RESUMEN
Plasmodium falciparum reticulocyte-binding protein homolog 5 (RH5) is the most advanced blood-stage malaria vaccine candidate and is being evaluated for efficacy in endemic regions, emphasizing the need to study the underlying antibody response to RH5 during natural infection, which could augment or counteract responses to vaccination. Here, we found that RH5-reactive B cells were rare, and circulating immunoglobulin G (IgG) responses to RH5 were short-lived in malaria-exposed Malian individuals, despite repeated infections over multiple years. RH5-specific monoclonal antibodies isolated from eight malaria-exposed individuals mostly targeted non-neutralizing epitopes, in contrast to antibodies isolated from five RH5-vaccinated, malaria-naive UK individuals. However, MAD8-151 and MAD8-502, isolated from two malaria-exposed Malian individuals, were among the most potent neutralizers out of 186 antibodies from both cohorts and targeted the same epitopes as the most potent vaccine-induced antibodies. These results suggest that natural malaria infection may boost RH5-vaccine-induced responses and provide a clear strategy for the development of next-generation RH5 vaccines.
RESUMEN
Many infections, including malaria, are associated with an increase in autoantibodies (AAbs). Prior studies have reported an association between genetic markers of susceptibility to autoimmune disease and resistance to malaria, but the underlying mechanisms are unclear. Here, we performed a longitudinal study of children and adults (n = 602) in Mali and found that high levels of plasma AAbs before the malaria season independently predicted a reduced risk of clinical malaria in children during the ensuing malaria season. Baseline AAb seroprevalence increased with age and asymptomatic Plasmodium falciparum infection. We found that AAbs purified from the plasma of protected individuals inhibit the growth of blood-stage parasites and bind P. falciparum proteins that mediate parasite invasion. Protected individuals had higher plasma immunoglobulin G (IgG) reactivity against 33 of the 123 antigens assessed in an autoantigen microarray. This study provides evidence in support of the hypothesis that a propensity toward autoimmunity offers a survival advantage against malaria.
Asunto(s)
Autoanticuerpos , Inmunoglobulina G , Malaria Falciparum , Plasmodium falciparum , Humanos , Plasmodium falciparum/inmunología , Autoanticuerpos/inmunología , Malaria Falciparum/inmunología , Malaria Falciparum/parasitología , Niño , Preescolar , Adulto , Inmunoglobulina G/inmunología , Inmunoglobulina G/sangre , Femenino , Malí , Masculino , Adolescente , Anticuerpos Antiprotozoarios/inmunología , Estudios Longitudinales , Lactante , Antígenos de Protozoos/inmunología , Adulto Joven , Autoantígenos/inmunología , Estudios Seroepidemiológicos , Persona de Mediana EdadRESUMEN
Immunity that controls parasitemia and inflammation during Plasmodium falciparum (Pf) malaria can be acquired with repeated infections. A limited understanding of this complex immune response impedes the development of vaccines and adjunctive therapies. We conducted a prospective systems biology study of children who differed in their ability to control parasitemia and fever following Pf infection. By integrating whole-blood transcriptomics, flow-cytometric analysis, and plasma cytokine and antibody profiles, we demonstrate that a pre-infection signature of B cell enrichment, upregulation of T helper type 1 (Th1) and Th2 cell-associated pathways, including interferon responses, and p53 activation associated with control of malarial fever and coordinated with Pf-specific immunoglobulin G (IgG) and Fc receptor activation to control parasitemia. Our hypothesis-generating approach identified host molecules that may contribute to differential clinical outcomes during Pf infection. As a proof of concept, we have shown that enhanced p53 expression in monocytes attenuated Plasmodium-induced inflammation and predicted protection from fever.
