Serologic evaluation of patients with primary and recurrent ocular toxoplasmosis for evidence of recent infection.

PURPOSE: To identify the frequency of recently acquired vs chronic systemic Toxoplasma gondii infections in patients with ocular toxoplasmosis. METHODS: Serum samples from 22 patients with primary ocular toxoplasmosis (not from scars) and 42 patients with recurrent ocular toxoplasmosis were tested for the presence of anti-T. gondii IgM, IgG, and IgA antibodies and compared with samples from 24 patients with other causes of uveitis. Intraocular production of anti-T. gondii IgG and IgA, and the presence of T. gondii DNA was determined in patient s and control subjects from whom ocular fluid was available. RESULTS: Serologic evidence of recently acquired infection was found for 11 (50%) of 22 patients with primary ocular toxoplasmosis and for one (2%) of 42 with recurrent ocular toxoplasmosis. In the uveitis control group, anti-T. gondii IgM antibodies could be detected in two (8%) of 24 patients, but anti-T. gondii IgA antibodies were not detectable. Patients with primary ocular toxoplasmosis and serologic markers of recently acquired systemic infection were significantly older than those with chronic infection (P = .008). Intraocular production of anti-T. gondii IgG was more frequently noted in patients with recurrent than primary ocular toxoplasmosis (81% vs 41%; P < .001), but intraocular T. gondii DNA was more frequently found in patients with primary ocular toxoplasmosis than in those with recurrent ocular toxoplasmosis (37% vs 4%; P < .01). CONCLUSIONS: Primary ocular toxoplasmosis can be seen in either recently acquired or chronic T. gondii infection. Patients with ocular disease and recently acquired infection were older and more likely to have T. gondii DNA in intraocular fluids.

Aetiological study of the presumed ocular histoplasmosis syndrome in the Netherlands.

AIM: To investigate whether presumed ocular histoplasmosis syndrome in the Netherlands is caused by Histoplasma capsulatum and whether other risk factors might play a role in the pathogenesis of this syndrome. METHODS: 23 patients were clinically diagnosed as having presumed ocular histoplasmosis syndrome based on the following criteria: peripapillary atrophy, punched out lesions, a macular disciform lesion or scar in one eye without vitritis. As controls, 66 sex and age matched healthy volunteers were used. Serum samples from both patients and controls were tested for the presence of antibodies against H capsulatum, Toxoplasma gondii, Toxocara canis et cati, Ascaris sp, and for the presence of antigens of Cryptococcus neoformans. Serum samples were also tested for the presence of autoantibodies against retinal or choroidal proteins. To investigate other risk factors, patients and controls were asked to fill in a health and travel related questionnaire. Ten patients with ocular toxoplasmosis were used as a disease control group. RESULTS: None of the patients with presumed ocular histoplasmosis syndrome or controls had circulating antibodies directed against H capsulatum. No risk factors could be identified and no indications for autoimmunity and no evidence for the role of the other infectious agents could be demonstrated. CONCLUSIONS: In a Dutch group of patients fulfilling the criteria of a disease currently named presumed ocular histoplasmosis syndrome, no risk factors or relation with the fungus H capsulatum could be detected.

Intraocular anti-Toxoplasma gondii IgA antibody production in patients with ocular toxoplasmosis.

PURPOSE: To investigate the immunoglobulin classes associated with the intraocular anti-Toxoplasma gondii antibody response during clinical ocular toxoplasmosis and to determine which immunoglobulin class is most helpful in the diagnosis of this disease. METHODS: Paired serum and intraocular fluid samples from 155 patients who had uveitis were tested for intraocular anti-T. gondii IgG, IgA, and IgM antibody production. The presence of T. gondii DNA was determined by polymerase chain reaction. Patients were divided into two groups, based on the initial clinical diagnosis; group 1 included 78 patients with presumed ocular toxoplasmosis, and group 2 included 77 patients with uveitis that was not clinically suspected to be ocular toxoplasmosis. Samples from 27 nonuveitis patients who underwent intraocular surgery were used as control subjects. The final diagnosis was based on the clinical course and interpretation of laboratory tests. RESULTS: A final diagnosis of ocular toxoplasmosis was made in 88 of 155 patients (group 1, 68; group 2, 20). Among these patients, 65% had intraocular IgG production, 52% had intraocular IgA production, 37.5% had both IgG and IgA production, 27% had IgG production only, and 15% had IgA production only. Of the 13 patients tested, only one had intraocular IgM production. Intraocular IgA could not be detected in patients who had final diagnoses other than ocular toxoplasmosis or in control subjects. A positive polymerase chain reaction combined with a test that was positive for intraocular IgG production had a sensitivity of 77%, which increased to 91% after the detection of intraocular IgA production was added. CONCLUSIONS: Immunoglobulin G is the major class involved in the humoral immune response against the T. gondii parasite, followed by IgA. The determination of IgA production is useful as an additional test in the diagnosis of ocular toxoplasmosis.

Analysis of immunoregulatory cytokines in ocular fluid samples from patients with uveitis.

