RESUMEN
BACKGROUND: Genetic diagnosis of inborn errors of immunity (IEI) is complex due to the large number of genes involved and their molecular features. Missense variants have been reported as the most common cause of IEI. However, the frequency of copy number variants (CNVs) may be underestimated since their detection requires specific quantitative techniques. At this point, the use of Next Generation Sequencing (NGS) is acquiring relevance. METHODS: In this article, we present our experience in the genetic diagnosis of IEI based on three diagnostic algorithms that allowed the detection of single nucleotide variants (SNVs) and CNVs. Following this approximation, 703 index cases were evaluated between 2014 and 2021. Sanger sequencing, MLPA, CGH array, breakpoint spanning PCR or a customized NGS-based multigene-targeted panel were performed. RESULTS: A genetic diagnosis was reached in 142 of the 703 index cases (20%), 19 of them presented deletions as causal variants. Deletions were also detected in 5 affected relatives and 16 healthy carriers during the family studies. Additionally, we compile, characterize and present all the CNVs detected by our diagnostic algorithms, representing the largest cohort of deletions related to IEI to date. Furthermore, three bioinformatic tools (LACONv, XHMM, VarSeq™) based on NGS data were evaluated. VarSeq™ was the most sensitive and specific bioinformatic tool; detecting 21/23 (91%) deletions located in captured regions. CONCLUSION: Based on our results, we propose a strategy to guide the molecular diagnosis that can be followed by expert and non-expert centres in the field of IEI.
Asunto(s)
Variaciones en el Número de Copia de ADN , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Variaciones en el Número de Copia de ADN/genética , Algoritmos , Masculino , Femenino , Polimorfismo de Nucleótido Simple , Niño , Mutación Missense/genéticaRESUMEN
OBJECTIVE: Wolf-Hirschhorn syndrome is a rare disease of genetic origin caused by the deletion of the distal end of chromosome 4, including at least the region p16.3. The objectives of this work were to determine the prevalence of the disease in the Spanish population, as well as to establish the geographical distribution of the syndrome throughout the Spanish geography, elucidating the age range in which there are more patients. METHODS: Patients diagnosed with the disease for nine years (2013-2021) throughout the Spanish territory were recruited for the research, thanks to agreements with the Spanish Association of Wolf-Hirschhorn Syndrome (AESWH). The clinical information of the patients was obtained from referring physicians using two standardized questionnaires completed with data from medical reports and the parent interview. The molecular characterization of the disease was made using SNP (single nucleotide polymorphism) microarrays (cytoSNP850K, Illumina, USA). The data were statistically processed using Microsoft Excel (Microsoft Corporation) and SPSS (IBM) software, using comparisons between two groups s with Student's t-test (for continuous variables) or with Chi-square tests (for categorical ones). For more than two groups, ANOVA analyses were performed (followed by Bonferroni or T3-Dunnett post hoc tests) for continuous variables and z-tests between column proportions for categorical variables. RESULTS: In Spain (until 2021) eighty people are diagnosed with this syndrome, estimating its prevalence at 1.69x10-4 per 10,000 inhabitants and / or 1/172,904 for each live newborn. This paper remarks the existence of important differences in prevalence between the different regions in Spain. The region with the most diagnosed patients was Madrid, although the highest prevalence was observed in Asturias. Significant differences have been established in terms of sex and disease (ratio of women to men of 2:1), and the mean age at diagnosis has been established at 7.20 years. CONCLUSIONS: The prevalence of this syndrome in Spain has been estimated well below the prevalence that is handled in scientific texts (1/50,000 newborns). In addition, we have determined that this prevalence shows large geographical differences, which allows us to affirm that this syndrome could be under-diagnosed in our country. Most of the patients included in this cohort are of paediatric age. It has not been possible to corroborate that mortality in this syndrome, in our population, occurs preferably during the first two years of life, as has been claimed.
