{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T23:15:40Z","timestamp":1740179740718,"version":"3.37.3"},"reference-count":1,"publisher":"Colegio Mexicano de Inmunologia Clinica y Alergia, A. C.","issue":"1","license":[{"start":{"date-parts":[[2024,4,23]],"date-time":"2024-04-23T00:00:00Z","timestamp":1713830400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0"}],"funder":[{"DOI":"10.13039\/501100013409","name":"Sistema General de Regal\u00edas de Colombia","doi-asserted-by":"publisher","award":["BPIN2020000100405"],"award-info":[{"award-number":["BPIN2020000100405"]}],"id":[{"id":"10.13039\/501100013409","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100013409","name":"Sistema General de Regal\u00edas de Colombia","doi-asserted-by":"publisher","award":["BPIN2020000100364"],"award-info":[{"award-number":["BPIN2020000100364"]}],"id":[{"id":"10.13039\/501100013409","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Rev Alerg Mex"],"abstract":"<jats:p>Introduction: The multiplex technique for cytokine measurement using flow cytometry offers the advantage of lower sample requirements than other methods such as enzyme-linked immunosorbent assay (ELISA). Although most of these assays are standardized on specific cytometers and software, it is important to assess their applicability to other equipment and open-source software, such as R, to expand their usage.\r\nObjectives: To quantify the production of Th1\/Th2\/Th17 cytokines induced by Ascaris lumbricoides antigens in peripheral blood mononuclear cells (PBMCs) using a multiplex technique.\r\nMethods: PBMCs were cultured from individuals with mild A. lumbricoides infection (n = 20) and uninfected individuals (n = 21) and stimulated with A. lumbricoides extract (ExtAscaris), a mix of anti-CD2\/CD3\/CD28 (CDmix) as a positive control, and only medium (negative control). Cytokines in the supernatants were measured using the BD\u2122 Cytometric Bead Array Human Th1\/Th2\/Th17 kit, to identify IFN-\u03b3, TNF, IL-10, IL-6, IL-4, IL-2, and IL-17A. Readings were taken on a spectral cytometer (Northern Lights, Cytek, USA), and analysis was performed using R software with packages \"tidyverse,\" \"beadplexr,\" \"flowCore,\" and \"arsenal.\" Cytokine concentrations were calculated using a 5-parameter logistic curve. The t-test was used to compare cases and controls, and statistical significance was set at p &lt; 0.05. The study was approved by the Ethics Committee of the University of Cartagena and the participants provided informed consent. This study was financially supported by the Colombian Sistema General de Regal\u00edas under the BPIN2020000100405 - BPIN2020000100364.\r\nResults: Efficient fluorescence intensity extraction for each cytokine was achieved using detection channel R8 and the \"mclust\" clustering model (Figure 1). No significant differences were found in the levels of the seven cytokines between cases and controls (Figure 2). Although the IFN-\u03b3 response to ExtAscaris was higher in cases than in controls (252.5 ng\/mL vs. 173.1 ng\/mL), the difference was not significant. IL-17A (detection limit: 18.9 pg\/mL) was more detectable in cases than controls (5 cases, 23% vs. 2 controls, 9.5%). IL-4 was only detected in the supernatants from CDmix-stimulated cultures but not with the Ascaris extract (Figure 2).\r\nConclusions: The multiplex technique using spectral flow cytometry combined with open-source software analysis proved applicable for quantifying cytokines induced by A. lumbricoides antigens in PBMCs. However, a more sensitive method is needed to evaluate IL-4 response in the context of ascariasis. The results did not reveal significant differences in cytokine production between cases and controls for the evaluated stimuli.<\/jats:p>","DOI":"10.29262\/ram.v71i1.1365","type":"journal-article","created":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T21:50:05Z","timestamp":1714168205000},"page":"64","source":"Crossref","is-referenced-by-count":0,"title":["Medici\u00f3n de citoquinas mediante una t\u00e9cnica multiplex por citometr\u00eda espectral: aplicaci\u00f3n en el estudio de la inmunobiolog\u00eda de la ascariasis"],"prefix":"10.29262","volume":"71","author":[{"given":"Ana","family":"Lozano","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carolina","family":"S\u00e1nchez-Marrugo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin","family":"Llin\u00e1s-Caballero","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlos","family":"Barrios","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathalie","family":"Acevedo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6007-5376","authenticated-orcid":false,"given":"Josefina","family":"Zakzuk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis","family":"Caraballo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"11808","published-online":{"date-parts":[[2024,4,23]]},"reference":[{"key":"56954","unstructured":"."}],"container-title":["Revista Alergia M\u00e9xico"],"original-title":[],"link":[{"URL":"https:\/\/revistaalergia.mx\/ojs\/index.php\/ram\/article\/download\/1365\/2171","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/revistaalergia.mx\/ojs\/index.php\/ram\/article\/download\/1365\/2172","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/revistaalergia.mx\/ojs\/index.php\/ram\/article\/download\/1365\/2171","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,26]],"date-time":"2024-04-26T21:50:20Z","timestamp":1714168220000},"score":1,"resource":{"primary":{"URL":"https:\/\/revistaalergia.mx\/ojs\/index.php\/ram\/article\/view\/1365"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,4,23]]},"references-count":1,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,4,23]]}},"URL":"https:\/\/doi.org\/10.29262\/ram.v71i1.1365","relation":{},"ISSN":["2448-9190","0002-5151"],"issn-type":[{"type":"electronic","value":"2448-9190"},{"type":"print","value":"0002-5151"}],"subject":[],"published":{"date-parts":[[2024,4,23]]}}}