RESUMEN
The analysis of ancient DNA (aDNA) from human skeletal remains can provide useful insights when investigating archaeological finds. One popular application of aDNA is to examine genealogical relationships between individuals recovered at the same archaeological site. For the reconstruction of genealogical relationships, several genetic markers are commonly used: autosomal STRs, mitochondrial lineages (based on SNP-analysis) and Y-chromosomal haplotypes (based on Y-STR-analysis). In this paper, we present the additional opportunities that X-STRs provide in aDNA kinship reconstruction, especially in deficiency cases and for the examination of father-daughter relationships. Possible applications are demonstrated on a range of different kinship reconstructions: confirmation of half-siblingship in the Lichtenstein cave (Germany), exclusion of two potential father-daughter relationships in Goslar (Germany), investigation of three siblingships in Boilstädt (Germany) as well as the confirmation of a father-daughter relationship in Stolpe (Germany). This study shows that the analysis of X-STRs can contribute to the investigation of relationship constellations otherwise difficult to approach (e.g. father-daughter relationships) and that X-STRs are useful to support and complement autosomal STRs, mtDNA and Y-STR data.
Asunto(s)
ADN Antiguo , ADN Mitocondrial , Humanos , Haplotipos/genética , ADN Mitocondrial/genética , Alemania , Restos Mortales , Repeticiones de Microsatélite/genética , Cromosomas Humanos Y/genéticaRESUMEN
Clostridium difficile infection (CDI), a leading cause of nosocomial infection, is a serious disease in North America, Europe, and Asia. CDI varies greatly from asymptomatic carriage to life-threatening diarrhea, toxic megacolon, and toxemia. The incidence of community-acquired infection has increased due to the emergence of hypervirulent antibiotic-resistant strains. These new strains contribute to the frequent occurrence of disease relapse, complicating treatment, increasing hospital stays, and increasing morbidity and mortality among patients. Therefore, it is critical to develop new therapeutic approaches that bypass the development of antimicrobial resistance and avoid disruption of gut microflora. Here, we describe the construction of a single heteromultimeric VHH-based neutralizing agent (VNA) that targets the two primary virulence factors of Clostridium difficile, toxins A (TcdA) and B (TcdB). Designated VNA2-Tcd, this agent has subnanomolar toxin neutralization potencies for both C. difficile toxins in cell assays. When given systemically by parenteral administration, VNA2-Tcd protected against CDI in gnotobiotic piglets and mice and to a lesser extent in hamsters. Protection from CDI was also observed in gnotobiotic piglets treated by gene therapy with an adenovirus that promoted the expression of VNA2-Tcd.
Asunto(s)
Anticuerpos Antibacterianos/uso terapéutico , Anticuerpos Neutralizantes/uso terapéutico , Antitoxinas/uso terapéutico , Clostridioides difficile/aislamiento & purificación , Infecciones por Clostridium/microbiología , Infecciones por Clostridium/terapia , Adenoviridae/genética , Animales , Proteínas Bacterianas/antagonistas & inhibidores , Toxinas Bacterianas/antagonistas & inhibidores , Modelos Animales de Enfermedad , Portadores de Fármacos , Evaluación Preclínica de Medicamentos , Enterotoxinas/antagonistas & inhibidores , Terapia Genética/métodos , Mesocricetus , Ratones Endogámicos C57BL , Porcinos , Resultado del TratamientoRESUMEN
Pulmonary tuberculosis (TB) is caused by Mycobacterium tuberculosis in susceptible humans. Here, we infected Diversity Outbred (DO) mice with â¼100 bacilli by aerosol to model responses in a highly heterogeneous population. Following infection, 'supersusceptible', 'susceptible' and 'resistant' phenotypes emerged. TB disease (reduced survival, weight loss, high bacterial load) correlated strongly with neutrophils, neutrophil chemokines, tumor necrosis factor (TNF) and cell death. By contrast, immune cytokines were weak correlates of disease. We next applied statistical and machine learning approaches to our dataset of cytokines and chemokines from lungs and blood. Six molecules from the lung: TNF, CXCL1, CXCL2, CXCL5, interferon-γ (IFN-γ), interleukin 12 (IL-12); and two molecules from blood - IL-2 and TNF - were identified as being important by applying both statistical and machine learning methods. Using molecular features to generate tree classifiers, CXCL1, CXCL2 and CXCL5 distinguished four classes (supersusceptible, susceptible, resistant and non-infected) from each other with approximately 77% accuracy using completely independent experimental data. By contrast, models based on other molecules were less accurate. Low to no IFN-γ, IL-12, IL-2 and IL-10 successfully discriminated non-infected mice from infected mice but failed to discriminate disease status amongst supersusceptible, susceptible and resistant M.