OBJETIVO: El Síndrome de Wolf-Hirschhorn es una enfermedad poco frecuente de origen genético causada por la deleción del extremo distal del cromosoma 4, que incluye preferentemente la región p16.3. Los objetivos de este trabajo fueron determinar la prevalencia de la enfermedad en la población española, así como establecer la distribución geográfica del síndrome a lo largo de la geografía nacional, dilucidando el rango de edad en el que existían más pacientes afectados. METODOS: Para la investigación se reclutaron 80 pacientes diagnosticados con el síndrome en el periodo 2013-2021, en todo el territorio español, gracias a los acuerdos con la Asociación Española de Síndrome Wolf-Hirschhorn (AESWH). La información clínica de los pacientes se obtuvo mediante dos cuestionarios estandarizados que fueron cumplimentados por médicos de referencia y los padres, siendo completados y corroborados con los distintos informes médicos de cada paciente y, al menos, una entrevista una entrevista a los padres. La caracterización molecular de la enfermedad se realizó mediante el uso de microarrays de SNP (del inglés, single nucleotide polymorphism) (CytoSNP 850K, Illumina). Los datos se trataron estadísiticamente utilizando los softwares Microsoft Excel (Microsoft Corporation) y SPSS (IBM), mediante las comparaciones entre dos grupos s con la prueba t de Student (para variables continuas) o con pruebas de Chi cuadrado (para las categóricas). Para más de dos grupos se realizó análisis ANOVA (seguido de las pruebas post hoc de Bonferroni o T3-Dunnett) para variables continuas y pruebas z entre proporciones de columna para variables categóricas. RESULTADOS: En España (hasta 2021) están diagnosticadas ochenta personas con este síndrome, estimándose su prevalencia en 1,69x10-4 por cada 10.000 habitantes y/o 1/172.904 por cada recién nacido vivo. En este trabajo se constató la existencia de importantes diferencias de prevalencia entre las comunidades autónomas de nuestro país. La comunidad con más pacientes diagnosticados fue Madrid, aunque la mayor prevalencia se observó en Asturias. Se establecieron diferencias estadísticamente significativas en cuanto al sexo y la enfermedad (proporción de mujeres sobre varones de 2:1), así como se estableció la edad media al diagnóstico en 7,20 años. CONCLUSIONES: La prevalencia de este síndrome en España se estima muy por debajo de la prevalencia que se maneja en los textos científicos (1 por cada 50.000 recién nacidos). Adicionalmente, hemos determinado que esta prevalencia muestra grandes diferencias geográficas, lo que nos permite afirmar que este síndrome podría encontrarse infra-diagnosticado en nuestro país. La mayor parte de los pacientes incluidos en esta cohorte se encuentran en edad pediátrica. No se ha podido corroborar que la mortalidad en este síndrome, en nuestra población, ocurra preferentemente durante los dos primeros años de vida, como se venía afirmando.
Asunto(s)
Síndrome de Wolf-Hirschhorn , Niño , Femenino , Humanos , Recién Nacido , Prevalencia , España/epidemiología , Síndrome de Wolf-Hirschhorn/diagnóstico , Síndrome de Wolf-Hirschhorn/epidemiología , Síndrome de Wolf-Hirschhorn/genéticaRESUMEN
FUNDAMENTOS: El Síndrome de Wolf-Hirschhorn es una enfermedad poco frecuente de origen genético causada por la delecióndel extremo distal del cromosoma 4, que incluye preferentemente la región p16.3. Los objetivos de este trabajo fueron determinar laprevalencia de la enfermedad en la población española, así como establecer la distribución geográfica del síndrome a lo largo de lageografía nacional, dilucidando el rango de edad en el que existían más pacientes afectados.MÉTODOS: Para la investigación se reclutaron 80 pacientes diagnosticados con el síndrome en el periodo 2013-2021, en todo elterritorio español, gracias a los acuerdos con la Asociación Española de Síndrome Wolf-Hirschhorn (AESWH). La información clínica de los pacientes se obtuvo mediante dos cuestionarios estandarizados que fueron cumplimentados por médicos de referencia y los padres, siendo completados y corroborados con los distintos informes médicos de cada paciente y, al menos, una entrevista una entrevista a los padres. La caracterización