RESUMEN
Introduction: Throughout the preclinical education curriculum, medical students learn numerous concepts that must be retained and added to over time. Previous studies have shown that utilizing drawing leads to improved retention of concepts. However, limited studies have investigated the use of drawing at the medical school level. The goal of this study was to utilize mechanism-based drawing aimed at presenting conceptual material rather than strict memorization before and after interrupted learning. Methods: Participants were randomly assigned to a drawing group or a text-only group and both groups received text #1 that explained a microbiology concept. The groups were instructed to read the text, but only the drawing group received a drawing prompt. The groups then completed post-test #1. During part #2 of the study, the groups were instructed to read text #2 with no drawing prompt. The two groups were instructed to complete post-test #2 which covered topics from text #1 and #2. p<0.05 was considered significant. Results: The drawing group performed significantly better on post-test #1 compared to the text-only group. There were no significant differences on overall performance on post-test #2. However, the drawing group performed significantly better on questions related to the material covered in text #1 on post-test #2. Conclusion: Results presented here demonstrate that students who draw perform significantly better when assessed on complex microbiology concepts, even after interrupted by the introduction of an unrelated concept. A future study should investigate the effectiveness of drawing after interruption by learning on long-term retention and performance.
RESUMEN
One of the commonest complications of Plasmodium falciparum malaria is the development of severe malarial anemia (SMA), which is, at least in part, due to malaria-induced suppression of erythropoiesis. Factors associated with suppression of erythropoiesis and development of SMA include accumulation of malarial pigment (hemozoin, PfHz) in bone marrow and altered production of inflammatory mediators, such as tumor necrosis factor (TNF)-α, and nitric oxide (NO). However, studies investigating the specific mechanisms responsible for inhibition of red blood cell development have been hampered by difficulties in obtaining bone marrow aspirates from infants and young children, and the lack of reliable models for examining erythroid development. As such, an in vitro model of erythropoiesis was developed using CD34+ stem cells derived from peripheral blood to examine the effects of PfHz, PfHz-stimulated peripheral blood mononuclear cell (PBMC)-conditioned media (CM-PfHz), TNF-α, and NO on erythroid cell development. PfHz only slightly suppressed erythroid cell proliferation and maturation marked by decreased expression of glycophorin A (GPA). On the other hand, CM-PfHz, TNF-α, and NO significantly inhibited erythroid cell proliferation. Furthermore, decreased proliferation in cells treated with CM-PfHz and NO was accompanied by increased apoptosis of erythropoietin-stimulated CD34+ cells. In addition, NO significantly inhibited erythroid cell maturation, whereas TNF-α did not appear to be detrimental to maturation. Collectively, our results demonstrate that PfHz suppresses erythropoiesis by acting both directly on erythroid cells, and indirectly via inflammatory mediators produced from PfHz-stimulated PBMC, including TNF-α and NO.
Asunto(s)
Anemia/etiología , Eritropoyesis , Hemoproteínas/metabolismo , Mediadores de Inflamación/metabolismo , Malaria/fisiopatología , Óxido Nítrico/fisiología , Factor de Necrosis Tumoral alfa/fisiología , Antígenos CD34/metabolismo , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Medios de Cultivo Condicionados , Eritropoyesis/efectos de los fármacos , Eritropoyetina/farmacología , Glicoforinas/metabolismo , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Hemoproteínas/aislamiento & purificación , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Malaria/complicaciones , Malaria/metabolismo , Malaria Falciparum/metabolismo , Donantes de Óxido Nítrico/farmacología , Pigmentos Biológicos/aislamiento & purificación , Pigmentos Biológicos/metabolismo , Proteínas RecombinantesRESUMEN
Plasmodium falciparum malaria is a leading global cause of infectious disease burden. In areas in which P. falciparum transmission is holoendemic, such as western Kenya, severe malarial anemia (SMA) results in high rates of pediatric morbidity and mortality. Although the pathophysiological basis of SMA is multifactorial, we recently discovered that suppression of unexplored hematopoietic growth factors that promote erythroid and myeloid colony development, such as stem cell growth factor (SCGF) (C-type lectin domain family member 11A [CLEC11A]), was associated with enhanced development of SMA and reduced erythropoietic responses. To extend these investigations, the relationships between a novel SCGF promoter variant (-539C/T, rs7246355), SMA (hemoglobin [Hb] < 6.0 g/dl), and reduced erythropoietic responses (reticulocyte production index [RPI], <2.0) were investigated with Kenyan children (n = 486) with falciparum malaria from western Kenya. Circulating SCGF was positively correlated with hemoglobin levels (r = 0.251; P = 0.022) and the reticulocyte production index (RPI) (r = 0.268; P = 0.025). Children with SMA also had lower SCGF levels than those in the non-SMA group (P = 0.005). Multivariate logistic regression analyses controlling for covariates demonstrated that individuals with the homologous T allele were protected against SMA (odds ratio, 0.57; 95% confidence interval [95% CI] 0.34 to 0.94; P = 0.027) relative to CC (wild-type) carriers. Carriers of the TT genotype also had higher SCGF levels in circulation (P = 0.018) and in peripheral blood mononuclear cell culture supernatants (P = 0.041), as well as an elevated RPI (P = 0.005) relative to individuals with the CC genotype. The results presented here demonstrate that homozygous T at -539 in the SCGF promoter is associated with elevated SCGF production, enhanced erythropoiesis, and protection against the development of SMA in children with falciparum malaria.
