Experimental paracoccidioidomycosis in the mouse. III. Histopathological and immunological findings after intravenous infection in the presence or absence of previous immunization

Fifty male white Swiss mice aged 4 weeks were inoculated with 5 x 10(5) viable yeast forms of Paracoccidioides brasiliensis (strain 18). Ten of these animals had been previously immunized with particulate P. brasiliensis antigenfor 4 weeks by intradermal injection. The controls consisted of 10 animals that were only immunized and 10 animals submitted to no treatment. The animals were sacrificed 2, 4, 7,11 and 16 weeks later. We studied: 1) the anti-P. brasiliensis delayed hypersensitivity response measured by the footpad test 24 hours prior to sacrifice; 2) the specific antibody production measured by double immunodiffusion in agar gel; 3) the histopathology of lungs, liver, spleen, adrenals and kidneys. We observed that: a) the immunized animals developed more intense cell-immune responses than the infected ones; b) infection reduced the cell- immune response of the immunized animals; c) intravenous infection of mice with P. brasiliensis was characterized by a systemic and progressive granulomatous inflammation. The animals infected after previous immunization showed less extensive lung inflammation, with smaller granulomas and fewer fungi. The results indicate that the present murine model mimics some findings of the human subacute form of paracoccidioidomycosis (systemic disease with depressed cellular immunity) and that the extrapulmonary immunization scheme was able to induce a certain degree of protection of the lung from infection with P. brasiliensis

O teste imunoenzimático elisa no líquido pericárdico: novo método para o diagnóstico post-mortem da doença de Chagas

Realizou-se o teste imunoenzimático ELISA, paralelamente à reação de imunofluorescência, para a detecção de anticorpos antí-Trypanosoma cruzi, em 137 amostras de líquidos pericárdicos humanos, colhidos na necropsia. Os resultados foram cotejados com os achados anatomopatológicos. Observou-se que: (1) os dois testes foram positivos em 30 casos e negativos em 105; (2) o teste ELISA foipositivo em 2 casos nos quais a immofluorescència revelou-se negativa; num desses casos, havia sinais morfológicos de doença de Chagas; (3) a média geométrica dos títulos obtidos com o teste ELISA foi significativamente maior que a da imunofluorescência; (4) o índice de concordância entre os dois testes apresentou o valor de 0,985. O presente relato parece-nos inédito quanto ao uso do teste imunoenzimático no líquidoperícárdicopara o diagnóstico post- mortem da doença de Chagas.

Antígenos solúveis liberados por tripomastigotas de Trypanosoma cruzi utilizados no teste de ELISA para detectar cura em pacientes chagásicos após tratamento específico

Dois antígenos solúveis de tripomastigotas do Trypanosoma cruzi, um obtido de sobrenadantes de culturas celulares (AgSb) e o outro excretado/secretado por essas formas em meio de cultura (AgES), foram avaliados em um teste de ELISA para o diagnóstico da infecção chagásica e controle de cura de pacientes tratados. Os pacientes tratados apresentavam testes de lise mediada pelo complemento e hemoculturas repetidamente negativos, apesar de permanecerem com a sorologia convencional positiva (pacientes dissociados). O teste de lise negativo indica que estes pacientes eliminaram a infecção. Entre os controles com infecção ativa, os AgSb e os AgES detectaram respectivamente 93 e 100% dos casos. No entanto, entre os pacientes dissociados, o teste de ELISA, utilizando os AgSb e AgES, foi positivo com 28% e 5% dos soros, respectivamente. Portanto, este teste com os AgES é indicado para o controle de curada doença de Chagas, podendo vir a substituir a reação de lise mediada pelo complemento no acompanhamento sorológico individual de pacientes tratados.

