Influence of tumor necrosis factor-alpha on the ability of monocytes and lymphocytes to destroy intraerythrocytic Plasmodium falciparum in vitro.

It has been shown that administration of TNF-alpha causes an increase of survival of plasmodium-infected mice. However, this anti-parasitic effect cannot be reproduced in vitro upon direct incubation of the cytokine with the parasite. This suggests that TNF-alpha may act through modulation of some plasmodicidal mechanism not yet clarified. We evaluated the effect of exogenous TNF-alpha on the phagocytosis of Plasmodium falciparum-infected erythrocytes by monocytes and its influence on the ability of monocytes and lymphocytes to inhibit parasite growth. The capacity of endogenous TNF-alpha to influence the ability of monocytes to inhibit the parasite was also verified. We found that addition of 33 ng TNF-alpha/mL to cultures of human monocytes and P. falciparum-infected erythrocytes increased the phagocytic index from 3.8 to 7.8 in the presence of serum containing P. falciparum antibody. TNF-alpha increased the capacity of monocyte plus lymphocyte to inhibit parasite growth by about 3 times at 0.5 and 5 ng/mL. Sera from severely ill P. falciparum-infected individuals inhibited the parasite growth, but addition of anti-TNF-alpha antibody was unable to modify this inhibition. These data show that TNF-alpha can increase the phagocytic capacity. This was probably due to an increased expression of Fc receptors on monocytes or to the modulation of Fc receptor signaling pathways by signals originating from the binding of TNF-alpha to its receptors. TNF-alpha also acted on lymphocytes plus monocytes by increasing the inhibition of P. falciparum by a mechanism not related to phagocytosis. These findings suggest that TNF-alpha has a pleiotropic anti-malaria effect and that this protective effect depends on the interplay of different factors, such as monocytes/macrophages, lymphocytes, and antibodies, in addition to other cells and molecules.

Coevolutionary networks: a novel approach to understanding the relationships of humans with the infectious agents.

Human organism is interpenetrated by the world of microorganisms, from the conception until the death. This interpenetration involves different levels of interactions between the partners including trophic exchanges, bi-directional cell signaling and gene activation, besides genetic and epigenetic phenomena, and tends towards mutual adaptation and coevolution. Since these processes are critical for the survival of individuals and species, they rely on the existence of a complex organization of adaptive systems aiming at two apparently conflicting purposes: the maintenance of the internal coherence of each partner, and a mutually advantageous coexistence and progressive adaptation between them. Humans possess three adaptive systems: the nervous, the endocrine and the immune system, each internally organized into subsystems functionally connected by intraconnections, to maintain the internal coherence of the system. The three adaptive systems aim at the maintenance of the internal coherence of the organism and are functionally linked by interconnections, in such way that what happens to one is immediately sensed by the others. The different communities of infectious agents that live within the organism are also organized into functional networks. The members of each community are linked by intraconnections, represented by the mutual trophic, metabolic and other influences, while the different infectious communities affect each other through interconnections. Furthermore, by means of its adaptive systems, the organism influences and is influenced by the microbial communities through the existence of transconnections. It is proposed that these highly complex and dynamic networks, involving gene exchange and epigenetic phenomena, represent major coevolutionary forces for humans and microorganisms.

[Effect of bed nets impregnated with deltamethrin on malaria morbidity in an area of the Brazilian Amazones]. / Ação de mosquiteiros impregnados com deltametrina sobre a morbidade da malária em uma área da Amazônia Brasileira.

In 1992 an investigation regarding the value of insecticide impregnated mosquito nets was conducted in the municipality of Costa Marques, Rondonia. Impregnated mosquito nets gave similar protection to those not impregnated, without modifying the incidence of infection during the season of low transmission. The multivariate analysis for age and antibody titre showed a significant protection of impregnated nets against the risk of infection only in the season of high transmission, when bed nets were used more correctly. There was no difference in the effect of both kinds of bed nets in the prevention of high parasitaemia. At the end of the study, there was a reduction of the prevalence of splenomegaly in both groups but hematocrit values rose to normal in the below 15 year olds using impregnated nets.