molecular de la enfermedad se realizó mediante el uso de microarrays de SNP(del inglés, single nucleotide polymorphism) (CytoSNP 850K, Illumina). Los datos se trataron estadísiticamente utilizando los softwaresMicrosoft Excel (Microsoft Corporation) y SPSS (IBM), mediante las comparaciones entre dos grupos s con la prueba t de Student (para variables continuas) o con pruebas de Chi cuadrado (para las categóricas). Para más de dos grupos se realizó análisis ANOVA (seguido de las pruebas post hoc de Bonferroni o T3-Dunnett) para variables continuas y pruebas z entre proporciones de columna para variables categóricas. RESULTADOS: En España (hasta 2021) están diagnosticadas ochenta personas con este síndrome, estimándose su prevalencia en1,69x10-4 por cada 10.000 habitantes y/o 1/172.904 por cada recién nacido vivo. En este trabajo se constató la existencia de importantesdiferencias de prevalencia entre las comunidades autónomas de nuestro país.(AU)
BACKGROUND: Wolf-Hirschhorn syndrome is a rare disease of genetic origin caused by the deletion of the distal end of chromo-some 4, including at least the region p16.3. The objectives of this work were to determine the prevalence of the disease in the Spanishpopulation, as well as to establish the geographical distribution of the syndrome throughout the Spanish geography, elucidating theage range in which there are more patients. METHODS: Patients diagnosed with the disease for nine years (2013-2021) throughout the Spanish territory were recruited for theresearch, thanks to agreements with the Spanish Association of Wolf-Hirschhorn Syndrome (AESWH). The clinical information of thepatients was obtained from referring physicians using two standardized questionnaires completed with data from medical reports andthe parent interview. The molecular characterization of the disease was made using SNP (single nucleotide polymorphism) microarrays (cytoSNP850K, Illumina, USA). The data were statistically processed using Microsoft Excel (Microsoft Corporation) and SPSS (IBM) software,using comparisons between two groups s with Students t-test (for continuous variables) or with Chi-square tests (for categorical ones).For more than two groups, ANOVA analyses were performed (followed by Bonferroni or T3-Dunnett post hoc tests) for continuous varia-bles and z-tests between column proportions for categorical variables. RESULTS: In Spain (until 2021) eighty people are diagnosed with this syndrome, estimating its prevalence at 1.69x10-4 per 10,000inhabitants and / or 1/172,904 for each live newborn. This paper remarks the existence of important differences in prevalence betweenthe different regions in Spain. The region with the most diagnosed patients was Madrid, although the highest prevalence was obser-ved in Asturias.(AU)
Asunto(s)
Humanos , Masculino , Femenino , Demografía , Prevalencia , Síndrome de Wolf-Hirschhorn , España , Salud Pública , Encuestas y CuestionariosRESUMEN
Pulmonary arterial hypertension (PAH) is a severe clinical condition characterized by an increase in mean pulmonary artery pressure, which leads to a right ventricular hypertrophy and potentially heart failure and death. In the last several years, many genes have been associated with PAH, particularly in idiopathic and heritable forms but also in associated forms. Here we described the identification of two unrelated families in which the AQP1 variant was found from a cohort of 300 patients. The variants were identified by whole exome sequencing (WES). In the first family, the variant was detected in three affected members from a hereditary PAH, and in the second family the proband had PAH associated with scleroderma. In addition, we have reviewed all cases published in the literature thus far of patients with PAH and AQP1 variants. Functional studies have led to some contradictory conclusions, and the evidence of the relationship of AQP1 and PAH is still limited. However, we describe two further families with PAH and variants in AQP1, expanding both the number of cases and the clinically associated phenotype. We provide further evidence of the association of AQP1 and the development of hereditary and associated forms of PAH.