Asunto(s)
Anemia/etiología , Anemia/genética , Regulación de la Expresión Génica/fisiología , Factores de Crecimiento de Célula Hematopoyética/genética , Lectinas Tipo C/genética , Malaria Falciparum/complicaciones , Preescolar , Femenino , Variación Genética , Genotipo , Humanos , Lactante , Masculino , Regiones Promotoras GenéticasRESUMEN
In areas of holoendemic Plasmodium falciparum transmission, severe malarial anemia (SMA) is a leading cause of pediatric morbidity and mortality. Although many soluble mediators regulate erythropoiesis, it is unclear how these factors contribute to development of SMA. Investigation of novel genes dysregulated in response to malarial pigment (hemozoin [PfHz]) revealed that stem cell growth factor (SCGF; also called C-type lectin domain family member 11A [CLEC11A]), a hematopoietic growth factor important for development of erythroid and myeloid progenitors, was one of the most differentially expressed genes. Additional experiments with cultured peripheral blood mononuclear cells (PBMCs) demonstrated that PfHz decreased SCGF/CLEC11A transcriptional expression in a time-dependent manner. Circulating SCGF levels were then determined for Kenyan children (n = 90; aged 3 to 36 months) presenting at a rural hospital with various severities of malarial anemia. SCGF levels in circulation (P = 0.001) and in cultured PBMCs (P = 0.004) were suppressed in children with SMA. Circulating SCGF also correlated positively with hemoglobin levels (r = 0.241; P = 0.022) and the reticulocyte production index (RPI) (r = 0.280; P = 0.029). In addition, SCGF was decreased in children with reduced erythropoiesis (RPI of <2) (P < 0.001) and in children with elevated levels of naturally acquired monocytic PfHz (P = 0.019). Thus, phagocytosis of PfHz promotes a decrease in SCGF gene products, which may contribute to reduced erythropoiesis in children with SMA.
Asunto(s)
Anemia/etiología , Factores de Crecimiento de Célula Hematopoyética/genética , Lectinas Tipo C/genética , Malaria/complicaciones , Preescolar , Eritropoyesis , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Factores de Crecimiento de Célula Hematopoyética/sangre , Factores de Crecimiento de Célula Hematopoyética/fisiología , Hemoproteínas/fisiología , Humanos , Lactante , Lectinas Tipo C/sangre , Lectinas Tipo C/fisiología , Malaria/metabolismo , Masculino , Fagocitosis , Reticulocitos/fisiologíaRESUMEN
Interleukin (IL)-1beta is a cytokine released as part of the innate immune response to Plasmodium falciparum. Because the role played by IL-1beta polymorphic variability in conditioning the immunopathogenesis of severe malarial anemia (SMA) remains undefined, relationships between IL-1beta promoter variants (-31C/T and -511A/G), SMA (hemoglobin [Hb] level <6.0 g/dL), and circulating IL-1beta levels were investigated in children with parasitemia (n= 566) from western Kenya. The IL-1beta promoter haplotype -31C/-511A (CA) was associated with increased risk of SMA (Hb level <6.0 g/dL; odds ratio [OR], 1.98 [95% confidence interval {CI}, 1.55-2.27]; P < .05) and reduced circulating IL-1beta levels (p <.05). The TA (-31T/-511A) haplotype was nonsignificantly associated with protection against SMA (OR, 0.52 [95% CI, 0.18-1.16]; p =.11) and elevated IL-1beta production ( p<.05). Compared with the non-SMA group, children with SMA had significantly lower IL-1beta levels and nonsignificant elevations in both IL-1 receptor antagonist (IL-1Ra) and the ratio of IL-1Ra to IL-1beta. The results presented demonstrate that variation in IL-1beta promoter conditions susceptibility to SMA and functional changes in circulating IL-1beta levels.