-tuberculosis-infected DO mice. Additional analyses identified CXCL1 as a promising peripheral biomarker of disease and of CXCL1 production in the lungs. From these results, we conclude that: (1) DO mice respond variably to M. tuberculosis infection and will be useful to identify pathways involving necrosis and neutrophils; (2) data from DO mice is suited for machine learning methods to build, validate and test models with independent data based solely on molecular biomarkers; (3) low levels of immunological cytokines best indicate a lack of exposure to M. tuberculosis but cannot distinguish infection from disease.
Asunto(s)
Pulmón/patología , Neutrófilos/metabolismo , Tuberculosis/sangre , Tuberculosis/patología , Animales , Biomarcadores/sangre , Quimiocina CXCL1/sangre , Quimiocina CXCL2/sangre , Quimiocina CXCL5/sangre , Quimiocinas/sangre , Citocinas/sangre , Modelos Animales de Enfermedad , Femenino , Predisposición Genética a la Enfermedad , Interferón gamma/sangre , Aprendizaje Automático , Ratones , Ratones Endogámicos C57BL , Mycobacterium tuberculosis , Necrosis , Factor de Necrosis Tumoral alfa/sangreRESUMEN
The increasing incidence and severity of infection by Clostridium difficile have stimulated attempts to develop new antimicrobial therapies. We report here the relative abilities of two antibiotics (metronidazole and vancomycin) in current use for treating C. difficile infection and of a third antimicrobial, surotomycin, to kill C. difficile cells at various stages of development and to inhibit the production of the toxin proteins that are the major virulence factors. The results indicate that none of the drugs affects the viability of spores at 8× MIC or 80× MIC and that all of the drugs kill exponential-phase cells when provided at 8× MIC. In contrast, none of the drugs killed stationary-phase cells or inhibited toxin production when provided at 8× MIC and neither vancomycin nor metronidazole killed stationary-phase cells when provided at 80× MIC. Surotomycin, on the other hand, did kill stationary-phase cells when provided at 80× MIC but did so without inducing lysis.
Asunto(s)
Antibacterianos/farmacología , Toxinas Bacterianas/biosíntesis , Clostridioides difficile/efectos de los fármacos , Clostridioides difficile/metabolismo , Enterotoxinas/biosíntesis , Lipopéptidos/farmacología , Péptidos Cíclicos/farmacología , Toxinas Bacterianas/genética , Pared Celular/efectos de los fármacos , Clostridioides difficile/genética , Enterotoxinas/genética , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Metronidazol/farmacología , Pruebas de Sensibilidad Microbiana , Mutación/genética , Esporas Bacterianas/efectos de los fármacos , Vancomicina/farmacología , Factores de Virulencia/metabolismoRESUMEN
Arboviruses are common agents of human febrile illness worldwide. In dengue-endemic areas illness due to other arboviruses have been misdiagnosed as dengue based only on clinical-epidemiological data. In this study we investigated the presence of Brazilian arboviruses in sera of 200 patients presenting acute febrile illness, during a dengue outbreak in Sinop, MT, Brazil. The results showed that 38 samples were positive to Dengue virus (DENV) type 1, two samples to DENV type 4, and six to Mayaro virus. These results indicate that arboviruses others than DENV are circulating in Sinop and the surrounding region, which are going undiagnosed. In addition, molecular and evolutionary analyses indicate that two MAYV genotypes are co-circulating in Mato Grosso, Brazil. Thus, a strong surveillance program must be implemented to evaluate and monitor the distribution and the true importance of non-dengue arboviruses in the etiology of acute febrile illnesses.
