Receptor Antagonist of IL-13 Exerts a Potential Negative Regulation During Early Infection of Human Schistosomiasis.

The pathology of schistosomiasis is associated with the formation of granulomas, and this process is associated with liver fibrosis. Studies indicate that Th1 cytokines reduce fibrosis in schistosomiasis, while Th2 cytokines play a part in the progression of fibrosis, and IL-13 has a critical role in this process. The IL-13Rα2 receptor, known as a 'receptor antagonist' binds with high affinity to IL-13, and studies have identified that this plays a part in reducing fibrosis and the size of granulomas. The objective of this study was to evaluate the function of IL-13Rα2 and cellular immune response in hepatic fibrosis. A negative correlation between IL-13Rα2 and IL-13 was found, suggesting an increase in cytokine in early fibrosis. Initially, a negative correlation between IFN-γ and IL-13 was found in patients without fibrosis, and subsequently, this correlation was found to be positive in patients with severe fibrosis, thereby highlighting a new mechanism for regulating the progress of periportal fibrosis. There was a positive correlation between the profiles of Th1 and Th2 cytokines, suggesting the presence of both responses, thus regulating the disease. The results contribute to a better understanding of the immune mechanisms that control the process of hepatic fibrogenesis in schistosomiasis in humans.

Immunological diagnosis of tuberculosis based on recombinant antigens ESAT-6 and CFP-10 in children from an endemic area in northeast Brazil.

Diagnostic tests for tuberculosis (TB) using interferon gamma (IFN-γ) responses produced by T lymphocytes after stimulation by early secretory antigen target 6 (ESAT-6), culture filtrate protein 10 (CFP-10) or purified protein derivate (PPD) were carried out using ELISA (enzyme-linked immunosorbent assay) in whole blood culture supernatants from children with suspected TB disease (n=21), latent TB infection (LTBI; n=17) and negative controls (NC; n=21) from Recife, Pernambuco, Brazil. The results were analysed using the ROC (receiver operating characteristic) curves and the areas under the curve (AUC) generated varied from 0.5 to 1.0 with higher values indicating increased discriminatory ability. Comparisons of AUCs were made using non-parametric assumptions, and the differences were considered significant if P<0.05. The ROC curve showed a statistical difference (P = 0.015) between the LTBI and NC groups with an AUC of 0.731, TB disease and NC (AUC=0.780; P=0.002) and a group with TB (latent infection+disease, n=38) and NC (AUC=0.758; P = 0.001) when the antigen used was ESAT-6. No statistical difference was found between the groups when CFP-10 or PPD was used. In conclusion, the ESAT-6 test may be the most appropriate for diagnosis of childhood TB, both LTBI and TB disease, when associated with epidemiological and clinical data, especially in endemic areas such as Brazil.

Immune factors and immunoregulation in tuberculosis.

Pathogens causing tuberculosis and other chronic infectious diseases of major public health importance commonly have complex mechanisms involved in their persistence in the host despite specific and sometimes strong immune responses. These diseases are also associated with the lack of efficient vaccines, difficult therapeutics and a high mortality rate among susceptible individuals. Here, we will review features of the host immune response that contribute to the occurrence of disease. In addition, we propose that the immune responses observed in tuberculosis cannot be interpreted solely on the basis of a Th1-Th2 counter-regulatory paradigm since there is growing evidence that natural regulatory T cells may play an important role in the regulation of host immune responses against Mycobacterium tuberculosis. Thus, the development of more effective vaccines against this bacterial disease should take into account the role of natural regulatory T cells in the progression to severe disease and persistence of infection. Finally, new treatments based on manipulation of regulatory T cells should be investigated.

Immune factors and immunoregulation in tuberculosis

Pathogens causing tuberculosis and other chronic infectious diseases of major public health importance commonly have complex mechanisms involved in their persistence in the host despite specific and sometimes strong immune responses. These diseases are also associated with the lack of efficient vaccines, difficult therapeutics and a high mortality rate among susceptible individuals. Here, we will review features of the host immune response that contribute to the occurrence of disease. In addition, we propose that the immune responses observed in tuberculosis cannot be interpreted solely on the basis of a Th1-Th2 counter-regulatory paradigm since there is growing evidence that natural regulatory T cells may play an important role in the regulation of host immune responses against Mycobacterium tuberculosis. Thus, the development of more effective vaccines against this bacterial disease should take into account the role of natural regulatory T cells in the progression to severe disease and persistence of infection. Finally, new treatments based on manipulation of regulatory T cells should be investigated.