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Linfocitos B/inmunología , Proteínas Sanguíneas/metabolismo , Inflamación/metabolismo , Malaria Falciparum/metabolismo , Plasmodium falciparum/fisiología , Células TH1/inmunología , Células Th2/inmunología , Proteína p53 Supresora de Tumor/metabolismo , Adolescente , Adulto , Animales , Anticuerpos Antiprotozoarios/metabolismo , Niño , Preescolar , Resistencia a la Enfermedad , Femenino , Perfilación de la Expresión Génica , Humanos , Lactante , Interferones/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Estudios Prospectivos , Receptores Fc/metabolismo , Transducción de Señal , Proteína p53 Supresora de Tumor/genética , Adulto JovenRESUMEN
BACKGROUND: Subcutaneous administration of the monoclonal antibody L9LS protected adults against controlled Plasmodium falciparum infection in a phase 1 trial. Whether a monoclonal antibody administered subcutaneously can protect children from P. falciparum infection in a region where this organism is endemic is unclear. METHODS: We conducted a phase 2 trial in Mali to assess the safety and efficacy of subcutaneous administration of L9LS in children 6 to 10 years of age over a 6-month malaria season. In part A of the trial, safety was assessed at three dose levels in adults, followed by assessment at two dose levels in children. In part B of the trial, children were randomly assigned, in a 1:1:1 ratio, to receive 150 mg of L9LS, 300 mg of L9LS, or placebo. The primary efficacy end point, assessed in a time-to-event analysis, was the first P. falciparum infection, as detected on blood smear performed at least every 2 weeks for 24 weeks. A secondary efficacy end point was the first episode of clinical malaria, as assessed in a time-to-event analysis. RESULTS: No safety concerns were identified in the dose-escalation part of the trial (part A). In part B, 225 children underwent randomization, with 75 children assigned to each group. No safety concerns were identified in part B. P. falciparum infection occurred in 36 participants (48%) in the 150-mg group, in 30 (40%) in the 300-mg group, and in 61 (81%) in the placebo group. The efficacy of L9LS against P. falciparum infection, as compared with placebo, was 66% (adjusted confidence interval [95% CI], 45 to 79) with the 150-mg dose and 70% (adjusted 95% CI, 50 to 82) with the 300-mg dose (P<0.001 for both comparisons). Efficacy against clinical malaria was 67% (adjusted 95% CI, 39 to 82) with the 150-mg dose and 77% (adjusted 95% CI, 55 to 89) with the 300-mg dose (P<0.001 for both comparisons). CONCLUSIONS: Subcutaneous administration of L9LS to children was protective against P. falciparum infection and clinical malaria over a period of 6 months. (Funded by the National Institute of Allergy and Infectious Diseases; ClinicalTrials.gov number, NCT05304611.).
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Anticuerpos Monoclonales Humanizados , Malaria Falciparum , Adulto , Niño , Femenino , Humanos , Masculino , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Enfermedades Endémicas/prevención & control , Inyecciones Subcutáneas , Estimación de Kaplan-Meier , Malaria Falciparum/tratamiento farmacológico , Malaria Falciparum/epidemiología , Malaria Falciparum/prevención & control , Malí/epidemiología , Plasmodium falciparum , Resultado del Tratamiento , Anticuerpos Monoclonales Humanizados/administración & dosificación , Anticuerpos Monoclonales Humanizados/efectos adversos , Anticuerpos Monoclonales Humanizados/uso terapéutico , Terapia por Observación Directa , Combinación Arteméter y Lumefantrina/administración & dosificación , Combinación Arteméter y Lumefantrina/uso terapéutico , Adulto Joven , Persona de Mediana EdadRESUMEN
BACKGROUND: CIS43LS is a monoclonal antibody that was shown to protect against controlled Plasmodium falciparum infection in a phase 1 clinical trial. Whether a monoclonal antibody can prevent P. falciparum infection in a region in which the infection is endemic is unknown. METHODS: We conducted a phase 2 trial to assess the safety and efficacy of a single intravenous infusion of CIS43LS against P. falciparum infection in healthy adults in Mali over a 6-month malaria season. In Part A, safety was assessed at three escalating dose levels. In Part B, participants were randomly assigned (in a 1:1:1 ratio) to receive 10 mg of CIS43LS per kilogram of body weight, 40 mg of CIS43LS per kilogram, or placebo. The primary efficacy end point, assessed in a time-to-event analysis, was the first P. falciparum infection detected on blood-smear examination, which was performed at least every 2 weeks for 24 weeks. At enrollment, all the participants received artemether-lumefantrine to clear possible P. falciparum infection. RESULTS: In Part B, 330 adults underwent randomization; 110 were assigned to each trial group. The risk of moderate headache was 3.3 times as high with 40 mg of CIS43LS per kilogram as with placebo. P. falciparum infections were detected on blood-smear examination in 39 participants (35.5%) who received 10 mg of CIS43LS per kilogram, 20 (18.2%) who received 40 mg of CIS43LS per kilogram, and 86 (78.2%) who received placebo. At 6 months, the efficacy of 40 mg of CIS43LS per kilogram as compared with placebo was 88.2% (adjusted 95% confidence interval [CI], 79.3 to 93.3; P<0.001), and the efficacy of 10 mg of CIS43LS per kilogram as compared with placebo was 75.0% (adjusted 95% CI, 61.0 to 84.0; P<0.001). CONCLUSIONS: CIS43LS was protective against P. falciparum infection over a 6-month malaria season in Mali without evident safety concerns. (Funded by the National Institute of Allergy and Infectious Diseases; ClinicalTrials.gov number, NCT04329104.).
