{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,12]],"date-time":"2026-02-12T13:11:14Z","timestamp":1770901874541,"version":"3.50.1"},"reference-count":39,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2005,10,14]],"date-time":"2005-10-14T00:00:00Z","timestamp":1129248000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2005,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Assessment of CD8<jats:sup>+<\/jats:sup> T\u2010cell activity is of significant importance for the evaluation of cellular immune responses to viral infections, especially in HIV. We present a new assay for the assessment of HIV\u2010specific cytotoxicity by multiparameter flow cytometry.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Target cells, pulsed with peptide pools (Gag or Nef), were stained with 5\u2010 (and \u20136)\u2010carboxyfluorescein diacetate succinimidyl ester (CFSE), cultured with specific or nonspecific effector cells, and finally stained with propidium iodide (PI). Determination of cytolysis is based on the enumeration of viable target cells (CFSE<jats:sup>hi<\/jats:sup>PI<jats:sup>\u2212<\/jats:sup>) in the test sample (target and specific effector cells) as compared with that of the viable target cells in the control sample (target and nonspecific effector cells). The <jats:sup>51<\/jats:sup>Cr\u2010release assay and IFN\u2010\u03b3 ELISpot were performed by standard procedures.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A comparison with the Cr\u2010release showed that the two assays were strongly correlated (<jats:italic>r<\/jats:italic> = 0.67; <jats:italic>P<\/jats:italic> &lt; 0.001) but the sensitivity of the flow cytometric assay was significantly higher (<jats:italic>P<\/jats:italic> &lt; 0.05), and the reproducibility good (CV, 7.7%). Good correlation was also found with the ELISpot assay (<jats:italic>r<\/jats:italic> = 0.66; <jats:italic>P<\/jats:italic> &lt; 0.01).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This new assay provides both specific and sensitive results when employed for the detection of HIV\u2010specific CTL and can be a valuable tool for the evaluation of cytolytic activity in vaccine trials or in HIV\u2010infected subjects, especially if such responses are present at low levels. \u00a9 2005 Wiley\u2010Liss, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.a.20189","type":"journal-article","created":{"date-parts":[[2005,10,14]],"date-time":"2005-10-14T18:17:12Z","timestamp":1129313832000},"page":"71-80","source":"Crossref","is-referenced-by-count":40,"title":["A novel assay for assessment of HIV\u2010specific cytotoxicity by multiparameter flow cytometry"],"prefix":"10.1002","volume":"68A","author":[{"given":"Karina","family":"Godoy\u2010Ramirez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbro","family":"M\u00e4kitalo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rigmor","family":"Thorstensson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric","family":"Sandstr\u00f6m","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gunnel","family":"Biberfeld","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hans","family":"Gaines","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,10,14]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/328345a0"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.9.6103-6110.1994"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.7.4650-4655.1994"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0065-2776(08)60745-2"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/nri911"},{"key":"e_1_2_6_7_2","first-page":"181","article-title":"Quantitative assay of the lytic action of immune lymphoid cells on 51\u2010Cr\u2010labelled allogeneic target cells in vitro; 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