Evaluation of SPf66 malaria vaccine efficacy in Brazil.

This paper reports the efficacy results of the randomized, placebo-controlled, field trial of SPf66 malaria vaccine in Costa Marques, Rondonia, Brazil. This region is characterized by the seasonal distribution of Plasmodium falciparum and P. vivax infections, and the recent occupation by migrants from nonendemic areas. A total of 800 individuals of both sexes, ranging in age from seven to 60 years, were included in the study. Of the initial cohort, 572 participants completed the vaccination schedule. Clinical and parasitologic evaluations were obtained by active and passive searches on a periodic basis. The overall protective efficacy against P. falciparum infections was -1.6% (-32.9% to 22.4%), and 14.1% (-17.0% to 36.9%) for the first episode. The overall protective efficacy for P. vivax infections was -19.7% (-44.8% to 1.03%), and -10.8% (-41.1% to 12.8%) for the first episode. No statistical evidence of an overall significant protective effect of SPf66 malaria vaccine against P. falciparum and P. vivax malaria was obtained in this trial.

PIP: The efficacy of the SPf66 malaria vaccine was evaluated in a randomized, placebo-controlled, field trial in Costa Marques, Rondonia, Brazil; a region characterized by seasonal Plasmodium falciparum and P. vivax infections and a recent influx of migrants from nonendemic areas. This vaccine is composed of sequences of 3 peptides from P. falciparum merozoite, erythrocytic stages, and a peptide from the circumsporozoite protein. Enrolled were 800 predominantly migrant men and women, 7-60 years of age, 572 of whom completed the vaccination schedule. Clinical and parasitologic evaluations were obtained by periodic active and passive searches up to Day 720. After the third dose, 5684 blood samples were collected and 967 (520 in the vaccine group and 447 in the placebo group) were positive. 212 P. falciparum infections (107 episodes in 76 participants in the vaccine group and 105 in 85 members of the placebo group) were confirmed after exclusion of relapses and treatment failures. A total of 427 P. vivax infections were detected (233 in 138 participants in the vaccine group and 194 in 127 participants in the placebo group). The overall protective efficacy against P. falciparum infections was -1.6% (95% confidence interval (CI), -32.9-22.4%) and 14.1% (95% CI, -17.0-36.9%) for the first episode. The overall protective efficacy for P. vivax infections was -19.7% (95% CI, -44.8-1.03%) and -10.8% (95% CI, -41.1-12.8%) for the first episode. These findings fail to provide evidence of an overall significant protective effect of the SPf66 malaria vaccine in this population.

Ação de mosquiteiros impregnados com deltametrina sobre a morbidade da malária em uma área da Amazônia Brasileira / Effect of bed nets impregnated with deltamethrin on malaria morbidity in an area of the Brazilian Amazones

Em 1992 foi feita uma investigação sobre o efeito protetor do uso de mosquiteiros impregnados com deltametrina, em uma população do município de Costa Marques, Rondônia, sujeita à transmissão malárica. Os mosquiteiros impregnados se comportaram de modo semelhante aos não impregnados, sem modificar os índices de infecção na época de baixa transmissão. A análise multivariada, por idade e títulos de anticorpos, mostrou uma proteção significante para o grupo com mosquiteiros impregnados contra o risco de infecção, apenas na estação de alta transmissão, quando os mosquiteiros foram usados mais regularmente. Não houve diferença no efeito de ambos os tipos de mosquiteiros na prevenção de elevadas parasitemias. Ao fim do estudo, ocorreu diminuição da prevalência de esplenomegalia em ambos os grupos, porém houve uma aparente recuperação da taxa normal de hematócrito em menores de 15 anos de idade em uso de mosquiteiros impregnados.

