Marcela Orsini

Combined diagnostic methods identify a remarkable proportion of asymptomatic Leishmania (Leishmania) chagasi carriers who present modulated cytokine profiles

Peripheral blood samples of 138 co-habitants from 25 families with recently diagnosed cases of visceral leishmaniasis in the Metropolitan Region of Belo Horizonte, Minas Gerais, Brazil, were analyzed by indirect fluorescent antibody test (IFAT), rK39 and Leishmania chagasi Enzyme Linked Immunosorbent Assay (ELISA), intradermal skin-test and Polymerase Chain Reaction (PCR) over a 12-month period. The cumulative positivity was significantly higher by PCR (29.7%) than by IFAT, rK39 ELISA, L. chagasi ELISA and intradermal skin-test (5.1%, 6.5%, 14.5% and 2.9%, respectively). In addition, the cytokine profile was measured in 16 of the 138 volunteers, of whom eight were asymptomatic carriers and eight were non-infected co-habitants. The innate immunity cells from asymptomatic carriers displayed, upon in vitro antigenic stimulation, a modulated increase in cytokine synthesis that was distinct from that observed in non-infected volunteers. This study suggests that the identification of a large proportion of asymptomatic carriers is facilitated when more than one diagnostic method is applied and that a mixed pattern of immune response is correlated with clinical status of asymptomatic individuals. These observations suggest also that asymptomatic infection by L. chagasi is a frequent event and that control programs could benefit by including this indicator in their interventions.

Antibody subclass profile against Leishmania braziliensis and Leishmania amazonensis in the diagnosis and follow-up of mucosal leishmaniasis

Sensitivities and specificities of IgG subclasses-ELISA and IgG-indirect fluorescent antibody test (IFAT) against Leishmania braziliensis (Lb) and L. amazonensis (La) antigens were determined in 17 patients with mucosal (ML) and 19 with muco-cutaneous (MCL) leishmaniasis. Using IFAT-IgG both antigens gave high sensitivities and were statistically similar, being 89.5% with La and 100% with Lb. Using ELISA, the highest sensitivity was achieved with total IgG for ML (94.7% with both antigens) and MCL (100% with both antigens). Cross-reactivity, observed with Chagas disease and malaria sera reduced the specificity of the IgG-based assays, being 50 to 70% with IFAT and 40 to 70% with ELISA. An increase in specificity was obtained with IgG1-ELISA (90% with Lb and 100% with La). Serum levels of anti-Lb-IgG and IgG3 dropped 90 days after treatment. IgG subclasses antibody detection constitute an valuable alternative to increase the efficiency of sorological diagnostics of ML/MCL.

Oral fluids for the immunodiagnosis of Schistosoma mansoni infection

Saliva and oral transudate were evaluated for their potential as human specimens in the detection of IgG antibodies against soluble Schistosoma mansoni egg antigen (SEA). Preliminary laboratory testing of 49 subjects, 37 with parasitological proven infection and 12 negative controls, displayed 100% sensitivity in ELISA using serum and oral transudate and 94.6% using saliva. The specificity of the ELISA with serum was 100% versus 91.7% with both oral fluids. Significant Spearman rank correlations of anti-SEA IgG levels with egg counts were observed for serum, oral transudate and saliva (P < 0.05). The sensitivity of dot-ELISA was 100% for serum, 89% for transudate and 81% for saliva, and specificity was 100% for all 3 samples. The immunodiagnostic value of ELISA for the detection of anti-SEA IgG antibodies in oral transudate was further evaluated in 197 individuals from an endemic area of Brazil. The ELISA using serum and oral transudate showed sensitivities of 98.8% and 100% respectively and specificities of 67.8% and 64.3% respectively. Use of oral fluids for the diagnosis of S. mansoni infection was equivalent to sera with respect to test efficacy, offering an alternative to blood collection.

Identification of Leishmania chagasi from skin in Leishmania/HIV co-infection: a case report

A case of HIV/Leishmania co-infection presenting both visceral and cutaneous manifestations is reported. Leishmania infection was confirmed by conventional methods (parasitological approach and serology) and by PCR. Leishmania chagasi isolated from the skin lesion was characterized by enzyme electrophoresis and by restriction fragment length polymorphism of the internal transcribed spacer of the ribosomal gene.

Low frequency of side effects following an incidental 25 times concentrated dose of yellow fever vaccine

In August/1999, a group of 14 adults from the staff of a private hospital in Contagem - Minas Gerais State, Brazil, received unintentionally a 25 times concentrated dose of the 17-DD yellow fever vaccine (Bio-Manguinhos), due to a mistake at the reconstitution step. All patients were clinically and laboratorially evaluated at days 5, 13 and 35 post vaccination. Frequency of side effects and clinical observations of this group of individuals were not different from the observed in recipients immunized with normal doses of the vaccine. At the second and third evaluation none of the subjects reported symptoms. None of the patients presented abnormalities at the physical examination at none of the time points and in all cases the blood examination was normal, except for a reduced number of platelets that was detected in one subject at the first and second evaluation and reverted to normal at third evaluation. At the first evaluation point, 8 subjects were serum negative and 6 serum positive for yellow fever at the plaque reduction neutralization test. In 5 subjects the observed titre was 10 times higher as the baseline of 2.36 Log10 mUI/ml. The samples collected at second and third evaluation (13th and 35th days) demonstrated that all subjects responded to the vaccination with the exception of one that did not present a positive result in any of the samples collected. This evaluation confirms the safety of the 17-DD yellow fever vaccine.