Asunto(s)
Anemia , Interleucina-1beta/genética , Malaria Falciparum , Polimorfismo Genético , Regiones Promotoras Genéticas/genética , Anemia/genética , Anemia/inmunología , Anemia/fisiopatología , Animales , Preescolar , Femenino , Genotipo , Haplotipos , Humanos , Lactante , Interleucina-1beta/biosíntesis , Interleucina-1beta/sangre , Malaria Falciparum/complicaciones , Malaria Falciparum/genética , Malaria Falciparum/inmunología , Malaria Falciparum/parasitología , Masculino , Parasitemia/complicaciones , Parasitemia/genética , Parasitemia/inmunología , Parasitemia/parasitología , Plasmodium falciparum , Índice de Severidad de la EnfermedadRESUMEN
Severe malarial anemia (SMA) is a leading cause of mortality among children in sub-Saharan Africa. Although the novel cytokine, interleukin (IL)-23, promotes anemia in chronic inflammatory diseases, the role of IL-23 in SMA remains undefined. Since IL-23 and IL-12 share the IL-12p40 subunit and IL-12Rbeta1 receptor, and are down-regulated by IL-10, relationships among these cytokines were explored in Kenyan children with varying severities of malarial anemia. Children with malarial anemia had increased circulating IL-23 and IL-10 and decreased IL-12 relative to healthy controls. Enhanced anemia severity and elevated parasitemia were associated with increased IL-10 relative to IL-23 and IL-12. Further exploration of the relationships among the cytokines using an in vitro model in which peripheral blood mononuclear cells were treated with synthetic hemozoin (sHz, malarial pigment) revealed that IL-12p35 and IL-23p19 transcripts had a sustained induction over 72 h, while IL-12p40 and IL-10 message peaked at 24 h, and rapidly declined thereafter. Taken together, results here show that IL-23 is elevated in children with malarial anemia, and that IL-10 and IL-12 appear to have important regulatory effects on IL-23 production during childhood malaria.
Asunto(s)
Anemia/inmunología , Interleucina-10/sangre , Interleucina-12/sangre , Interleucina-23/sangre , Malaria Falciparum/inmunología , Anemia/sangre , Anemia/parasitología , Animales , Preescolar , Femenino , Hemoproteínas/inmunología , Hemoglobinas/análisis , Humanos , Lactante , Recién Nacido , Interleucina-10/inmunología , Interleucina-12/inmunología , Interleucina-23/inmunología , Kenia , Leucocitos Mononucleares/inmunología , Malaria Falciparum/parasitología , Masculino , Parasitemia , Plasmodium falciparum/inmunología , Plasmodium falciparum/fisiología , Subunidades de Proteína/inmunología , Subunidades de Proteína/metabolismoRESUMEN
Plasmodium falciparum malaria is one of the leading causes of childhood morbidity and mortality in sub-Saharan Africa. The host immune response to P. falciparum is a critical determinant of malarial pathogenesis and disease outcomes. Macrophage migration inhibitory factor (MIF) is a central regulator of innate immune responses to bacterial and parasitic infections. Our recent investigations demonstrated that peripheral blood MIF production was suppressed in children with severe malaria. Because examination of MIF production in children with active disease does not account for the inherent ability of the host to generate MIF, basal circulating MIF and peripheral blood mononuclear cell (PBMC) MIF transcript levels were determined in healthy children with a history of either mild or severe malaria. Children with prior mild malaria had higher plasma MIF levels and PBMC MIF transcripts than children with an identical number of previous episodes of severe malaria. These results suggest that increased basal MIF production may be important in generating immune responses that protect against the development of severe malaria.
