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PubMed1984

An anti-human immunoglobulin M monoclonal antibody for detection of antibodies to Toxoplasma gondii.

An anti-human mu-chain monoclonal antibody, Tibi 82, was produced and tested for specificity by radioimmunoassay. Its reliability in detecting IgM antibodies to Toxoplasma gondii was tested by two reverse immunosorbent methods (IgM-ISAGA and IgM-SPIHA) and the IgM fluorescent antibody test (IgM-IFA) on 400 sera. Whereas the results obtained with Tibi 82 and with two polyclonal reagents were highly correlated, the third commercial polyclonal reagent provided many false negative results. By standardizing IgM binding, Tibi 82 allowed the comparison of IgM-ISAGA with IgM-SPIHA on 100 sera: 17% of the sera tested showed discrepancies due to the different toxoplasma antigens used. Although Tibi 82 facilitated the reading of results and enhanced sensitivity and specificity of the double-sandwich IgM-IFA method, the latter was still less sensitive than IgM-ISAGA with Tibi 82. Tests with the monoclonal antibody were consistently superior to tests with polyclonal antibodies.

Antibodies, Anti-Idiotypic↗
PubMed1995

Diagnosis of Toxoplasma gondii infection in AIDS patients by a tissue-culture technique.

Toxoplasma gondii infection was investigated in 14 AIDS patients with neurological involvement who showed clinical and computer tomographic scan signs suggestive of toxoplasmic encephalitis or extraneuronal localization suggestive of toxoplasmic infection. Blood, lung secretion (bronchoalveolar lavage and/or induced sputum sample) and cerebral spinal fluid (CSF) samples were cultured, and the results compared with the results of direct examination by Giemsa and by immunofluorescence of lung secretions and CSF. Toxoplasma gondii were observed directly in only four patients by immunofluorescence in bronchoalveolar lavage, induced sputum and CSF samples, and in three of these patients by Giemsa staining of bronchoalveolar lavage and CSF smears. In contrast, parasites were detected after 48 h in blood cultures from 11 of the 14 AIDS patients, in CSF cultures from eight of them and also in cultures of bronchoalveolar lavage and induced sputum from the six patients with respiratory and radiological features indicative of lung tissue damage. The findings indicate the value of tissue culture for diagnosis of toxoplasmosis as well as for monitoring the effects of treatment and also indicate that culture of induced sputum could be a powerful tool in establishing the incidence of pulmonary toxoplasmosis in AIDS patients.

AIDS-Related Opportunistic Infections↗
PubMed1981

Toxoplasma infection and abortion in dairy goats.

Serological investigation by the indirect haemagglutination test for Toxoplasma antibodies was carried out in a flock of dairy goats. Antibody titres ranging from 1:4 to 1:1,024 were found in 34% of 371 apparently healthy animals. High antibody titres of 1:256 suggestive of recent infection were noted in 18(4.9%) animals including 13 nannies. The occurrence of a number of non-brucella abortions associated with high serological titres and the isolation of the parasite from the brain tissues of an aborted foetus suggested Toxoplasma as a probable aetiologic factor. Sera having significantly levels of haemagglutinating antibodies (greater than or equal to 1:128) were also found positive to the card agglutination test. Epidemiological aspects and the public health potential of subclinical toxoplasmosis in dairy goats are discussed.

Abortion, Veterinary↗
PubMed1988

Cytotoxic activity of monocytes against Toxoplasma gondii in acute, chronic and reactivated murine toxoplasmosis.

The phagocytic activity and cytotoxicity of peripheral blood monocytes (against toxoplasma tachyzoites) was studied in acute, chronic and reactivated toxoplasma infected Swiss albino mice. During acute infection, a low phagocytic activity was observed on the 4th day post infection (dpi) (P less than 0.01) and a low monocyte cytotoxicity was noticed after the 2nd dpi (P less than 0.01) which further decreased till the 8th dpi. In contrast, both the parameters were significantly increased during chronic infection. Increase in monocytic cytotoxicity was manifested on the 3rd dpi (P less than 0.001) whereas phagocytosis showed an increase on the 12th dpi (P less than 0.05). The reactivated group showed no change in both the parameters when compared with the control immunosuppressed group (P greater than 0.05).

Animals↗
PubMedDate not supplied

Status of toxoplasma antibodies in recurrent fetal loss in U.A.E. women.