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Anticuerpos Monoclonales Humanizados , Antimaláricos , Malaria Falciparum , Adulto , Humanos , Antimaláricos/efectos adversos , Antimaláricos/uso terapéutico , Combinación Arteméter y Lumefantrina/uso terapéutico , Malaria Falciparum/diagnóstico , Malaria Falciparum/tratamiento farmacológico , Malaria Falciparum/prevención & control , Malí , Plasmodium falciparum , Anticuerpos Monoclonales Humanizados/efectos adversos , Anticuerpos Monoclonales Humanizados/uso terapéutico , Cefalea/inducido químicamenteRESUMEN
In malaria-naïve children and adults, Plasmodium falciparum-infected red blood cells (Pf-iRBCs) trigger fever and other symptoms of systemic inflammation. However, in endemic areas where individuals experience repeated Pf infections over many years, the risk of Pf-iRBC-triggered inflammatory symptoms decreases with cumulative Pf exposure. The molecular mechanisms underlying these clinical observations remain unclear. Age-stratified analyses of uninfected, asymptomatic Malian individuals before the malaria season revealed that monocytes of adults produced lower levels of inflammatory cytokines (IL-1ß, IL-6 and TNF) in response to Pf-iRBC stimulation compared to monocytes of Malian children and malaria-naïve U.S. adults. Moreover, monocytes of Malian children produced lower levels of IL-1ß and IL-6 following Pf-iRBC stimulation compared to 4-6-month-old infants. Accordingly, monocytes of Malian adults produced more IL-10 and expressed higher levels of the regulatory molecules CD163, CD206, Arginase-1 and TGM2. These observations were recapitulated in an in vitro system of monocyte to macrophage differentiation wherein macrophages re-exposed to Pf-iRBCs exhibited attenuated inflammatory cytokine responses and a corresponding decrease in the epigenetic marker of active gene transcription, H3K4me3, at inflammatory cytokine gene loci. Together these data indicate that Pf induces epigenetic reprogramming of monocytes/macrophages toward a regulatory phenotype that attenuates inflammatory responses during subsequent Pf exposure. Trial Registration: ClinicalTrials.gov NCT01322581.
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Malaria Falciparum/inmunología , Malaria/inmunología , Monocitos/metabolismo , Fenotipo , Adulto , Niño , Preescolar , Citocinas/metabolismo , Eritrocitos/metabolismo , Humanos , Lactante , Inflamación/inmunología , Inflamación/metabolismo , Macrófagos/metabolismo , Malaria/sangre , Malaria Falciparum/sangre , Monocitos/inmunología , Plasmodium falciparum/inmunología , Plasmodium falciparum/metabolismoRESUMEN
BACKGROUND: There is currently no specific equation for estimating glomerular filtration rate (GFR) in Chinese children with chronic kidney disease (CKD). The commonly used equations are less robust than expected; we therefore sought to derive more appropriate equations for GFR estimation. METHODS: A total of 751 Chinese children with CKD were divided into 2 groups, training group (n = 501) and validation group (n = 250). In the training group, a univariate linear regression model was used to calculate predictability of variables associated with GFR. Residuals were compared to determine multivariate predictability of GFR in the equation. Standard regression techniques for Gaussian data were used to determine coefficients of GFR-estimating equations after logarithmic transformation of measured GFR (iGFR), height/serum creatinine (height/Scr), cystatin C, blood urea nitrogen (BUN), and height. These were compared with other well-known equations using the validation group. RESULTS: Median 99mTc-DTPA GFR was 90.1 (interquartile range: 67.3-108.6) mL/min/1.73 m2 in training dataset. Our CKD equation, eGFR (mL/min/1.73 m2) = 91.021 [height(m)/Scr(mg/dL)/2.7]0.443 [1.2/Cystatin C(mg/L)]0.335 [13.7/BUN (mg/dL)]-0.095 [ 0.991male] [height(m)/1.4]0.275, was derived. This was further tested in the validation group, with percentages of eGFR values within 30% and 15% of iGFR (P30 and P15) of 76.00% and 48.40%, respectively. For centres with no access to cystatin C, a creatinine-based equation, eGFR (mL/min/1.73 m2) = 89.674 [height(m)/Scr(mg/dL)/2.7]0.579 [ 1.007male] [height(m)/1.4]0.187, was derived, with P30 and P15 73.60% and 49.20%, respectively. These were significantly higher compared to other well-known equations (p < 0.05). CONCLUSION: We developed equations for GFR estimation in Chinese children with CKD based on Scr, BUN and cystatin C. These are more accurate than commonly used equations in this population.