Asunto(s)
Insuficiencia Cardíaca , Hipertensión Pulmonar , Hipertensión Arterial Pulmonar , Acuaporina 1/genética , Hipertensión Pulmonar Primaria Familiar/genética , Humanos , Hipertensión Pulmonar/genética , Hipertensión Arterial Pulmonar/genética , Secuenciación del ExomaRESUMEN
Data on convalescent plasma (CP) treatment in COVID-19 outpatients are scarce. We aimed to assess whether CP administered during the first week of symptoms reduced the disease progression or risk of hospitalization of outpatients. Two multicenter, double-blind randomized trials (NCT04621123, NCT04589949) were merged with data pooling starting when <20% of recruitment target was achieved. A Bayesian-adaptive individual patient data meta-analysis was implemented. Outpatients aged ≥50 years and symptomatic for ≤7days were included. The intervention consisted of 200-300mL of CP with a predefined minimum level of antibodies. Primary endpoints were a 5-point disease severity scale and a composite of hospitalization or death by 28 days. Amongst the 797 patients included, 390 received CP and 392 placebo; they had a median age of 58 years, 1 comorbidity, 5 days symptoms and 93% had negative IgG antibody-test. Seventy-four patients were hospitalized, 6 required mechanical ventilation and 3 died. The odds ratio (OR) of CP for improved disease severity scale was 0.936 (credible interval (CI) 0.667-1.311); OR for hospitalization or death was 0.919 (CI 0.592-1.416). CP effect on hospital admission or death was largest in patients with ≤5 days of symptoms (OR 0.658, 95%CI 0.394-1.085). CP did not decrease the time to full symptom resolution. TRIAL REGISTRATION: Clinicaltrials.gov NCT04621123 and NCT04589949. REGISTRATION: NCT04621123 and NCT04589949 on https://www. CLINICALTRIALS: gov.
Asunto(s)
COVID-19 , Teorema de Bayes , COVID-19/terapia , Humanos , Inmunización Pasiva , Persona de Mediana Edad , Estudios Multicéntricos como Asunto , Pacientes Ambulatorios , Ensayos Clínicos Controlados Aleatorios como Asunto , SARS-CoV-2 , Resultado del Tratamiento , Sueroterapia para COVID-19RESUMEN
La osteoporosis es un trastorno para tener en cuenta en niños con patologías crónicas graves o con algunas enfermedades genéticas que predisponen al incremento de la fragilidad ósea. La osteoporosis primaria es una entidad con etiologías emergentes y puede ocurrir en forma sindrómica. La asociación con pliegues retinianos congénitos debe orientar al diagnóstico de osteoporosis-pseudoglioma (OMIM 259770), síndrome poco frecuente (prevalencia de 1/2000000), que se origina por la pérdida de función de la proteína LRP5 (low-density lipoprotein receptor-related protein 5) y compromete la vía de señalización de Wnt/ß-catenina. Se presenta el caso de un niño con pliegues retinianos congénitos, ceguera progresiva y múltiples fracturas cuyo estudio clínico, bioquímico y genético confirmó el diagnóstico de osteoporosis primaria debido a una nueva variante inactivante en el gen LRP5 en homocigosis
Osteoporosis should be considered in children with severe chronic diseases or in association with some genetic diseases that bear an increased risk of bone fragility. Primary osteoporosis is an entity in which emerging aetiologies are being recognized. Its association with congenital retinal folds should guide the diagnosis to the Osteoporosis-Pseudoglioma syndrome (OMIM 259770), a rare disease (prevalence of 1/2000000), caused by the loss of function of the protein LRP5 (low-density lipoprotein receptor-related protein 5) resulting in the alteration of the Wnt/ß-catenin signalling pathway. We report the case of a child with congenital retinal folds, progressive loss of vision and multiple fractures whose clinical, biochemical and genetic studies confirmed the diagnosis of primary osteoporosis due to a novel homozygous inactivating variant in LRP5
Asunto(s)
Humanos , Masculino , Niño , Osteoporosis/diagnóstico , Osteoporosis/terapia , Ceguera , Fracturas MúltiplesRESUMEN
Osteoporosis should be considered in children with severe chronic diseases or in association with some genetic diseases that bear an increased risk of bone fragility. Primary osteoporosis is an entity in which emerging aetiologies are being recognized. Its association with congenital retinal folds should guide the diagnosis to the Osteoporosis-Pseudoglioma syndrome (OMIM 259770), a rare disease (prevalence of 1/2 000 000), caused by the loss of function of the protein LRP5 (low-density lipoprotein receptor-related protein 5) resulting in the alteration of the Wnt/ß-catenin signalling pathway. We report the case of a child with congenital retinal folds, progressive loss of vision and multiple fractures whose clinical, biochemical and genetic studies confirmed the diagnosis of primary osteoporosis due to a novel homozygous inactivating variant in LRP5.