Preliminary results on the effects of CD40/CD40L interactions and SAC-induction on IFN-gamma expression in human schistosomiasis.

In this communication the authors analyzed the pattern of expression of IFN-gamma as a surrogate type 1 response in different clinical forms of schistosomiasis in response to stimulation involving T-cell dependent and T-cell independent pathways, to investigate which pathways were functional in human schistosomiasis, and to further characterize the nature of Th1 response impairment in this parasitic disease.

Specific situations related to acute schistosomiasis in Pernambuco, Brazil.

The present work reports on two epidemiological episodes resulting in acute schistosomiasis involving wealthy persons living in the State of Pernambuco, Brazil. The authors discuss the epidemiological, clinical and serologic characteristics of the acute infections and also the way in which the conditions for transmission occurred.

[An outbreak of acute schistosomiasis at Porto de Galinhas beach, Pernambuco, Brazil]. / Epidemia de esquistossomose aguda na praia de Porto de Galinhas, Pernambuco, Brasil.

We recently confirmed several cases of acute schistosomiasis in Porto de Galinhas beach, Northeast Brazil. A total of 662 patients were diagnosed by parasitological and clinical examinations. The infection likely occurred during the September 7 national holiday, when heavy rainfall flooded the Ipojuca River and people were infected when the water covered their yards. Families were continuously exposed to infection for a period of three weeks until the water had completely dried up. Previous investigation suggests that snail vectors were introduced as a result of landfill in marshy areas. The swamp-flooding of such areas facilitated the emergence of slums surrounded by snail breeding sites. Heavy rainfall caused open-air sewage ditches to overflow, allowing for infection of snails by Schistosoma mansoni. Thus, continuous floods were responsible for the spread of human infection. Clinical and laboratory results identified 62% of acute cases of S. mansoni. Complementary studies are being conducted to define the impact and epidemiological meaning of the acute schistosomiasis outbreak.

Mapping of the N terminus of the Schistosoma mansoni tegumental antigen Sm15 to its predicted precursor protein.

Sm15 is a major Schistosoma mansoni 15 kDa tegumental antigen, resulting from the proteolytic processing of a larger precursor. The amino terminus of Sm15 was identified by direct amino acid sequencing, and the antigen was tentatively mapped to the segment spanning amino acids 362-497 of the precursor. This will allow subsequent studies to elucidate the possible immunological role of proteolytic processing in schistosomiasis.

Onset and evolution of nephropathy in rats with spontaneous diabetes mellitus.

The occurrence of renal diabetic complications was studied in diabetic nonobese IIM/FmeSS (eSS) rats. The results were compared with eumetabolic Wistar rats paired by sex and age. Between 6 and 12 months of age, eSS male rats had higher fructosamine values and glucose intolerance as well as increasing proteinuria and uremia. Enhancement in water, calcium and phosphorus fractional excretion with a concomitant lower sodium excretion, was observed from 12 months of age on. 18- and 21-month-old eSS rats exhibited fasting hyperglycaemia and rising values of fructosamine, glucose intolerance and glycosuria. Simultaneously, a notorious worsening of proteinuria as well as alterations in glomerular filtration were verified. Optic microscopy of 12-month-old eSS rat kidneys showed areas of tubular dilatation with protein cylinders. In 21-month-old eSS animals, kidneys appeared overtly damaged. Increased capsular, glomerular and Henle's thin loop diameters were verified in 12- and 21-month-old eSS rats. Glomeruli showed diffuse hypertrophy of mesangial tissue and thickening of the basement membrane. Areas of markedly atrophic and dilated tubules containing acidophilic proteinaceous material were observed. At age of 21 months, kidneys of eumetabolic Wistar control rats presented foci of interstitial and pielic inflammatory infiltrates.

Cytokine production in acute versus chronic human Schistosomiasis mansoni: the cross-regulatory role of interferon-gamma and interleukin-10 in the responses of peripheral blood mononuclear cells and splenocytes to parasite antigens.