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Cistatina C , Insuficiencia Renal Crónica , Niño , Masculino , Humanos , Tasa de Filtración Glomerular , Pueblos del Este de Asia , CreatininaRESUMEN
BACKGROUND: Plasmodium falciparum causes the majority of malaria cases worldwide and children in sub-Saharan Africa are the most vulnerable group affected. Non-sterile clinical immunity that protects from symptoms develops slowly and is relatively short-lived. Moreover, current malaria vaccine candidates fail to induce durable high-level protection in endemic settings, possibly due to the immunomodulatory effects of the malaria parasite itself. Because dendritic cells play a crucial role in initiating immune responses, the aim of this study was to better understand the impact of cumulative malaria exposure as well as concurrent P. falciparum infection on dendritic cell phenotype and function. METHODS: In this cross-sectional study, the phenotype and function of dendritic cells freshly isolated from peripheral blood samples of Malian adults with a lifelong history of malaria exposure who were either uninfected (n = 27) or asymptomatically infected with P. falciparum (n = 8) was assessed. Additionally, plasma cytokine and chemokine levels were measured in these adults and in Malian children (n = 19) with acute symptomatic malaria. RESULTS: With the exception of lower plasmacytoid dendritic cell frequencies in asymptomatically infected Malian adults, peripheral blood dendritic cell subset frequencies and HLA-DR surface expression did not differ by infection status. Peripheral blood myeloid dendritic cells of uninfected Malian adults responded to in vitro stimulation with P. falciparum blood-stage parasites by up-regulating the costimulatory molecules HLA-DR, CD80, CD86 and CD40 and secreting IL-10, CXCL9 and CXCL10. In contrast, myeloid dendritic cells of asymptomatically infected Malian adults exhibited no significant responses above the uninfected red blood cell control. IL-10 and CXCL9 plasma levels were elevated in both asymptomatic adults and children with acute malaria. CONCLUSIONS: The findings of this study indicate that myeloid dendritic cells of uninfected adults with a lifelong history of malaria exposure are able to up-regulate co-stimulatory molecules and produce cytokines. Whether mDCs of malaria-exposed individuals are efficient antigen-presenting cells capable of mounting an appropriate immune response remains to be determined. The data also highlights IL-10 and CXCL9 as important factors in both asymptomatic and acute malaria and add to the understanding of asymptomatic P. falciparum infections in malaria-endemic areas.
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Citocinas/sangre , Células Dendríticas/parasitología , Malaria Falciparum/sangre , Adulto , Infecciones Asintomáticas , Quimiocinas/sangre , Niño , Preescolar , Estudios Transversales , Eritrocitos/parasitología , Femenino , Humanos , Malaria/sangre , Masculino , Malí , Persona de Mediana Edad , Fenotipo , Plasmodium falciparum/fisiologíaRESUMEN
Natural killer (NK) cells are key effector cells of innate resistance capable of destroying tumors and virus-infected cells through cytotoxicity and rapid cytokine production. The control of NK cell responses is complex and only partially understood. PD-1 is an inhibitory receptor that regulates T cell function, but a role for PD-1 in regulating NK cell function is only beginning to emerge. Here, we investigated PD-1 expression on NK cells in children and adults in Mali in a longitudinal analysis before, during, and after infection with Plasmodium falciparum malaria. We found that NK cells transiently upregulate PD-1 expression and interleukin-6 (IL-6) production in some individuals during acute febrile malaria. Furthermore, the percentage of PD-1 expressing NK cells increases with age and cumulative malaria exposure. Consistent with this, NK cells of malaria-naive adults upregulated PD-1 following P. falciparum stimulation in vitro Additionally, functional in vitro studies revealed that PD-1 expression on NK cells is associated with diminished natural cytotoxicity but enhanced antibody-dependent cellular cytotoxicity (ADCC). These data indicate that PD-1+ NK cells expand in the context of chronic immune activation and suggest that PD-1 may contribute to skewing NK cells toward enhanced ADCC during infections such as malaria.
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Células Asesinas Naturales/inmunología , Malaria Falciparum/inmunología , Plasmodium falciparum/patogenicidad , Receptor de Muerte Celular Programada 1/metabolismo , Adulto , Factores de Edad , Animales , Citotoxicidad Celular Dependiente de Anticuerpos , Antígeno CD56/metabolismo , Línea Celular , Niño , Proteínas Ligadas a GPI/metabolismo , Humanos , Interleucina-6/metabolismo , Células K562 , Estudios Longitudinales , Malaria/inmunología , Ratones , Receptores de IgG/metabolismoRESUMEN
Many chronic infections, including malaria and HIV, are associated with a large expansion of CD21-CD27- 'atypical' memory B cells (MBCs) that exhibit reduced B cell receptor (BCR) signaling and effector functions. Little is known about the conditions or transcriptional regulators driving atypical MBC differentiation. Here we show that atypical MBCs in malaria-exposed individuals highly express the transcription factor T-bet, and that T-bet expression correlates inversely with BCR signaling and skews toward IgG3 class switching. Moreover, a longitudinal analysis of a subset of children suggested a correlation between the incidence of febrile malaria and the expansion of T-bethi B cells. The Th1-cytokine containing supernatants of malaria-stimulated PBMCs plus BCR cross linking induced T-bet expression in naïve B cells that was abrogated by neutralizing IFN-γ or blocking the IFN-γ receptor on B cells. Accordingly, recombinant IFN-γ plus BCR cross-linking drove T-bet expression in peripheral and tonsillar B cells. Consistent with this, Th1-polarized Tfh (Tfh-1) cells more efficiently induced T-bet expression in naïve B cells. These data provide new insight into the mechanisms underlying atypical MBC differentiation.