Asunto(s)
Factores Inhibidores de la Migración de Macrófagos/biosíntesis , Malaria Falciparum/inmunología , Plasmodium falciparum/inmunología , Animales , Niño , Preescolar , Estudios de Cohortes , Femenino , Gabón , Humanos , Estudios Longitudinales , Factores Inhibidores de la Migración de Macrófagos/sangre , Factores Inhibidores de la Migración de Macrófagos/genética , Factores Inhibidores de la Migración de Macrófagos/inmunología , Masculino , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , ARN Mensajero/sangre , ARN Mensajero/genéticaRESUMEN
Severe malarial anemia (SMA), caused by Plasmodium falciparum infections, is one of the leading causes of childhood mortality in sub-Saharan Africa. Although the molecular determinants of SMA are largely undefined, dysregulation in host-derived inflammatory mediators influences disease severity. Macrophage migration inhibitory factor (MIF) is an important regulator of innate inflammatory responses that has recently been shown to suppress erythropoiesis and promote pathogenesis of SMA in murine models. To examine the role of MIF in the development of childhood SMA, peripheral blood MIF production was examined in Kenyan children (aged <3 years, n = 357) with P. falciparum malarial anemia. All children in the study were free from bacteremia and human immunodeficiency virus type 1. Since deposition of malarial pigment (hemozoin [Hz]) contributes to suppression of erythropoiesis, the relationship between MIF concentrations and monocytic acquisition of Hz was also examined in vivo and in vitro. Circulating MIF concentrations declined with increasing severity of anemia and significantly correlated with peripheral blood leukocyte MIF transcripts. However, MIF concentrations in peripheral blood were not significantly associated with reticulocyte production. Multivariate regression analyses, controlling for age, gender, and parasitemia, further revealed that elevated levels of pigment-containing monocytes (PCM) was associated with SMA and decreased MIF production. In addition, PCM levels were a better predictor of hemoglobin and MIF concentrations than parasite density. Additional experiments in malaria-naive individuals demonstrated that hemozoin caused both increased and decreased MIF production in cultured peripheral blood mononuclear cells (PBMC) in a donor-specific manner, independent of apoptosis. However, PBMC MIF production in children with acute malaria progressively declined with increasing anemia severity. Results presented here demonstrate that acquisition of hemozoin by monocytes is associated with suppression of peripheral blood MIF production and enhanced severity of anemia in childhood malaria.
Asunto(s)
Anemia/sangre , Anemia/etiología , Hemoproteínas/metabolismo , Factores Inhibidores de la Migración de Macrófagos/sangre , Malaria Falciparum/complicaciones , Monocitos/metabolismo , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lactante , Kenia , Masculino , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Severe malarial anemia (MA) is the primary manifestation of severe malaria among children in areas of holoendemic Plasmodium falciparum transmission. Although overproduction of inflammatory-derived cytokines are implicated in the immunopathogenesis of severe MA, chemokines such as regulated on activation, normal T-cell expressed and secreted (RANTES, CCL5) are largely unexplored in childhood malaria. We found that RANTES is decreased during severe MA (p<0.01), and associated with suppression of erythropoiesis (p<0.05) and malaria-induced thrombocytopenia (p<0.05). These findings suggest that thrombocytopenia may be a source of reduced RANTES which may contribute, at least in part, to suppression of erythropoiesis in children with malarial anemia.
Asunto(s)
Quimiocina CCL5/antagonistas & inhibidores , Quimiocina CCL5/sangre , Malaria Falciparum/sangre , Plasmodium falciparum , Animales , Quimiocina CCL5/biosíntesis , Niño , Femenino , Humanos , Lactante , Masculino , Trombocitopenia/sangreRESUMEN
Severe malarial anemia (SMA) is a primary cause of morbidity and mortality in immune-naïve infants and young children residing in areas of holoendemic Plasmodium falciparum transmission. Although the immunopathogenesis of SMA is largely undefined, we have previously shown that systemic interleukin-12 (IL-12) production is suppressed during childhood blood-stage malaria. Since IL-10 and tumor necrosis factor alpha (TNF-alpha) are known to decrease IL-12 synthesis in a number of infectious diseases, altered transcriptional regulation of these inflammatory mediators was investigated as a potential mechanism for IL-12 down-regulation. Ingestion of naturally acquired malarial pigment (hemozoin [PfHz]) by monocytes promoted the overproduction of IL-10 and TNF-alpha relative to the production of IL-12, which correlated with an enhanced severity of malarial anemia. Experiments with cultured peripheral blood mononuclear cells (PBMC) and CD14(+) cells from malaria-naïve donors revealed that physiological concentrations of PfHz suppressed IL-12 and augmented IL-10 and TNF-alpha by altering the transcriptional kinetics of IL-12p40, IL-10, and TNF-alpha, respectively. IL-10 neutralizing antibodies, but not TNF-alpha antibodies, restored PfHz-induced suppression of IL-12. Blockade of IL-10 and the addition of recombinant IL-10 to cultured PBMC from children with SMA confirmed that IL-10 was responsible for malaria-induced suppression of IL-12. Taken together, these results demonstrate that PfHz-induced up-regulation of IL-10 is responsible for the suppression of IL-12 during malaria.
