J.L. Guillén

Larval distribution of Toxocara canis in BALB/c mice at nine weeks and one year post-inoculation.

The migratory route of Toxocara canis in the mouse includes two phases: a hepato-pulmonary phase and a myotropic-neurotropic phase. A study was made of the migratory behaviour of larvae one year post-inoculation (p.i.) comparing the results to those obtained at the end of an initial experiment (day 63 p.i). The larval distribution per organ was already definitive at 9 weeks p.i. The total parasitic load showed a reduction when the experiment was prolonged for one year.

Bilateral ocular toxocariasis demonstrated by aqueous humor enzyme-linked immunosorbent assay.

PURPOSE/METHODS A 14-year-old girl with a history of contact with puppies and chronic bilateral panuveitis was examined. The fundus of the right eye showed an inferotemporal pars plana exudate. The fundus of the left eye showed an elevated posterior mass. RESULTS/CONCLUSION Toxocara antibodies in the aqueous humor detected by enzyme-linked immunosorbent assay had a Goldmann-Witmer coefficient of 8.63 in the right eye and 8.94 in the left eye. Toxocariasis may be a cause of bilateral panuveitis.

Cross-reactions of sera from Toxascaris leonina and Ascaris suum infected mice with Toxocara canis, Toxascaris leonina and Ascaris suum antigens

The ELISA method using larval ES products and homogenized Toxocara canis, Toxascaris leonina and Ascaris suum adult worms extract, was used to determine the possible cross-reactions in BALB/c and C57BL/10 mice inoculated with embryonated eggs or adult worms extract of T. leonina of T. leonina or A. suum in single and multiple doses. When we used sera of mice infected with embryonated eggs of T. leonina against different heterologous Ag, no cross-reactions against T. canis ES and A. suum ES Ag were observed using a single dose. Similarly, in multiple doses no response against T. canis ES Ag was observed. In mice inoculated with adult worms extract of T. leonina cross-reactions with T. canis ES and A. suum ES Ag did not occur. Sera from BALB/c mice infected with embryonated eggs of A. suum, was tested using ES Ag from both A. suum and T. canis and no reactions were observed. This fact confirmed the resistance of this murine strain to A. suum embryonated eggs. When we used sera of susceptible C57BL/10 mice infected against different heterologous Ag, we observed no cross-reactions against T. canis ES Ag. In the case of both BALB/c and C57BL/10 and C57BL/10 mice immunized with a single dose of A. suum adult crude extract no cross-reactions were seen against ES T. canis Ag and with sera from C57BL/10 mice against ES T. leonina. These facts confirmed the specificity of the ES T. canis Ag.

Comparative study of assays detecting circulating immune complexes and specific antibodies in patients infected with Toxocara canis

A sandwich ELISA method using previously described E/S antigen-specific monoclonal antibodies has been developed to detect circulating immune complexes in patients infected with Toxocara canis. This technique could be used for the study of the dynamics of the parasite-host relationship, as we believe the detection of immune complexes and/or soluble antigen to be an improvement over detection of antibodies only. In this parasitosis, antibodies may be present in residual levels for prolonged periods after active infection.

Seroprevalence of toxocariasis in children and adults in Madrid and Tenerife, Spain

A study on the seroprevalence of toxocariasis, using ELISA with Toxocara larval excretory-secretory antigens, was carried out on human populations in two regions of Spain. Sera from a population of 195 children from Madrid and 143 children from Santa Cruz de Tenerife (Canary Isles), showed a prevalence of 0% and 4.2% respectively. Sera from a population of 272 adults from Madrid and 803 adults from Santa Cruz de Tenerife showed a prevalence of 3.6% and 17.4%. Reasons for these differences in the seroprevalence of Toxocara in the different age groups from the two regions are discussed.

Persistence of immune response in human toxocariasis as measured by ELISA

The antibody titer was followed in a group of patients, clinically diagnosed with toxocariasis, during a 5 year period. We observed that larvae can survive for at least 5 years in humans. Antigenic stimulation was enough to keep high levels of immunoglobulins over this period. Antibody levels decreased slowly and this pattern is similar to that shown by animal models.