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Linfocitos B/citología , Linfocitos B/inmunología , Diferenciación Celular/inmunología , Regulación de la Expresión Génica/inmunología , Memoria Inmunológica/inmunología , Interferón gamma/biosíntesis , Malaria/inmunología , Adolescente , Adulto , Niño , Preescolar , Femenino , Proteínas Fetales/metabolismo , Humanos , Lactante , Malaria/metabolismo , Masculino , Receptores de Antígenos de Linfocitos B/metabolismo , Proteínas de Dominio T Box/metabolismo , Adulto JovenRESUMEN
Background: Chronic asymptomatic Plasmodium falciparum infections are common in endemic areas and are thought to contribute to the maintenance of malaria immunity. Whether treatment of these infections increases the subsequent risk of clinical episodes of malaria is unclear. Methods: In a 3-year study in Mali, asymptomatic individuals with or without P. falciparum infection at the end of the 6-month dry season were identified by polymerase chain reaction (PCR), and clinical malaria risk was compared during the ensuing 6-month malaria transmission season. At the end of the second dry season, 3 groups of asymptomatic children were identified: (1) children infected with P. falciparum as detected by rapid diagnostic testing (RDT) who were treated with antimalarials (n = 104), (2) RDT-negative children whose untreated P. falciparum infections were detected retrospectively by PCR (n = 55), and (3) uninfected children (RDT/PCR negative) (n = 434). Clinical malaria risk during 2 subsequent malaria seasons was compared. Plasmodium falciparum-specific antibody kinetics during the dry season were compared in children who did or did not harbor asymptomatic P. falciparum infections. Results: Chronic asymptomatic P. falciparum infection predicted decreased clinical malaria risk during the subsequent malaria season(s); treatment of these infections did not alter this reduced risk. Plasmodium falciparum-specific antibodies declined similarly in children who did or did not harbor chronic asymptomatic P. falciparum infection during the dry season. Conclusions: These findings challenge the notion that chronic asymptomatic P. falciparum infection maintains malaria immunity and suggest that mass drug administration during the dry season should not increase the subsequent risk of clinical malaria.
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Malaria Falciparum/epidemiología , Plasmodium falciparum , Adolescente , Adulto , Anticuerpos Antiprotozoarios/sangre , Anticuerpos Antiprotozoarios/inmunología , Antimaláricos/farmacología , Antimaláricos/uso terapéutico , Infecciones Asintomáticas , Niño , Preescolar , Enfermedad Crónica , Femenino , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Lactante , Malaria Falciparum/tratamiento farmacológico , Malaria Falciparum/parasitología , Malaria Falciparum/transmisión , Masculino , Malí/epidemiología , Plasmodium falciparum/efectos de los fármacos , Plasmodium falciparum/genética , Plasmodium falciparum/inmunología , Vigilancia de la Población , Riesgo , Estaciones del Año , Adulto JovenRESUMEN
In malaria-naïve individuals, Plasmodium falciparum infection results in high levels of parasite-infected red blood cells (iRBCs) that trigger systemic inflammation and fever. Conversely, individuals in endemic areas who are repeatedly infected are often asymptomatic and have low levels of iRBCs, even young children. We hypothesized that febrile malaria alters the immune system such that P. falciparum re-exposure results in reduced production of pro-inflammatory cytokines/chemokines and enhanced anti-parasite effector responses compared to responses induced before malaria. To test this hypothesis we used a systems biology approach to analyze PBMCs sampled from healthy children before the six-month malaria season and the same children seven days after treatment of their first febrile malaria episode of the ensuing season. PBMCs were stimulated with iRBC in vitro and various immune parameters were measured. Before the malaria season, children's immune cells responded to iRBCs by producing pro-inflammatory mediators such as IL-1ß, IL-6 and IL-8. Following malaria there was a marked shift in the response to iRBCs with the same children's immune cells producing lower levels of pro-inflammatory cytokines and higher levels of anti-inflammatory cytokines (IL-10, TGF-ß). In addition, molecules involved in phagocytosis and activation of adaptive immunity were upregulated after malaria as compared to before. This shift was accompanied by an increase in P. falciparum-specific CD4+Foxp3- T cells that co-produce IL-10, IFN-γ and TNF; however, after the subsequent six-month dry season, a period of markedly reduced malaria transmission, P. falciparum-inducible IL-10 production remained partially upregulated only in children with persistent asymptomatic infections. These findings suggest that in the face of P. falciparum re-exposure, children acquire exposure-dependent P. falciparum-specific immunoregulatory responses that dampen pathogenic inflammation while enhancing anti-parasite effector mechanisms. These data provide mechanistic insight into the observation that P. falciparum-infected children in endemic areas are often afebrile and tend to control parasite replication.