In 1992 an investigation regarding the value of insecticide impregnated mosquito nets was conducted in the municipality of Costa Marques, Rondonia. Impregnated mosquito nets gave similar protection to those not impregnated, without modifying the incidence of infection during the season of low transmission. The multivariate analysis for age and antibody titre showed a significant protection of impregnated nets against the risk of infection only in the season of high transmission, when bed nets were used more correctly. There was no difference in the effect of both kinds of bed nets in the prevention of high parasitaemia. At the end of the study, there was a reduction of the prevalence of splenomegaly in both groups but hematocrit values rose to normal in the below 15 year olds using impregnated nets.

Factors influencing phagocytosis of Salmonella typhimurium by macrophages in murine schistosomiasis.

Macrophages from Schistosoma mansoni-infected mice showed depressed capacity to increase the phagocytosis in the presence of a high bacterial load, due to a reduced involvement of these cells in phagocytosis and to a deficient ability to increase the number of phagocytosed bacteria. Normal and Salmonella-infected mice increased their phagocytic capacity when exposed to a high bacterial load. Antibody to Salmonella increased the phagocytic capacity of macrophages from Schistosoma-infected mice due to an increase in the number of bacteria phagocytosed but caused no modification in the number of macrophages engaged in phagocytosis. Our data indicate that macrophages from Schistosoma-infected mice work close to their functional limit, since no increase in phagocytosis was observed after increasing the bacterial load. Specific antibodies can improve their phagocytic capacity and, therefore, could help clearing concurrent infection.

The effects of anesthesia with thiopental on T lymphocyte responses to antigen and mitogens in vivo and in vitro.

In this study we show that antigen-specific lymphocyte proliferation and interleukin (IL)-2 production by peripheral blood lymphocytes from patients under thiopental anesthesia are significantly depressed. In contrast, mitogen-induced lymphocyte proliferation and IL-2 secretion are not depressed. We have also shown that tetanus toxoid (TT) specific CD4+ T cell clones, with a known cytokine production profile, were sensitive to the inhibitory effects of thiopental and exhibited decreased proliferation to TT as well as decreased secretion of IL-2. We observed no difference regarding IL-4 production by these clones. The data suggest that the immunosuppressive effect of thiopental is confined to antigen-specific responses. In addition, we have shown that whereas IL-2 and interferon-gamma production is dramatically impaired by the drug, IL-4 production is not significantly altered. This last finding has important implications regarding the type of immune response that is most affected by this anesthetic agent. In spite of the transient decrease in antigen-driven IL-2 synthesis, no clinical evidence of infection was noted in any healthy patient.

Safety evaluation of SPf66 malaria vaccine in Brazil

The frequency and description of side effects secondary to the subcutaneous application of SPf66 malaria vaccine and placebo are reported for each dose of application in the participants of the vaccine efficacy trial in Brazil. Side effects evaluated two hours after each application were detected in 8.0 per cent, 30.2 per cent and 8.8 per cent, for the 1st, 2nd and 3rd dose, respectively, in the SPf66 group, and in 7.0 per cent, 8.5 per cent and 2.9 per cent in the placebo group. Local reactions such as mild inflammation, nodule and pain or erythema frequently accompanied by pruritus were the most common reactions detected in both groups (3.8 per cent, 29.1 per cent and 8.5 per cent in the SPf66 group and 4.0 per cent, 7.6 per cent and 2.5 per cent in the placebo group). Among vaccinees, local side effects after the 2nd dose were more frequent in females. Systemic side effects were expressed mainly through general symptoms referred by the participants and were most frequent after the 1st dose in both groups (4.3 per cent in the SPf66 group and 3.0 per cent in the placebo group). Muscle aches and fever were referred by few participants. No severe adverse reactions were detected for either dose of application or group.

Safety evaluation of SPf66 malaria vaccine in Brazil.