Asunto(s)
Infecciones por Alphavirus/epidemiología , Dengue/epidemiología , Brotes de Enfermedades , ARN Viral/análisis , Adolescente , Adulto , Alphavirus/genética , Infecciones por Alphavirus/virología , Arbovirus , Brasil/epidemiología , Dengue/virología , Virus del Dengue/genética , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto JovenRESUMEN
BACKGROUND: Clostridium difficile is a primary cause of antibiotic-associated diarrhea that typically develops when gut microbiota is altered. Conventional treatment for C. difficile infection (CDI) is additional antimicrobial administration, which further disrupts normal intestinal microbiota, often resulting in poor treatment outcomes. METHODS: A pregnant dairy cow was repeatedly immunized with recombinant mutants of toxins A and B produced by C. difficile, and the resultant hyperimmune bovine colostrum (HBC) was evaluated for therapeutic efficacy in gnotobiotic piglets with diarrhea due to CDI. Control piglets received nonimmune colostrum. To determine the impact of HBC on gut microbiota, 1 of 2 groups of piglets transplanted with normal human gut microbiota was treated with HBC. RESULTS: Nonimmune colostrum-treated piglets developed moderate to severe diarrhea and colitis. In contrast, HBC-treated piglets had mild or no diarrhea and mild or no colitis. Lyophilization had no detectable impact on HBC efficacy. HBC had no discernible effect on the composition of normal human gut microbiota in the porcine intestinal tract. CONCLUSIONS: HBC provides an oral, cost-effective, and safe alternative to antibiotic therapy for CDI. By preserving intestinal microbiota, HBC may be more efficacious than antibiotics. Additional studies are warranted to establish HBC as a viable immunotherapeutic agent for human use against CDI.
Asunto(s)
Clostridioides difficile/inmunología , Infecciones por Clostridium/inmunología , Infecciones por Clostridium/terapia , Calostro/inmunología , Anciano , Animales , Antibacterianos/inmunología , Bovinos , Colitis/inmunología , Colitis/microbiología , Colitis/terapia , Diarrea/inmunología , Diarrea/microbiología , Femenino , Humanos , Factores Inmunológicos/inmunología , Enfermedades Intestinales/inmunología , Intestinos/inmunología , Intestinos/microbiología , Masculino , Persona de Mediana Edad , Embarazo , PorcinosRESUMEN
The use of anti-toxin human monoclonal antibodies (HMab) as treatment for C. difficile infection has been investigated in animal models and human clinical trials as an alternative to or in combination with traditional antibiotic therapy. While HMab therapy appears to be a promising option, how systemically administered IgG antibodies protect the colonic mucosa during Clostridium difficile infection is unknown. Using the gnotobiotic piglet model of Clostridium difficile infection, we administered a mixture of anti-TcdA and anti-TcdB HMabs systemically to piglets infected with either pathogenic or non-pathogenic C. difficile strains. The HMabs were present throughout the small and large intestinal tissue of both groups, but significant HMabs were present in the lumen of the large intestines only in the pathogenic strain-infected group. Similarly, HMabs measured in the large intestine over a period of 2-4 days following antibody administration were not significantly different over time in the gut mucosa among the groups, but concentrations in the lumen of the large intestine were again consistently higher in the pathogenic strain-infected group. These results indicate that systemically administered HMab IgG reaches the gut mucosa during the course of CDI, protecting the host against systemic intoxication, and that leakage through the damaged colon likely protects the mucosa from further damage, allowing initiation of repair and recovery.
