{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T15:04:07Z","timestamp":1781103847055,"version":"3.54.1"},"reference-count":0,"publisher":"National Academy of Sciences","issue":"8","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1984,4]]},"abstract":"<jats:p>The effectiveness of the hapten-gelatin antigen-affinity fractionation technique for selection of hapten-specific B cells activatable by \"T-cell-dependent\" (TD) stimuli was assessed. Normal adult murine spleen cells were fractionated on fluorescein (Flu)-gelatin layers and the adherent cells were cultured singly or in small numbers with various sources of syngeneic keyhole limpet hemocyanin (KLH)-primed T lymphocytes. Conditions were defined under which the addition of Flu-KLH caused optimal clonal proliferation and differentiation of B cells into anti-Flu directly hemolytic plaque-forming cells (pfc). It was found that 3-5% of the Flu-specific B cells could be activated, versus 1 in 5000 unfractionated spleen cells. The mean enrichment factor for fractionation was 179, almost identical to that seen when the stimulus is \"T-cell-independent\" (TI), showing that the method is capable of isolating B cells responsive to antigenic stimuli requiring specific T-cell help. Efforts were made to determine whether TD B cells constituted a separate population from TI B cells by determining clone frequencies using Flu-KLH, the TI antigen Flu-polymerized flagellin (Flu-POL), or a mixture of both for stimulation. With Flu-POL alone and with the mixed stimulus 2-3 times more pfc clones were produced than with Flu-KLH, yet evidence for separate B-cell subsets was not obtained because of strong \"bystander\" stimulation due to the presence of the carrier-primed T cells in a confined volume of 10 microliters.<\/jats:p>","DOI":"10.1073\/pnas.81.8.2479","type":"journal-article","created":{"date-parts":[[2006,5,31]],"date-time":"2006-05-31T05:39:40Z","timestamp":1149053980000},"page":"2479-2483","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":9,"title":["Single-cell studies on hapten-specific B cells: response to T-cell-dependent antigens."],"prefix":"10.1073","volume":"81","author":[{"given":"G S","family":"Hebbard","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B L","family":"Pike","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G J","family":"Nossal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1984,4]]},"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.81.8.2479","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T12:02:33Z","timestamp":1649851353000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.81.8.2479"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1984,4]]},"references-count":0,"journal-issue":{"issue":"8","published-print":{"date-parts":[[1984,4]]}},"alternative-id":["10.1073\/pnas.81.8.2479"],"URL":"https:\/\/doi.org\/10.1073\/pnas.81.8.2479","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[1984,4]]},"assertion":[{"value":"1984-04-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}