{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T04:11:55Z","timestamp":1747195915340,"version":"3.40.5"},"reference-count":39,"publisher":"Public Library of Science (PLoS)","issue":"11","license":[{"start":{"date-parts":[[2020,11,25]],"date-time":"2020-11-25T00:00:00Z","timestamp":1606262400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Cluster of differentiation 4 (CD4) molecule expressed on the leukocytes is known to function as a co-receptor for class II major histocompatibility complex (MHC) binding to T cell receptor (TCR) on helper T cells. We previously identified two CD4 alleles (CD4.A and CD4.B) in a Microminipig population based on nucleotide sequencing and PCR detection of their gene sequences. However, CD4.B protein expression was not examined because of the unavailability of a reactive antibody to a CD4.B epitope. In this study, we have produced two swine-specific monoclonal antibodies (mAbs) against CD4.B molecules, one that recognizes only CD4.B (b1D7) and the other that recognizes both the CD4.A and CD4.B alleles (x1E10) and that can be used to distinguish CD4 T cell subsets by flow cytometry and immunohistochemistry. Using these two mAbs, we identified CD4.A and CD4.B allele-specific proteins on the surface of CD4.A (+\/+) and CD4.B (+\/+) T cells at a similar level of expression. Moreover, stimulation of peripheral blood mononuclear cells (PBMCs) derived from CD4.A (+\/+) and CD4.B (+\/+) swine with toxic shock syndrome toxin-1 (TSST-1)<jats:italic>in vitro<\/jats:italic>similarly activated both groups of cells that exhibited a slight increase in the CD4\/CD8 double positive (DP) cell ratio. A large portion of the DP cells from the allelic CD4.A (+\/+) and CD4.B (+\/+) groups enhanced the total CD4 and class I swine leukocyte antigen (SLA) expression. The x1E10 mAb delayed and reduced the TSST-1-induced activation of CD4 T cells. Thus, CD4.B appears to be a functional protein whose expression on activated T cells is analogous to CD4.A.<\/jats:p>","DOI":"10.1371\/journal.pone.0242572","type":"journal-article","created":{"date-parts":[[2020,11,25]],"date-time":"2020-11-25T18:35:26Z","timestamp":1606329326000},"page":"e0242572","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Preparation and characterization of monoclonal antibodies recognizing two CD4 isotypes of 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Eguchi-Ogawa","year":"2018","journal-title":"Anim Biotechnol"},{"issue":"3","key":"pone.0242572.ref012","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1007\/s11033-010-0272-2","article-title":"Association of goat (Capra hircus) CD4 gene exon 6 polymorphisms with ability of sperm internalizing exogenous DNA","volume":"38","author":"L Wang","year":"2011","journal-title":"Mol Biol Rep"},{"issue":"5","key":"pone.0242572.ref013","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1089\/aid.2014.0303","article-title":"The CD4 C868T Polymorphism and Its Correlation with HIV-1 Infection in a Chinese Population","volume":"31","author":"Y Lu","year":"2015","journal-title":"AIDS Res Hum Retroviruses"},{"issue":"9","key":"pone.0242572.ref014","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1086\/597616","article-title":"A Common CD4 Gene Variant Is Associated with an Increased Risk of HIV-1 Infection in Kenyan Female Commercial Sex Workers","volume":"199","author":"J 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Immunopathol"},{"issue":"1","key":"pone.0242572.ref018","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1186\/s12917-016-0856-8","article-title":"Identification and characterization of two CD4 alleles in Microminipigs","volume":"12","author":"T Matsubara","year":"2016","journal-title":"BMC Veterinary Research"},{"issue":"2","key":"pone.0242572.ref019","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1254\/jphs.10R16FM","article-title":"Microminipig, a non-rodent experimental animal optimized for life science research: preface","volume":"115","author":"N Kaneko","year":"2011","journal-title":"J Pharmacol Sci"},{"issue":"1","key":"pone.0242572.ref020","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1177\/0023677214557169","article-title":"Body and major organ sizes of young mature microminipigs determined by computed tomography","volume":"49","author":"M Takasu","year":"2015","journal-title":"Lab 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Kametani","year":"2016","journal-title":"PLoS One"},{"key":"pone.0242572.ref024","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1111\/j.1365-2052.2009.01860.x","article-title":"Molecular characterization of swine leucocyte antigen class I genes in outbred pig populations","volume":"40","author":"C Ho","year":"2009","journal-title":"Animal Genetics"},{"issue":"4","key":"pone.0242572.ref025","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1111\/j.1365-2052.2010.02019.x","article-title":"Molecular characterization of swine leucocyte antigen class II genes in outbred pig populations","volume":"41","author":"C