La osteoporosis es un trastorno para tener en cuenta en niños con patologías crónicas graves o con algunas enfermedades genéticas que predisponen al incremento de la fragilidad ósea. La osteoporosis primaria es una entidad con etiologías emergentes y puede ocurrir en forma sindrómica. La asociación con pliegues retinianos congénitos debe orientar al diagnóstico de osteoporosis-pseudoglioma (OMIM 259770), síndrome poco frecuente (prevalencia de 1/2 000 000), que se origina por la pérdida de función de la proteína LRP5 (low-density lipoprotein receptor-related protein 5) y compromete la vía de señalización de Wnt/ß-catenina. Se presenta el caso de un niño con pliegues retinianos congénitos, ceguera progresiva y múltiples fracturas cuyo estudio clínico, bioquímico y genético confirmó el diagnóstico de osteoporosis primaria debido a una nueva variante inactivante en el gen LRP5 en homocigosis.
Asunto(s)
Osteogénesis Imperfecta/diagnóstico , Niño , Marcadores Genéticos , Pruebas Genéticas , Homocigoto , Humanos , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad/genética , Masculino , Mutación , Osteogénesis Imperfecta/genéticaRESUMEN
In 31 participants who started first-line antiretroviral therapy in the NEAT 001/ANRS 143 clinical trial, we found after 96 weeks a statistically significant increase in blood telomere length (TL) of 0.04 (T/S Ratio) (p = 0.03). This increase was positively correlated with both the change in the percentage of CD4+ T-cells and with the decrease of CD38+ molecules on Central Memory CD8+ and negatively correlated with the change in the percentage of CD4+ Effector Memory cells. Increase in TL could be an expression of immune reconstitution and the associated decrease in immune activation. We acknowledge for the low statistical power due to the small sample size and the potential for false positive results due to multiple testing. Hence, further studies are needed to confirm these observations.
Asunto(s)
Antirretrovirales/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Infecciones por VIH/inmunología , Subgrupos de Linfocitos T/inmunología , Telómero/inmunología , ADP-Ribosil Ciclasa 1/inmunología , Adulto , Antirretrovirales/uso terapéutico , Terapia Antirretroviral Altamente Activa/métodos , Recuento de Linfocito CD4/métodos , Femenino , Infecciones por VIH/tratamiento farmacológico , Seropositividad para VIH/inmunología , VIH-1/efectos de los fármacos , VIH-1/inmunología , Humanos , Memoria Inmunológica/inmunología , Inmunofenotipificación/métodos , Activación de Linfocitos/inmunología , Masculino , Persona de Mediana Edad , Carga Viral/inmunologíaRESUMEN
BACKGROUND: IGF1 is a key factor in fetal and postnatal growth. To date, only three homozygous IGF1 gene defects leading to complete or partial loss of IGF1 activity have been reported in three short patients born small for gestational age. We describe the fourth patient with severe short stature presenting a novel homozygous IGF1 gene mutation. RESULTS: We report a boy born from consanguineous parents at 40 weeks of gestational age with intrauterine growth restriction and severe postnatal growth failure. Physical examination revealed proportionate short stature, microcephaly, facial dysmorphism, bilateral sensorineural deafness and mild global developmental delay. Basal growth hormone (GH) fluctuated from 0.2 to 29 ng/mL, while IGF1 levels ranged from -1.15 to 2.95 SDS. IGFBP3 was normal-high. SNP array delimited chromosomal regions of homozygosity, including 12q23.2 where IGF1 is located. IGF1 screening by HRM revealed a homozygous missense variant NM_000618.4(IGF1):c.322T>C, p.(Tyr108His). The change of the highly conserved Tyr60 in the mature IGF1 peptide was consistently predicted as pathogenic by multiple bioinformatic tools. Tyr60 has been described to be critical for IGF1 interaction with type 1 IGF receptor (IGF1R). In vitro, HEK293T cells showed a marked reduction of IGF1R phosphorylation after stimulation with serum from the patient as compared to sera from age-matched controls. Mutant IGF1 was also less efficient in inducing cell growth. CONCLUSION: The present report broadens the spectrum of clinical and biochemical presentation of homozygous IGF1 defects and underscores the variability these patients may present depending on the IGF/IGF1R pathway activity.