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Inmunidad Adaptativa , Linfocitos T CD4-Positivos/inmunología , Citocinas/inmunología , Malaria Falciparum/inmunología , Plasmodium falciparum/inmunología , Adolescente , Linfocitos T CD4-Positivos/metabolismo , Niño , Preescolar , Citocinas/sangre , Femenino , Humanos , Inflamación/sangre , Inflamación/inmunología , Inflamación/parasitología , Malaria Falciparum/sangre , Malaria Falciparum/epidemiología , Masculino , Plasmodium falciparum/metabolismoRESUMEN
BACKGROUND: Plasmodium falciparum reticulocyte-binding protein homologue 5 (PfRH5) is a blood-stage parasite protein essential for host erythrocyte invasion. PfRH5-specific antibodies raised in animals inhibit parasite growth in vitro, but the relevance of naturally acquired PfRH5-specific antibodies in humans is unclear. METHODS: We assessed pre-malaria season PfRH5-specific immunoglobulin G (IgG) levels in 357 Malian children and adults who were uninfected with Plasmodium. Subsequent P. falciparum infections were detected by polymerase chain reaction every 2 weeks and malaria episodes by weekly physical examination and self-referral for 7 months. The primary outcome was time between the first P. falciparum infection and the first febrile malaria episode. PfRH5-specific IgG was assayed for parasite growth-inhibitory activity. RESULTS: The presence of PfRH5-specific IgG at enrollment was associated with a longer time between the first blood-stage infection and the first malaria episode (PfRH5-seropositive median: 71 days, PfRH5-seronegative median: 18 days; P = .001). This association remained significant after adjustment for age and other factors associated with malaria risk/exposure (hazard ratio, .62; P = .02). Concentrated PfRH5-specific IgG purified from Malians inhibited P. falciparum growth in vitro. CONCLUSIONS: Naturally acquired PfRH5-specific IgG inhibits parasite growth in vitro and predicts protection from malaria. These findings strongly support efforts to develop PfRH5 as an urgently needed blood-stage malaria vaccine. CLINICAL TRIALS REGISTRATION: NCT01322581.
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Anticuerpos Antiprotozoarios/inmunología , Proteínas Portadoras/inmunología , Malaria Falciparum/inmunología , Plasmodium falciparum/inmunología , Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Inmunoglobulina G/inmunología , Lactante , Vacunas contra la Malaria/inmunología , Malaria Falciparum/parasitología , Malaria Falciparum/transmisión , Masculino , Placenta/inmunología , Placenta/parasitología , Embarazo , Reticulocitos/inmunología , Reticulocitos/parasitología , Adulto JovenRESUMEN
BACKGROUND: As public health efforts seek to eradicate malaria, there has been an emphasis on eliminating low-density parasite reservoirs in asymptomatic carriers. As such, diagnosing submicroscopic Plasmodium infections using PCR-based techniques has become important not only in clinical trials of malaria vaccines and therapeutics, but also in active malaria surveillance campaigns. However, PCR-based quantitative assays that rely on nucleic acid extracted from dried blood spots (DBS) have demonstrated lower sensitivity than assays that use cryopreserved whole blood as source material. METHODS: The density of Plasmodium falciparum asexual parasites was quantified using genomic DNA extracted from dried blood spots (DBS) and the sensitivity of two approaches was compared: quantitative real-time PCR (qPCR) targeting the P. falciparum 18S ribosomal RNA gene, either with an initial conventional PCR amplification prior to qPCR (nested qPCR), or without an initial amplification (qPCR only). Parasite densities determined by nested qPCR, qPCR only, and light microscopy were compared. RESULTS: Nested qPCR results in 10-fold higher sensitivity (0.5 parasites/µl) when compared to qPCR only (five parasites/ul). Among microscopy-positive samples, parasite densities calculated by nested qPCR correlated strongly with microscopy for both asymptomatic (Pearson's r=0.58, P<0.001) and symptomatic (Pearson's r=0.70, P<0.0001) P. falciparum infections. CONCLUSION: Nested qPCR improves the sensitivity for the detection of P. falciparum blood-stage infection from clinical DBS samples. This approach may be useful for active malaria surveillance in areas where submicroscopic asymptomatic infections are prevalent.