Asunto(s)
Anemia/parasitología , Hemoproteínas/metabolismo , Interleucina-10/metabolismo , Interleucina-12/genética , Malaria Falciparum/inmunología , Plasmodium falciparum , Subunidades de Proteína/genética , Anemia/genética , Anemia/inmunología , Animales , Anticuerpos/farmacología , Células Cultivadas , Preescolar , Regulación hacia Abajo , Femenino , Hemoproteínas/farmacología , Humanos , Lactante , Recién Nacido , Interleucina-10/antagonistas & inhibidores , Interleucina-10/sangre , Interleucina-12/sangre , Interleucina-12/metabolismo , Subunidad p40 de la Interleucina-12 , Receptores de Lipopolisacáridos/análisis , Malaria Falciparum/genética , Masculino , Monocitos/efectos de los fármacos , Monocitos/inmunología , Fagocitosis , Transcripción Genética , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Protective immunity against Plasmodium falciparum is partially mediated through binding of malaria-specific IgG antibodies to Fcgamma receptors. Polymorphic variability in Fcgamma RIIa (H/R-131) is associated with differential binding of IgG subtypes and malaria disease outcomes. However, the role of Fcgamma RIIa-131 variability in conditioning susceptibility to severe malarial anemia, the primary manifestation of severe malaria in holoendemic P. falciparum transmission areas, is largely undefined. Thus, Fcgamma RIIa-H131R polymorphism was investigated in 493 children who came to a hospital with acute malaria. Variation in Fcgamma RIIa-131 was not significantly associated with severe malarial anemia (hemoglobin [Hb] < 6.0 g/dL) or malaria anemia (Hb < 8.0 g/dL). However, relative to the heterozygous genotype, homozygotes for the R131 alleles were protected against high-density parasitemia (>or= 10,000 parasites/microL; odds ratio [OR] = 0.58, 95% confidence interval [CI] = 0.37-0.92, P = 0.02), while homozygotes for the H131 alleles were mildly protective (OR = 0.71, 95% CI = 0.45-1.13, P = 0.14). Additional multivariate analyses showed that infection with human immunodeficiency virus type 1 did not influence the associations between FcgammaRIIa-H131R polymorphism and malaria disease outcomes. Genotypic results presented here parallel data illustrating that parasite density is unrelated to the severity of anemia in children with acute malaria. Thus, although homozygosity for the R131 allele protects against high-density parasitemia, FcgammaRIIa-131 polymorphism does not protect against malaria anemia.
Asunto(s)
Anemia/inmunología , Antígenos CD/genética , Malaria Falciparum/inmunología , Parasitemia/inmunología , Receptores de IgG/genética , Anemia/sangre , Anemia/epidemiología , Anemia/patología , Estudios Transversales , Femenino , Humanos , Lactante , Recién Nacido , Kenia/epidemiología , Malaria Falciparum/sangre , Malaria Falciparum/epidemiología , Malaria Falciparum/patología , Masculino , Parasitemia/sangre , Parasitemia/epidemiología , Parasitemia/patología , Polimorfismo Genético , Prevalencia , Índice de Severidad de la EnfermedadRESUMEN
Cytokines and effector molecules are important immunoregulatory molecules in human malaria. Tumor necrosis factor (TNF)-alpha limits malaria parasitemia but also promotes pathogenesis at high concentrations, whereas prostaglandin E2 (PGE2) inhibits TNF-alpha production and is reduced in childhood malaria, at least in part, through suppression of cyclooxygenase (COX)-2 following the ingestion of Plasmodium falciparum hemozoin (pfHz; malarial pigment) by peripheral blood mononuclear cells (PBMCs). Although molecular interactions between TNF-alpha and PGE2 are largely unexplored in human malaria, results presented here show that pfHz-induced suppression of PBMC COX-2 gene products induces overproduction of TNF-alpha. Moreover, addition of exogenous PGE2 to pfHz-treated PBMCs dose-dependently decreased TNF-alpha production, whereas experimental COX inhibitors and antipyretics used during human malaria generated increased TNF-alpha production. Healthy, malaria-exposed children had elevated levels of circulating bicyclo-PGE2/TNF-alpha, compared with children with malarial anemia (P<.01), with systemic bicyclo-PGE2 and TNF-alpha significantly associated with hemoglobin concentrations (r=0.745; P<.01). The results of the present study illustrate that pfHz-induced suppression of PGE2 promotes overproduction of TNF-alpha, which is associated with enhanced malarial anemia.
