Functional convergence of a germline-encoded neutralizing antibody response in rhesus macaques immunized with HCV envelope glycoproteins.

Human IGHV1-69-encoded broadly neutralizing antibodies (bnAbs) that target the hepatitis C virus (HCV) envelope glycoprotein (Env) E2 are important for protection against HCV infection. An IGHV1-69 ortholog gene, VH1.36, is preferentially used for bnAbs isolated from HCV Env-immunized rhesus macaques (RMs). Here, we studied the genetic, structural, and functional properties of VH1.36-encoded bnAbs generated by vaccination, in comparison to IGHV1-69-encoded bnAbs from HCV patients. Global B cell repertoire analysis confirmed the expansion of VH1.36-derived B cells in immunized animals. Most E2-specific, VH1.36-encoded antibodies cross-neutralized HCV. Crystal structures of two RM bnAbs with E2 revealed that the RM bnAbs engaged conserved E2 epitopes using similar molecular features as human bnAbs but with a different binding mode. Longitudinal analyses of the RM antibody repertoire responses during immunization indicated rapid lineage development of VH1.36-encoded bnAbs with limited somatic hypermutation. Our findings suggest functional convergence of a germline-encoded bnAb response to HCV Env with implications for vaccination in humans.

Broadly Neutralizing Antibodies Targeting New Sites of Vulnerability in Hepatitis C Virus E1E2.

Increasing evidence indicates that broadly neutralizing antibodies (bNAbs) play an important role in immune-mediated control of hepatitis C virus (HCV) infection, but the relative contribution of neutralizing antibodies targeting antigenic sites across the HCV envelope (E1 and E2) proteins is unclear. Here, we isolated thirteen E1E2-specific monoclonal antibodies (MAbs) from B cells of a single HCV-infected individual who cleared one genotype 1a infection and then became persistently infected with a second genotype 1a strain. These MAbs bound six distinct discontinuous antigenic sites on the E1 protein, the E2 protein, or the E1E2 heterodimer. Three antigenic sites, designated AS108, AS112 (an N-terminal E1 site), and AS146, were distinct from previously described antigenic regions (ARs) 1 to 5 and E1 sites. Antibodies targeting four sites (AR3, AR4-5, AS108, and AS146) were broadly neutralizing. These MAbs also displayed distinct patterns of relative neutralizing potency (i.e., neutralization profiles) across a panel of diverse HCV strains, which led to complementary neutralizing breadth when they were tested in combination. Overall, this study demonstrates that HCV bNAb epitopes are not restricted to previously described antigenic sites, expanding the number of sites that could be targeted for vaccine development.IMPORTANCE Worldwide, more than 70 million people are infected with hepatitis C virus (HCV), which is a leading cause of hepatocellular carcinoma and liver transplantation. Despite the development of potent direct acting antivirals (DAAs) for HCV treatment, a vaccine is urgently needed due to the high cost of treatment and the possibility of reinfection after cure. Induction of multiple broadly neutralizing antibodies (bNAbs) that target distinct epitopes on the HCV envelope proteins is one approach to vaccine development. However, antigenic sites targeted by bNAbs in individuals with spontaneous control of HCV have not been fully defined. In this study, we characterize 13 monoclonal antibodies (MAbs) from a single person who cleared an HCV infection without treatment, and we identify 3 new sites targeted by neutralizing antibodies. The sites targeted by these MAbs could inform HCV vaccine development.

Zika-specific neutralizing antibodies targeting inter-dimer envelope epitopes.

Zika virus (ZIKV) is an emerging pathogen that causes devastating congenital defects. The overlapping epidemiology and immunologic cross-reactivity between ZIKV and dengue virus (DENV) pose complex challenges to vaccine design, given the potential for antibody-dependent enhancement of disease. Therefore, classification of ZIKV-specific antibody targets is of notable value. From a ZIKV-infected rhesus macaque, we identify ZIKV-reactive B cells and isolate potent neutralizing monoclonal antibodies (mAbs) with no cross-reactivity to DENV. We group these mAbs into four distinct antigenic groups targeting ZIKV-specific cross-protomer epitopes on the envelope glycoprotein. Co-crystal structures of representative mAbs in complex with ZIKV envelope glycoprotein reveal envelope-dimer epitope and unique dimer-dimer epitope targeting. All four specificities are serologically identified in convalescent humans following ZIKV infection, and representative mAbs from all four groups protect against ZIKV replication in mice. These results provide key insights into ZIKV-specific antigenicity and have implications for ZIKV vaccine, diagnostic, and therapeutic development.