Asunto(s)
Antitoxinas/administración & dosificación , Clostridioides difficile/inmunología , Colon/patología , Enterocolitis Seudomembranosa/patología , Enterocolitis Seudomembranosa/prevención & control , Inmunoglobulina G/administración & dosificación , Mucosa Intestinal/patología , Animales , Anticuerpos Monoclonales/administración & dosificación , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/inmunología , Toxinas Bacterianas/antagonistas & inhibidores , Toxinas Bacterianas/inmunología , Colon/inmunología , Modelos Animales de Enfermedad , Enterocolitis Seudomembranosa/mortalidad , Enterotoxinas/antagonistas & inhibidores , Enterotoxinas/inmunología , Humanos , Mucosa Intestinal/inmunología , PorcinosRESUMEN
The incidence of Clostridium difficile infection (CDI) and associated mortality have increased rapidly worldwide in recent years. Therefore, it is critical to develop new therapies for CDI. In this study, we generated a novel, potently neutralizing, tetravalent, and bispecific antibody composed of 2 heavy-chain-only VH (VHH) binding domains against both TcdA and TcdB (designated "ABA") that reverses fulminant CDI in mice infected with an epidemic 027 strain after a single injection of the antibody. We demonstrated that ABA bound to both toxins simultaneously and displayed a significantly enhanced neutralizing activity both in vitro and in vivo. Additionally, ABA was able to broadly neutralize toxins from clinical C. difficile isolates that express both TcdA and TcdB but failed to neutralize the toxin from TcdA(-)TcdB(+) C. difficile strains. This study thus provides a rationale for the development of multivalent VHHs that target both toxins and are broadly neutralizing for treating severe CDI.
Asunto(s)
Anticuerpos Antibacterianos/uso terapéutico , Proteínas Bacterianas/inmunología , Toxinas Bacterianas/inmunología , Clostridioides difficile/inmunología , Enterocolitis Seudomembranosa/prevención & control , Enterotoxinas/inmunología , Animales , Sitios de Unión de Anticuerpos/inmunología , Enterocolitis Seudomembranosa/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Ratones , Pruebas de NeutralizaciónRESUMEN
Malglycemia is a temporary problem that has significant negative sequelae. This article attempts to clarify and educate oncology nurses about the impact and management of steroid-induced malglycemia on patients with cancer receiving treatment. A management algorithm is provided to aid in evaluation and treatment decisions.
Asunto(s)
Glucemia/metabolismo , Neoplasias/terapia , Esteroides/efectos adversos , Humanos , Neoplasias/complicaciones , Neoplasias/enfermeríaRESUMEN
BACKGROUND: Dengue is a serious public health problem in numerous countries. The ability to rapidly diagnosis dengue is important for patient triage and management. Detection of dengue viral protein, NS1, represents a new approach to dengue diagnosis. OBJECTIVE: The present study aims to evaluate if there are false negative results using the NS1 Ag rapid assay (Panbio(®) Dengue Early ELISA) in two different epidemiological situations (epidemic and non-epidemic). STUDY DESIGN: 220 serum samples from patients with clinical symptoms of classical dengue fever were tested by NS1 antigen capture ELISA and Multiplex-Nested-PCR. RESULTS: In samples collected in a non-epidemic period we found a 100% agreement of ELISA and RT-PCR in dengue negative samples and 85% agreement of ELISA and RT-PCR in dengue positive samples. But when we tested samples during an epidemic period (large DENV-4 outbreak) we found 15% false negative results (p<0.05) in dengue negative samples. CONCLUSIONS: Due to false negative results for DENV-4, the sole use of the Panbio(®) Dengue Early ELISA assay as a screening method for monitoring circulating dengue serotypes must be reevaluated.