A retrospective analysis to determine the status of toxoplasma (IgG & IgM) antibodies in UAE women with recurrent fetal loss was done using immunofluorescence assay. Two thousand three hundred and fourty three patients with one or more fetal loss were studied over a period of five years. In patients with fetal loss, the range of toxoplasma IgG seropositivity varied from 24.2-30.6%. There were 3 patients with IgM positive. Only in a single patient one of her two abortions could be attributed to acute toxoplasmosis. Habitual fetal loss cannot be attributed to chronic toxoplasmosis. Also, 67.2% of the women of child bearing age group in U.A.E were found to be seronegative, highlighting the need for routine antenatal screening to detect primary acute toxoplasmosis.

Abortion, Habitual↗
PubMed2005

Polymerase chain reaction for detection of Toxoplasma gondii in human biological samples.

Using the polymerase chain reaction (PCR), Toxoplasma gondii from gene TGR1E with primers TGR1E-1, TGR1E-2 (standard PCR), and from B1 gene with primers TM1, TM2, TM3 (hemi-nested PCR) was detected in biological samples from 347 individuals (441 biological materials). Of the total of 441 biological materials, T. gondii DNA was detected in 5.2 %; it was positive in the following samples: blood (n = 6), blood from newborns (2), biopsies (2) and samples of progenitor cells (2) (from candidates for bone marrow transplantation). DNA of T. gondii was also revealed in 11 samples (8.3 %) of 120 cases of pregnant women during prenatal examinations. A positive result in the blood was also found in two cases of newborn babies from mothers who were infected in later pregnancy. The positive PCR examination was confirmed by serological methods (ELISA and complement fixation test). Agreement of PCR results and the detection of antibodies against toxoplasma was found in 83.3 %. Rapid PCR examination for the confirmation of acute parasitemia T. gondii is particularly important for the patients in whom the infection may cause serious consequences (e.g., for fetus in pregnant women or for patients suffering from imunosuppression).

Adult↗
PubMed2000

Biochemical characterization of mitogen-activated protein (MAP) kinase activity in Toxoplasma gondii.

Mitogen-activated protein (MAP) kinase or extracellular signal-regulated kinase (ERK) are activated by many extracellular stimuli. In this study, we investigated whether MAP kinase and tyrosine kinases were involved in transducing signals in Toxoplasma gondii. Using anti-phosphotyrosine and anti-active ERK antibodies, we identified several phosphorylated proteins in Toxoplasma. In particular, phosphorylation of a 47 kDa and a 43 kDa protein increased strongly after calcium influx. MAP kinase activity, caused by calcium influx, was determined using either a specific synthetic peptide, or an in gel kinase assay. Conversely, calcium chelators (BAPTA and EGTA) and a calcium channel blocker (nifedipine) inhibited this activation. Also, a specific inhibitor of MAP kinase kinase (PD 098059) blocked MAP kinase activity. Three specific anti-MAP kinase antibodies recognized the 47 kDa and 43 kDa proteins, which were putatively identified as ERK1- and ERK2-homologs, respectively. These findings provide early evidence of signal transduction involving members of the MAP kinase family in T. gondii.

Animals↗
PubMed2002

Mast cells in the eyes of Calomys callosus(Rodentia: Cricetidae) infected by Toxoplasma gondii.

The mast cell is a powerful effector cell for the innate immune system, acting through the secretion of several distinct mediators. Few studies have demonstrated the relationship between mast cells and toxoplasmosis. In this study, mast cells were investigated in two experimental Toxoplasma infections using Calomys callosus (Rodentia: Cricetidae) as the host. Animals were inoculated either intraperitoneally or via the conjunctiva with tachyzoites of Toxoplasma gondii (RH strain) and sacrificed after 5 days or 24 h, respectively. Enucleated eyes were processed for histological and ultrastructural analysis. Neither experimental infection altered the localization of mast cells compared to control eyes, but they did lead to an accumulation in some tissues as well as to their activation. There was a significant increase in the number of mast cells within 5 days and 24 h after infection. The ocular lesions were characterized by the presence of tachyzoites, inflammatory cells and vasodilatation in the iris and retina. In conclusion, mast cells were mobilized in these experimental infections, suggesting that they play an important role in the host inflammatory response after infection with T. gondii.

Animals↗
PubMed2002

Regulation of tumor necrosis factor alpha and its specific receptors during Toxoplasma gondii infection in human monocytic cells.