Cross-reactions of sera from Toxocara canis-infected mice with Toxascaris leonina and Ascaris suum antigens

The ELISA method using larval excretory-secretory (E/S) products and homogenized Toxocara canis, Toxascaris leonina and Ascaris suum adult worm extract were used to determine possible cross-reactions in BALB/c and C57BL/10 mice, inoculated with embryonated eggs or adult worm extract of T. canis in single and multiple doses. When we used sera of mice infected with embryonated eggs of T. canis against different heterologous antigens, we observed no cross-reactions in BALB/c mice against A. suum E/S and adult worm extract antigens with a single dose. In multiple doses this was absent too against T. leonina adult worm extract in BALB/c mice, and in both strains against A. suum E/S and adult worm extract. In BALB/c mice inoculated with adult worm extract of T. canis we did not observe cross-reactions with A. suum E/S antigen with both inoculation doses. In the remainder of the experiments, we observed cross-reactions of different intensities.

Isotype specific immune responses in murine experimental toxocariasis

In this work, a murine experimental model of toxocariasis has been developed in BALB/c, C57BL/10 and C3H murine strains orally inoculated with 4,000 Toxocara canis embryonated eggs, in order to investigate the isotype-specific immune responses against excretory-secretory antigens from larvae. T. canis specific IgG+M, IgM, IgG, IgA, IgG1, IgG2a and IgG3 were tested by ELISA. The dynamics of the specific immunoglobulins (IgG+IgM) production showed a contrasting profile regarding the murine strain. Conversely to the results obtained with the IgM isotype, the IgG antibody class showed similar patterns to those obtained with IgG+IgM antibodies, only in the case of the BALB/c strain, being different and much higher than the obtained with IgG+IgM antibodies, when the C3H murine strain was used. The antibodies IgG+IgM tested in BALB/c and C57BL/10 were both of the IgM and IgG isotypes. Conversely, in the C3H strain only IgG specific antibody levels were detected. The IgG1 subclass responses showed a similar profile in the three murine strains studied, with high values in BALB/c, as in the case of the IgG responses.

Serological evidence of toxocariasis in patients from Spain with a clinical suspicion of visceral larva migrans.

Sera from patients with clinical characteristics of toxocariasis were assayed using the ELISA method and larval excretory-secretory antigen. Four hundred and seven samples of Toxocara serology were received at the laboratory of Ciudad Sanitaria Juan Canalejo Hospital of Corunna, Spain, from 1984 to 1989. Of these, 30 were from adults, 332 from children and 45 from patients of unknown age, resulting in Toxocara seroprevalences of 23.3%, 32.8% and 17.7% respectively. The reasons for these serological differences in the rural and urban areas of Galicia, Spain are discussed.

Evaluation of chemotherapy in experimental toxocarosis by determination of specific immune complexes

Parasitism by the larval phase of Toxocara canis is a chronic process in which the larvae survive in the tissues, resulting in the constant stimulation of the immune system. As a result, the detection of specific antibodies may not reflect the active state of the parasite. We have studied the dynamics of the production of specific immune complexes by ELISA with the monoclonal antibody TC-1 in rabbits inoculated with single and multiple doses of T. canis eggs. We also compared this with the production of specific antibodies and their possible modification after treatment with mebendazole. The specific antibodies against excretory-secretory antigen were detected with peaks at 10 and 12 weeks depending on the dose and remained positive during the entire experiment (62 weeks). Treatment caused an increase in the level of detectable antibodies dropping to similar levels to the controls. Specific immune complexes were detected only in multiple doses, and were then positive during the entire experiment. From the beginning of treatment the values of immune complexes fell quickly, remaining at undetectable levels during the rest of the experiment. For this reason the detection of specific immune complexes is a valid technique for monitoring the efficiency of treatment.