Asunto(s)
Dengue/diagnóstico , Dengue/virología , Ensayo de Inmunoadsorción Enzimática/métodos , Brasil , Virus del Dengue/genética , Virus del Dengue/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática/normas , Reacciones Falso Negativas , Humanos , Reacción en Cadena de la Polimerasa , ARN Viral/sangre , Juego de Reactivos para Diagnóstico/normas , Proteínas no Estructurales Virales/sangreRESUMEN
The American/Asian genotype of Dengue virus type 2 (DENV-2) was introduced into the Americas in the 80's. Although there is no data showing when this genotype was first introduced into Brazil, it was first detected in Brazil in 1990. After which the virus spread throughout the country and major epidemics occurred in 1998, 2007/08 and 2010. In this study we sequenced 12 DENV-2 genomes obtained from serum samples of patients with dengue fever residing in São José do Rio Preto, São Paulo (SJRP/SP), Brazil, in 2008. The whole open reading frame or envelope sequences were used to perform phylogenetic, phylogeographic and evolutionary analyses. Isolates from SJRP/SP were grouped within one lineage (BR3) close to isolates from Rio de Janeiro, Brazil. Isolates from SJRP were probably introduced there at least in 2007, prior to its detection in the 2008 outbreak. DENV-2 circulation in Brazil is characterized by the introduction, displacement and circulation of three well-defined lineages in different times, most probably from the Caribbean. Thirty-seven unique amino acid substitutions were observed among the lineages, including seven amino acid differences in domains I to III of the envelope protein. Moreover, we dated here, for the first time, the introduction of American/Asian genotype into Brazil (lineage BR1) to 1988/89, followed by the introduction of lineages BR2 (1998-2000) and BR3 (2003-05). Our results show a delay between the introduction and detection of DENV-2 lineages in Brazil, reinforcing the importance and need for surveillance programs to detect and trace the evolution of these viruses. Additionally, Brazilian DENV-2 differed in genetic diversity, date of introduction and geographic origin and distribution in Brazil, and these are important factors for the evolution, dynamics and control of dengue.
Asunto(s)
Virus del Dengue/genética , Filogenia , Brasil , Virus del Dengue/clasificación , Variación Genética/genética , Genoma Viral/genética , Genotipo , Sistemas de Lectura Abierta/genéticaRESUMEN
Hyperimmune bovine colostrum (HBC), produced by vaccination of a cow during gestation, is rich in targeted immunoglobulins, and can be used to treat a variety of diseases. The published history of HBC use for treating gastrointestinal infections in humans has developed over the past several decades and demonstrates the promise of this type of therapeutic for GI infectious disease. HBC, or purified derivative products, have been used successfully for treatment or prevention of cryptosporidiosis, shigellosis, rotavirus, enterotoxigenic E. coli, and C. difficile infection (CDI). Given the positive results of previous studies using HBC for treatment of CDI, we have produced HBC with antibodies against the two most important virulence factors of C. difficile, TcdA and TcdB, using a novel recombinant vaccine. Our preliminary results demonstrate efficacy of the HBC product for treatment of CDI in the gnotobiotic piglet model, and warrant more thorough investigation. HBC may provide an effective treatment alternative to antibiotics, which can spare the normal gut microflora, and reduce rates of recurrence and antibiotic resistance.
Asunto(s)
Proteínas Bacterianas/inmunología , Toxinas Bacterianas/inmunología , Clostridioides difficile/inmunología , Calostro/inmunología , Enterocolitis Seudomembranosa/terapia , Enterotoxinas/inmunología , Animales , Bovinos , Criptosporidiosis/inmunología , Criptosporidiosis/prevención & control , Criptosporidiosis/terapia , Disentería Bacilar/inmunología , Disentería Bacilar/prevención & control , Disentería Bacilar/terapia , Enterocolitis Seudomembranosa/inmunología , Enterocolitis Seudomembranosa/prevención & control , Escherichia coli Enterotoxigénica , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/prevención & control , Infecciones por Escherichia coli/terapia , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/prevención & control , Infecciones por Helicobacter/terapia , Helicobacter pylori/inmunología , Humanos , Factores Inmunológicos/inmunología , Infecciones por Rotavirus/inmunología , Infecciones por Rotavirus/prevención & control , Infecciones por Rotavirus/terapia , Vacunas Sintéticas/inmunologíaRESUMEN
Following successive outbreaks of dengue fever caused predominantly by dengue virus (DENV) 2 and 3, DENV-1 is now the primary serotype circulating in Brazil. We sequenced and analyzed Brazilian DENV-1 genomes and found that all isolates belong to genotype V and are subdivided into three lineages, which were introduced during four different events. The first introduction occurred in 1984-85, the second in 1997-99, and the third and fourth occurred from 2004 to 2007. These events were associated with an increase in genetic diversity but not with positive selection. Moreover, a potential new recombinant strain derived from two distinct lineages was detected. We demonstrate that the dynamics of DENV-1 in Brazil is characterized by introduction, movement, local evolution, and lineage replacement. This study strengthens the relevance of genotype surveillance in order to identify, trace, and control virus populations circulating in Brazil and Latin America.
