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[Seroprevalence of cytomegalovirus and Toxoplasma gondii in healthy subjects under 30 years old in Santiago, Chile].

BACKGROUND: Infections by Cytomegalovirus and Toxoplasma gondii are endemic in Chile and only a low proportion of infected individuals have clinical manifestations. AIM: To study the prevalence of infection by Cytomegalovirus and Toxoplasma gondii in Chile. SUBJECTS AND METHODS: The prevalence of IgG antibodies against Cytomegalovirus and Toxoplasma gondii were studied in 560 subjects under 30 years old, using an ELISA technique. Age, socioeconomic level, breast feeding, assistance to nurseries and number of family members were considered as risk factors for these infections. RESULTS: Infection by Cytomegalovirus had a global prevalence of 60%. It showed an epidemiological pattern of late acquisition in high socioeconomic levels and a pattern of early infection in medium and low socioeconomic levels. Eighty to 90% of sera were positive for the infection in adult subjects of the three socioeconomic levels. There was a positive correlation between the duration of breast feeding and the frequency of Cytomegalovirus infection. Infection by Toxoplasma gondii had a global prevalence of 24.6%. The rates of susceptible individuals were 80 and 50% in high and medium-low socioeconomic levels respectively. CONCLUSIONS: The knowledge about the frequency of these infections in high risk populations such as women during their reproductive years and immunodepressed individuals, will allow the implementation of preventive measures.

Adolescent↗

Prevalence of antibody to Toxoplasma gondii in Hyogo Prefecture, Japan: comparison at a 10-year interval.

We have previously surveyed antibody to Toxoplasma gondii among Japanese populations. In this report, we studied the current situation of antibody prevalence and compared it with the situation approximately 10 years before. A total of 1,800 human serum samples were collected at the Central Laboratory of Kobe University Hospital from June through September, 1995, and were examined for Toxoplasma antibody using enzyme-linked immunosorbent assay. The overall prevalence was found to be 16.4% with similar seropositivities in males (16.8%) and females (15.9%). Prevalence of antibody increased with age, reaching 41.0% in males and 59.0% in females at 80 years or more. Comparison with data obtained at the same hospital in 1981 to 1982 indicated that seropositivities at 30-39 years significantly decreased in both males and females. A positivity currently obtained at a certain age group was similar to a positivity previously obtained at the corresponding age group (10 years younger), indicating that the previous age prevalence curve shifted to the current position. A serological survey among stray cats collected in Hyogo Prefecture in 1990-1991 indicated that 44 (19.0%) of 231 cats were positive for Toxoplasma antibody as determined by the latex agglutination test. These results suggested less transmission of Toxoplasma to humans in Hyogo Prefecture at least for the last 10 years comparing with the situation in the past, although the transmission cycle among cats was still active in nature.

Adolescent↗

Toxoplasma serology in Zambian and Ugandan patients infected with the human immunodeficiency virus.

In the USA and Europe, toxoplasmosis is well recognized as an important cause of morbidity and mortality among immunocompromised individuals. Toxoplasma gondii has been shown to be a common opportunistic infection in patients infected with the human immunodeficiency virus (HIV) in the USA and Europe with published estimates ranging from 20% to 80%. The importance of Toxoplasma infection in East Africa has not yet been defined. The seroprevalence rates of toxoplasmosis in Zambian and Ugandan patients were determined using the dye test (DT) and the latex agglutination test (LAT). The geographical variation in seroprevalence rates noted in western countries was also found in these African countries, with Zambia showing significantly lower rates than Uganda. 34% of Ugandan (64/186) and 4% of Zambian (8/187) patients infected with HIV, compared with 27% of Ugandan (26/93) and 11% of Zambian (20/189) HIV-negative persons, had anti-Toxoplasma immunoglobulin G antibodies. With the LAT, 13% of the Ugandan and 7% of the Zambian sera gave a false positive result. The relevance of Toxoplasma serology in Africa is discussed.

Antibodies, Protozoan↗

Development of genetic systems for Toxoplasma gondii.

The protozoan parasite Toxoplasma gondii has recently emerged as an important opportunistic pathogen in humans. Toxoplasma also shares a number of biological features with Plasmodium and Eimeria, which are important pathogens of humans and animals. Because o f the ease o f experimental use, David Sibley, Elmer Pfefferkom and John Boothroyd have undertaken the development of genetics in Toxoplasma as a model intracellular parasite. Toxoplasma is presently the only parasitic protozoan where both classical and molecular genetics are feasible. The recent advances in this system are highlighted here, along with potential applications of genetics for understanding intracellular parasitism.