Sites of vulnerability in HCV E1E2 identified by comprehensive functional screening.

The E1 and E2 envelope proteins of hepatitis C virus (HCV) form a heterodimer that drives virus-host membrane fusion. Here, we analyze the role of each amino acid in E1E2 function, expressing 545 individual alanine mutants of E1E2 in human cells, incorporating them into infectious viral pseudoparticles, and testing them against 37 different monoclonal antibodies (MAbs) to ascertain full-length translation, folding, heterodimer assembly, CD81 binding, viral pseudoparticle incorporation, and infectivity. We propose a model describing the role of each critical residue in E1E2 functionality and use it to examine how MAbs neutralize infection by exploiting functionally critical sites of vulnerability on E1E2. Our results suggest that E1E2 is a surprisingly fragile protein complex where even a single alanine mutation at 92% of positions disrupts its function. The amino-acid-level targets identified are highly conserved and functionally critical and can be exploited for improved therapies and vaccines.

Potent Zika and dengue cross-neutralizing antibodies induced by Zika vaccination in a dengue-experienced donor.

Zika virus (ZIKV) has caused significant disease, with widespread cases of neurological pathology and congenital neurologic defects. Rapid vaccine development has led to a number of candidates capable of eliciting potent ZIKV-neutralizing antibodies (reviewed in refs. 1-3). Despite advances in vaccine development, it remains unclear how ZIKV vaccination affects immune responses in humans with prior flavivirus immunity. Here we show that a single-dose immunization of ZIKV purified inactivated vaccine (ZPIV)4-7 in a dengue virus (DENV)-experienced human elicited potent cross-neutralizing antibodies to both ZIKV and DENV. Using a unique ZIKV virion-based sorting strategy, we isolated and characterized multiple antibodies, including one termed MZ4, which targets a novel site of vulnerability centered on the Envelope (E) domain I/III linker region and protects mice from viremia and viral dissemination following ZIKV or DENV-2 challenge. These data demonstrate that Zika vaccination in a DENV-experienced individual can boost pre-existing flavivirus immunity and elicit protective responses against both ZIKV and DENV. ZPIV vaccination in Puerto Rican individuals with prior flavivirus experience yielded similar cross-neutralizing potency after a single vaccination, highlighting the potential benefit of ZIKV vaccination in flavivirus-endemic areas.

ATP-Binding Cassette Transporter A1 Deficiency in Human Induced Pluripotent Stem Cell-Derived Hepatocytes Abrogates HDL Biogenesis and Enhances Triglyceride Secretion.

Despite the recognized role of the ATP-binding Cassette Transporter A1 (ABCA1) in high-density lipoprotein (HDL) metabolism, our understanding of ABCA1 deficiency in human hepatocytes is limited. To define the functional effects of human hepatocyte ABCA1 deficiency, we generated induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (HLCs) from Tangier disease (TD) and matched control subjects. Control HLCs exhibited robust cholesterol efflux to apolipoprotein A-I (apoA-I) and formed nascent HDL particles. ABCA1-deficient HLCs failed to mediate lipid efflux or nascent HDL formation, but had elevated triglyceride (TG) secretion. Global transcriptome analysis revealed significantly increased ANGPTL3 expression in ABCA1-deficient HLCs. Angiopoietin-related protein 3 (ANGPTL3) was enriched in plasma of TD relative to control subjects. These results highlight the required role of ABCA1 in cholesterol efflux and nascent HDL formation by hepatocytes. Furthermore, our results suggest that hepatic ABCA1 deficiency results in increased hepatic TG and ANGPTL3 secretion, potentially underlying the elevated plasma TG levels in TD patients.