The frequency and description of side effects secondary to the subcutaneous application of SPf66 malaria vaccine and placebo are reported for each dose of application in the participants of the vaccine efficacy trial in Brazil. Side effects evaluated two hours after each application were detected in 8.0%, 30.2% and 8.8%, for the 1st, 2nd and 3rd dose, respectively, in the SPf66 group, and in 7.0%, 8.5% and 2.9% in the placebo group. Local reactions such as mild inflammation, nodule and pain or erythema frequently accompanied by pruritus were the most common reactions detected in both groups (3.8%, 29.1% and 8.5% in the SPf66 group and 4.0%, 7.6% and 2.5% in the placebo group). Among vaccinees, local side effects after the 2nd dose were more frequent in females. Systemic side effects were expressed mainly through general symptoms referred by the participants and were most frequent after the 1st dose in both groups (4.3% in the SPf66 group and 3.0% in the placebo group). Muscle aches and fever were referred by few participants. No severe adverse reactions were detected for either dose of application or group.

Antibody response to Salmonella typhi in 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. This diminished ability to mount an immune response towards typhoid antigens during schistosomiasis may interfere with the clearance of the bacteria from blood stream and, therefore, play a role in the prolonged survival of salmonella as observed in some patients with chronic salmonellosis associated with schistosomiasis.

SPf66 vaccine trial in Brazil: conceptual framework study design and analytical approach

This paper describes the study population and the study design of the phase III field trial of the SPf66 vaccine in Brazil. Assessment of validity and precision principles necessary for the appropriate evaluation of the protective effect of the vaccine are discussed, as well as the results of the preliminary analyses of the gathered data. The analytical approach for the estimation of the protective effect of the vaccine is presented. This paper provides the conceptual framework for future publications.

SPf66 vaccine trial in Brazil: conceptual framework study design and analytical approach.

This paper describes the study population and the study design of the phase III field trail of the SPf66 vaccine in Brazil. Assessment of validity and precision principles necessary for the appropriate evaluation of the protective effect of the vaccine are discussed, as well as the results of the preliminary analyses of the gathered data. The analytical approach for the estimation of the protective effect of the vaccine is presented. This paper provides the conceptual framework for future publications.

[Pulmonary involvement in malaria (review)]. / Comprometimento pulmonar na malária (revisão).

Pulmonary involvement occurs in 3 to 10% of the cases of Plasmodium falciparum malaria and represents the most serious complication of this infection, with a lethality of about 70%. The understanding of its pathogenesis is still very fragmentary, however it is recognized that activation of the immune system by antigens released by the parasite plays an important role in the induction and worsening of lung damage. Capillary endothelial cells, which control the flux of fluids to the interstitial space, appear to be the most involved structure. These cells are activated by cytokines, produced by lymphocytes and macrophages during the immune response, and express receptors and molecules of adhesion, allowing for sequestration of parasitized erythrocytes and adherence of cells, which will produce locally inflammatory mediators. The inflammatory reaction and lesion of endothelial cells that ensue, together with the hemodynamic alterations induced by the capillary blockade due to the sequestration of parasitized erythrocytes and leukocytes, cause alterations of the vascular permeability and transfer of liquid to intertitial space and alveoles. Severe cases are clinically expressed by a picture of Adult Respiratory Distress Syndrome. The clinical manifestations of pulmonary involvement may start suddenly at any time during the course of malaria, even after disappearance of circulating parasites. The inducing factors are unknown. Hyperparasitemia, renal failure and pregnancy are predisposing factors. The prognosis will depend on how fast the diagnosis is established and convenient treatment initiated. If parasites are present they shall be treated with schizonticidal drugs, hemodynamic parameters continuously evaluated, preferably through a Swam-Ganz catheter. Appropriate oxygen supply and fluid balance have to be warranted. Other complications of malaria, frequently associated to the pulmonary involvement, need special attention and proper treatment. A better understanding of the pathogenesis of lung damage associated to malaria will certainly help to improve treatment and reduce morbidity and mortality.

Phagocytic and bactericidal function of mouse macrophages to Salmonella typhimurium in schistosomiasis mansoni.

