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Value of PCR for detection of Toxoplasma gondii in aqueous humor and blood samples from immunocompetent patients with ocular toxoplasmosis.

Toxoplasma gondii infection is an important cause of chorioretinitis in the United States and Europe. Most cases of Toxoplasma chorioretinitis result from congenital infection. Patients are often asymptomatic during life, with a peak incidence of symptomatic illness in the second and third decades of life. Diagnosis is mainly supported by ophthalmological examination and a good response to installed therapy. However, establishment of a diagnosis by ophthalmological examination alone can be difficult in some cases. To determine the diagnostic value of PCR for the detection of T. gondii, 56 blood and 56 aqueous humor samples from 56 immunocompetent patients were examined. Fifteen patients with a diagnosis of ocular toxoplasmosis had increased serum anti-T. gondii immunoglobulin G levels but were negative for anti-T. gondii immunoglobulin M (group 1), and 41 patients were used as controls (group 2). Samples were taken before antiparasitic therapy was initiated, and only one blood sample and one aqueous humor sample were obtained for each patient. Single nested PCRs and Southern blot hybridization were performed with DNA extracted from these samples. The results obtained showed sensitivity and specificity values of 53. 3 and 83%, respectively. Interestingly, among all patients with ocular toxoplasmosis, a positive PCR result with the aqueous humor sample was accompanied by a positive PCR result with the blood sample. This result suggests that ocular toxoplasmosis should not be considered a local event, as PCR testing of blood samples from patients with ocular toxoplasmosis yielded the same result as PCR testing of aqueous humor samples. PCR testing may be useful for discriminating between ocular toxoplasmosis and other ocular diseases, and also can avoid the problems associated with ocular puncture.

Adolescent↗

Immunoblotting can help the diagnosis of ocular toxoplasmosis.

AIMS: To determine whether IgG immunoblotting can improve the diagnosis of ocular toxoplasmosis. METHODS: Samples of serum were tested from patients with ocular lesions that could be caused by toxoplasmosis. All such samples from Scotland and Northern Ireland are usually referred to the Scottish Toxoplasma Reference Laboratory. From questionnaires filled out by the clinicians, two groups of sera were identified: ocular toxoplasmosis (active and quiescent), n = 54 (group 1); and eye disease as a result of other causes, n = 36 (group 2). Control groups were made up of sera from patients with no eye disease and a normal dye test result (< or = 125 IU/ml), n = 16 (group 3); and toxoplasma seronegative, cytomegalovirus (CMV) positive, and herpes simplex virus (HSV) positive sera (group 4), n = 18. RESULTS: Immunoblots with an active pattern could be identified (IgG antibodies against at least four antigens with molecular weight of 6, 20, 22, 23, 25, and 36 kDa). Significantly more of this pattern was found in group 1 (33 of 54; 61.1%) compared with group 2 (nine of 36; 25%) or group 3 (six of 16; 37.5%). Within group 1, significantly more sera with an active pattern had dye test results > or = 65 IU/ml compared with those without. More sera from patients < 30 years of age were found with the active pattern in group 1 compared with group 2. No group 4 sera had active immunoblot patterns. CONCLUSIONS: The immunoblot result adds more support to the diagnosis of ocular toxoplasmosis. In cases where the clinical diagnosis is difficult, immunoblots are particularly indicated; if negative, other causes of eye disease should be sought.

Adult↗

Experimental ocular toxoplasmosis in primates.

The rabbit serves as the only existing experimental model for ocular toxoplasmosis. This model has many deficiencies. Our technique for the production of acute toxoplasmic retinitis in monkeys is described herein. This primate model should be more representative than the rabbit model of human ocular toxoplasmosis.

Animals↗

The role of hypersensitivity reactions to toxoplasma antigens in experimental ocular toxoplasmosis in nonhuman primates.

To assess the role of ocular hypersensitivity reactions to Toxoplasma antigens in previously sensitized ocular tissues, we used a nonhuman primate model of ocular toxoplasmosis. Each eye of eight monkeys was inoculated with living Toxoplasma organisms into the inner retinal layers. All eyes developed necrotizing retinochoroiditis. Healing of the lesion occurred with the formation of a retinochoroiditic scar. Four months later, Toxoplasma antigens were injected through the right internal carotid artery. Four weeks after this, Toxoplasma antigens were inoculated into the left retina. Iritis, vitritis, and retinal edema occurred in response to the administration of the antigens, but no recurrent necrotizing retinochoroiditis was produced in this model. These findings suggested that hypersensitivity to Toxoplasma antigens does not play a major role in triggering recurrences of toxoplasmic retinochoroiditis in nonhuman primates.

