negative individuals with neurological manifestations (group 3). Serum and cerebrospinal fluid were examined for the presence of anti-T. gondii IgG, by indirect immunofluorescence. In 72 of them, the total amounts of these antibodies were determined in order to assess local production of anti-T. gondii antibodies in the central nervous system and to correlate their titers with infection activity in patients with AIDS and neurotoxoplasmosis. IgG titers > or = 1/64 in cerebrospinal fluid reached 100% specificity for the diagnosis of neurotoxoplasmosis in AIDS. Evidence of local synthesis of these antibodies was detected in 42.8% of patients of group 1, in 29.1% of patients of group 2 and in no patient of group 3. The test showed 70.8% specificity and therefore was not useful in our study for the differential diagnosis of neurotoxoplasmosis in patients with AIDS.

AIDS-Related Opportunistic Infections↗