The contribution of interleukin (IL)-10 and interferon (IFN)-gamma to the regulation of type 1 and type 2 cytokine responses was investigated in Brazilians with different clinical forms of schistosomiasis mansoni. Cells from members of a family with acute intestinal schistosomiasis responded to schistosomal soluble egg antigen (SEA) or soluble adult worm antigen preparation (SWAP) with greater amounts of IFN-gamma than did cells from several patients with chronic intestinal schistosomiasis; IL-10 levels were similar. Neutralization of IL-10 had no effect on the SEA-specific IFN-gamma response in patients with acute infection, whereas SWAP-induced IFN-gamma was increased in both groups. Anti-IL-10 also up-regulated SEA-specific IFN-gamma protein and mRNA responses in most splenocyte cultures from hepatosplenic schistosomiasis patients but had no effect on antigen-specific IL-4 or IL-5 production. Neutralization of IFN-gamma resulted in a comparable increase in SWAP-specific IL-10 and IL-5, while IL-4 was not affected. These studies demonstrate that early disease in schistosomiasis is associated with a significant IFN-gamma response and that IL-10 contributes to the suppression of that response during both early and chronic infection.

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.

Vaccines against human parasitic diseases: an overview.

There are no vaccines currently available to control the major human parasitic diseases, although there is evidence of acquired immunity and resistance to reinfection in most of the parasitic infections. The present manuscript concentrates on the vaccines for parasitic diseases that are in the most advanced stages of development: malaria, leishmaniasis and schistosomiasis. Vaccines for malaria and leishmaniasis have been taken to clinical trials while vaccines for schistosomiasis are being considered for Phase I (assessment of safety and immune responsiveness in volunteers). We have attempted to give a factual account of the present status of knowledge of vaccines against human parasitic diseases, emphasizing both the successes and setbacks. We do not intend to cover the enormous literature in the field but have concentrated on the most promising antigenic preparations. Finally, some new approaches for the development of vaccines are discussed including nucleic acid vaccines and the use of cytokines as adjuvants.

Influence of gonadectomy in eSS diabetic rats.

The influence of gonadectomy on some variables related to the diabetic syndrome was studied in the eSS line of rats, a nonobese model of spontaneous non-insulin-dependent diabetes, whose biochemical and histopathological manifestations are more severe in males than in females. Rats were gonadectomized at 90 days of age. Spayed animals showed higher body weight, impaired intolerance to glucose at 9 and 12 months of age, lower insulinemia and a decreased number of large pancreatic islets. Castrated rats revealed lower body weight when compared with controls. However, those males did not evidence impairment in the intolerance to glucose, changes insulinemia or remarkable modifications in endocrine pancreas histology. In kidneys, a lower cellular area in superficial proximal convoluted tubules was noticed. Despite the lower biomass registered in orchidectomized animals, their diabetic evolution was not modified. Conversely, ovariectomy appeared to be a worsening factor.

Expression of recombinant antigens in Escherichia coli: application on immunochemical studies of Schistosoma mansoni tegumental antigens.

Sm15 and Sm13 are recognized by antibodies from mice protectively vaccinated with tegumental membranes, suggesting a potential role in protective immunity. In order to raise antibodies for immunochemical investigations, the genes for these antigens were expressed in pGEX and pMal vectors so that comparisons could be made among different expression systems and different genes. The fusion proteins corresponding to several parts of the gene for the precursor of Sm15 failed in producing antibodies recognizing the parasite counterpart. On the other hand, antibodies raised against Sm13 MBP-fusion proteins recognized the 13 kDa tegumental protein. Thus the peculiarities of the gene of interest are important and the choice of the expression system must sometimes be decided on an empirical basis.

Factors involved in Schistosoma mansoni infection in rural areas of northeast Brazil.

Two contiguous villages in Tracunhaém county (State of Pernambuco), endemic for schistosomiasis, were studied: Itapinassu (138 inhabitants) and São Joaquim (91 inhabitants). Agriculture predominates in the former region while ceramics is the main activity in the latter. Although no statistical difference was found regarding prevalence, severe infection (> 400 epg) predominated in Itapinassu, probably related to the kind of occupation. No association was found between parasite burden and severity of disease, in spite of the high infection rates for Schistosoma mansoni in both communities (approx. 60%). Typical epidemiological features of schistosomiasis such as age-related prevalences and intensities of infection (high in children, low in adults) were also mutual characteristics. Nutritional status determined through anthropometric evaluation was carried out by measuring specific anthropometric indicators. A deficit of energy intake, as well as vitamin A and riboflavin deficiencies were detected. The prevalence of moderate or severe undernutrition in patients under 18 years old was 21.9% in Itapinassu and 24.1% in São Joaquim. In this group an association was found between prevalence of schistosomiasis and chronic undernutrition. Similarly, for patients over 18 year old the prevalence of undernutrition was higher than 20%. However, in this case no association between nutritional status and either prevalence of schistosomiasis or parasite burden could be detected. The two communities had not been treated for eight years.