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ADN Protozoario/sangre , Pruebas con Sangre Seca/métodos , Malaria Falciparum/diagnóstico , Plasmodium falciparum/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Humanos , Malaria Falciparum/parasitología , Microscopía , Parasitemia , Plasmodium falciparum/aislamiento & purificaciónRESUMEN
Degradation of acetamiprid in wastewater was studied in a dielectric barrier discharge (DBD) reactor. This reactor produces ultraviolet light and reactive species like ozone (O3) can be used for the treatment of wastewater. We examined the factors that could affect the degradation process, including the discharge power, and the initial concentrations of acetamiprid, and O3 which is generated by the DBD reactor. We also investigated the effect of adding Na2B4O7 as a radical scavenger to probe the role of hydroxyl radical in the reaction. The results indicated that acetamiprid could be removed from aqueous solution effectively and hydroxyl radicals played an important role during the degradation by the low temperature plasma. The degradation process of acetamiprid fits the first-order kinetics. The degradation efficiency was 83.48 percent at 200 min when the discharge power was 170 W and the initial acetamiprid concentration was 50 mg/L. The removal efficiency of acetamiprid decreased with the increasing concentration of Na2B4O7 because B4O7(2-) is an excellent radical scavenger that inhibited the generation of OH during the DBD process. The removal efficiency of acetamiprid improved in the presence of O3. The main reason was that O3 can oxidize certain organic compounds directly or indirectly by generating hydroxyl radicals. The degradation products of acetamiprid were characterized qualitatively and quantitatively using high performance liquid chromatography, mass spectrometry and UV-vis spectroscopy.
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Frío , Ozono/farmacología , Piridinas/metabolismo , Aguas Residuales/química , Biodegradación Ambiental , Boratos/farmacología , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Neonicotinoides , Ozono/análisis , Piridinas/análisis , Piridinas/química , Rayos Ultravioleta , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/metabolismoRESUMEN
Malaria is a major public health problem, but many of the factors underlying the pathogenesis of this disease are not well understood. Here, we demonstrate in Malian children that susceptibility to febrile malaria following infection with Plasmodium falciparum is associated with the composition of the gut microbiome prior to the malaria season. Gnotobiotic mice colonized with the fecal samples of malaria-susceptible children had a significantly higher parasite burden following Plasmodium infection compared to gnotobiotic mice colonized with the fecal samples of malaria-resistant children. The fecal microbiome of the susceptible children was enriched for bacteria associated with inflammation, mucin degradation, gut permeability and inflammatory bowel disorders (e.g., Ruminococcus gauvreauii, Ruminococcus torques, Dorea formicigenerans, Dorea longicatena, Lachnoclostridium phocaeense and Lachnoclostridium sp. YL32). However, the susceptible children also had a greater abundance of bacteria known to produce anti-inflammatory short-chain fatty acids and those associated with favorable prognosis and remission following dysbiotic intestinal events (e.g., Anaerobutyricum hallii, Blautia producta and Sellimonas intestinalis). Metabolomics analysis of the human fecal samples corroborated the existence of inflammatory and recovery-associated features within the gut microbiome of the susceptible children. There was an enrichment of nitric oxide-derived DNA adducts (deoxyinosine and deoxyuridine) and long-chain fatty acids, the absorption of which has been shown to be inhibited by inflamed intestinal epithelial cells, and a decrease in the abundance of mucus phospholipids. Nevertheless, there were also increased levels of pseudouridine and hypoxanthine, which have been shown to be regulated in response to cellular stress and to promote recovery following injury or hypoxia. Overall, these results indicate that the gut microbiome may contribute malaria pathogenesis and suggest that therapies targeting intestinal inflammation could decrease malaria susceptibility.