Asunto(s)
Trastornos del Crecimiento/genética , Pérdida Auditiva Sensorineural/genética , Factor I del Crecimiento Similar a la Insulina/deficiencia , Mutación Missense/genética , Anomalías Múltiples/genética , Proliferación Celular , Biología Computacional , Simulación por Computador , Retardo del Crecimiento Fetal/genética , Células HEK293 , Homocigoto , Humanos , Lactante , Factor I del Crecimiento Similar a la Insulina/genética , Masculino , Linaje , Polimorfismo de Nucleótido Simple/genética , Receptor IGF Tipo 1 , Receptores de Somatomedina/genética , Tirosina/genéticaRESUMEN
El síndrome de Wolf-Hirschhorn es una entidad polimalformativa debida a la microdeleción en la región distal del brazo corto del cromosoma 4 (4p16.3), el cual produce una serie de manifestaciones clínicas, que pueden variar dependiendo del tipo y tamaño del defecto genético en este síndrome de genes contiguos. Se presentan cinco pacientes, tres de ellos de sexo femenino, todos con los hallazgos clínicos primordiales, con rasgo facial característico de "apariencia en casco de guerrero griego", retraso en el crecimiento y del desarrollo psicomotor. Además de la deleción parcial en la región distal del brazo corto del cromosoma 4, en dos pacientes, se encontraron alteraciones genéticas adicionales, mediante el uso de microarrays de polimorfismos de nucleótido único. Se resaltan las características clínicas del síndrome de Wolf-Hirschhorn con la finalidad de orientar el diagnóstico, brindar una atención médica interdisciplinaria y, a través de su confirmación, brindar un adecuado asesoramiento genético familiar.
Wolf-Hirschhorn syndrome is a polymalformative entity due to the microdeletion in the distal region of the short arm of chromosome 4 (4p16.3), which produces a series of clinical manifestations that can vary depending on the type and size of the genetic defect in this contiguous gene syndrome. Five patients are presented, three of them female, all with the primary clinical findings, characterized by "Greek warrior helmet appearance" facial feature, growth retardation and psychomotor development delay. In addition to the partial deletion in the distal region of the short arm of chromosome 4, two additional genetic alterations were found in two patients, through the use of single nucleotide polymorphism arrays. The clinical characteristics of Wolf-Hirschhorn syndrome are highlighted in order to guide the diagnosis, provide interdisciplinary medical care and, through its confirmation, provide adequate family genetic counseling.
Asunto(s)
Humanos , Masculino , Femenino , Lactante , Síndrome de Wolf-Hirschhorn , Grupo de Atención al Paciente , Anomalías Múltiples , Análisis por Micromatrices , Asesoramiento GenéticoRESUMEN
Wolf-Hirschhorn syndrome is a polymalformative entity due to the microdeletion in the distal region of the short arm of chromosome 4 (4p16.3), which produces a series of clinical manifestations that can vary depending on the type and size of the genetic defect in this contiguous gene syndrome. Five patients are presented, three of them female, all with the primary clinical findings, characterized by "Greek warrior helmet appearance" facial feature, growth retardation and psychomotor development delay. In addition to the partial deletion in the distal region of the short arm of chromosome 4, two additional genetic alterations were found in two patients, through the use of single nucleotide polymorphism arrays. The clinical characteristics of Wolf-Hirschhorn syndrome are highlighted in order to guide the diagnosis, provide interdisciplinary medical care and, through its confirmation, provide adequate family genetic counseling.
