Functional characterization of the turkey macrophage migration inhibitory factor

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dc.contributor.authorPark, Myeongseonen
dc.contributor.authorKim, Sungwonen
dc.contributor.authorFetterer, Raymond H.en
dc.contributor.authorDalloul, Rami A.en
dc.contributor.departmentAnimal and Poultry Sciencesen
dc.date.accessioned2020-04-21T14:50:31Zen
dc.date.available2020-04-21T14:50:31Zen
dc.date.issued2016-08en
dc.description.abstractMacrophage migration inhibitory factor (MIF) is a soluble protein that inhibits the random migration of macrophages and plays a pivotal immunoregulatory function in innate and adaptive immunity. The aim of this study was to clone the turkey MIF (TkMIF) gene, express the active protein, and characterize its basic function. The full-length TkMIF gene was amplified from total RNA extracted from turkey spleen, followed by cloning into a prokaryotic (pET11a) expression vector. Sequence analysis revealed that TkMIF consists of 115 amino acids with 12.5 kDa molecular weight. Multiple sequence alignment revealed 100%, 65%, 95% and 92% identity with chicken, duck, eagle and zebra finch MIFs, respectively. Recombinant TkMIF (rTkMIF) was expressed in Escherichia coli and purified through HPLC and endotoxin removal. SDS-PAGE analysis revealed an approximately 13.5 kDa of rTkMIF monomer containing T7 tag in soluble form. Western blot analysis showed that anti-chicken MIF (ChMIF) polyclonal antisera detected a monomer form of TkMIF at approximately 13.5 kDa size. Further functional analysis revealed that rTkMIF inhibits migration of both mononuclear cells and splenocytes in a dose-dependent manner, but was abolished by the addition of anti-ChMIF polyclonal antisera. qRT-PCR analysis revealed elevated transcripts of pro inflammatory cytokines by rTkMIF in LPS-stimulated monocytes. rTkMIF also led to increased levels of IFN-gamma and IL-17F transcripts in Con A-activated splenocytes, while IL-10 and IL-13 transcripts were decreased. Overall, the sequences of both the turkey and chicken MIF have high similarity and comparable biological functions with respect to migration inhibitory activities of macrophages and enhancement of pro-inflammatory cytokine expression, suggesting that turkey and chicken MIFs would be biologically cross-reactive.en
dc.description.adminPublic domain – authored by a U.S. government employeeen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1016/j.dci.2016.04.005en
dc.identifier.eissn1879-0089en
dc.identifier.issn0145-305Xen
dc.identifier.pmid27062968en
dc.identifier.urihttp://hdl.handle.net/10919/97856en
dc.identifier.volume61en
dc.language.isoenen
dc.rightsCC0 1.0 Universalen
dc.rights.urihttp://creativecommons.org/publicdomain/zero/1.0/en
dc.subjectMacrophage migration inhibitory factoren
dc.subjectTurkeyen
dc.subjectChemotaxisen
dc.subjectCytokinesen
dc.titleFunctional characterization of the turkey macrophage migration inhibitory factoren
dc.title.serialDevelopmental and Comparative Immunologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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