Asunto(s)
Virus del Dengue/clasificación , Virus del Dengue/genética , Dengue/virología , Variación Genética , ARN Viral/genética , Brasil , Virus del Dengue/aislamiento & purificación , Evolución Molecular , Genotipo , Humanos , Recombinación Genética , Análisis de Secuencia de ADNRESUMEN
HIV infection results in a decrease in circulating CD4(+) T-cell and naive T-cell numbers. If such losses were associated with an erosion of T-cell receptor (TCR) repertoire diversity in the peripheral T-cell pool, this might exacerbate the state of persistent immunodeficiency. Existing methods for the analysis of the TCR repertoire have demonstrated skewed distributions of TCR genes in HIV-infected subjects but cannot directly measure TCR diversity. Here we used AmpliCot, a quantitative assay based on DNA hybridization kinetics, to measure TCR diversity in a cross-sectional comparison of 19 HIV-infected persons to 18 HIV-uninfected controls. HIV-infected persons had a 10-fold decrease in total TCR repertoire diversity in 1.5 mL of blood compared with uninfected controls, with decreased diversity correlating most closely with a lower CD4(+) T-cell percentage. Nonetheless, the TCR repertoire diversity of sort-purified T-cell subpopulations in HIV-infected and HIV-uninfected subjects was comparable. These observations suggest that the TCR repertoire diversity changes in whole blood during HIV disease progression are primarily the result of changes in the number and proportion of T-cell subpopulations and that most HIV-infected persons may retain a sufficiently diverse TCR repertoire to permit immune reconstitution with antiretroviral therapy alone, without thymopoiesis.
Asunto(s)
Variación Genética/fisiología , Infecciones por VIH/inmunología , VIH-1/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Adulto , Anciano , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Estudios de Cohortes , Progresión de la Enfermedad , Regulación hacia Abajo , Femenino , Estudios de Seguimiento , Reordenamiento Génico de Linfocito T/genética , Reordenamiento Génico de Linfocito T/fisiología , Infecciones por VIH/sangre , Infecciones por VIH/genética , Humanos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismoRESUMEN
Cryptosporidium parvum induces the formation of an actin-dense plaque which is essential for the successful invasion of epithelial cells. Host molecules that are involved in the regulation of this cytoskeleton reorganization are unknown. Here we identified that calcium-dependent thiol protease calpain is critical for regulating parasite-induced actin polymerization. C. parvum invasion induced activation of calpain. Inhibition of calpain activity by overexpression of the endogenous inhibitor calpastatin diminished the formation of the actin-dense plaque and decreased the initial invasion of parasites. Our data indicates a key role of calpain activity of host cell in C. parvum infection via regulating cytoskeleton reorganization.