El síndrome de Wolf-Hirschhorn es una entidad polimalformativa debida a la microdeleción en la región distal del brazo corto del cromosoma 4 (4p16.3), el cual produce una serie de manifestaciones clínicas, que pueden variar dependiendo del tipo y tamaño del defecto genético en este síndrome de genes contiguos. Se presentan cinco pacientes, tres de ellos de sexo femenino, todos con los hallazgos clínicos primordiales, con rasgo facial característico de "apariencia en casco de guerrero griego", retraso en el crecimiento y del desarrollo psicomotor. Además de la deleción parcial en la región distal del brazo corto del cromosoma 4, en dos pacientes, se encontraron alteraciones genéticas adicionales, mediante el uso de microarrays de polimorfismos de nucleótido único. Se resaltan las características clínicas del síndrome de Wolf-Hirschhorn con la finalidad de orientar el diagnóstico, brindar una atención médica interdisciplinaria y, a través de su confirmación, brindar un adecuado asesoramiento genético familiar.
Asunto(s)
Polimorfismo de Nucleótido Simple , Síndrome de Wolf-Hirschhorn/genética , Femenino , Humanos , Lactante , Masculino , Análisis por Micromatrices , FenotipoRESUMEN
The transmission of HIV can be prevented by the application of neutralizing monoclonal antibodies and lectins. Traditional recombinant protein manufacturing platforms lack sufficient capacity and are too expensive for developing countries, which suffer the greatest disease burden. Plants offer an inexpensive and scalable alternative manufacturing platform that can produce multiple components in a single plant, which is important because multiple components are required to avoid the rapid emergence of HIV-1 strains resistant to single microbicides. Furthermore, crude extracts can be used directly for prophylaxis to avoid the massive costs of downstream processing and purification. We investigated whether rice could simultaneously produce three functional HIV-neutralizing proteins (the monoclonal antibody 2G12, and the lectins griffithsin and cyanovirin-N). Preliminary in vitro tests showed that the cocktail of three proteins bound to gp120 and achieved HIV-1 neutralization. Remarkably, when we mixed the components with crude extracts of wild-type rice endosperm, we observed enhanced binding to gp120 in vitro and synergistic neutralization when all three components were present. Extracts of transgenic plants expressing all three proteins also showed enhanced in vitro binding to gp120 and synergistic HIV-1 neutralization. Fractionation of the rice extracts suggested that the enhanced gp120 binding was dependent on rice proteins, primarily the globulin fraction. Therefore, the production of HIV-1 microbicides in rice may not only reduce costs compared to traditional platforms but may also provide functional benefits in terms of microbicidal potency.
Asunto(s)
Fármacos Anti-VIH , Anticuerpos Monoclonales , Endospermo , Anticuerpos Anti-VIH , Proteína gp120 de Envoltorio del VIH/antagonistas & inhibidores , VIH-1/química , Oryza , Plantas Modificadas Genéticamente , Fármacos Anti-VIH/química , Fármacos Anti-VIH/metabolismo , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/genética , Endospermo/química , Endospermo/genética , Endospermo/metabolismo , Anticuerpos Anti-VIH/biosíntesis , Anticuerpos Anti-VIH/química , Anticuerpos Anti-VIH/genética , Proteína gp120 de Envoltorio del VIH/química , Proteína gp120 de Envoltorio del VIH/genética , Proteína gp120 de Envoltorio del VIH/metabolismo , Oryza/química , Oryza/genética , Oryza/metabolismo , Plantas Modificadas Genéticamente/química , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismoRESUMEN
This paper reports on the combined solar photo-Fenton/biological treatment of an industrial effluent (initial total organic carbon, TOC, around 500mgL(-1)) containing a non-biodegradable organic substance (alpha-methylphenylglycine at 500mgL(-1)), focusing on pilot plant tests performed for design of an industrial plant, the design itself and the plant layout. Pilot plant tests have demonstrated that biodegradability enhancement is closely related to disappearance of the parent compound, for which a certain illumination time and hydrogen peroxide consumption are required, working at pH 2.8 and adding Fe(2+)=20mgL(-1). Based on pilot plant results, an industrial plant with 100m(2) of CPC collectors for a 250L/h treatment capacity has been designed. The solar system discharges the wastewater (WW) pre-treated by photo-Fenton into a biotreatment based on an immobilized biomass reactor. First, results of the industrial plant are also presented, demonstrating that it is able to treat up to 500Lh(-1) at an average solar ultraviolet radiation of 22.9Wm(-2), under the same conditions (pH, hydrogen peroxide consumption) tested in the pilot plant.