{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,9]],"date-time":"2025-11-09T07:33:19Z","timestamp":1762673599710},"reference-count":45,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2005,12,7]],"date-time":"2005-12-07T00:00:00Z","timestamp":1133913600000},"content-version":"vor","delay-in-days":3050,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Eur J Immunol"],"published-print":{"date-parts":[[1997,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Major histocompatibility complex (MHC) class II molecules bind to numerous peptides and display these on the cell surface for T cell recognition. In a given immune response, receptors on T cells recognize antigenic peptides that are a minor population of MHC class II\u2010bound peptides. To control which peptides are presented to T cells, it may be desirable to use recombinant MHC molecules with covalently bound antigenic peptides. To study T cell responses to such homogenous peptide\u2010MHC complexes, we engineered an HLA\u2010DR1 cDNA coding for influenza hemagglutinin, influenza matrix, or HIV p24 gag peptides covalently attached via a peptide spacer to the N terminus of the DR1 \u03b2 chain. Co\u2010transfection with DR \u03b1 cDNA into mouse L cells resulted in surface expression of HLA\u2010DR1 molecules that reacted with monoclonal antibodies (mAb) specific for correctly folded HLA\u2010DR epitopes. This suggested that the spacer and peptide did not alter expression or folding of the molecule. We then engineered an additional peptide spacer between the C terminus of a truncated \u03b2 chain (without transmembrane or cytoplasmic domains) and the N terminus of full\u2010length DR \u03b1 chain. Transfection of this cDNA into mouse L cells resulted in surface expression of the entire covalently linked heterotrimer of peptide, \u03b2 chain, and \u03b1 chain with the expected molecular mass of approximately 66 kDa. These single\u2010chain HLA\u2010DR1 molecules reacted with mAb specific for correctly folded HLA\u2010DR epitopes, and identified one mAb with [MHC + peptide] specificity. Affinity\u2010purified soluble secreted single\u2010chain molecules with truncated \u03b1 chain moved in electrophoresis as compact class II MHC dimers. Cell surface two\u2010chain or single\u2010chain HLA\u2010DR1 molecules with a covalent HA peptide stimulated HLA\u2010DR1\u2010restricted HA\u2010specific T cells. They were immunogenic <jats:italic>in vitro<\/jats:italic> for peripheral blood mononuclear cells. The two\u2010chain and single\u2010chain HLA\u2010DR1 molecules with covalent HA peptide had reduced binding for the bacterial superantigens staphylococcal enterotoxin A and B and almost no binding for toxic shock syndrome toxin\u20101. The unique properties of these engineered HLA\u2010DR1 molecules may facilitate our understanding of the complex nature of antigen recognition and aid in the development of novel vaccines with reduced superantigen binding.<\/jats:p>","DOI":"10.1002\/eji.1830270817","type":"journal-article","created":{"date-parts":[[2007,3,2]],"date-time":"2007-03-02T05:38:11Z","timestamp":1172813891000},"page":"1933-1941","source":"Crossref","is-referenced-by-count":12,"title":["A recombinant single\u2010chain human class II MHC molecule (HLA\u2010DR1) as a covalently linked heterotrimer of \u03b1 chain, \u03b2 chain, and antigenic peptide, with immunogenicity <i>in vitro<\/i> and reduced affinity for bacterial superantigens"],"prefix":"10.1002","volume":"27","author":[{"given":"Xiaojie","family":"Zhu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sina","family":"Bavari","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Ulrich","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Scheherazade","family":"Sadegh\u2010Nasseri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Soldano","family":"Ferrone","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Louise","family":"McHugh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Mage","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,12,7]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-5699(94)90220-8"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.182.5.1591"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.182.5.1579"},{"key":"e_1_2_1_5_2","doi-asserted-by":"crossref","first-page":"3039","DOI":"10.4049\/jimmunol.157.7.3039","volume":"157","author":"Oukka M.","year":"1996","journal-title":"J. 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