PIP: Patterns of schistosomiasis infection were compared in two contiguous endemic villages in Northeast Brazil's Tracunhaem County (Pernambuco State): Itapinassu (138 inhabitants) and Sao Joaquim (91 inhabitants). The overall prevalence of schistosomiasis in Tracunhaem State was 58.7%; this rate was 61.6% in Itapinassu and 54.2% in Sao Joaquim. Severe infection (400 epg) was more prevalent in Itapinassu (35.1%) than Sao Joaquim (13.3%) and ultrasound revealed more severe pathologic changes (e.g., periportal fibrosis, right liver lobe shrinkage, left lobe and spleen enlargement) in the former village. The higher prevalence of severe infection in Itapinassu is likely related to the predominance of agricultural occupations; in Sao Joaquim, most residents are engaged in ceramics. Schistosomiasis prevalence was significantly positively associated with increasing age, male sex, residence in the village for more than 5 years, daily water contact, fishing, laundering, less than a 10 m distance from an infected stream, lack of cesspools, and chronic undernutrition. A deficit of energy intake, as well as vitamin A and riboflavin deficiencies, was detected in both villages. The role of each of these factors (especially nutritional status) will be analyzed further in order to develop an integrated model for local control of the disease.

Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection. II. Corynebacterium parvum.

The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied. The C. parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks), thirty days before (prophylactic treatment) or after infection (curative treatment). The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0.018), resulting in 44% protection. The "in vivo" leukocyte response in both prophylactic and curative groups was higher as compared to the infected/non treated group (p = 0.009 and p = 0.003, respectively). Tissue reactions were described in the experimental and control groups, but there were not remarkable differences among them. The possible biological implications and relevance of the findings for the defensive response of the host and control of schistosomiasis are discussed.

Effects of dietary sucrose option on the diabetic syndrome of the eSS rat.

The eSS rat is a model of human spontaneous non-insulin-dependent diabetes. Male eSS rats were divided at the age of 4 months into two groups (eSSA and eSSB), both receiving the usual commercial balanced diet with sucrose also made available to eSSA. Sucrose intake did not imply a higher caloric diet, and no differences were found between groups in body weight and plasma triglyceride levels from 6 to 12 months of age. Sucrose option resulted in lower protein, lipid and carbohydrate intakes in eSSA animals. Plasma glucose values were higher in eSSA at different times of the tolerance curve. Likewise, eSSA kidneys showed significantly higher capsular and glomerular diameters and there was a discrete PAS-positive thickening of their basement membrane. We conclude that prolonged ad libitum sucrose intake, without weight gain, causes a moderate metabolic impairment and renal lesions in the eSS diabetic rat.

"In vivo" leukocyte chemotaxis in experimental mice Schistosoma mansoni infection.

The "in vivo" chemotaxis was studied in C57Bl/10 mice 10, 30, 50 and 60 days after a Schistosoma mansoni infection in comparison to a control group (uninfected mice). Staphylococcal protein A was injected into a connective tissue air pouch of control and experimental mice and the leukocyte chemotaxis was counted. A decrease in polymorphonuclear (PMN) leukocyte response was found in infected mice in comparison to the control group (p < 0.05). The 10 day infected mice showed a decreased PMN leukocyte response respecting the control group (p < 0.05) and this finding became more evident 30 and 50 days post-infection. Although the PMN leukocyte response of 60 day infected mice increased in comparison to 50 day infected animals, it was still significantly lower the control response. The mononuclear leukocyte response was not significantly different between infected or uninfected mice.

Standardization of the dot enzyme-linked immunosorbent assay (dot-ELISA) for experimental plague.

A dot enzyme linked immunosorbent assay (dot-ELISA) was previously developed to detect specific antibodies in rabbits sera immunized against F1A protein obtained from Yersinia pestis. This antigen was covalently linked onto the surface of dacron (polyethyleneterephthalate). Here, standard conditions are described for the optimization of this procedure: an amount of 20 ng of F1A protein was fixed onto dacron; anti-rabbit IgG peroxidase conjugate diluted 1:8,000 and 30% non-fat instant milk as blocking substance were used throughout the method. This procedure was compared with that employing nitrocellulose as solid-phase which showed to be more sensitive. However, the method based on dacron did not show false positive reactions against non-immunized rabbits sera at low antigen amount and diluted anti-IgG peroxidase conjugate.