Evaluation of a direct immunofluorescent antibody (difma) test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis

A direct immunofluorescent antibody (DIFMA) test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL) in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

Diagnostic tests for amoebic liver abscess: comparison of enzyme - linked immunosorbent assay (Elisa) and counterimmunoelectrophoresis (CIE)

The liver abscess is the most frequent extraintestinal complication of intestinal amoebiasis: its diagnosis is suggested by the clinical picture but it must be confirmed by paraclinic tests. Themost stringent diagnosis requires identification of E. histolytica. But this is possible only in a few cases. Serological tests greatly improve the diagnosis of this severe complication of amoebiasis. We compared the Enzyme Linfed Immunosorbent Assay and the Counterimmunoeletrophoresis techniques. Both techniques were used to detect amoebic antibodies in 50 control patients, 30 patients with liver abscess and 30 patients with intestinal amoebiasis. All the sera from control patients gave negative results iin both techniques. When analysing the sera from patients with intestinal amoebiasis, 10% of them were positive by ELISA but non by CIE. The sera of patients with liver abscess, we found that 90% were positive by the ELISA method and 66.6% by the CIE technique. In patients with amoebic liver abscess, the results showed that the ELISA was more sensitive than the CIE, as it presented a higher sensitivity (100%) than that of the CIE technique (66%).

The FML (Fucose Mannose Ligand) of Leishmania donovani: a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar

The Fucose-Mannose Ligand (FML) of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, identifying patients with overt kala-azar (p < 0.001, when compared to normal sera), and subjects with subclinical infection. More than 20% apparently healthy subjects with positive reaction to FML developed overt kala-azar during the following 10 months. In the screening of human blood donnors, a prevalence of 5% of sororeactive subjects was detected, attaining 17% in a single day. The GP36 glycoprotein of FHL is specifically reconized by human kala-azar sera. The immunoprotective effect of FML on experimental L. donovanii infection was tested in swiss albino mice. The protection scheemes included three weekly doses of FML, supplemented or not with saponin by the subcutaneous or intraperitoneal routes and challenge with 2x 10(7) amastigotes of Leishmania donovani. An enhancement of 80.0 % in antibody response (p<0.001) and reduction of 85.5 % parasite liver burden (p<0.001) was detected in animals immunized with FML saponin, unrespectivety of the immunization route.

Antibody response to Salmonella typhi lw human Schistosomiasis mansoni

Antibody response to Salmonella typhi O and H antigens was evaluated in 24 individuals with either hepatointestinal or hepatosplenic schistosomiasis mansoni before and after typhoid vaccination, and compared with that of non-infected controls. Before vaccination, Schistosoma-infected patients showed a higher frequency of positive antibody to O antigen and the same frequency to H antigen when compared with that of healthy individuals. However, those with hepatosplenic schistosomiasis showed higher titres of antibody to H antigen than those with hepatointestinal disease or healthy individuals. Infected subjects, particularly those with hepatointestinal disease, showed a decreased response after typhoid vaccine. Tins diminished ability to mount an immune response towards typhoid antigens dining schistosomiasis may interfere ivith the clearance of the bacteria from blood stream and, therefore, play a role in the prolonged survival of salmonella as obsewed in some patients with chronic salmonellosis associated with schistosomiasis.

Human immune response to triatomine embryo extract

Dipetalogaster maximus embryo extracts were used to stimulate peripheral blood mononuclear cells (PBMC) and in ELISA with sera either from Trypanosoma cruzi infected or non-infected individuals. The results showed that there was significant proliferative response and high antibody titers in sera of chagasic patients.

Effect of a booster-dose of rabies vaccine on the duration of virus neutralizing antibody titers in bovines

Humoral immune response using inactivated rabies vaccine was studied in 35 nelore cross-bred bovines of western region of São Paulo state. Ninety days after vaccination, 13 (92.8%) animals presented titers 30.5IU/ml, through mouse neutralization test. After 180 days, 9 (64.3%) sera showed titers 30.5IU/ml, after 270 days, only one (7.1%) showed a titer of 0.51IU/ml, and after 360 days, all animals showed titers < 0.5IU/ml. Group of animals receiving booster dose 30 days after vaccination presented, two months after, all with titers > 0.5IU/ml. At 180 days, 17 (80.9%) sera presented titers > 0.5IU/ml; at 270 days, 15 (71.4%), with titers 30.5IU/ml and at 360 days, 4 (19.0%), with titers 30.5IU/ml. Booster-dose ensured high levels of neutralizing antibodies for at least three months, and 240 days after revaccination, 71.4% of animals were found with titers 30.5IU/ml.

