{"id":26030,"date":"2022-02-21T17:27:14","date_gmt":"2022-02-21T09:27:14","guid":{"rendered":"https:\/\/www.accscicn.com\/?p=26030"},"modified":"2022-10-26T15:40:08","modified_gmt":"2022-10-26T07:40:08","slug":"app-mantis-22022101","status":"publish","type":"post","link":"https:\/\/www.accscicn.com\/es\/vision\/aplicacion\/app-mantis-22022101\/","title":{"rendered":"Cribado automatizado y reproducible de anticuerpos HLA en el trasplante de \u00f3rganos mediante el dispensador MANTIS"},"content":{"rendered":"<div data-elementor-type=\"wp-post\" data-elementor-id=\"26030\" class=\"elementor elementor-26030\" data-elementor-post-type=\"post\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-77fd721b elementor-section-full_width elementor-section-stretched elementor-section-height-default elementor-section-height-default\" data-id=\"77fd721b\" data-element_type=\"section\" data-settings=\"{&quot;stretch_section&quot;:&quot;section-stretched&quot;,&quot;background_background&quot;:&quot;classic&quot;}\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-60b31c21\" data-id=\"60b31c21\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-14400749 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"14400749\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-11dc6372\" data-id=\"11dc6372\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-44f993ae elementor-widget elementor-widget-heading\" data-id=\"44f993ae\" data-element_type=\"widget\" data-widget_type=\"heading.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.21.0 - 20-05-2024 *\/\n.elementor-heading-title{padding:0;margin:0;line-height:1}.elementor-widget-heading .elementor-heading-title[class*=elementor-size-]>a{color:inherit;font-size:inherit;line-height:inherit}.elementor-widget-heading .elementor-heading-title.elementor-size-small{font-size:15px}.elementor-widget-heading .elementor-heading-title.elementor-size-medium{font-size:19px}.elementor-widget-heading .elementor-heading-title.elementor-size-large{font-size:29px}.elementor-widget-heading .elementor-heading-title.elementor-size-xl{font-size:39px}.elementor-widget-heading .elementor-heading-title.elementor-size-xxl{font-size:59px}<\/style><h1 class=\"elementor-heading-title elementor-size-default\"><b>Cribado automatizado y reproducible de anticuerpos HLA en el trasplante de \u00f3rganos mediante el dispensador MANTIS<\/b><\/h1>\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-3b7efc4e elementor-widget elementor-widget-text-editor\" data-id=\"3b7efc4e\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.21.0 - 20-05-2024 *\/\n.elementor-widget-text-editor.elementor-drop-cap-view-stacked .elementor-drop-cap{background-color:#69727d;color:#fff}.elementor-widget-text-editor.elementor-drop-cap-view-framed .elementor-drop-cap{color:#69727d;border:3px solid;background-color:transparent}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap{margin-top:8px}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap-letter{width:1em;height:1em}.elementor-widget-text-editor .elementor-drop-cap{float:left;text-align:center;line-height:1;font-size:50px}.elementor-widget-text-editor .elementor-drop-cap-letter{display:inline-block}<\/style>\t\t\t\t<p><strong>Fondo<\/strong><\/p><p>El sistema de ant\u00edgenos leucocitarios humanos (HLA) forma parte de nuestro sistema inmunitario adaptativo y desempe\u00f1a un papel importante tanto en la defensa contra las enfermedades como en el trasplante de \u00f3rganos. Los donantes con marcadores HLA compatibles tienen menos probabilidades de rechazar un \u00f3rgano.<\/p><p>Investigadores del Laboratorio de Inmunolog\u00eda M\u00e9dica (LMI) del Centro M\u00e9dico de la Universidad Radboud (Pa\u00edses Bajos) realizan un cribado para detectar anticuerpos espec\u00edficos HLA en el suero de posibles receptores de \u00f3rganos. La detecci\u00f3n de anticuerpos contra marcadores HLA es necesaria para evitar el rechazo hiperagudo del \u00f3rgano donado. El rechazo puede deberse a transfusiones de sangre, trasplantes o embarazos. El laboratorio realiza cribados trimestrales para detectar estos anticuerpos nocivos en el suero de los pacientes.<\/p><p>Estos anticuerpos pueden detectarse cruzando el suero de un paciente con una suspensi\u00f3n de m\u00e1s de 60 linfocitos \u00fanicos de diferentes individuos con distintos marcadores HLA. Esto tambi\u00e9n se conoce como prueba de citotoxicidad dependiente del complemento (CDC).<\/p><p>En este estudio, se utiliz\u00f3 el procesador de l\u00edquidos MANTIS\u00ae de FORMULATRIX\u00ae para dispensar los linfocitos aislados en las placas Terasaki preparadas.