Asunto(s)
Anemia/inmunología , Dinoprostona/metabolismo , Malaria Falciparum/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Adulto , Analgésicos no Narcóticos/administración & dosificación , Analgésicos no Narcóticos/farmacología , Anemia/tratamiento farmacológico , Animales , Estudios de Casos y Controles , Niño , Preescolar , Ciclooxigenasa 2/sangre , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Dinoprostona/sangre , Femenino , Expresión Génica , Hemoproteínas/farmacología , Humanos , Leucocitos Mononucleares/metabolismo , Estudios Longitudinales , Malaria Falciparum/tratamiento farmacológico , Masculino , Fagocitosis , Plasmodium falciparum/metabolismo , Estudios Prospectivos , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Malarial anemia (MA) is a multifactorial disease for which the complex etiological basis is only partially defined. The association of clinical, nutritional, demographic, and socioeconomic factors with parasitemia, anemia, and MA was determined for children presenting at a hospital in a holoendemic area of Plasmodium falciparum transmission in western Kenya. Parasitemia was not associated with malaria disease severity. In univariate logistic regression, fever was significantly associated with parasitemia, and wasting was associated with increased presentation of MA. Caretaker's level of education and occupation were significantly correlated with parasitemia, anemia, and MA. Housing structure was also significantly associated with parasitemia and anemia. Bed net use was protective against parasitemia but not anemia or MA. Multivariate logistic regression models demonstrated that fever, mother's occupation, and bed net use were associated with parasitemia. In the current study, none of the factors were associated with anemia or MA in the multivariate models.
Asunto(s)
Anemia/parasitología , Enfermedades Endémicas , Malaria Falciparum/complicaciones , Parasitemia/parasitología , Plasmodium falciparum/crecimiento & desarrollo , Anemia/sangre , Anemia/epidemiología , Animales , Preescolar , Estudios Transversales , Femenino , Hemoglobinas/metabolismo , Vivienda , Humanos , Lactante , Kenia/epidemiología , Malaria Falciparum/sangre , Malaria Falciparum/epidemiología , Malaria Falciparum/parasitología , Masculino , Estado Nutricional , Parasitemia/sangre , Parasitemia/complicaciones , Parasitemia/epidemiología , Población Rural , Clase SocialRESUMEN
OBJECTIVE: Since the primary hematological complication in both pediatric HIV-1 and malaria is anemia, co-infection with these pathogens may promote life-threatening severe malarial anemia (SMA). The primary objective of the study was to determine if HIV-1 exposure [HIV-1(exp)] and/or HIV-1 infection [HIV-1(+)] increased the prevalence of SMA in children with acute malaria. DESIGN: The effect of HIV-1 exposure and HIV-1 infection on the prevalence of SMA (hemoglobin < 6.0 g/dl), parasitemia (parasites/microl), and high-density parasitemia (HDP, >or= 10 000 parasites/mul) was investigated in children Asunto(s)
Anemia/parasitología
, Anemia/virología
, Infecciones por VIH/complicaciones
, VIH-1/aislamiento & purificación
, Malaria Falciparum/complicaciones
, Enfermedad Aguda
, Anemia/sangre
, Preescolar
, Países en Desarrollo
, Femenino
, Hematócrito
, Hemoglobinas/análisis
, Humanos
, Lactante
, Kenia
, Masculino
, Parasitemia/complicaciones
, Salud Rural/estadística & datos numéricos
RESUMEN
Plasmodium falciparum malaria remains one of the most frequently lethal diseases affecting children in sub-Saharan Africa, yet the immune mediators that regulate pathogenesis are only partially defined. Since macrophage migration inhibitory factor (MIF) is important for regulating innate immunity in bacterial and parasitic infections, circulating MIF and peripheral blood mononuclear cell (PBMC) MIF transcripts were investigated in children with acute falciparum malaria. Peripheral blood levels of MIF-regulatory cytokines and effector molecules, including interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-12, IL-10, transforming growth factor (TGF)-beta1, bicyclo-prostaglandin (PG) E2, and nitric oxide synthase activity were also determined. Circulating MIF and PBMC MIF mRNA were significantly lower in children with acute malaria relative to healthy, malaria-exposed children. Peripheral blood MIF levels showed no association with either parasitemia or hemoglobin concentrations. Circulating MIF was, however, significantly associated with IL-12 and TGF-beta1. Multiple regression analyses revealed that IFN-gamma was the most significant predictor of peripheral blood MIF concentrations. These findings suggest that reduced MIF production may promote enhanced disease severity in children with falciparum malaria.
