Genetic, epidemiological and biological analysis of interleukin-10 promoter single-nucleotide polymorphisms suggests a definitive role for-819C/T in leprosy susceptibility

Leprosy is a complex infectious disease influenced by genetic and environmental factors. The genetic contributing factors are considered heterogeneous and several genes have been consistently associated with susceptibility like PARK2, tumor necrosis factor (TNF), lymphotoxin-alpha (LTA) and vitamin-D receptor (VDR). Here, we combined a case-control study (374 patients and 380 controls), with meta-analysis (5 studies; 2702 individuals) and biological study to test the epidemiological and physiological relevance of the interleukin-10 (IL-10) genetic markers in leprosy. We observed that the -819T allele is associated with leprosy susceptibility either in the case-control or in the meta-analysis studies. Haplotypes combining promoter single-nucleotide polymorphisms also implicated a haplotype carrying the -819T allele in leprosy susceptibility (odds ratio (OR) = 1.40; P = 0.01). Finally, we tested IL-10 production in peripheral blood mononuclear cells stimulated with Mycobacterium leprae antigens and found that -819T carriers produced lower levels of IL-10 when compared with noncarriers. Taken together, these data suggest that low levels of IL-10 during the disease outcome can drive patients to a chronic and unprotective response that culminates with leprosy.

In vitro and skin lesion cytokine profile in Brazilian patients with borderline tuberculoid and borderline lepromatous leprosy

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Uso de células mononucleares da medula óssea no tratamento de tendinites induzidas experimentalmente em equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização histomorfológica da atrofia inflamatória proliferativa na próstata canina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Periodontal Tissue Engineering After Tooth Replantation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Human pulp responses to in-office tooth bleaching

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Correlation between antigenemia of Paracoccidioides brasiliensis and inhibiting effects of plasma in patients with paracoccidioidomycosis

Metabolites produced by pathogenic fungi may be involved in the pathogenesis of fungal infections consequently altering the defence mechanisms of the host. In this study the levels of Paracoccidioides brasiliensis antigens detected in the plasma of patients with paracoccidioidomycosis correlated with the suppression index detected by the low mitogenic response of peripheral blood mononuclear cells (PBMC) to phytohaemaglutinin (PHA). This inhibitory effect on lymphoproliferation was observed in the plasma of 58% of the patients, suggesting the presence of inhibitory factors. Plasma samples from paracoccidioidomycosis patients having or not having inhibitory factors showed no significant effect on chromosomes of lymphocytes from healthy individuals, However, these plasmas had a suppressive activity on the blastogenic response of these lymphocytes stimulated with PHA, that was independent of a cytotoxic effect. P. brasiliensis antigens added to the proliferative response of PBMC from healthy individuals stimulated or not stimulated with PHA showed a dose-dependent suppressor effect, reproducing the inhibitory effect of patients' plasma. We suggest that the antigens of P, brasiliensis present in the plasma of patients, even at low concentrations, can play an important role in the reduction of the cellular immune response and in the genesis of the immuneregulatory disturbances observed in paracoccidioidomycosis.

Enhanced airways responsiveness in rats depleted of sensory neuropeptides by neonatal capsaicin treatment

This study aimed to investigate the in vivo and in vitro reactivity of airway smooth muscle in rats depleted of sensory neuropeptides by treatment with capsaicin at neonatal stage. Wistar rats were neonatally injected with either capsaicin (50 mg/kg, s.c., 2nd day of life) or its vehicle (10% ethanol and 10% Tween 80, in 0.9% w/v NaCl solution) and used at adult ages (60-70 days later). Analysis of the lungs showed a higher number of infiltrating neutrophils, eosinophils and mononuclear cells into the peribronchiolar regions of capsaicin-pretreated rats compared to vehicle group. This was associated with a higher contraction index of bronchiolar wall in the capsaicin group. The in vitro tracheal reactivity in response to methacholine (full muscarinic agonist) and pilocarpine (partial muscarinic agonist) was also significantly higher in capsaicin-pretreated rats compared to vehicle group. In conclusion, the neuropeptide depletion by capsaicin neonatal treatment lead to marked contraction of the rat airways at adult age, suggesting a protective role for C fibers in the lungs. (C) 2003 Elsevier B.V. Ireland Ltd. All rights reserved.

Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion

Estudou-se sequencialmente, à microscopia eletrônica de transmissão, a interação entre Paracoccidioides brasiliensis (Pb) e células inflamatórias em hamsters inoculados por via intratesticular. Seis horas após inoculações havia predominância de neutrófilos, estando presentes algumas células mononucleares e eosinófilos. Os neutrófilos foram progressivamente substituídos por células mononucleares. Fungos viáveis apresentavam-se fagocitados ou circunscritos por células inflamatórias, geralmente com ampla interface hospedeiro-parasita. Fungos mortos ou degenerados eram acompanhados de interfase estreita. A camada externa da parede do Pb era às vezes quebrada quando em contacto com neutrófilos, em vários pontos, sendo os fragmentos dessa parede descamados e fagocitados. Células fúngicas pequenas com um único núcleo se relacionavam com ampla interfase enquanto as células maiores e multinucleadas apresentavam paredes irregulares, por vezes, contendo lomasoma e/ou estrutura semelhante à mielina. Diferentes padrões de interação do Pb com células do hospedeiro podem ser decorrentes do a fluxo de células inflamatórias funcionalmente diferentes ao local de inoculação ou à idade dos fungos ou ambos os fatores.

Histopathology of tick-bite lesions in naturally infested capybaras (Hydrochoerus hydrochaeris) in Brazil

In the present work features of tick-bite lesions were evaluated in capybaras naturally infested with Amblyomma cajennense and Amblyomma dubitatum ticks. Gross appearance of tick bite site was characterized by a mild swelling and erythema. Microscopic examination revealed the cement cone, a tube-like homogenous eosinophilic mass penetrating deep into the dermis. This structure was surrounded in the dermis by a cellular infiltrate and free eosinophilic granules and was associated to edema of variable intensity. Necrosis was a common feature deep in the dermis particularly at the far end of the eosinophilic tube. Hyperplasia, cellular edema and occasionally necrosis of keratinocytes could be seen at both sides of the ruptured epidermis. Cellular infiltrate was constituted overwhelmingly by polymorphonuclear leukocytes with eosinophilic granules. In capybaras cells with such features can be either eosinophils or heterophils (pseudoeosinophils), the latter being the equivalent of neutrophils of other mammals. Ultrastructural analysis of the cellular infiltrate revealed the predominance of heterophils over eosinophils. Mononuclear cells and mast cells and, in lesser numbers, basophils were also seen at skin attachment sites. The presence of heterophils in the reaction of capybaras against Amblyomma ticks is an outstanding feature but its role in the reaction to the tick is not known. It is however speculated that capybara heterophils might be associated with a more permissive environment for tick feeding and pathogen transmission as already shown for the equivalent cell type, the neutrophil, in the reaction of the dog against the Rhipicephalus sanguineus tick.

Correlation among immune response, morphogenesis of the granulomatous reaction and spleen lymphoid structure in murine experimental paracoccidioidomycosis

We studied the correlation among cellular immune response, the pattern of lung granulomatous lesions and alterations in spleen lymphoid structure in Swiss mice inoculated intravenously with Paracoccidioides brasiliensis strain 18. The animals were evaluated at 24, 48 and 96 h after infection and further studied weekly for 18 weeks by: (i) the macrophage migration inhibition test with phytohemagglutinin (PHA) and P. brasiliensis antigen (PbAg); and (ii) histopathology of the lung and spleen lesions. One group of animals was gamma -irradiated (8 Gy), infected under the same conditions and evaluated for the pattern of lung granulomatous lesions and spleen lymphoid structure at 24, 48 and 96 h after infection. During the first week of infection, the non-irradiated animals presented a positive response to PHA and PbAg, compact granulomas in the lungs and a typical hyperplasia of the spleen white pulp. However, from weeks 2 to 5, a depression of the cell-mediated immunity (CMI) response to PHA and PbAg was observed in association with granulomas presenting only large mononuclear cells and lacking both giant cells and a peripheral halo of small mononuclear cells. This pattern of granuloma formation was similar to that seen in gamma -irradiated animals, whose cells involved in CMI were absent. After week 7, the non-irradiated animals showed granulomas characterized by the presence of giant cells and a peripheral halo of small mononuclear cells. This type of granuloma was formed concomitantly with recovery of the CMI and of the lymphoid structure of the spleen. The results showed a correlation among granulomas composed of large mononuclear cells, hypoplasia of the splenic tissue and impaired CMI. This correlation indicated that although granuloma morphogenesis per se does not depend on the activation of CMI, this response is important at later stages during modulation of the cellular composition of the granulomas.

Effect of age on the gastrointestinal-associated mucosal immune response of humans

Age-related changes in gastrointestinal-associated mucosal immune response have not been well studied. Thus, we investigated the effect of age on this response and compared these responses to those of peripheral immune cells. Saliva, blood, and intestinal biopsies were collected from young and old healthy subjects to determine immunoglobulin (Ig) levels and to isolate peripheral blood mononuclear cells, intraepithelial lymphocytes (IELs), and lamina propria lymphocytes (LPLs). Although subject age did not influence the level of total IgA found in saliva, IgA levels in serum increased (p < .05) with age. Older subjects' peripheral blood mononuclear cell proliferation and IL-2 production were significantly lower than those of young subjects. LPLs from older subjects produced significantly less IL-2 in response to all stimuli than did that from the young. IEL's ability to proliferate and produce IL-2 was not affected by subject age. Thus, LPL but not IEL demonstrated an age-related decline in immune function similar to that seen in peripheral lymphocytes.