Journal Article↗

Comparison of four different immunoassays for detection of Toxoplasma-specific immunoglobulin G.

Serum specimens from 246 women of childbearing age were tested for immunoglobulin G (IgG) antibodies to Toxoplasma gondii by four different commercial assays: Abbott IMx microparticle enzyme immunoassay (EIA), Mercia Toxo-G EIA, Bartels Prima EIA, and bioMerieux Vitek (VIDAS) enzyme-linked fluorescent assay. A total of 27 specimens were initially positive with all four assays, 202 specimens were negative, and 17 specimens were discrepant (disagreement among the assays). After repeating tests for the 17 discrepant samples, five resolved (one was positive and four were negative). The 12 remaining discrepant samples were tested by an indirect fluorescent antibody (IFA) assay for Toxoplasma IgG antibodies; five specimens were positive for Toxoplasma IgG by IFA. The resolved sensitivities of the various kits ranged from 88% (bioMerieux VIDAS) to 94% (Abbott IMx), and the specificities were all 98%-99%. These results show that the four serologic tests used for detection of Toxoplasma IgG give very similar results and can all be readily used by clinical laboratories for screening purposes.

Adolescent↗

The investigation of toxoplasma infection associated with pregnancy.

An audit was undertaken to determine case compliance with prenatal testing and investigation of infants for toxoplasma infection. Subsequently, the effect of enhanced reference unit Intervention was studied. The proportion of cases of toxoplasma infection associated with pregnancy completing an investigation programme was calculated. The effect of continued and short-term additional intervention was assessed and reasons for failure to comply were sought. The status of the child was established in 30% of cases when acute maternal toxoplasma infection was detected. Continuous reference unit intervention significantly improved case compliance to 45% over a 3-year period, but the effect was lost when the additional measures were withdrawn. Failure to complete the investigation procedure was associated with loss of patient-clinician contact and clinician/laboratory error. Enhanced intervention did not result in a significant improvement in compliance with the investigation programme for babies with clinical abnormality. The benefits of testing for toxoplasma infection associated with pregnancy are limited by failure to complete necessary investigations.

Adult↗

Pathology of Toxoplasma gondii infection in the nude rat. An experimental model of toxoplasmosis in the immunocompromised host?

Genetically athymic nu/nu (nude) rats, deficient in T cells, die from infection with various Toxoplasma gondii strains, including RH and Prugniaud strains. In contrast, these strains cause chronic infections without apparent symptoms in immunocompetent non-nude rats. We show here that nude rats die in two to three weeks after RH infection and three to four weeks after Prugniaud infection. Histological examination of brains from nude rats at different time points after infection, revealed an absence of lesions after RH strain infection and cysts with usually no inflammation after Prugniaud infection. Lungs from nude rats developed a fibrin alveolitis using either strain, whereas myocarditis with focal areas of necrosis were observed only after Prugniaud infection. Cysts and, in some cases, tachyzoites in the necrotic lesions were easily identifiable. The two strains of T. gondii elicited in nude rats a granulomatous hepatitis that only differed in intensity. Spleen and mesenteric lymph nodes appeared totally non reactive in both cases. This model allows immunological and parasitological studies by comparison with immunocompetent rat infection. Published data concerning toxoplasma pathology in AIDS, therefore, suggest that acute toxoplasma infection in nude rats may be a useful model for studying disseminated forms of toxoplasma infection found in AIDS patients.

Animals↗

Modelling toxoplasma incidence from longitudinal seroprevalence in Stockholm, Sweden.

Age-stratified data on toxoplasma seroprevalence in pregnant women in Stockholm, Sweden for the years 1969, 1979 and 1987 provide the basis for an analysis of temporal patterns of Toxoplasma gondii infection, and estimation of the risk of maternal toxoplasmosis, in this population. A catalytic infection model, in which the rate or force of infection is assumed to be a function of time (and not, as is more usual, age), was employed to describe the observed changes in levels of toxoplasma seropositivity. A range of simple incidence functions (up to 3 parameters) were fitted using a method of maximum likelihood. The data were significantly better described by a linear or an exponential decay in the rate of infection through time compared with a constant level. More complex incidence functions gave no better data description. Thus, whilst there is strong evidence for declining incidence in Stockholm over the past 4-5 decades, the data do not allow discrimination between different possibilities for the nature of this decline. Based on these modelling results, best estimates of the force of infection in 1987 acting on susceptible women are within the range 0 to 0.0045/susceptible/year (95% confidence limits), yielding a possible risk of maternal toxoplasmosis of between 0 and 2.7 cases/1000 pregnancies. These values are shown to be significantly lower than estimates based upon an assumption of temporal stability in toxoplasma incidence, which may be of practical significance to public health policy.