Patients infected with schistosomes may develop a clinical picture of chronic salmonellosis. We have investigated the altered function of macrophages capable of playing a role in the development of chronic salmonellosis associated with Schistosoma mansoni in an experimental model. The capacity of mouse peritoneal macrophages to ingest and kill Salmonella was assessed in mice infected with S. mansoni with or without concurrent Salmonella typhimurium infection. Schistosomiasis was associated with a significant decrease in the phagocytic index of macrophages, due to the reduced number of cells engaged in phagocytosis. However, the number of bacteria ingested by these cells was comparable to that of the control group. The bactericidal capacity of macrophages from S. mansoni-infected mice was also significantly lower than that of cells from normal mice. Macrophages from animals infected only with Salmonella typhimurium showed an increased phagocytic capacity. It was concluded that S. mansoni infection alters phagocytosis and intracellular destruction of salmonellae. This demonstration of a novel mechanism of survival of salmonellae represents a step forward in understanding the pathogenesis and management of chronic septicemic salmonellosis.

Cytokines and dysregulation of the immune response in human malaria.

The dysregulation of the immune response by malaria parasite has been considered as a possible constraint to the effectiveness of malaria vaccination. In spite of the important role interleukin-1 (IL-1) plays on the immunoregulation, and its ability to mimic various features of clinical malaria, reports on IL-1 in malaria are lacking. We found that only 2 out of 35 subjects with acute malaria showed increased levels of serum IL-1 alpha by enzyme immunoassay. To assess whether IL-1 could interfere with T-lymphocyte responses, blood mononuclear cells from patients infected with Plasmodium falciparum, P. vivax, or healthy subjects were cultured with phytohemagglutinin, and lymphocyte proliferation measured 72 h later by 3H-thymidine incorporation. Our data showed that T-lymphocyte responses are depressed both in P. falciparum (10,500 +/- 2,900) and P. vivax malaria (13,000 +/- 3,300), as compared to that of healthy individuals (27,000 +/- 3,000). Addition of IL-1 partially reversed depression of malaria lymphocytes, but had no effect on normal cells. On the other hand, T-lymphocytes from malaria infected-subjects presented a minimal decrease in proliferation, when cultured in the presence of exogenous PGE2. These data indicate the occurrence of two defects of immunoregulation in malaria: a deficiency of IL-1 production by monocytes/macrophages, and an increased resistance of lymphocytes to the antiproliferative effect of PGE2.

Specific treatment of hepatosplenic schistosomiasis can increase T-lymphocyte reactivity.

It has been recognized that Schistosoma mansoni infection causes depression of T-cell responsiveness. In this study we have evaluated whether immunodepression associated to schistosomiasis could be reverted by specific treatment. T-cell immune response was assessed by means of intradermal tests using recall antigens in a group of 22 patients with hepatosplenic schistosomiasis, one year after treatment with oxamniquine and compared with a group of untreated hepatosplenic patients. Only 27% of treated patients presented complete anergy to all tested antigens, in marked contrast to 80% unresponsiveness showed by hepatosplenic patients without treatment. Although most of the treated individuals showed some response to the tested antigens, in some individuals this unresponsiveness still persisted after treatment. Anergy was not found in any normal individual of the control group. It was concluded that Schistosoma mansoni infected patients may recover their normal immune responsiveness after the elimination of the worm by treatment.

T cell-dependent immunodepression in vivo in Schistosoma mansoni infected patients.

T-cell function was evaluated in 29 patients with either hepatointestinal or hepatosplenic schistosomiasis by intradermal tests to recall antigens. Immunodepression was detected in 26% of the subjects with hepatointestinal schistosomiasis and in 50% of those with the hepatosplenic form. Cellular immunodepression was related to worm load and spleen size. This non specific T-cell immunodepression may represent a serious constraint to the elimination of intracellular pathogens both in hepatosplenic or hepatointestinal schistosomiasis.