Asunto(s)
Regulación hacia Abajo/inmunología , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Factores Inhibidores de la Migración de Macrófagos/antagonistas & inhibidores , Malaria Falciparum/inmunología , Plasmodium falciparum/inmunología , ARN Mensajero/antagonistas & inhibidores , Animales , Niño , Preescolar , Citocinas/metabolismo , Humanos , Leucocitos Mononucleares/parasitología , Factores Inhibidores de la Migración de Macrófagos/biosíntesis , Factores Inhibidores de la Migración de Macrófagos/sangre , Factores Inhibidores de la Migración de Macrófagos/genética , Malaria Falciparum/sangre , Malaria Falciparum/diagnóstico , ARN Mensajero/biosíntesis , ARN Mensajero/sangre , Índice de Severidad de la Enfermedad , Transcripción Genética/inmunologíaRESUMEN
BACKGROUND: The molecular immunological interactions between HIV and malaria are largely undefined. Since tumor necrosis factor (TNF)-alpha is elevated during acute malaria and increases with HIV-1 disease progression, TNF-alpha production may be an important mediator for interactions between malaria and HIV-1. METHODS: To examine the stage-specific immunological interactions between HIV and malaria, peripheral blood mononuclear cells (PBMC) and CD14 cells were isolated and cultured from rhesus macaques at different stages of SIV infection. Cultures were stimulated with lipopolysaccharide (LPS) and interferon (IFN)-gamma in the presence of Plasmodium falciparum-derived hemozoin (Hz) or synthetic Hz (sHz). TNF-alpha transcripts and soluble protein were examined by real time reverse transcription-PCR and ELISA, respectively. The effects of Hz on viral replication were determined by measurement of p27 antigen with varying concentrations of TNF-alpha neutralizing antibodies. RESULTS: Hz and sHz significantly increased LPS- and IFN-gamma-induced TNF-alpha protein and transcripts in PBMC from animals with late stage SIV infection (i.e., AIDS). Hz and sHz also induced high levels of sustained TNF-alpha transcripts in PBMC from the AIDS group. During the late stage of disease, CD14 cells were the primary source of TNF-alpha production. Stimulation of PBMC with Hz and sHz significantly increased viral replication that was dose-dependently reduced by the addition of TNF-alpha neutralizing antibodies. CONCLUSIONS: Hz promotes high levels of TNF-alpha production from PBMC during AIDS and increases viral replication in SIV-infected animals.
Asunto(s)
VIH-1/fisiología , Hemoproteínas/farmacología , Leucocitos Mononucleares/inmunología , Plasmodium falciparum/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Replicación Viral/fisiología , Animales , Leucocitos Mononucleares/parasitología , Leucocitos Mononucleares/virología , Lipopolisacáridos/farmacología , Macaca mulatta , Malaria Falciparum/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunologíaRESUMEN
Chemokines regulate the host immune response to a variety of infectious pathogens. Since the role of chemokines in regulating host immunity in children with Plasmodium falciparum malaria has not previously been reported, circulating levels of beta-chemokines (MIP-1alpha, MIP-1beta, and RANTES) and their respective transcriptional profiles in ex vivo peripheral blood mononuclear cells (PBMCs) were investigated. Peripheral blood MIP-1alpha and MIP-1beta levels were significantly elevated in mild and severe malaria, while RANTES levels decreased with increasing disease severity. Beta-chemokine gene expression profiles in blood mononuclear cells closely matched those of circulating beta-chemokines, illustrating that PBMCs are a primary source for the observed pattern of beta-chemokine production during acute malaria. Statistical modeling revealed that none of the chemokines was significantly associated with either parasitemia or anemia. Additional investigations in healthy children with a known history of malaria showed that children with prior severe malaria had significantly lower baseline RANTES production than children with a history of mild malaria, suggesting inherent differences in the ability to produce RANTES in these two groups. Baseline MIP-1alpha and MIP-1beta did not significantly differ between children with prior severe malaria and those with mild malaria. Additional in vitro experiments in PBMCs from healthy, malaria-naïve donors revealed that P. falciparum-derived hemozoin (Hz; malarial pigment) and synthetic Hz (beta-hematin) promote a similar pattern of beta-chemokine gene expression. Taken together, the results presented here demonstrate that children with severe malaria have a distinct profile of beta-chemokines characterized by increased circulating levels of MIP-1alpha and MIP-1beta and decreased RANTES. Altered patterns of circulating beta-chemokines result, at least in part, from Hz-induced changes in beta-chemokine gene expression in blood mononuclear cells.