Adolescent↗

The effect of congenital and adult-acquired Toxoplasma infections on the motor performance of mice.

Motor performance was assessed in three groups of mice infected with Toxoplasma. One group was infected when adult. Two groups were infected congenitally: the first was born to dams infected during gestation and the second to dams which were chronically infected prior to mating. All mice were placed individually on a rotating cylinder and the number of falls from it noted over a two-minute period. Infected mice fell significantly more often than uninfected controls. The difference was independent of emotionality (as measured by defaecation) and general body health (as measured by body weight and a subjectively assessed health rating). There was no significant difference in motor performance between the two congenitally infected groups. However, the offspring of mice infected during pregnancy fell significantly more often than mice infected when adult. There were no significant correlations between motor performance and the actual number of Toxoplasma tissue cysts in the brains (or in separate defined sectors of the brains) of infected mice. We suggest that differences between infected and uninfected mice result from pathological changes caused by proliferating toxoplasms in the brains of infected mice. An immunopathological reaction due to the presence of the tissue cysts may also be involved. Other possible factors contributing to observed deficits in motor performance of infected mice are discussed. We suggest that such interference with the motor performance of Toxoplasma infected mice may render them more susceptible to predation by the domestic cat, the definitive host of Toxoplasma.

Animals↗

The effect of congenital and adult-acquired Toxoplasma infections on activity and responsiveness to novel stimulation in mice.

Activity and responsiveness to novel stimulation were assessed in three groups of mice infected with Toxoplasma. One group was infected when adult; two groups were infected congenitally, one born to dams infected during gestation, the other to dams chronically infected prior to mating. Each mouse was tested in a box, the floor of which was marked off into 16 equal squares, and its activity was measured over ten minutes by counting the number of times the mouse entered each square. Infected mice were more active. In addition, infected mice showed a smaller relative preference for the more novel central area of the box, especially towards the end of the observation period. These differences were independent of emotionality (as measured by defaecation counts), general health (as measured by subjective health ratings and body weight) and the number of Toxoplasma tissue cysts in specified brain regions. We suggest that differences arise from pathological changes caused by proliferating toxoplasms in the brains of the infected mice; an immunopathological reaction due to the presence of tissue cysts in the brain may also be involved. Other possible factors contributing to observed deficits in behaviour are also discussed. We suggest that such deficits may render Toxoplasma-infected mice more susceptible to predation by the domestic cat, the definitive host of Toxoplasma.

Animals↗

Toxoplasmosis in sheep. I. Effect of a killed vaccine on lambing losses caused by experimental challenge with Toxoplasma gondii.

A total of 50 two-tooth ewes were immunized pre-tupping using a killed vaccine of disintegrated Toxoplasma tachyzoites with Freunds incomplete adjuvant. At mid-pregnancy, the ewes were challenged intravenously with 1 x 10(4) live T. gondii tachyzoites. There was no difference in the lambing percentage between ewes vaccinated once and unvaccinated ewes. The lambing percentage for ewes vaccinated twice was significantly lower (p<0.05 normal score) than unvaccinated ewes. The serological response to vaccination and challenge was measured by the Toxoplasma dye test. The level of dye test antibodies at the time of challenge suggests that effective immunity to Toxoplasma n abortion is not dependent on dye test antibody titre. These results suggest that a killed vaccine would Toxoplasma on fertility and lambing performance in not be an effective tool to estimate the effects of commercial flocks.

Journal Article↗

A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Toxoplasma gondii.

An enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of Toxoplasma gondii infections on single serum dilutions was developed. This test system is a standardized kit designed to detect circulating specific antibodies to Toxoplasma gondii in human sera. It consists of Toxoplasma gondii soluble antigen-coated microtitration multiwell plates, specific immunoglobulin-enzyme conjugate and other required reagents. In a clinical trial performed on sera from 1,035 clinically suspected toxoplasmosis cases, the Sabin Feldman Dye Test (SFDT) and this ELISA system agreed closely. Relative to the SFDT, the sensitivity and specificity of the latter was 98.0% and 97.6% respectively with a correlation coefficient of 0.97. In a further study of 121 sera, the Indirect Fluorescent Antibody Test (IFAT), the Indirect Haemagglutination Test (IHAT) and this ELISA procedure showed over 90% agreement, with correlation coefficient of 0.98 and 0.95 respectively. Within the working concentration of specific antibody to T. gondii in human serum, there was a linear relationship between the ELISA values and the WHO international standard for human anti-Toxoplasma serum.

Animals↗

Activity of roxithromycin against Toxoplasma gondii in murine models.

Investigations into the activity of roxithromycin against murine toxoplasma infections are reviewed. Roxithromycin is an active drug against murine toxoplasmosis after intraperitoneal challenge with the RH strain of Toxoplasma gondii. Roxithromycin protected 100% of mice after five daily doses of 540 mg per kg administered by gavage. The cure rate after treatment of peritoneal infections seemed to be related to the length of the therapy. Roxithromycin also decreased the number of toxoplasma cysts, after intracerebral infection with the C56 strain and showed synergistic activity when combined with gamma interferon. Thus, roxithromycin could be a worthwhile alternative to current therapy against toxoplasma infections. Clinical studies on its activity and safety, especially in pregnancy, are warranted.

Animals↗

Meta-analysis of prophylactic treatments against Pneumocystis carinii pneumonia and toxoplasma encephalitis in HIV-infected patients.

In a meta-analysis, we examined the efficacy of aerosolized pentamidine, trimethoprim-sulfamethoxazole, and dapsone or dapsone/pyrimethamine for the prevention of Pneumocystis carinii pneumonia and toxoplasma encephalitis in patients with HIV infection. Of 22 trials, 13 compared trimethoprim-sulfamethoxazole with aerosolized pentamidine, nine compared dapsone alone or in combination with pyrimethamine with aerosolized pentamidine, and eight compared trimethoprim-sulfamethoxazole with dapsone/pyrimethamine. In total, 1484 patients were treated with trimethoprim-sulfamethoxazole, 1548 patients with dapsone/pyrimethamine or dapsone, and 1800 patients with aerosolized pentamidine. For dapsone/pyrimethamine versus aerosolized pentamidine, the risk ratio for P. carinii pneumonia was 0.90 (95% confidence interval [CI], 0.71-1.15), and for toxoplasma encephalitis it was 0.72 (95% CI, 0.54-0.97). For trimethoprim-sulfamethoxazole versus aerosolized pentamidine, the risk ratio of P. carinii pneumonia was 0.59 (95% CI, 0.45-0.76), and for toxoplasma encephalitis it was 0.78 (95% CI, 0.55-1.11). For trimethoprim-sulfamethoxazole versus dapsone/pyrimethamine, the risk ratio of P. carinii pneumonia was 0.49 (95% CI, 0.26-0.92), and for toxoplasma encephalitis it was 1.17 (95% CI, 0.68-2.04). Although current evidence does not allow a definitive recommendation, administration of trimethoprim-sulfamethoxazole for prophylaxis of P. carinii pneumonia and toxoplasmosis in patients with HIV infection is consistent with the available data.

AIDS-Related Opportunistic Infections↗

Production and properties of immune interferon from spleen cell cultures of Toxoplasma-infected mice.

In response to antigenic stimulation, spleen cells from Toxoplasma-infected mice produce a factor showing inhibitory activity against vesicular stomatitis virus infection in L cell cultures. When BALB/C and ICR mice were inoculated intraperitoneally with the low-virulent S-273 strain of T. gondii, such activity was first detected in 4 and 7 days and reached maximum levels at 10 and 14 days respectively, and retained these levels for at least three weeks. However, BALB/C mice, which are considerably more sensitive to Toxoplasma infection than ICR mice, produced significantly smaller amounts of interferon (IF) after challenge with the high virulent strain. The IF produced in this system possessed certain known properties of immune (type II) IF and was not neutralized by rabbit antiserum against mouse type I IF. The immune IF preparation also inhibited multiplication of Toxoplasma within nonphagocytic L cells in an IF-like fashion, whereas Newcastle disease virus-induced (type I) IF had no effect on this parasite. The antiviral and anti-Toxoplasma activity in immune IF preparations could not be distinguished solely on the bases of their molecular weight and isoelectric point. The experiments with anti-theta serum plus complement and with nylon wool column effluent cells strongly suggest that immune IF was produced by T lymphocytes and required the assistance of macrophages.

