{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T13:02:52Z","timestamp":1753880572226,"version":"3.41.2"},"reference-count":53,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2024,8,6]],"date-time":"2024-08-06T00:00:00Z","timestamp":1722902400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,8,6]],"date-time":"2024-08-06T00:00:00Z","timestamp":1722902400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R01 AI146257"],"award-info":[{"award-number":["R01 AI146257"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2024,8,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Diarrheal diseases with infectious etiology remain a major cause of death globally, particularly in low-income countries.\n              <jats:italic>Entamoeba histolytica<\/jats:italic>\n              is a pathogenic protozoan parasite that is the causative agent of amebiasis. Amebiasis has a wide presentation in clinical severity with many factors, including the bacterial microbiota, contributing to this variation. The innate immune response also plays a critical role in regulating the severity of\n              <jats:italic>E. histolytica<\/jats:italic>\n              infection, with neutrophils reported to have a protective role. Despite this, the precise mechanism of how neutrophils mediate amebic killing is poorly understood. Thus, modern platforms that allow for inquiry of granulocyte\u2013ameba interactions will increase our understanding of this disease. Herein, we describe an assay for neutrophil killing of\n              <jats:italic>E. histolytica<\/jats:italic>\n              by utilizing high-dimensional spectral flow cytometry. Neutrophils were isolated from wild-type 5-week-old C57BL\/6 mice and co-cultured with\n              <jats:italic>E. histolytica<\/jats:italic>\n              at various multiplicity of infections (MOIs). After co-culture, neutrophils and\n              <jats:italic>E. histolytica<\/jats:italic>\n              were stained for spectral flow cytometry. Cell populations were identified using surface markers and fluorescence minus one (FMO) controls. We have previously shown that animals colonized with a component of the human microbiota,\n              <jats:italic>Clostridium scindens<\/jats:italic>\n              , were protected from\n              <jats:italic>E. histolytica<\/jats:italic>\n              . This protection was associated with elevated neutrophil count. Here, we explored amebic killing capacity and observed that neutrophils from animals with\n              <jats:italic>C. scindens<\/jats:italic>\n              possessed heightened amebic killing compared with controls. Thus, this study establishes a novel platform that can provide an in-depth analysis of granulocyte\u2013parasite interactions in various contexts, including during alteration of the intestinal microbiota.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                The tools for studying host immune cell\n                <jats:italic>\u2013E. histolytica<\/jats:italic>\n                interactions are limited. Factors, such as parasite heterogeneity, infectivity, and difficulties with culture systems and animal models, make interrogation of these interactions challenging. Thus,\n                <jats:italic>Entamoeba<\/jats:italic>\n                researchers can benefit from next-generation models that allow for the analysis of both host and parasite cells. Here, we demonstrate the use of a novel platform that allows for the determination of parasite\u2013host cell interactions and customizable high-dimensional phenotyping of both populations. Indeed, spectral flow cytometry can approach &gt;40 markers on a single panel and can be paired with custom-developed parasite antibodies that can be conjugated to fluorochromes via commercially available kits. This platform affords researchers the capability to test highly precise hypotheses regarding host\u2013parasite interactions.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.00472-24","type":"journal-article","created":{"date-parts":[[2024,6,18]],"date-time":"2024-06-18T13:01:56Z","timestamp":1718715716000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["A high-dimensional platform for observing neutrophil\u2013parasite interactions"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2148-4086","authenticated-orcid":false,"given":"Brandon A.","family":"Thompson","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases and International Health, Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julio","family":"Revilla","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and International Health, Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9055-9057","authenticated-orcid":false,"given":"Savannah","family":"Brovero","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and International Health, Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0408-0805","authenticated-orcid":false,"given":"Stacey L.","family":"Burgess","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and International Health, Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(12)60560-1"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S2214-109X(23)00496-5"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2022.2158656"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-091313-103550"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1172\/jci110377"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/158.2.398"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1155\/2013\/890603"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1982.tb01250.x"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1992.tb05432.x"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0600623103"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra022710"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/14651858.CD006085.pub3"},{"key":"e_1_3_3_14_2","first-page":"96","article-title":"Practical murine hematopathology: a comparative review and implications for research","volume":"65","author":"O\u2019Connell KE","year":"2015","unstructured":"O\u2019Connell KE, Mikkola AM, Stepanek AM, Vernet A, Hall CD, Sun CC, Yildirim E, Staropoli JF, Lee JT, Brown DE. 2015. 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