<\/p><p>\u00a0<\/p><p><strong>Materiales y m\u00e9todos<\/strong><\/p><p>En primer lugar, se dispens\u00f3 1 \u03bcl de suero del paciente en una placa Terasaki (Greiner) utilizando un dispensador de suero ART Robbins (Sunnyvale, CA). Los linfocitos del donante se aislaron de la sangre utilizando el m\u00e9todo de gradiente de densidad Lymphoprep (Elitech Group) o nitr\u00f3geno l\u00edquido, despu\u00e9s se cultivaron y se lavaron con colorante CFDA C195 (Invitrogen). A continuaci\u00f3n se prepar\u00f3 una suspensi\u00f3n de linfocitos de 4 x 106 c\u00e9lulas por ml.<\/p><p>El dispensador MANTIS\u00ae se utiliza para dispensar 1 \u03bcL de linfocitos aislados en placas Terasaki preparadas. Los pasos posteriores incluyen la adici\u00f3n de suspensiones de complemento de conejo (Immucore, Norcross, GA) y EDTA (Merck, KGaA, Darmstadt, Alemania), hemoglobina y yoduro de propidio, y la incubaci\u00f3n.<\/p><p>Tras el \u00faltimo paso de adici\u00f3n, las microplacas se observaron con un microscopio invertido de fluorescencia (Leitz Leica Diavert Inverted Microscope withfluorescent light illuminator (480-525nm) Oculair 10x) para determinar si hab\u00eda una reacci\u00f3n positiva o negativa entre el suero del paciente y los linfocitos del donante. La presencia de una reacci\u00f3n positiva o negativa entre el suero del paciente y los linfocitos del donante.<\/p><p>El porcentaje de destrucci\u00f3n (% kill) se determin\u00f3 a partir de los resultados de la tinci\u00f3n fluorescente (v\u00e9ase la figura 1). El porcentaje de destrucci\u00f3n es el porcentaje de c\u00e9lulas positivas (rojas) respecto al n\u00famero total de c\u00e9lulas positivas y negativas (verdes). Las puntuaciones porcentuales se muestran en la Tabla 1.<\/p><p><strong><img fetchpriority=\"high\" decoding=\"async\" class=\"size-full wp-image-26032 alignnone\" src=\"https:\/\/www.accscicn.com\/wp-content\/uploads\/2022\/02\/1.png\" alt=\"\" width=\"194\" height=\"291\" \/><\/strong><\/p><p>Cuadro 1: Porcentaje de muertes<\/p><p><strong><img decoding=\"async\" class=\"size-full wp-image-26033 alignnone\" src=\"https:\/\/www.accscicn.com\/wp-content\/uploads\/2022\/02\/2.png\" alt=\"\" width=\"557\" height=\"178\" srcset=\"https:\/\/www.accscicn.com\/wp-content\/uploads\/2022\/02\/2.png 557w, https:\/\/www.accscicn.com\/wp-content\/uploads\/2022\/02\/2-300x96.png 300w\" sizes=\"(max-width: 557px) 100vw, 557px\" \/><\/strong><\/p><p>Fig. 1: Muestra de linfocitos obviamente observable (A) Reacci\u00f3n negativa, verde. (B) Reacci\u00f3n positiva, rojo. (C) Reacci\u00f3n fuertemente positiva, rojo.<\/p><p>\u00a0<\/p><p><strong>Resultados y conclusiones<\/strong><\/p><p>Como el equipo existente ya no era compatible, el laboratorio tuvo que encontrar un dispensador autom\u00e1tico alternativo para a\u00f1adir linfocitos preparados a las placas Terasaki. La dispensaci\u00f3n manual introduce una cantidad variable de variabilidad durante el experimento, lo que resulta inaceptable para el investigador.<\/p><p>Un n\u00famero cada vez mayor de laboratorios que ya no utilizan equipos compatibles para el cribado de HLA tendr\u00e1n que sustituir esta tecnolog\u00eda obsoleta, por lo que el equipo del LMI ha accedido a compartir los resultados de su evaluaci\u00f3n en beneficio de la comunidad acad\u00e9mica m\u00e1s amplia implicada en esta importante labor.<\/p><p>El instrumento MANTIS de FORMULATRIX se considera una t\u00e9cnica superior para a\u00f1adir linfocitos en comparaci\u00f3n con la dosificaci\u00f3n manual con pipeta y se ha utilizado con \u00e9xito en flujos de trabajo rutinarios de laboratorio. Proporciona resultados en menos tiempo y con mayor consistencia y precisi\u00f3n.<\/p><p>\u00a0<\/p><p><strong>Agradecimientos y reconocimientos<\/strong><\/p><p>FORMULATRIX\u00ae desea agradecer a Yvonne van Berkel (analista senior Cellulaire Immunologie en Histocompatibiliteit) y al equipo de LMI su trabajo en la evaluaci\u00f3n del flujo de trabajo HLA del dispensador MANTIS\u00ae y los datos proporcionados en esta nota de aplicaci\u00f3n. Los resultados de los datos proporcionados en esta nota de aplicaci\u00f3n. Los otros miembros del equipo fueron Siham Akdimi, Gaby Derksen, Suzanne Hendriks y Jeroen Slager.<\/p><p>Fuente: @BostonFummerle<\/p><p>Fecha: 2022.02.21<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-322c4bf2\" data-id=\"322c4bf2\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-294f015d elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"294f015d\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.21.0 - 20-05-2024 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