Dot enzyme-linked immunosorbent assay (dot-ELISA) for schistosomiasis diagnosis using dacron as solid-phase

Dacron and nitrocellulose were evaluated as matrices for the dot enzyme linked immunosorbent assay (dot-ELISA) for schistosomiasis and compared to indirect immunofluorescence (IMF). Titration of sera from 18 schistosomiasis patients against soluble worm antigen preparation (SWAP) was carried out and sera from healthy individuals from non-endemic areas were used as controls. The IMF was less sensitive than the dot-ELISAs, although the difference was not statistically significant (p > 0.05). The dot-ELISA based on nitrocellulose was as sensitive as that using dacron. Stability did not differ between nitrocellulose and dacron. Specificity was lower when dacron was used than when nitrocellulose was used, although the difference was not statistically significant (p > 0.05). In conclusion, this work showed that nitrocellulose and dacron performed similarly in dot-ELISA, suggesting that they may be used alternatively in population surveillance in endemic areas.

Experimental visceral leishmaniasis in high and low antibody - producer mice (selection IV-A)

Leishmaniasis is a typical parasite infection whose protective immunity depends on macrophage activation. Susceptibility to Leishmania donovani infection was compared in H (high antibody responder) and L (low antibody responder) mice from selection IV-A. H mice infected intravenously with 10(7) amastigotes of L. donovani were more susceptible to infection than their L counterparts. This higher susceptibility was characterized by a higher splenic and hepatic parasite burden. An increased splenic index was observed in both lines after sixty days of infection. This splenomegaly was caused, at least partially, by an increase in the number of splenic cells as determined by direct counts of cells from spleen. The results show that selection IV-A is susceptible to visceral leishmaniasis, with the H line being more susceptible than the L line.

Hepatitis B virus surface antigen (HBsAg) and antibody (anti-HBs) forming immune complexes in fulminant hepatitis

This paper reports an unusual pattern of serological HBV markers and the presence of HBsAg/anti-HBs immune complexes in serum samples from two patients with fulminant hepatitis from the Brazilian Western Amazon Basin. The diagnosis was made by both serologic tests and demonstration of antigen/antibody complexes by transmission electron microscopy. Concurrent Delta virus superinfection is also discussed.

Relação entre o diagnóstico sorológico (ELISA) e a gravidade da tuberculose pulmonar na infância

Foram estudadas 48 crianças de 0 a 13 anos através da realização do ensaio imunoenzimático ligado a enzima (ELISA) para pesquisa de anticorpos da classe IgG antiPPD, visando estabelecer correlação entre a resposta imune humoral medida pela sorologia e a gravidade da tuberculose, segundo formas radiológicas (leve, moderada e grave). A amostra foi composta de 29 crianças com tuberculose e 19 sem tuberculose comunicantes de tuberculose). Os valores médios (medianas) da densidade óptica do teste ELISA foram, respectivamente: 0,098 na forma gânglio-pulmonar (leve), 0,092 na forma pneumônica (moderada) e 0,134 na tuberculose miliar (grave). Nas crianças não tuberculosas com radiografia de tórax normal, o ELISA foi igual a 0,020. Os achados evidenciam valores mais elevados do teste sorológico relacionados à maior gravidade da doença (p= 0,0007).

Toxoplasma antibody and stool parasites in public school children, Rolândia, Paraná, Brazil

The occurrence of toxoplasmosis and enteroparasitosis was studied in 434 children from elementary schools in the rural and urban areas of Rolândia, Paraná State, Brazil. Sera and fecal samples from all the students were submitted to IFA for Toxoplasma gondii and coproparasitological tests, respectively. The children were tested by Amsler grid and 72 of them were examined for the presence of lesions compatible with ocular toxoplasmosis. Some variables were tested but none showed increased risk for toxoplasmosis. The distribution according to sex and age and also same other variables are presented and discussed. Correlations between Amsler's grid test, toxoplama RIFI, occurrence of eyes lesions and enteroparasitosis are also considered.