Title Učinak tumorskog glikoproteina 72 na decidualne dendritičke stanice rane trudnoće in vitro : doktorski rad Title (english) The effects of tumor associated glycoprotein 72 on dendritic cells in vitro in early pregnancy Author Arnela Redžović Mentor Gordana Laškarin (mentor)Committee member Ines Mrakovčić-Šutić (predsjednik povjerenstva)Committee member Sabina Rabatić (član povjerenstva)Committee member Neda Smiljan Severinski (član povjerenstva)Committee member Gordana Laškarin (član povjerenstva)Granter University of Rijeka Defense date and country 2012-01-01, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 612 - Physiology Abstract Cilj istraživanja: Ispitali smo fenotipska i funkcijska obilježja subpopulacija decidualnih CD1a+ i CD83+ dendritičkih stanica stimuliranih i/ili nestimuliranih s TAG-72 (od eng. Tumor associated glycoprotein 72), te njihovo međudjelovanje s autolognim decidualnim stanicama NK i limfocitima T.
Materijali i metode: Enzimatskom razgradnjom tkiva dobili smo decidue prvog tromjesečja trudnoće uredna tijeka te sluznice jajovoda s mjesta ektopične implantacije i iz mjesta udaljenije od implantacije,
... More te sluznice materišta žena s ektopičnom trudnoćom. Protočnu citometriju koristili smo u metodama jednostruke i dvostruke imunofluorescencije kojom smo istražili fenotip, stupanj sazrijevanja, unutarstaničnog izražavanja citokina i kemokina, sposobnost vezanja i internalizacije glikoproteina TAG-72 u decidualnim CD1a+ i/ili CD83+ stanicama.
Rezultati: CD1a+ stanice su nezrelije od CD83+ stanica zbog slabijeg izražavanja CD83, HLA-DR i kostimulacijskih molekula, dok obiluju endocitoznim receptorima (CD206 i CD209), IL-15, IFN- γ te izražavaju CD195 receptor za proupalne kemokine. Samo CD1a+ stanice povećavaju proliferaciju i izražavanje citotoksičnih posrednika u autolognim stanicama NK, ali smanjuju njihovo izražavanje IL-15 i NKp46. Decidualne CD56+ stanice učinkovitije ubijaju CD83+ stanice. TAG-72 se veže i internalizira na CD1a+ stanice putem domene za prepoznavanje ugljikohidrata na CD206 i CD209 receptore. TAG-72 smanjuje izražavanje proupalnih citokina i kemokina, a povećava izražavanje D6 i IL-1RII na CD1a+ stanicama. CD1a+ stanice stimulirane s TAG-72, smanjuju IFN-γ u autolognim singeničnim i alogeničnim limfocitima T, čak i u prisusutvu lipopolisaharida, a povećavaju proliferaciju limfocita T. TAG-72 je utvrđen u jajovodu na mjestu implantacije, ali nije u sluznici materišta istih žena ili žena s eutopičnom urednom ili patološkom trudnoćom. U jajovodu ektopične trudnoće CD1a+ stanice izražavaju manje IL-15 i IFN-γ nego iste u sluznici materišta.
Zaključak: Uklanjanje TAG-72 s majčino-fetalne plohe uredne trudnoće vjerojatno doprinosi boljoj kontroli prodiranja trofoblasta posredstvom stanica NK i limfocita T, jer ne utječe na sazrijevanje CD1a+ stanica bez protuupalnih svojstava, koja inače nastaju u prisustvu TAG-72. Less Abstract (english) Objectives: We investigated the phenotype and functional characteristics of decidual CD1a+ and CD83+ dendritic cells stimulated with or without TAG-72 (Tumor Associated Glycoprotein 72) and their interactions with autologous decidual NK cells and T cells.
Methods of the study: The first trimestar decidua of normal human pregnancy, mucosa of the fallopian tubes with ectopic implantation and away from the place of implantation, and the endometrium of women with ectopic pregnancy were obtained
... More by collagenase digestion. We used flow cytometry to analyze the expression of phenotype, degree of activation, intracellular expression of cytokines, chemokines, binding assay and FITC–dextran andocytosis by CD1a+ or/and CD83+ dendritic cells. Magnetic sepration was used for purification of decidual subpopulations. We visualised isolated, refined decidual CD1a+ cells by immunocytochemistry. Presence and distribution of CD1a+ , CD209+ and TAG-72+ cells at the feto-maternal interface were analyzed by immunohistology.
Results: Decidual CD1a+ dendritic cells have immature phenotype, due to low expression of CD83, HLA-DR and costimulatory molecules, but they express high amount of endocytic receptors (CD206 and CD209). Contrary to CD83+ cells, CD1a+ cells expressed more IL-15, IFN-γ and CD195 chemokine receptor for inflammatory chemokines. The interaction between CD1a+ cells and autologous NK cells lead to reduced expression of IL-15 and NKp46 receptor and increased expression of cytotoxic mediators in CD56+ cells. Furthermore, CD1a+ dendritic cells efficiently stimulate proliferation of CD56+ cells compared to CD83+ dendritic cells. CD56+ cells significantly killed CD83+ cells.
TAG-72 was bound and internalized by carbohydrate recognition domain to CD206 and CD209 receptors expressed on CD1a+ cells in a dose dependent manner. TAG-72 in vitro decreased the expression of CD83 and CD91 molecules, IL-15, IFN-γ and CCL17, whereas it increased the expression of D6 and IL-1RII on CD1a+ dendritic cells. TAG-72 stimulated CD1a+ dendritic cells decreased the expression of IFN-γ in autologous syngenic and allogenic cells from umbilical cord blood, even in the presence of lipopolysaccharide. TAG-72 enhanced also proliferation of T cells isolated from umbilical cord blood in a dose dependent manner. Intracellular expression of IL-15 and IFN-γ in CD1a+ dendritic cells at the site of implantation and distant from implantation site in tubal pregnancy, where the TAG-72 was present, were very low, in compare to the lining of the uterus in ectopic pregnancy, where the TAG-72 missing.
Conclusion: Removal of TAG-72 from feto-maternal interface, probably contributes to a better control of trophoblast invasion through NK and T cells, because it does not affect the maturation of CD1a+ cells without anti-inflammatory properties that otherwise occur when is TAG-72 present. Less Keywords
Dendritičke stanice
Stanice NK
Limfociti T
C–tip lektinskih receptora (DC-SIGN)
Tumorski glikoprotein (TAG-)72
Imunosna tolerancija
Keywords (english)
Dendritic cells
NK cells
T cells
C–tip lectin receptor (DC-SIGN)
Tumor associated glycoprotein (TAG-)72
Immunological tolerance
Language croatian URN:NBN urn:nbn:hr:188:009732 Study programme Title: Biomedicine Postgraduate (doctoral) study programme Catalog URL http://libraries.uniri.hr/cgi-bin/ucat/unilib.cgi?form=D1121130082 Type of resource Text Extent 247 str; 30 cm File origin Born digital Access conditions Closed access Terms of use Created on 2017-01-19 19:21:29