RESUMEN
BACKGROUND: In experimental models of human and mouse malaria, sterilizing liver stage immunity that blocks progression of Plasmodium infection to the symptomatic blood stage can be readily demonstrated. However, it remains unclear whether individuals in malaria-endemic areas acquire such immunity. METHODS: In Mali, 251 healthy children and adults aged 4-25 years who were free of blood-stage Plasmodium infection by polymerase chain reaction (PCR) were enrolled in a longitudinal study just prior to an intense 6-month malaria season. Subsequent clinical malaria episodes were detected by weekly active surveillance and self-referral. Asymptomatic P. falciparum infections were detected by blood-smear microscopy and PCR analysis of dried blood spots that had been collected every 2 weeks for 7 months. RESULTS: As expected, the risk of clinical malaria decreased with increasing age (log-rank test, P = .0038). However, analysis of PCR data showed no age-related differences in P. falciparum infection risk (log-rank test, P = .37). CONCLUSIONS: Despite years of exposure to intense P. falciparum transmission, there is no evidence of acquired, sterile immunity to P. falciparum infection in this population, even as clinical immunity to blood-stage malaria is clearly acquired. Understanding why repeated P. falciparum infections do not induce sterile protection may lead to insights for developing vaccines that target the liver stage in malaria-endemic populations.
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Inmunidad Adaptativa , Malaria Falciparum/epidemiología , Malaria Falciparum/inmunología , Plasmodium falciparum/inmunología , Adulto , Niño , Estudios de Cohortes , Humanos , Estudios Longitudinales , Malaria Falciparum/prevención & control , Malí/epidemiologíaRESUMEN
The dielectric barrier discharge (DBD) multi-component system containing plasma, α-Fe2O3/FeVO4, and peroxymonosulfate (PMS) with high catalytic activity was successfully constructed. Thereinto, α-Fe2O3/FeVO4 was loaded on the honeycomb ceramic plate (HCP) surface (α-Fe2O3/FeVO4/HCP) and placed under the water surface below the discharge area. The catalytic activity was evaluated by the removal rate of gatifloxacin (GAT), and the DBD+α-Fe2O3/FeVO4+PMS system exhibited the optimal catalytic activity. The enhanced catalytic activity can be attributed to the fact that the occurrence of synergistic catalysis that simultaneously includes plasma oxidation, photocatalysis, PMS oxidation, O3 catalysis, and Fenton reaction. The effect of various initial degradation parameters including input power, PMS dosage, pH, etc. On GAT removal was investigated. DBD+α-Fe2O3/FeVO4+PMS system has a significant increase in the concentration of H2O2 and O3, and the role played in the multi-component system was analyzed. The identification and analysis of organic matters during GAT degradation were visualized with the help of 3D EEMs. HPLC-MS and theoretical calculations identified the major intermediates and further deduced the possible GAT degradation pathways. Additionally, the acute toxicity of the major intermediates was predicted by the QSAR model. Finally, the possible mechanisms of synergistic catalysis to enhance catalytic activity were discussed based on the characteristics of several advanced oxidation processes (AOPs) and the results of experimental and characterization. This work provides a feasible technical route and theoretical basis for wastewater treatment by plasma combined with other AOPs.
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Peróxido de Hidrógeno , Peróxidos , Gatifloxacina , Peróxidos/química , CatálisisRESUMEN
IMPORTANCE: There is increasing evidence that microbes residing within the intestines (gut microbiota) play important roles in the well-being of humans. Yet, there are considerable challenges in determining the specific role of gut microbiota in human diseases owing to the complexity of diverse internal and environmental factors that can contribute to diseases. Mice devoid of all microorganisms (germ-free mice) can be colonized with human stool samples to examine the specific contribution of the gut microbiota to a disease. These approaches have been primarily focused on stool samples obtained from individuals in Western countries. Thus, there is limited understanding as to whether the same methods used to colonize germ-free mice with stool from Western individuals would apply to the colonization of germ-free mice with stool from non-Western individuals. Here, we report the results from colonizing germ-free mice with stool samples of Malian children.
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Microbioma Gastrointestinal , Intestinos , Niño , Humanos , Animales , Ratones , Modelos Animales de Enfermedad , Vida Libre de Gérmenes , HecesRESUMEN
The development of clinical immunity to Plasmodium falciparum malaria is thought to require years of parasite exposure, a delay often attributed to difficulties in developing protective antibody levels. In this study, we evaluated several P. falciparum vaccine candidate antigens, including apical membrane antigen 1 (AMA-1), circumsporozoite protein (CSP), erythrocyte binding antigen 175 (EBA-175), and the 19-kDa region of merozoite surface protein 1 (MSP1(19)). After observing a more robust antibody response to MSP1(19), we evaluated the magnitude and longevity of IgG responses specific to this antigen in Peruvian adults and children before, during, and after P. falciparum infection. In this low-transmission region, even one reported prior infection was sufficient to produce a positive anti-MSP1(19) IgG response for >5 months in the absence of reinfection. We also observed an expansion of the total plasmablast (CD19(+) CD27(+) CD38(high)) population in the majority of individuals shortly after infection and detected MSP1-specific memory B cells in a subset of individuals at various postinfection time points. This evidence supports our hypothesis that effective antimalaria humoral immunity can develop in low-transmission regions.