{"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\/en\/insight\/application\/app-mantis-22022101\/","title":{"rendered":"Automated, reproducible HLA antibody screening in organ transplants using the MANTIS\u00ae Dispenser"},"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>Automated, reproducible HLA antibody screening in organ transplants using the MANTIS\u00ae Dispenser<\/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>Background<\/strong><\/p><p>The human leukocyte antigen (HLA) system is part of our adaptive immune system and plays an important role in both disease defense and organ transplantation. Donors with matching HLA markers are less likely to reject organs.<\/p><p>Researchers at the Laboratory of Medical Immunology (LMI) at Radboud University Medical Center in the Netherlands screen for HLA-specific antibodies from the sera of potential organ recipients. Detection of antibodies against HLA markers is necessary to avoid hyperacute rejection of the donor organ. Rejection may be caused by blood transfusion, transplantation or pregnancy. The laboratory performs quarterly screenings to detect these harmful antibodies in patient sera.<\/p><p>These antibodies can be detected by cross-matching a patient's serum with a suspension of more than 60 unique lymphocytes from different individuals with different HLA markers. This is also called a complement-dependent cytotoxicity (CDC) test.<\/p><p>In this study, the MANTIS\u00ae Liquid Processor from FORMULATRIX\u00ae was used to dispense the isolated lymphocytes into the prepared Terasaki plates.<\/p><p>\u00a0<\/p><p><strong>Materials and Methods<\/strong><\/p><p>First, 1 \u03bcl of patient serum was dispensed into a Terasaki plate (Greiner) using an ART Robbins (Sunnyvale, CA) serum dispenser. Donor lymphocytes are isolated from the blood using the density gradient Lymphoprep (Elitech Group) or liquid nitrogen method, then cultured and washed with CFDA C195 (Invitrogen) dye. A lymphocyte suspension of 4 x 106 cells per ml is then prepared.<\/p><p>The MANTIS\u00ae Dispenser is used to dispense 1 \u03bcL of isolated lymphocytes into the prepared Terasaki plates. Subsequent steps include the addition of suspensions of rabbit complement (Immucore, Norcross, GA) and EDTA (Merck, KGaA, Darmstadt, Germany), hemoglobin and propidium iodide, and incubation.<\/p><p>After the final addition step, microplates were observed under an inverted fluorescent microscope (Leitz Leica Diavert Inverted Microscope withfluorescent light illuminator (480-525nm) Oculair 10x) to determine between patient serum and donor lymphocytes whether there is a positive or negative reaction between patient serum and donor lymphocytes.<\/p><p>The percentage kill (% kill) measurement was determined from the fluorescent staining results (see Figure 1). The percentage kill is the percentage of positive cells (red) to the total number of positive and negative cells (green). The percentage scores are shown in Table 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>Table 1: Percentage kill score<\/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>Figure 1: Obviously observable lymphocyte samples (A) Negative reaction, green. (B) Positive reaction, red. (C) Strongly positive reaction, red.<\/p><p>\u00a0<\/p><p><strong>Results and Conclusion<\/strong><\/p><p>Since the existing equipment is no longer supported, laboratories must find an alternative automated dispenser for adding prepared lymphocytes to Terasaki plates. Manual dispensing introduces a variable amount of variability during the experiment, which is unacceptable for the investigator.<\/p><p>A growing number of laboratories using no longer supported devices for HLA screening will need to replace this outdated technology, so the team at LMI has agreed to share their evaluation for the benefit of the broader academic community involved in this important work.<\/p><p>The MANTIS instrument from FORMULATRIX is considered a superior technique for adding lymphocytes compared to manual pipette spiking and has been successfully used in routine laboratory workflows. It provides results in less time and with higher consistency and accuracy.<\/p><p>\u00a0<\/p><p><strong>Thanks and Acknowledgements<\/strong><\/p><p>FORMULATRIX\u00ae would like to thank Yvonne van Berkel (senior analyst Cellulaire Immunologie en Histocompatibiliteit) and the LMI team for their work in evaluating the HLA workflow of the MANTIS\u00ae dispenser and for the data provided in this application note. The results of the data provided in this application note. Other members of the team are Siham Akdimi, Gaby Derksen, Suzanne Hendriks and Jeroen Slager.<\/p><p>Source: @BostonFummerle<\/p><p>Time: 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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