Asunto(s)
Calpaína/antagonistas & inhibidores , Calpaína/metabolismo , Criptosporidiosis/inmunología , Criptosporidiosis/parasitología , Cryptosporidium parvum , Línea Celular Tumoral , Cryptosporidium parvum/fisiología , HumanosRESUMEN
BACKGROUND: Dengue fever is one of the most significant re-emerging tropical diseases, despite our expanding knowledge of the disease, viral tropism is still not known to target heart tissues or muscle. METHODS: A prospective pediatric clinical cohort of 102 dengue hemorrhagic fever patients from Colombia, South America, was followed for 1 year. Clinical diagnosis of myocarditis was routinely performed. Electrocardiograph and echocardiograph analysis were performed to confirm those cases. Immunohistochemistry for detection of dengue virus and inflammatory markers was performed on autopsied heart tissue. In vitro studies of human striated skeletal fibers (myotubes) infected with dengue virus were used as a model for myocyte infection. Measurements of intracellular Ca2+ concentration as well as immunodetection of dengue virus and inflammation markers in infected myotubes were performed. RESULTS: Eleven children with dengue hemorrhagic fever presented with symptoms of myocarditis. Widespread viral infection of the heart, myocardial endothelium, and cardiomyocytes, accompanied by inflammation was observed in 1 fatal case. Immunofluorescence confocal microscopy showed that myotubes were infected by dengue virus and had increased expression of the inflammatory genes and protein IP-10. The infected myotubes also had increases in intracellular Ca2+ concentration. CONCLUSIONS: Vigorous infection of heart tissues in vivo and striated skeletal cells in vitro are demonstrated. Derangements of Ca2+ storage in the infected cells may directly contribute to the presentation of myocarditis in pediatric patients.
Asunto(s)
Virus del Dengue/fisiología , Corazón/virología , Músculo Esquelético/virología , Dengue Grave/patología , Tropismo Viral , Calcio/análisis , Células Cultivadas , Niño , Preescolar , Estudios de Cohortes , Colombia , Citosol/química , Virus del Dengue/patogenicidad , Ecocardiografía , Electrocardiografía , Femenino , Humanos , Inmunohistoquímica , Lactante , Mediadores de Inflamación/análisis , Masculino , Microscopía , Músculo Esquelético/patología , Miocardio/patología , Estudios ProspectivosRESUMEN
Serotonin is a monoamine acting as a neuromediator in the central and peripheral nervous system. Recently, serotonin has also been shown to influence T- and B-cell function. The serotonin transporter is central in the regulation of the serotonergic system and widely expressed on cells of the immune system. A functional length polymorphism in the promoter of the serotonin transporter gene (5-HTTLPR) has been implicated in the genetic background of depression. Psoriasis is a complex disease with a polygenetic inheritance. In light of the role of T-cell mediated inflammation in psoriasis and the increased prevalence of depression in psoriatic patients, we analyzed the 5-HTTLPR polymorphism in 309 patients with psoriasis vulgaris and 315 healthy control individuals. No significant differences in genotype distribution and allele frequencies were found. There was also no difference in the score of the Hamilton Rating Scale for Depression in patients with psoriasis (n = 137) characterized by carriage of different 5-HTTLPR genotypes. These findings argue against a major contribution of the 5-HTTLPR polymorphism to psoriasis susceptibility and the occurrence of depressive symptoms among psoriatic patients.
Asunto(s)
Psoriasis/genética , Proteínas de Transporte de Serotonina en la Membrana Plasmática/genética , Adulto , Estudios de Casos y Controles , Depresión , Progresión de la Enfermedad , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo Genético , Psoriasis/inmunología , Psoriasis/fisiopatología , Psoriasis/psicología , Proteínas de Transporte de Serotonina en la Membrana Plasmática/inmunología , Proteínas de Transporte de Serotonina en la Membrana Plasmática/metabolismo , Índice de Severidad de la EnfermedadRESUMEN
Dengue virus (DENV) is a mosquito-borne flavivirus that causes an acute febrile disease in humans, characterized by musculoskeletal pain, headache, rash and leukopenia. The cause of myalgia during DENV infection is still unknown. To determine whether DENV can infect primary muscle cells, human muscle satellite cells were exposed to DENV in vitro. The results demonstrated for the first time high-efficiency infection and replication of DENV in human primary muscle satellite cells. Changes in global gene expression were also examined in these cells following DENV infection using Affymetrix GeneChip analysis. The differentially regulated genes belonged to two main functional categories: cell growth and development, and antiviral type I interferon (IFN) response genes. Increased expression of the type I IFN response genes for tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), melanoma-derived antigen 5 (MDA-5), IFN-gamma-inducible protein 10 (IP-10), galectin 3 soluble binding protein (LGals3BP) and IFN response factor 7 (IRF7) was confirmed by quantitative RT-PCR. Furthermore, higher levels of cell-surface-bound intracellular adhesion molecule-1 (ICAM-1) and soluble ICAM-1 in the cell-culture medium were detected following DENV infection. However, DENV infection impaired the ability of the infected cells in the culture medium to upregulate cell-surface expression of MHC I molecules, suggesting a possible mechanism of immune evasion by DENV. The findings of this study warrant further clinical research to identify whether muscle cells are targets for DENV infection during the acute stage of the disease in vivo.
