It is clear that leukotrienes mediate inflammatory response; new aspects of leukotriene function have recently been described. In this study, we demonstrate that leukotrienes are key chemical mediators in the control of parasite burdens in mice infected with Strongyloides venezuelensis. High leukotriene levels were detected in the lungs and small intestines of Swiss mice. In infected Swiss mice treated with MK886, a leukotriene synthesis inhibitor, numbers of adult worms, and eggs/g/feces were greater than in infected-only animals. The MK886 treatment inhibited leukotriene B4 production in the lungs and small intestines, albeit on different postinfection days. Similarly, parasite burdens and eggs/g/feces were greater in 5-lipoxygenase−/− mice than in wild-type animals. These observation were confirmed by histopathological study of the duodena. We subsequently observed significant lower numbers of eosinophils and mononuclear cells in the blood, peritoneal cavity fluid, and bronchoalveolar lavage fluid of Swiss mice treated with MK886. In the lung parenchyma of infected animals, MK886 significantly inhibited synthesis of IL-5 at the beginning of infection, whereas levels of IL-12 increased progressively throughout the postinfection period. However, levels of leukotriene C4, PGE2, TNF-α, IL-3, IL-4, IFN-γ, and IL-10 were comparable between the treated and untreated groups. Nevertheless, IgE and IgG1 (but not IgG2a) synthesis was also significantly inhibited by MK886 administration. Therefore, in S. venezuelensis-infected mice, adult worm and egg burdens are leukotriene dependent. These findings indicate potential immunostimulatory strategies involving leukotriene administration, and may serve as an alert to physicians treating Strongyloides stercoralis-infected patients presenting asthma-like symptoms because use of 5-lipoxygenase inhibitors may worsen the infection.

To characterize the physicochemical properties of the mesangial IgA in primary IgA nephropathy, acid-eluates from percutaneous renal biopsies of 20 patients were examined. The acid-eluates were obtained from 1287 +/- 498 glomerular sections. The IgA content (mean 15 +/- 10 ng) represented 0.4% of the total eluted proteins. To analyze the molecular weight and the charge of eluted IgA, 11 eluates were subjected to high pressure liquid chromatography (at pH 6.8 and/or pH 3.5) and five eluates to isoelectric focusing on agarose. IgA was detected in the fractions by an IgA-RIA. Comparison of the elution profiles at different pH showed a statistically significant decrease of the excluded IgA peak (greater than or equal to 1,000,000 daltons), and a significant increase of polymeric IgA peaks (1,000,000-320,000 and 320,000 daltons) in acidic chromatography, as compared to non-dissociating conditions. Under acidic conditions, polymeric IgA represent 64% of total eluted IgA. Secretory component binding to polymeric IgA was demonstrated in four out of eight eluates tested. The isoelectric point (pI) of eluted IgA ranged from 4.5 to 5.6, contrasting with the broader and more neutral pI of normal serum IgA (4.5 to 6.8). This study shows that the multimeric nature of IgA, the formation of IgA complexes, and the anionic charge of IgA are likely to be involved in the mesangial IgA deposition in idiopathic IgA nephropathy.

In the present work we demonstrate that hamsters infected with L. donovani eliminate large quantities of immunoglobulins in the urine. This alteration is clearly a consequence of a conspicuous immune complex glomerulonephritis readily detectable 7 days after the beginning of infection. L. donovani antigens and hamsters immunoglobulins (Igs) were revealed as granular deposits in the mesangial areas and contiguous loops of the glomeruli. Histopathological alterations such as focal mesangial proliferation with progression to diffuse proliferation were observed in the first 3 weeks of infection. From day 35 onwards, all diseased animals presented large deposits of amyloid material of predominantly glomerular localization. In consonance with these alterations, Igs were detected in the urine by day 21 of infection and their concentration increased substantially with the progression of disease. In contrast, serum Igs increased until day 21, when their concentration dropped steadily.

Background The thermo-dimorphic fungi Paracoccidioides spp. are the etiological agents of paracoccidioidomycosis. Although poorly studied, paracoccin (PCN) from P. brasiliensis has been shown to harbor lectinic, enzymatic, and immunomodulatory properties that impact disease development. Methods Mutants of P. brasiliensis overexpressing PCN (ov-PCN) were constructed by Agrobacterium tumefaciens-mediated transformation. Ov-PCN strains were analyzed and inoculated intranasally or intravenously to mice. Fungal burden, lung pathology, and survival were monitored to evaluate virulence. Electron microscopy was used to evaluate the size of chito-oligomer particles released by ov-PCN or wild-type strains to growth media. Results ov-PCN strains revealed no differences in cell growth and viability, although PCN overexpression favored cell separation, chitin processing that results in the release of smaller chito-oligomer particles, and enhanced virulence. Our data show that PCN triggers a critical effect in the cell wall biogenesis through the chitinase activity resulting from overexpression of PCN. As such, PCN overexpression aggravates the disease caused by P. brasiliensis. Conclusions Our data is consistent with a model in which PCN modulates the cell wall architecture via its chitinase activity. These findings highlight the potential for exploiting PCN function in future therapeutic approaches.