Inflammatory response to different endodontic irrigating solutions

Aim the aim of this study was to evaluate the inflammatory response to irrigating solutions injected into the peritoneal cavity of mice.Methodology Sixty mice received intra-peritoneal injections of 0.3 mL of 0.5% sodium hypochlorite, 2.0% chlorhexidine digluconate or phosphate buffered saline (PBS, control). Five animals of each group were sacrificed at 4, 24, 48 h and 7 days after the injection. Liquid from the peritoneal cavity of each animal was collected for the total and differential counting of inflammatory cells and protein leakage.Results the 0.5% sodium hypochlorite solution group had greater migration of neutrophils and mononuclear cells to the peritoneal cavity from 48 to 168 h (P < 0.05). There was a significant increase in protein leakage to the peritoneal cavity after 4 up to 48 h in the 0.5% sodium hypochlorite group compared to the control group. Protein leakage was similar in all groups at 168 h. The 2.0% chlorhexidine group had similar results to the control group at all time periods.Conclusions the 0.5% sodium hypochlorite solution induced an inflammatory response, however, the 2.0% chlorhexidine digluconate solution did not induce a significant inflammatory response.

SUPPORT OF PARACOCCIDIOIDES-BRASILIENSIS MULTIPLICATION BY HUMAN MONOCYTES OR MACROPHAGES - INHIBITION BY ACTIVATED PHAGOCYTES

The interaction of human monocytes or monocyte-derived macrophages and yeast-form Paracoccidioides brasiliensis was studied in vitro. Yeast cells were readily ingested by adherent monocytes or macrophages. Multiplication of P. brasiliensis, measured by growth as colony forming units (cfu) on a supplemented medium with good plating efficiency, was greater in monocyte co-cultures compared to the number of cfu obtained from complete tissue-culture medium (CTCM). Multiplication increased with time in macrophage cocultures, e.g., from two-six-fold in 24 h to nine-fold in 72 h. Microscopic observations indicated that ingested yeast cells multiplied inside macrophages. When monocytes were treated with supernate cytokines (CK) from concanavalin-A-stimulated mononuclear cells, then co-cultured with P. brasiliensis, multiplication was significantly inhibited compared with control monocyte co-cultures. Treatment of macrophages-derived from monocytes by culture in vitro for 3 days-for a further 3 days with CK resulted in maximal inhibition of multiplication over the subsequent 72 h. Similarly, when monocyte-derived macrophages (after culture for 7 days) were treated for 3 days with recombinant human gamma-interferon (IFN; 300 U/ml) or CK they restricted multiplication of P. brasiliensis by 65% and 95%, respectively, compared with control macrophages, Antibody to IFN abrogated the effect of IFN or CK treatment. These findings show that ingested P. brasiliensis can multiply in human monocytes or macrophages and that this multiplication can be restricted by activated monocytes or macrophages.

DELAYED-TYPE HYPERSENSITIVITY RESPONSE IN AN ISOGENIC MURINE MODEL OF PARACOCCIDIOIDOMYCOSIS

The specific delayed-type hypersensitivity (DTH) response was evaluated in resistant (A/SN) and susceptible (B10.A) mice intraperitoneally infected with yeasts from a virulent (Pb18) or from a non-virulent (Pb265) Paracoccidioides brasiliensis isolates. Both strains of mice were footpad challenged with homologous antigens. Pb18 infected A/SN mice developed an evident and persistent DTH response late in the course of the disease (90th day on) whereas B10.A animals mounted a discrete and ephemeral DTH response at the 14th day post-infection. A/SN mice infected with Pb265 developed cellular immune responses whereas B10.A mice were almost always anergic. Histological analysis of the footpads of infected mice at 48 hours after challenge showed a mixed infiltrate consisting of predominantly mononuclear cells. Previous infection of resistant and susceptible mice with Pb18 did not alter their DTH responses against heterologous unrelated antigens (sheep red blood cells and dinitrofluorobenzene) indicating that the observed cellular anergy was antigen-specific. When fungal related antigens (candidin and histoplasmin) were tested in resistant mice, absence of cross-reactivity was noted. Thus, specific DTH responses against P. brasiliensis depend on both the host's genetically determined resistance and the virulence of the fungal isolate.