Large, Diverse Population Cohorts of hiPSCs and Derived Hepatocyte-like Cells Reveal Functional Genetic Variation at Blood Lipid-Associated Loci.

Genome-wide association studies have struggled to identify functional genes and variants underlying complex phenotypes. We recruited a multi-ethnic cohort of healthy volunteers (n = 91) and used their tissue to generate induced pluripotent stem cells (iPSCs) and hepatocyte-like cells (HLCs) for genome-wide mapping of expression quantitative trait loci (eQTLs) and allele-specific expression (ASE). We identified many eQTL genes (eGenes) not observed in the comparably sized Genotype-Tissue Expression project's human liver cohort (n = 96). Focusing on blood lipid-associated loci, we performed massively parallel reporter assays to screen candidate functional variants and used genome-edited stem cells, CRISPR interference, and mouse modeling to establish rs2277862-CPNE1, rs10889356-DOCK7, rs10889356-ANGPTL3, and rs10872142-FRK as functional SNP-gene sets. We demonstrated HLC eGenes CPNE1, VKORC1, UBE2L3, and ANGPTL3 and HLC ASE gene ACAA2 to be lipid-functional genes in mouse models. These findings endorse an iPSC-based experimental framework to discover functional variants and genes contributing to complex human traits.

Efectividad de un programa de rehabilitación respiratoria en pacientes con esclerosis múltiple / Effectiveness of a respiratory rehabilitation program on patients with multiple sclerosis

Las personas con esclerosis múltiple presentan alteraciones de la comunicación oral relacionadas con una insuficiencia respiratoria, lo que obliga a los especialistas en logopedia a trabajar en este problema desde la óptica de su especialidad. Objetivo Evaluar la efectividad de un programa intensivo de rehabilitación respiratoria en personas con esclerosis múltiple, para aumentar su capacidad respiratoria y mejorar la coordinación fonorrespiratoria. Métodos Estudio de intervención cuasi experimental en un grupo de 30 personas adultas de ambos sexos con esclerosis múltiple. Todos recibieron el programa durante cuatro semanas con una hora diaria de ejercitación. Para valorar la efectividad del programa, se realizó una evaluación inicial y otra final de la capacidad vital inspiratoria y espiratoria forzada y de la coordinación fonorrespiratoria, por personal externo. El deterioro neurológico se midió mediante la Escala Expandida de Discapacidad de Kurtzke. Resultados En la muestra hubo predominio de mujeres, una edad promedio de 40,43 ± 11,46 años y 13,40 ± 7,76 años de evolución de la enfermedad. Se encontró esclerosis múltiple progresiva en 22 pacientes y 8 tenían formas de brote-remisión. El deterioro neurológico fue de 5,8 (±1,51) como promedio, lo que habla a favor de una discapacidad moderada en la muestra. Se encontró un aumento de la capacidad vital inspiratoria y espiratoria forzada y un aumento en el tiempo máximo de fonación, en la emisión de series de palabras bisílabas y en la emisión de series de números. Conclusiones El programa intensivo de rehabilitación respiratoria, contribuye al aumento de la capacidad respiratoria de las personas con esclerosis múltiple, por lo que se recomienda su aplicación en el proceso de neurorrehabilitación

People with multiple sclerosis present with altered oral communication related to respiratory failure, which forces the speech therapists to work on this problem within their range of specialty. Objective To evaluate the effectiveness la efectividad) of an intensive respiratory rehabilitation program. Methods Quasi-experimental interventional study carried out in a group of 30 adults of both sexes suffering from multiple sclerosis. All these patients were included in the program for 4 weeks, having one-hour training every day. For assessing the effectiveness of the program, an initial and a final evaluation of the forced vital inspiratory and expiratory capacity and of the phonorespiratory coordination was made by outside experts. The neurological deterioration was measured according the Kurtzkeïs extended disability scale. Results Women predominated in the sample; the average age was 40.43 ± 11.46 years and progression of disease was 13.40 ± 7.76 years. Progressive multiple sclerosis was found in 22 patients and the onset-remission forms in 8 patients. The neurological deterioration amounted to 5.8 (±1.51) as an average, which speaks for the moderate disability rate present in the simple. There was observed increased vital forced inspiratory and expiratory capacities and increase in maximum phonation length and in pronouncing series of two-syllable words and series of numbers. Conclusions The intensive respiratory rehabilitation program helps to increase the respiratory capacity of the patients with multiple sclerosis, hence, its implementation in the neurorehabilitation process is recommended