Animals↗

Correlation between increased susceptibility to primary Toxoplasma gondii infection and depressed production of gamma interferon in pregnant mice.

To explore a possible mechanism of pregnancy-associated suppression of T cell-mediated immunity to Toxoplasma gondii, acquired resistance and gamma interferone (IFN-gamma) production in pregnant mice were compared with those in virgin mice after infection with the S-273 strain of this protozoan parasite. The 50% lethal dose of this strain was less than 200 tachyzoites for pregnant mice and 2,800 organisms for virgin controls. Toxoplasma-induced production of both IFN-alpha and IFN-gamma in the bloodstream of pregnant mice was significantly depressed as compared with that in virgin controls. The administration of recombinant murine IFN-gamma (rMuIFN-gamma) resulted in a significant decrease of mortality and parasitic growth in the organs of pregnant mice infected with a lethal dose of S-273 strain tachyzoites. Thus, the impairment of T cell-mediated immune responses was evident in pregnant mice from the impaired IFN-gamma-generating capacity and poor survival rate after primary infection with Toxoplasma. When mice with chronic Toxoplasma infection were injected with specific antigen, the resultant production of IFN-gamma was also significantly suppressed during pregnancy. However, there was no direct correlation between the serum levels of IFN-gamma and susceptibility to reinfection, since the mortality rate of chronically infected pregnant mice after the challenge with the high virulent RH strain was not significantly higher than that of virgin controls.

Animals↗

Development of granulomatous common variable immunodeficiency subsequent to infection with Toxoplasma gondii.

Common variable immunodeficiency (CVID) is a heterogeneous immunodeficiency that is accompanied by granulomatous lesions in 5-10% of cases. Why some patients develop granulomatous disease remains unclear. Here we describe a 12-year-old previously healthy girl who presented with pancytopenia and granulomatous lymphoproliferation subsequent to infection with Toxoplasma gondii. Loosely arranged non-fibrosing granulomas were observed in the liver, lymph nodes and lung, but no Toxoplasma tachyzoites could be demonstrated and polymerase chain reaction (PCR) and culture were negative for Toxoplasma and a wide range of other pathogens. While the patient had a normal peripheral B cell status at presentation, the development of CVID could be observed during the following months, leading to a loss of memory B cells. This was accompanied by an increasingly activated CD4(+) T cell compartment and high serum levels of angiotensin-converting enzyme (ACE), tumour necrosis factor (TNF) and sCD25. Steroid therapy reduced pancytopenia, granulomatous lymphoproliferation and cytokine elevations, but did not improve the B cell status. This is the first report of an association of Toxoplasma infection with granulomatous CVID and provides one of the rare examples where the onset of CVID could be documented subsequent to an infectious disease.

Acute Disease↗

Immunological Study on the Host Cell Penetration Factor of Toxoplasma gondii.

The host cell penetration factor (PEF) of Toxoplasma gondii was studied by biochemical and immunological techniques. Sephadex gel filtration of an ammonium sulfate-precipitated PEF yielded two components with different molecular weight, but both having penetration-enhancing activity. The methods for purification removed at least 99.9% of extraneous protein. For demonstration of a significant enhancement of penetration, 0.001 mug of protein was sufficient. Biochemically, they appeared to be acid proteins with the same electrophoretic mobility. Both components showed maximal enhancement of penetration at pH 7.6 and 37 C. PEF antisera reduced the penetrative capacity of Toxoplasma parasites. The penetration-enhancing effect of the two components of PEF was inhibited by antiserum against any of them. Moreover, immunologically identical immunoprecipitates were obtained when antiserum reacted with the two components. The results thus indicated that the two components of PEF were immunogenically identical and that the difference in molecular weights might result from aggregation. Immunofluorescence indicated that PEF was related to cytoplasmic structures located in the anterior end of Toxoplasma. A possible relation between these structures and the paired organelle or the convoluted tubes was discussed. The number of parasites with immunofluorescence was low shortly after host cell penetration and increased during the intracellular life of the parasites after kinetics, previously observed for synthesis of PEF as well as for lysosomal activity of Toxoplasma.

Journal Article↗