Animals↗

Reactivation of ocular toxoplasmosis after LASIK.

PURPOSE: To report a reactivation of ocular toxoplasmosis after LASIK. METHODS: Case report of a 34-year-old man who underwent bilateral LASIK. The posterior segment examination revealed an old toxoplasmosis scar in the retinal periphery of the right eye. RESULTS: Uncorrected visual acuity improved postoperatively, and the patient was satisfied. However, 52 days after the procedure, he complained of loss of visual acuity in his right eye. Examination revealed signs of anterior uveitis, vitreitis, and active chorioretinal lesion satellite of the old toxoplasmosis scar. The patient was treated with a multidrug regiment with resolution of the vitreous and lesion activity. CONCLUSIONS: Toxoplasmosis reactivation may develop after LASIK.

Adult↗

Experimental ocular toxoplasmosis induced in naive and preinfected mice by intracameral inoculation.

We have developed a murine model to investigate the pathogenesis of acquired ocular toxoplasmosis. Tachyzoites of PLK strain of Toxoplasma gondii were intracamerally inoculated under anesthesia into the right eyes of naive or perorally preinfected C57BL/6 and MRL-MpJ mice. Clinical and histopathological observations of responses to intraocular infection were analyzed. Ocular inflammation from Toxoplasma gondii is dose-dependent in both strains of mice. After inoculation of fifty parasites, no evidence of inflammation was observed in the eyes of naive mice. The eyes of naive mice that received 500 or 5,000 parasites developed inflammatory changes by day 6 post challenge. By day 8, the changes progressed to moderate to severe intraocular inflammation. Histologic analysis of the ocular lesions demonstrated mononuclear cell infiltration and necrosis predominantly in the anterior segment of the eyes of the naive mice. Inoculation of 50,000 tachyzoites induced a destructive ocular inflammatory response and was uniformly lethal to the mice by approximately one week after challenge. In contrast, eyes from mice previously orally infected with Toxoplasma gondii and that received a 50 or 500 parasite intracameral challenge revealed no inflammation, but the eyes receiving 5,000 parasites demonstrated necrotic focal retinochoroiditis with vitreitis by day 8 after challenge. The murine model reproduces some features of ocular toxoplasmosis in humans and may be suitable for large-scale controlled studies of the pathogenesis and therapeutics of acquired ocular toxoplasmosis as well as for study of the mechanisms of immune privilege in the eye.

Animals↗

An update on current practices in the management of ocular toxoplasmosis.

PURPOSE: To update information that was published by the AMERICAN JOURNAL OF OPHTHALMOLOGY in 1991 about treatment practices for ocular toxoplasmosis by uveitis specialists. DESIGN: Physician survey. METHODS: A written questionnaire was distributed to all physician-members (n = 147) of the American Uveitis Society. The questionnaire was modeled after a similar device used to survey uveitis specialists in 1991. Information contained on 96 returned questionnaires was tabulated. RESULTS: Among 79 respondents who evaluate and manage patients with ocular toxoplasmosis, 15% treat all cases regardless of clinical findings (in contrast to 6% in 1991). The major indications for treatment among other respondents were severe inflammatory responses and proximity of retinal lesions to the fovea and optic disk. The majority of clinical factors considered in five categories (vision, lesion location, lesion size, lesion characteristics, and vitreous inflammatory reaction) were identified to be relative or absolute indications for treatment by a greater proportion of respondents in the current survey than in the 1991 survey. A total of nine drugs (or commercially available combinations) were used in 24 different regimens as treatments of choice for typical cases of recurrent toxoplasmic retinochoroiditis, with the combination of pyrimethamine, sulfadiazine, and prednisone being the most commonly used regimen (29% of respondents). CONCLUSIONS: Uveitis specialists appear to be more likely to treat patients with ocular toxoplasmosis in 2001 than in 1991. Although the majority of survey respondents adhere to a traditional approach to the management of toxoplasmic retinochoroiditis (a discrete course of systemic drug treatment during active disease using multiple antiparasitic drugs with or without corticosteroids), there is still no consensus regarding the choice of antiparasitic agents for treatment regimens. Survey results provide useful information for treating physicians and for clinical investigators interested in therapy.

Adult↗

Experimental ocular toxoplasmosis in genetically susceptible and resistant mice.