Tratamiento de la disgrafia fonológica cinestésica en un paciente con apraxia oral post accidente cerebro vascular / Treatment of kinesthetic phonological disgraphia in patients with oral apraxia sequelar a stroke

Introducción: los problemas en la ejecución de la escritura dificultan las actividades de la vida diaria. El conocimiento de las características de esta habilidad, en personas que han padecido de una lesión del sistema nervioso, es imprescindible para la estruturación de un abordage terapéutico con vista a un mejor desempeño en su medio social. Objetivo: describir las características de la escritura de un paciente con apraxia oral y comprobar la influencia de un sistema de actividades para compensar la disgrafia fonológica cinestésica. Metodología: se presentó un paciente (FBZ) que presentava apraxia oral y disgrafia fonológica. Durante dos meses recibió por parte de los especialistas un sistema de actividades que tenía como objetivo mejorar la percepción y diferenciación de los sonidos del habla por su posición articulatoria con vista a compensar el transtorno que presentava en la escritura. Como protocolo de evaluación se le aplicó al inicio y final del tratamiento el capítulo V referente al diagnóstico de la escritura del test Boston. Resultados: mejoró la composición y orden de los grafemas dentro de la palabra y su inteligibilidad en el plano oral. Conclusiones: el sistema de actividades aplicado influyó positivamente en la corrección de la disgrafia fonológica.

Introduction: problems in the execution of the deed hinder activities of daily life. Knowledge of the characteristics of this ability, in people who have suffered injury to the nervous system, it is essencial to structure a therapeutic approach with a view to improved performance in their social environment. Objective: to discribe the characteristics of the writing of a patient with oral apraxia and check the influence of a system to compensate knesthetic activities phonological dysgraphia. Methodology: A patient (FBZ) presenting oral apraxia and phonological dysgraphia presented. For two months he received from specialists a system of activities aimed to improve the reception and differentiation of speech sounds by the articulatory position overlooking compensate disorder presenting in writing. As evaluation protocol was applied at the beginning and end of treatment Chapter V concerning the diagnostic test script Boston. Results: Improved composition and order of graphemes within the word and inteligibility planus. Conclusions: The applied system positively influenced the activities in the correction of phonoaudiological dysgraphia.

Papel de la terapia logopédica en pacientes con lesión medular cervical / Speech therapy in the patients with spinal cord injury

En este estudio preliminar exponemos la experiencia de la aplicación del entrenamiento motriz-respiratorio -vocálico en pacientes con patología raqui-medular con nivel de afectación cervical.Esta terapia está indicada para los casos con disminución de la intesidad vocal por déficit respiratorio ; también los pacientes con lesiones medulares cervicales padecen con frecuencia deficiencias para toser, expectorar y respirar con profundidad, además de en la emissión de la voz con fuerza, todo lo cual justifica esta terapia logopédica. La estrategia terapéutica está encaminada a lograr la recuperación de la fuerza muscular diafragmática e intercostal. Aquí se expone el resultado obtenido en la aplicación de la terapia, con tres pacientes, en el año 1998, en el Servicio de Rehabilitación Logopédica del CIREN. Nuestros resultados evidencian la efectividad del entrenamiento motriz-respiratorio-vocálico en lesionados medulares cervicales como un método efectivo para el logro de la recuperación funcional de la fortaleza de la tos,la expectoración, la respiración y la voz

Logopedia: rehabilitar las funciones del habla / Logopedics: to rehabilitate speech functions

La rehabilitación logopédica es un servicio sui géneris en el CIREN, institución dodne la neurorrehabilitación, en general, se realiza en un sistema multifactorial de forma intensiva y sistemática