Ho","year":"2010","journal-title":"Anim Genet"},{"issue":"1","key":"pone.0242572.ref026","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.ab.2013.08.009","article-title":"Modified S\/MAR episomal vectors for stably expressing fluorescent-protein tagged transgenes with small cell-to-cell fluctuations","volume":"443","author":"A Mizutani","year":"2013","journal-title":"Anal Biochem"},{"key":"pone.0242572.ref027","first-page":"6540","article-title":"W J Gadella Jr T. Cyan and yellow super fluorescent proteins with improved brightness, protein folding, and FRET F\u00f6rster radius","volume":"45","author":"G-J Kremers","year":"2006","journal-title":"Biochemistry"},{"key":"pone.0242572.ref028","doi-asserted-by":"crossref","first-page":"308","DOI":"10.5582\/bst.2014.01084","article-title":"Differentiation ability of multipotent hematopoietic stem\/progenitor cells detected by a porcine specific anti-CD117 monoclonal antibody","volume":"8","author":"S Ohshima","year":"2014","journal-title":"Biosci Trends"},{"issue":"2","key":"pone.0242572.ref029","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/0022-2836(92)90964-L","article-title":"Accurate modeling of protein conformation by automatic segment matching","volume":"226","author":"M Levitt","year":"1992","journal-title":"J Mol Biol"},{"key":"pone.0242572.ref030","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S0022-2836(77)80200-3","article-title":"The Protein Data Bank: A Computer-based Archival File For Macromolecular Structures","volume":"112","author":"F Bernstein","year":"1977","journal-title":"J Mol Biol"},{"issue":"4919","key":"pone.0242572.ref031","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1126\/science.2549633","article-title":"The MHC-binding and gp120-binding functions of CD4 are separable","volume":"245","author":"D Lamarre","year":"1989","journal-title":"Science"},{"issue":"3","key":"pone.0242572.ref032","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1073\/pnas.0911004107","article-title":"Structure of HIV-1 gp120 with gp41-interactive region reveals layered envelope architecture and basis of conformational mobility","volume":"107","author":"M Pancera","year":"2010","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"16","key":"pone.0242572.ref033","doi-asserted-by":"crossref","first-page":"6423","DOI":"10.1073\/pnas.87.16.6423","article-title":"Molecular characteristics of recombinant human CD4 as deduced from polymorphic crystals","volume":"87","author":"P Kwong","year":"1990","journal-title":"Proc Natl Acad Sci U S A"},{"key":"pone.0242572.ref034","first-page":"500","article-title":"Functional and phenotypic analysis of porcine peripheral blood CD4\/CD8 double-positive T cells","volume":"87","author":"F Zuckermann","year":"1996","journal-title":"Immunology"},{"issue":"2","key":"pone.0242572.ref035","doi-asserted-by":"crossref","first-page":"53","DOI":"10.3390\/pathogens7020053","article-title":"Manipulation of Innate and Adaptive Immunity by Staphylococcal Superantigens","volume":"7","author":"S Tuffs","year":"2018","journal-title":"Pathogens"},{"key":"pone.0242572.ref036","doi-asserted-by":"crossref","first-page":"1101","DOI":"10.1080\/07391102.1998.10509004","article-title":"Spatial structure model of the CD4 receptor-binding site of the HIV envelope protein gp120","volume":"15","author":"A Andrianov","year":"1998","journal-title":"J Biomol Struct Dyn"},{"issue":"6","key":"pone.0242572.ref037","doi-asserted-by":"crossref","first-page":"e38491","DOI":"10.1371\/journal.pone.0038491","article-title":"An improved protocol for efficient engraftment in NOD\/LTSZ-SCIDIL-2R\u03b3null mice allows HIV replication and development of anti-HIV immune responses","volume":"7","author":"M Singh","year":"2012","journal-title":"PLoS ONE"},{"issue":"3","key":"pone.0242572.ref038","doi-asserted-by":"crossref","first-page":"799","DOI":"10.4049\/jimmunol.1801085","article-title":"Human Anti-HIV-1 gp120 Monoclonal Antibodies with Neutralizing Activity Cloned from Humanized Mice Infected with HIV-1","volume":"202","author":"M Gawron","year":"2019","journal-title":"J Immunol"},{"issue":"7778","key":"pone.0242572.ref039","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1038\/s41586-019-1545-0","article-title":"Mapping 123 million neonatal, infant and child deaths between 2000 and 2017","volume":"574","author":"R Burstein","year":"2019","journal-title":"Nature"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0242572","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,18]],"date-time":"2024-08-18T01:58:59Z","timestamp":1723946339000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0242572"}},"subtitle":[],"editor":[{"given":"Karen L.","family":"Elkins","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2020,11,25]]},"references-count":39,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2020,11,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0242572","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2020,11,25]]}}}