PURPOSE: To investigate the T-helper cell cytokine profiles in two well-defined clinical uveitis entities caused by an infectious mechanism. METHODS: Cytokines (interleukin [IL]-2, IL-4, IL-6, IL-10, and interferon [IFN]-gamma) were measured in ocular fluid samples obtained from patients with herpes simplex- or varicella-zoster virus-induced acute retinal necrosis (ARN; n = 17) and toxoplasma chorioretinitis (n = 27) using enzyme-linked immunosorbent assay techniques. The data were compared with data for 51 control samples taken during cataract surgery (n = 10), vitrectomy in diabetic retinopathy (n = 10), eye bank eyes (n = 10) and with samples from patients with "autoimmune" uveitis (n = 21). RESULTS: Interleukin-6 was detected in 44 of 51 control samples and 43 of 44 eyes of patients with uveitis. The highest levels in the control samples were detected in 9 of 10 vitreous samples from patients with diabetic retinopathy (mean, 648 pg/ml). In 8 of 10 samples taken from patients during cataract surgery and in 7 of 10 eye bank eyes the amount of IL-6 was significantly lower (mean, 10 pg/ml and 136 pg/ml, respectively). Interleukin-6 levels in patients with ARN (mean, 1436 pg/ml) were significantly higher than in those with toxoplasma chorioretinitis (mean, 272 pg/ml). Interleukin-2 was detected in one of the samples from patients with toxoplasma chorioretinitis (1105 pg/ml) and in three samples from the control subjects suffering from Fuchs' heterochromic anterior uveitis (mean, 752 pg/ml). No IL-4 (<2 pg/ml) was detected either in patient or control samples. Interferon-gamma could be detected in 7 of 17 ARN patients (range, 277-3483 pg/ml), in 13 of 27 samples from patients with toxoplasma chorioretinitis (range, 12-250 pg/ml), and in 1 of 21 of the samples from control subjects with uveitis (31 pg/ml) but was absent in nonuveitic control samples. Interleukin-10 was detected in 10 of 17 ARN patients (range, 29-3927 pg/ml), in 13 of 27 samples from patients with toxoplasma chorioretinitis (range, 4-67 pg/ml), and in only 3 of 51 control samples (6 pg/ml, 16 pg/ml, and 20 pg/ml). CONCLUSIONS: Various immunoregulatory cytokines (IL-6, IL-10, and IFN-gamma) were detected in ocular fluid samples from patients with uveitis. A separate role for either a T-helper type 1 or T-helper type 2 response in the pathogenesis of clinical uveitis could not be proven.

Differences between intraocular and serum antibody responses in patients with ocular toxoplasmosis.

PURPOSE: To investigate the antigen specificity of the intraocular anti-Toxoplasma gondii antibody response in patients with ocular toxoplasmosis. METHODS: Paired ocular fluid and serum samples were collected from 13 patients with active ocular toxoplasmosis. Serum IgM anti-T. gondii antibodies were tested to distinguish recently-acquired from chronic infection. Anti-T. gondii IgG specificity was analyzed by immunoblotting using a crude T. gondii extract. RESULTS: Two of the 13 patients tested were IgM positive and considered to have acquired ocular toxoplasmosis. The antibody specificity in ocular fluid and serum of these two patients was similar, whereas in the patients with presumed chronic disease, marked differences could be observed. Most ocular fluid samples contained antibodies that stained a 28-kD antigen more intensely than did antibodies from paired serum samples. Using absorption and elution experiments, we demonstrated that this 28-kD protein was identical to the GRA-2 antigen, which is expressed in both the tachyzoite and the bradyzoite stages of the parasite. CONCLUSIONS: Our results show that the intraocular T. gondii antibody response of patients with recurrent ocular toxoplasmosis differs from the systemic response. This finding may have implications for our understanding of the immunopathogenesis of ocular toxoplasmosis and could be employed to improve diagnosis of the disease.

Increased presence of Epstein-Barr virus DNA in ocular fluid samples from HIV negative immunocompromised patients with uveitis.

AIMS: To investigate whether routine testing for Epstein-Barr virus (EBV) is necessary in the examination of a patient with uveitis. METHODS: Intraocular EBV DNA was determined in 183 ocular fluid samples taken from patients with AIDS and uveitis, HIV negative immunocompromised uveitis, acute retinal necrosis, toxoplasma chorioretinitis, intraocular lymphoma, anterior uveitis, and miscellaneous uveitis of unknown cause. In 82 samples from this group of patients paired serum/ocular fluid analysis was performed to detect local antibody production against EBV. Controls (n = 46) included ocular fluid samples taken during surgery for diabetic retinopathy, macular pucker, or cataract. RESULTS: Serum antibody titres to EBV capsid antigen proved to be significantly increased in HIV negative immunocompromised patients with uveitis (p < 0.01) compared with controls. Local antibody production revealed only three positive cases out of 82 patients tested, two results were borderline positive and one patient had uveitis caused by VZV. EBV DNA was detected in three out of 46 control ocular fluid samples. In the different uveitis groups EBV DNA was noted, but was not significantly higher than in the controls, except in six out of 11 HIV negative immunocompromised patients (p = 0.0008). In four out of these six cases another infectious agent (VZV, HSV, CMV, or Toxoplasma gondii) had previously been identified as the cause of the uveitis. CONCLUSIONS: When comparing various groups of uveitis patients, EBV DNA was found more often in HIV negative immunocompromised patients with uveitis. Testing for EBV does not have to be included in the routine management of patients with uveitis, since indications for an important role of this virus were not found in the pathogenesis of intraocular inflammation.