Asunto(s)
Virus del Dengue/inmunología , Perfilación de la Expresión Génica , Antígenos de Histocompatibilidad Clase I/biosíntesis , Células Musculares/virología , Células Cultivadas , Regulación de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Growth hormone (GH) is an underappreciated but important regulator of T cell development that can reverse age-related declines in thymopoiesis in rodents. Here, we report findings of a prospective randomized study examining the effects of GH on the immune system of HIV-1-infected adults. GH treatment was associated with increased thymic mass. In addition, GH treatment enhanced thymic output, as measured by both the frequency of T cell receptor rearrangement excision circles in circulating T cells and the numbers of circulating naive and total CD4(+) T cells. These findings provide compelling evidence that GH induces de novo T cell production and may, accordingly, facilitate CD4(+) T cell recovery in HIV-1-infected adults. Further, these randomized, prospective data have shown that thymic involution can be pharmacologically reversed in humans, suggesting that immune-based therapies could be used to enhance thymopoiesis in immunodeficient individuals.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/fisiopatología , Hormona del Crecimiento/uso terapéutico , VIH-1 , Timo/efectos de los fármacos , Síndrome de Inmunodeficiencia Adquirida/inmunología , Adulto , Anciano , Recuento de Linfocito CD4 , Estudios Cruzados , Hormona del Crecimiento/efectos adversos , Humanos , Factor I del Crecimiento Similar a la Insulina/análisis , Linfopoyesis/efectos de los fármacos , Persona de Mediana Edad , Estudios Prospectivos , Timo/fisiopatologíaRESUMEN
Variations in the melanocortin-1 receptor (MC1R) and in the glutathione-S transferase genes mu1 (GSTM1) and theta 1 (GSTT1) have been reported to influence UV sensitivity and melanoma risk. MC1R is one of the major genes that determine skin pigmentation because the melanocortin-1 receptor regulates eumelanin synthesis. GSTT1 and GSTM1 are enzymes expressed in the skin that detoxify products of oxidative stress reactions caused by UV irradiation. In this study variations in the MC1R, GSTM1 and T1 genes were analyzed in 347 healthy subjects and 322 patients with cutaneous malignant melanoma by direct cycle sequencing, RFLP and multiplex PCR. Important phenotypic characteristics of the study participants were obtained to assess whether genetic associations occurred independently of phenotypic risk factors for melanoma. We found an association of the MC1R D84E and R151C polymorphisms with melanoma (odds ratios for carriage of the rare allele 4.96, 95% CI [1.06-23.13], P = 0.032, and 1.69, 95% CI [1.12-2.55], P = 0.013, respectively). Melanoma risk increased with the number of variant MC1R alleles carried by an individual (P = 0.003). In a multivariate model, however, only the D84E polymorphism influenced melanoma risk independently of the risk factors fair skin type, high nevus count and high age (P = 0.047). There was no effect of homozygous GST M1 or T1 deletions on melanoma risk. In contrast to previous data, there was no evidence that GSTM1 deficiency influences melanoma risk in the subgroup of individuals with red or blond hair.