Severe forms of paracoccidioidomycosis (Pcm) are accompanied by intense immunological involvement characterized by depression of the cell-mediated immune response and by high levels of antibodies in serum with no protective function. These changes can be reversed by antifungal treatment. It has been suggested that antigens of Paracoccidioides brasiliensis released into the circulation during the active phase of the disease may be involved in the genesis of the changes in the immune response. In the present study, we evaluated the antigenemia of patients with Pcm using a competitive enzyme-linked immunosorbent assay (ELISA-c) capable of detecting 6 ng of antigen per ml of serum. Twenty-seven of 88 serum samples tested gave positive results, with the highest frequency of positivity being detected in patients with the severe acute form of the disease; these patients had the highest antigen levels (0.03 to 3.4 micrograms/ml). Follow-up of one case showed a correlation between antigen levels in serum and evolution of the disease. False-positive reactions were observed in sera from patients with histoplasmosis, aspergillosis, and cryptococcosis. The results indicate that the described method has potential for clinical application, especially with respect to the evaluation of disease activity. Quantification of fungal antigens in the serum of patients with active Pcm represents an objective parameter for the study of the physiopathology of the disease.

Monocyte migration into tissues, an important event in inflammation, requires an intricate interplay between determinants on cell surfaces and extracellular matrix (ECM). Galectin-3 is able to modulate cell-ECM interactions and is an important mediator of inflammation. In this study, we sought to investigate whether interactions established between galectin-3 and ECM glycoproteins are involved in monocyte migration, given that the mechanisms by which monocytes move across the endothelium and through the extravascular tissue are poorly understood. Using the in vitro transwell system, we demonstrated that monocyte migration was potentiated in the presence of galectin-3 plus laminin or fibronectin, but not vitronectin, and was dependent on the carbohydrate recognition domain of the lectin. Only galectin-3-fibronectin combinations potentiated the migration of monocyte-derived macrophages. In binding assays, galectin-3 did not bind to fibronectin, whereas both the full-length and the truncated forms of the lectin, which retains carbohydrate binding ability, were able to bind to laminin. Our results show that monocytes migrate through distinct mechanisms and selective interactions with the extracellular matrix driven by galectin-3. We suggest that the lectin may bridge monocytes to laminin and may also activate these cells, resulting in the positive regulation of other adhesion molecules and cell adhesion to fibronectin.

Algal lectins induced neutrophil migration, which was inhibited by a monosaccharide, contrasting with the view that they only recognize complex oligosaccharides. Neutrophil chemotaxis assays are appropriate to study low molecular mass lectins containing a single carbohydrate recognition domain, as is the case of some lectins from algae and mammals.

SUMMARYIn il previous report analysing kidney sections by immunofluorescence we showed that hamsters infecied wilh L. di>nt)ntnidc\clop a glomerulonephrilis (GN) associated wilh deposition of hatiister immunoglobulins and parasite antigens in the glomcruli. In this study we characterize these immune cotiiponents eluted from the kidneys. The eluted immunoglobulins showed specificity for L. donovani antigens and hamster imtnunoglobulins (rheumatoid factor-like aclivity). The lour isotypcs IgGl. lgG2. IgA and IgM wcrcdcicclcd. Several /., tloncnini anugcns were detected in the renal eluates by Western blot and itTimuneprecipitation using '-^l-labellcd cluatcs. Proteins with tnol. wt of 134, 82, 52. 31, and 26 kD were detected by Western blot and proteins with 134, 110, 93, 89 and 48 kD were detected by imtnunoprccipitallon. With ihe exception ofthe 134 kD protein which was recognized by both rabbit anti-promastigote ;md rabbit anti-imiastigotc sera all the others were recognized only by the an ti-Limastigolc scrum. The 134 kD protein was the only one isolated from the kidneys of infected hamster immunocotnplexcd with IgG and was the only one detected iti a protnastigole lysate using IgG from L. donomni-'mfccied hamsters.Keywords Leishmania donovani glomerulonephritis experimental kala-azur immune complexes IMRODUCTION Kala-azar or visceral leishmaniasis is a disease caused by /.. donovani. an obligatory intracellular protozoan, which infccls e.vclusiveiy the macrophages of the rcticulocndolliclial system, predominantly in ihc spleen, liver ;itid bone miirrow. The infection is clinically characterised by lever. hcp;ilosplenoniegaly, anaemia and Ieucopenia sis [15][16][17] ;md in olher protozoan-induced infections such as malaria [!S|. schistosomiasis [19] and trypanosomiasis [20], A common trait observed in renal lesions resulting from protozoan infection is the presence of immunodeposits occurring in a granular pattern. The characicri/ation ofthe molecules found iti these deposits can help lo understand why they accumulate in the renal structures and how they cause renal injury, Thehamster is a valuable animal model for kala-azar. In this animal the disease results in hepatosplenomegaly, anaemia [2l[ heightened serum globulin levels [22j, imnuinosiippression [23j. polyclonal B cell activalion [24.24a] and icnal involvctncnt [25,25al, In previous reports we described that hamsters infected with /... donovani developed a conspieuous itnmunc complex glomerulonephrilis (GN) dclined by ihc presence ofgranular deposits of /,. dtinovani antigens and hamster imniLinoglobuiins in the mcsangium and glomeruli loops .In this study we characterize the imtnunoglobulins and the L. donovani antigens eluted from the kidneys of iniected hatTisters,