Genetic factors determining the pathogenesis and course of ocular toxoplasmosis are poorly understood. In this study, we explored the development of experimental ocular pathogenesis in genetically dissimilar mice infected with either the RH strain, the PLK strain, or the immunodominant surface antigen 1 (SAG1 [P30])-deficient mutant of the RH strain of Toxoplasma gondii. At 11 days postinfection, ocular infection of C57BL/6 mice with all of the strains of parasites resulted in severe inflammatory lesions and high numbers of parasites in eye tissue; less severe ocular lesions at earlier histopathology and prolonged survival were observed in this mouse strain infected with either the major surface antigen 1-deficient SAG1(-/-) strain or the less virulent PLK strain compared with RH infection. In contrast, both BALB/c and CBA/J mice had less severe lesions and low numbers of parasites in their eye tissue, and infection developed into the chronic stage in these mice. There were significantly higher serum levels of gamma interferon and tumor necrosis factor alpha in C57BL/6 mice than in BALB/c and CBA/J mice following ocular infection. These observations confirm earlier reports on systemic immunity to these parasites that the route of Toxoplasma infection markedly influences survival of mice. Our data indicate that genetic factors of the host as well as the parasite strain are critical in determining susceptibility to experimental ocular toxoplasmosis in murine models.

Animals↗

Vascular anastomoses in ocular toxoplasmosis.

Toxoplasmosis is a frequent cause of uveitis seen in clinical practice and fundus scars typical of ocular toxoplasmosis are common. It has been reported that vascular anastomoses between the retinal and choroidal circulation can occur through the damaged Bruch's membrane in fundus scars resulting from ocular toxoplasmosis. Although it has been stated that these vascular anastomoses are a rare occurrence, it has also been suggested that they are relatively common. In order to determine the prevalence rate of patients with vascular anastomoses in toxoplasmic fundus scars, 3,850 consecutive optometry patient files were studied retrospectively. Seventy-four patients (1.92%) had a clinical diagnosis of ocular toxoplasmosis with typical fundus scars, and two of these patients (2.70%) had documented vascular anastomoses.

Chorioretinitis↗

[Ocular toxoplasmosis. Comparison between two biological methods to study aqueous humor].

To diagnose ocular toxoplasmosis with certainty is often difficult and requires anti Toxoplasma gondii antibodies screening in the aqueous humour. We evaluated the results obtained in 30 controls and 5 patients investigated, using the ELISA and indirect immunofluorescence techniques to determine Desmonts's C coefficient and the sera/aqueous humour densities ratio obtained with ELISA. None of the chorioretinitis-free controls showed a false positive result. Negative serum antibodies levels indicated an absence of ocular toxoplasmosis. The sensitivity of this method depends on several biological parameters, notably the extent of ocular inflammation. Evaluating the C coefficient by the ELISA technique increases this sensitivity.

Albumins↗

Socioeconomic conditions as determining factors in the prevalence of systemic and ocular toxoplasmosis in Northeastern Brazil.

OBJECTIVE: To determine the prevalence of systemic and ocular toxoplasmosis among 1024 students in the city of Natal, Northeastern Brazil, and correlate it with demographic, socioeconomic and epidemiological risk factors. METHODS: The study population was randomly selected, asked to fill out a questionnaire, provide a blood sample for IgG and IgM (MEIA) serology and a hemogram, and undergo an eye examination. RESULTS: The seroprevalence for IgG was 46% (95% CI = 42.9-49.2%) and that for IgM was 1.4% (95% CI = 0.8-2.4%). The prevalence of ocular lesions was 1.15% (95% CI = 0.6-2.0%). In the univariate analyses, confirmed by multivariate analysis, the socioeconomic conditions were determinants in the prevalence of systemic and ocular toxoplasmosis (mother's schooling = literacy/OR = 2.9 and p < 0.001). CONCLUSIONS: The prevalence of systemic toxoplasmosis, although high, was lower than that found in studies performed in the South and Southeast of Brazil, and the incidence of ocular lesions was totally different, being lower by a factor varying from 5 to 17. Although important epidemiological variables, such as owning a cat, drinking unfiltered water or having had contact with lakes or rivers, were found to be correlated with toxoplasmosis in the preliminary analysis, they lost their influence when included in the logistic model. However, further studies must be undertaken to identify the reasons for these findings, including the determination of the strains of Toxoplasma gondii encountered in different regions of the country and the sources of the water utilized by these populations.

Adolescent↗

A prospective, randomized trial of pyrimethamine and azithromycin vs pyrimethamine and sulfadiazine for the treatment of ocular toxoplasmosis.