The coccidian Toxoplasma gondiiis an obligate intracellular parasite which can infect all cell types. Among the cytokines elicited by the immune response to Toxoplasma, tumor necrosis factor alpha (TNF-alpha) acts synergistically with gamma interferon (IFN-gamma) and plays a major role in host cell resistance. We have previously reported that TNF-alpha production induced by IFN-gamma/LPS decreases after T. gondii infection of human myelomonocytic THP-1 cells. Here, we investigated the regulation of TNF-alpha and its specific receptors during T. gondii infection of THP-1 cells. We found that TNF-alpha production was regulated at a post-transcriptional level and that TNF receptor expression was regulated at a pretranscriptional level. The TNF receptor I shedding and the fall in TNF-alpha levels observed after T. gondii infection would thus be induced by a parasite component with serine protease activity. These findings indicate that T. gondii participates not only in controlling the cytotoxic effects of TNF-alpha during the infection process, but also in signal transduction mediated mainly by TNF receptor I.

Animals↗
PubMed2002

Fetal Toxoplasma infection after oocyst inoculation of pregnant rats.

Six groups totalling 53 Wistar rats were fed 10(4)oocysts from one of six different Toxoplasma strains at 15 days of pregnancy. The overall transplacental transmission rate was 51%. This varied between 10% and 80%, dependent on the strain used. The strains of Toxoplasma which are more pathogenic for mice were transmitted transplacentally more frequently than the strains of intermediate or low pathogenicity. There were no statistically significant differences in the rate of congenital transmission of Toxoplasmain rats fed oocysts (present work) or cysts (previous work).

Animals↗
PubMed2003

Immunodiagnosis of primary Toxoplasma gondii infection in sheep by the use of a P30 IgG avidity ELISA.

Toxoplasma gondii is a protozoan parasite with a worldwide distribution. In both sheep and humans, if the parasite is encountered during pregnancy, fetal infection and abortion can occur. Therefore, Toxoplasma infection in sheep has a major economic impact upon sheep farming. Clinically, there is a need to distinguish recent (acute) infections from longstanding (chronic) infections. However, current serological techniques, such as detection of anti-T. gondii IgG, cannot discriminate between acute and chronic infections. Increasing immunoglobulin avidity is a good determining factor of how recent an infection is. In this study, we describe the application and validation of a T. gondii IgG avidity ELISA, based on the use of an affinity-purified, native T. gondii P30 antigen. The assay was used to examine sera from eight sheep experimentally infected with T. gondii and found that all seroconverted within 21 days post-infection (p.i.), beginning with avidities that were initially low but that increased over time, with all sheep reaching high IgG avidity within 10 weeks p.i. In addition, sera from clinically healthy but T. gondii-seropositive lambs and ewes and seropositive ewes with a history of abortion were also subjected to a preliminary serological investigation. High IgG avidities were found in 80% of the seropositive lambs, in 90% of the clinically healthy ewes and in 97% of the ewes with abortion problems. These findings indicate that the animals had most likely contacted the parasite a longer time ago.

Abortion, Veterinary↗
PubMed2005

Identification of proteins interacting with Toxoplasma SRCAP by yeast two-hybrid screening.

Toxoplasma gondii is an opportunistic protozoan parasite that differentiates into latent cysts (bradyzoite) that can be reactivated during immunosuppression. TgSRCAP (Toxoplasma gondii Snf2-related CBP activator protein) is a SWI2/SNF2 family chromatin remodeler whose expression increases during cyst development. Identifying the proteins associating with TgSRCAP during the pre-cyst stage (tachyzoite) will increase our understanding of how parasite differentiation is initiated. We employed the yeast two-hybrid system to identify proteins that may interact directly with TgSRCAP. A stretch of 1,060 amino acids between ATPase subdomains IV and V of TgSRCAP was chosen as "bait" since the corresponding region in human SRCAP interacts with other proteins, including CREB binding protein. We have identified several novel parasite-specific transcription factors predicted to be in the T. gondii genome. Metabolic enzymes that may participate in cyst development were also identified as interacting with TgSRCAP.

Animals↗
PubMed2006

Usefulness of Toxoplasma gondii recombinant antigens (GRA1, GRA7 and SAG1) in an immunoglobulin G avidity test for the serodiagnosis of toxoplasmosis.

The precise diagnosis of an acute and recent Toxoplasma infection in pregnant women and the newborn child is important before treatment. This study describes a new Toxoplasma gondii IgG avidity test based on a combination of recombinant GRA1, GRA7 and SAG1 antigens and shows that this test is useful for diagnostic purposes and may replace the lysed, whole-cell antigens. Although more sera need to be tested, the results obtained here suggest that the IgG avidity test performed with rec-antigens correlated more with the stage of a T. gondii infection than the IgG avidity results obtained with the lysed, whole-cell antigen test, the VIDAS Toxo IgG avidity (bioMérieux).