Asunto(s)
Quimiocina CCL5/genética , Proteínas Inflamatorias de Macrófagos/genética , Malaria Falciparum/inmunología , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/sangre , Niño , Preescolar , Femenino , Perfilación de la Expresión Génica , Hemoproteínas/farmacología , Hemoglobinas/análisis , Humanos , Proteínas Inflamatorias de Macrófagos/sangre , Malaria Falciparum/sangre , Malaria Falciparum/parasitología , MasculinoRESUMEN
Molecular immunologic determinants of disease severity during Plasmodium falciparum malaria are largely undetermined. Our recent investigations showed that peripheral blood mononuclear cell (PBMC) cyclooxygenase-2 (COX-2) gene expression and plasma prostaglandin E(2) (PGE(2)) production are suppressed in children with falciparum malaria relative to healthy, malaria-exposed children with partial immunity. Furthermore, decreased COX-2/PGE(2) levels were significantly associated with increased plasma interleukin-10 (IL-10), an anti-inflammatory cytokine that inhibits the expression of COX-2 gene products. To determine the mechanism(s) responsible for COX-2-derived PGE(2) suppression, PBMCs were cultured from children with falciparum malaria. PGE(2) production was suppressed under baseline and COX-2-promoting conditions (stimulation with lipopolysaccharide [LPS] and interferon [IFN]-gamma) over prolonged periods, suggesting that an in vivo-derived product(s) was responsible for reduced PGE(2) biosynthesis. Ingestion of hemozoin (malarial pigment) by PBMC was investigated as a source of COX-2/PGE(2) suppression in PBMCs from healthy, malaria-naive adults. In addition, synthetically prepared hemozoin, beta-hematin, was used to investigate the effects of the core iron component of hemozoin, ferriprotoporphyrin-IX (FPIX). Physiologic concentrations of hemozoin or b-hematin suppressed LPS- and IFN-gamma-induced COX-2 mRNA in a time- and dose-dependent manner, resulting in decreased COX-2 protein and PGE(2) production. Suppression of COX-2/PGE(2) by hemozoin was not due to decreased cell viability as evidenced by examination of mitochondrial bioactivity. These data illustrate that ingestion of FPIX by blood mononuclear cells is responsible for suppression of COX-2/PGE(2). Although hemozoin induced overproduction of IL-10, neutralizing IL-10 antibodies failed to restore PGE(2) production. Thus, acquisition of hemozoin by blood mononuclear cells is responsible for suppression of PGE(2) in malaria through inhibition of de novo COX-2 transcripts via molecular mechanisms independent of increased IL-10 production.
Asunto(s)
Dinoprostona/metabolismo , Leucocitos Mononucleares/metabolismo , Malaria Falciparum/metabolismo , Prostaglandina-Endoperóxido Sintasas/biosíntesis , Estudios de Casos y Controles , Células Cultivadas , Niño , Preescolar , Ciclooxigenasa 2 , Dinoprostona/sangre , Regulación Enzimológica de la Expresión Génica , Hemoproteínas/farmacología , Humanos , Interferón gamma , Interleucina-10/biosíntesis , Interleucina-10/sangre , Leucocitos Mononucleares/enzimología , Lipopolisacáridos , Malaria Falciparum/sangre , Malaria Falciparum/inmunología , Proteínas de la Membrana , Prostaglandina-Endoperóxido Sintasas/genética , ARN Mensajero/biosíntesisRESUMEN
Experiments outlined here investigate the role of nitric oxide (NO) in the pathogenesis of Plasmodium falciparum-induced malarial anemia (MA). The results show that ex vivo and in vitro NO synthase (NOS) activity in peripheral blood mononuclear cells (PBMCs) is significantly elevated in children with MA and inversely associated with hemoglobin levels. Additional experiments using PBMCs from non-malaria-exposed donors demonstrate that physiologic amounts of P. falciparum-derived hemozoin augment NOS type 2 (NOS2) transcripts and NO production. Results of these experiments illustrate that elevated NO production in children with MA is associated with decreased hemoglobin concentrations and that hemozoin can induce NOS2-derived NO formation in cultured blood mononuclear cells.