OBJECTIVE: To compare the effects of two treatment regimens, one of which included azithromycin, for the treatment of sight-threatening (near optic disk or fovea) ocular toxoplasmosis. DESIGN: Prospective, randomized open-labeled multicenter study, masked in part with regard to evaluation. METHODS: PARTICIPANTS TOTAL ENROLLMENT: 46 patients with sight-threatening ocular toxoplasmosis; pyrimethamine and azithromycin group: 24 patients; pyrimethamine and sulfadiazine group: 22 patients. INTERVENTION: Patients were randomized into two treatment regimens. Group 1 was treated with pyrimethamine and azithromycin complemented with folinic acid and the addition of prednisone from day 3. Group 2 was treated with pyrimethamine and sulfadiazine complemented with folinic acid and the addition of prednisone from day 3. Patients used study medications daily for 4 weeks. Ocular and laboratory examinations were performed at least weekly during the observation period. The study was masked in part with regard to evaluation. MAIN OUTCOME MEASURES: An assessment was made of the time to resolution of the intraocular inflammatory activity, the size of the retinochoroidal lesion, and visual acuity before and after the treatment as well as all adverse effects of treatments. RESULTS: Adverse effects were more frequent in the pyrimethamine/sulfadiazine group (P <.04), and three patients in this group had to discontinue treatment. The time to resolution of inflammatory activity, decrease in size of retinochoroidal lesions, and optimal visual acuity did not differ between the two treatment groups. The number of patients who developed recurrences during the first year after treatment was similar for both groups. CONCLUSIONS: The efficacy of the multidrug regimen with pyrimethamine and azithromycin was similar to the standard treatment with pyrimethamine and sulfadiazine. However, the frequency and severity of adverse effects was significantly lower with a regimen containing pyrimethamine and azithromycin. Multidrug therapy with the combination of pyrimethamine and azithromycin appears to be an acceptable alternative for treatment of sight-threatening ocular toxoplasmosis.

Adolescent↗

Ocular toxoplasmosis and Hodgkin's disease: report of two cases.

Isolated ocular toxoplasmosis developed in two patients with Hodgkin's disease. The diagnosis was made by ophthalmoscopy of the fundus, serologic tests, and evolution. This raises the possibility that Toxoplasma infection and the localization of retinochoroiditic lesions on the macula could be the results of the immunosuppression associated with Hodgkin's disease.

Adolescent↗

The antibody response in experimental ocular toxoplasmosis.

BACKGROUND: The dynamics of the humoral immune response in ocular toxoplasmosis (OT) are poorly understood. We therefore investigated this process in a rabbit model of the disease. MATERIALS AND METHODS: Of 24 infection-naive adult rabbits, 12 were left untreated and 12 were systematically infected with 5,000 tachyzoites of the non-cystforming BK strain of Toxoplasma gondii. Three months later, all rabbits were inoculated transvitreally with 5,000 tachyzoites of Toxoplasma gondii. Paired samples of aqueous humor and serum were analyzed temporally for their total and specific IgG contents. RESULTS: In infection-naïve rabbits with primary OT, specific IgG reached detectable levels in the inoculated eyes between 5 and 15 days after inoculation. In infection-immunized rabbits with secondary OT, a significant increase in specific IgG was regularly detected after 5 days. The antibody ratio C was diagnostic (> or =3) from day 15 onward in primary OT and from day 21 onward in secondary OT. In the uninfected partner eyes, the antibody ratio C was found sporadically diagnostic from day 15 onward in primary OT, but at no time in secondary OT. Specific IgG persisted both locally and in the serum until the end of the monitoring period (100 days). CONCLUSION: Our findings relating to the rabbit model of OT reveal three features of clinical relevance: a diagnostic window precedes the establishment of a humoral immune response; specific antibodies persist long after the cessation of disease activity; and in primary OT, the antibody ratio C may also increase in the uninfected partner eye.

Animals↗

Ocular toxoplasmosis: in the storm of the eye.

Ocular toxoplasmosis (OT) can occur in the children of mothers infected with Toxoplasma gondii during pregnancy. It is not limited to the congenitally infected, but can also occur following adult-acquired infection or as a result of disease reactivation in immune-compromised and pregnant individuals. Many aspects of immune privilege in the eye, including constitutive TGF-beta expression and reduced MHC class 1 expression, would appear at first to favour parasite survival. Conversely, many of the mechanisms that control parasite multiplication in other anatomical sites, such as nitric oxide expression, IFN-gamma and TNF-alpha, are known to disrupt immune privilege and are associated with ocular damage. Taking into account the opposing needs of limiting parasite multiplication and minimizing tissue destruction we review the pathogenesis of OT in the murine model.

Animals↗