Animals↗
PubMed2000

The taxonomic importance of obligate heteroxeny: distinction of Hammondia hammondi from Toxoplasma gondii--another opinion.

We enumerate identical and divergent findings concerning the obligate heteroxenous Hammondia hammondi and the facultatively homoxenous or heteroxenous Toxoplasma gondii. Differences exist in life-cycles, transmission, and host range, especially transmissibility to birds and mammals other than rodents, in ultrastructural morphology, immunity and serology in cats and to lesser degree in rodents, in DNA sequences and in isoenzymes. Because the recognition of obligate heteroxeny is essential to study these organisms and to recognize them as taxa, it is advantageous to give heteroxeny a generic rather than a specific value. Characterization of organisms with the life-cycle patterns of Hammondia, Sarcocystis, Frenkelia, and Toxoplasma is best achieved by means of the genera presently used.

Animals↗
PubMed2001

Towards the Toxoplasma gondii proteome: position of 13 parasite excretory antigens on a standardized map of two-dimensionally separated tachyzoite proteins.

High resolution two-dimensional separation of Toxoplasma gondii tachyzoite lysate revealed up to 224 distinct protein spots in Coomassie-stained gel. Computional matching of 14 digitized gels yielded a standard two-dimensional proteome map. The excretory T. gondii dense granule proteins GRA1-GRA8, S16/acid phosphatase, nucleoside triphosphate hydrolase, and H4 were identified by Western blotting of both total gel and isolated protein spots. In addition, two excretory antigens defined by parasite-specific monoclonal T cells, p36 and p40, were mapped by a novel T-cell blotting technique based on electroeluting single protein spots and testing the eluates for antigenic activity against the T-cell clones. In summary, these results represent a first step in Toxoplasma proteome analysis.

Animals↗
PubMed2000

Neospora caninum: is it really different from Hammondia heydorni or is it a strain of Toxoplasma gondii? An opinion.

The published data concerning Toxoplasma gondii, Hammondia hammondi, H. heydorni and Neospora caninum on one side and between T. gondii on the other were neglected by most authors. As conclusion we are convinced that there are only two valid species: Isospora (Toxoplasma) gondii and Hammondia heydorni. The first includes as a strain H. hammondi and the latter N. caninum. In any case there is absolutely no reason (with respect to general Zoological nomenclature) to create new genera!

Animals↗
PubMed1996

Toxoplasma gondii strain-specific transcript levels of SAG1 and their association with virulence.

Toxoplasma gondii isolates can be classified into mouse-virulent and -avirulent strains. Since the major surface antigen of Toxoplasma gondii, SAG1, has been shown to be important for the invasion process and might thus be involved in mouse virulence as well, we analyzed the corresponding gene of mouse-virulent and -avirulent strains. In addition to eight mismatches, mouse-virulent strains harbored five copies of an up-stream 27-bp repeat in the promotor region of SAG1 as compared with four copies in avirulent strains. Reverse-transcriptase polymerase chain reaction revealed that SAG1 expression levels in the mouse-virulent T. gondii strains were at least 4-fold those in the avirulent strains. Since SAG1 seems to mediate invasion, it is suggested that the possibly higher steady-state expression of SAG1 in mouse-virulent strains of T. gondii is associated with virulence and facilitates faster invasion by these strains in comparison with avirulent strains.

Animals↗
PubMed1997

Toxoplasma gondii virulence markers identified by random amplified polymorphic DNA polymerase chain reaction.

Genomic DNAs from 35 Toxoplasma gondii strains were amplified by random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) using 18 arbitrary 10-mer primers. At least four primers were found to generate DNA fragments that discriminate the 35 T. gondii strains into a genotype of virulent strains and a genotype of avirulent strains. Primer B12 was found to generate a virulence-specific fragment and primers B5, C8, and C20 were found to generate avirulence-specific fragments, which in all cases clearly identified either the virulence phenotype or the avirulence phenotype, respectively. In addition, the DNA polymorphic bands detected were analyzed by parsimony and distance analysis. A similar genetic relationship among the T. gondii strains was determined by the two phylogenetic methods, which use completely different assumptions. Consistent with the division of the 35 strains into a genotype of virulent strains and a genotype of avirulent strains, both analyses revealed 2 clonal lineages directly correlated with murine virulence. These results strongly support the hypothesis that the genus Toxoplasma may actually contain two clonal lineages correlated with virulence, which have evolved independently following their initial separation.

Animals↗