Multisubunit cullin-RING ubiquitin ligase 4 (CRL4)-DCAF12 recognizes the C-terminal degron containing acidic amino acid residues. However, its physiological roles and substrates are largely unknown. Purification of CRL4-DCAF12 complexes revealed a wide range of potential substrates, including MOV10, an "ancient" RNA-induced silencing complex (RISC) complex RNA helicase. We show that DCAF12 controls the MOV10 protein level via its C-terminal motif in a proteasome- and CRL-dependent manner. Next, we generated Dcaf12 knockout mice and demonstrated that the DCAF12-mediated degradation of MOV10 is conserved in mice and humans. Detailed analysis of Dcaf12-deficient mice revealed that their testes produce fewer mature sperms, phenotype accompanied by elevated MOV10 and imbalance in meiotic markers SCP3 and γ-H2AX. Additionally, the percentages of splenic CD4+ T and natural killer T (NKT) cell populations were significantly altered. In vitro, activated Dcaf12-deficient T cells displayed inappropriately stabilized MOV10 and increased levels of activated caspases. In summary, we identified MOV10 as a novel substrate of CRL4-DCAF12 and demonstrated the biological relevance of the DCAF12-MOV10 pathway in spermatogenesis and T cell activation.
- Klíčová slova
- C-terminal degron, DCAF12, MOV10, T cell activation, WDR40A, spermatogenesis,
- MeSH
- aktivace lymfocytů fyziologie MeSH
- antigeny nádorové metabolismus MeSH
- buněčné linie MeSH
- CD4-pozitivní T-lymfocyty metabolismus MeSH
- HCT116 buňky MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- NKT buňky metabolismus MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- RNA-helikasy metabolismus MeSH
- spermatogeneze fyziologie MeSH
- ubikvitin metabolismus MeSH
- ubikvitinligasy metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny nádorové MeSH
- DCAF12 protein, human MeSH Prohlížeč
- IL17RB protein, human MeSH Prohlížeč
- Mov10 protein, human MeSH Prohlížeč
- proteasomový endopeptidasový komplex MeSH
- RNA-helikasy MeSH
- ubikvitin MeSH
- ubikvitinligasy MeSH
Recognition of glycosylation patterns is one of the basic features of innate immunity. Ability of C-type lectin-like receptors such as NKR-P1 to bind saccharide moieties has become recently a controversial issue. In the present study, binding assay with soluble fluorescently labeled recombinant rat NKR-P1A and mouse NKR-P1C proteins revealed apparently no affinity to the various neoglycoproteins. Lack of functional linkage between NKR-P1 and previously described saccharide binder was supported by the fact, that synthetic N-acetyl-D-glucosamine octabranched dendrimer on polyamidoamine scaffold (GN8P) did not change gene expression of NKR-P1 isoforms in C57BL/6 and BALB/c mice divergent in the NK gene complex (both in vitro and in vivo). Surprisingly, N-acetyl-D-glucosamine-coated tetrabranched polyamido-amine dendrimer specifically binds to NKT cells and macrophages but not to NK cells (consistently with changes in cytokine patterns). Despite the fact that GN8P has been tested as an immunomodulator in anti-cancer treatment animal models for many years, surprisingly no changes in cytokine profiles in serum relevant to anti-cancer responses using B16F10 and CT26 harboring mouse strains C57BL/6 and BALB/c are observed. Our results indicate possible indirect involvement of NK cells in GN8P mediated immune responses.
- Klíčová slova
- Anti-tumor immunity, C-type lectin related protein, Carbohydrate dendrimer, Clr, GN4P-A: GlcNAc4-PAMAM-ATTO 565, GN4P-NH(2)-GlcNAc(4)-PAMAM, GN4P: GlcNAc4-PAMAM, GN8P: GlcNAc8-PAMAM, GlcNAc, Gzmb, Macrophages, N-acetyl-d-glucosamine, N-acetyl-d-glucosamine-coated octabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer fluorescently labeled with ATTO 565, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer with free NH(2) group, NK cells, NKG2D, NKR-P1, NKR-P1 receptors, NKT cells, PAMAM dendrimer, PMA, Prf, SBA, SMC, granzyme B, natural killer group 2, member D, natural killer receptor protein 1, perforin, phorbol 12-myristate 13-acetate, polyamidoamine dendrimer, soybean agglutinin, spleen mononuclear cell,
- MeSH
- acetylglukosamin imunologie metabolismus MeSH
- buňky NK imunologie metabolismus MeSH
- dendrimery metabolismus MeSH
- experimentální nádory farmakoterapie genetika imunologie MeSH
- exprese genu účinky léků imunologie MeSH
- glykokonjugáty imunologie metabolismus farmakologie MeSH
- interferon gama krev genetika imunologie MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- lektinové receptory NK-buněk - podrodina B genetika imunologie metabolismus MeSH
- lektiny typu C genetika imunologie metabolismus MeSH
- makrofágy imunologie metabolismus MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- NKT buňky imunologie metabolismus MeSH
- oligosacharidy imunologie metabolismus MeSH
- polyaminy imunologie metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protein - isoformy genetika imunologie metabolismus MeSH
- průtoková cytometrie MeSH
- slezina cytologie imunologie metabolismus MeSH
- TNF-alfa krev genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- acetylglukosamin MeSH
- dendrimery MeSH
- glykokonjugáty MeSH
- interferon gama MeSH
- lektinové receptory NK-buněk - podrodina B MeSH
- lektiny typu C MeSH
- oligosacharidy MeSH
- Poly(amidoamine) MeSH Prohlížeč
- polyaminy MeSH
- protein - isoformy MeSH
- TNF-alfa MeSH
CD4(+)CD25(+)Foxp3(+) T regulatory cells (Tregs) and CD1d-restricted invariant natural killer T (iNKT) cells are two cell types that are known to regulate immune reactions. Depletion or inactivation of Tregs using specific anti-CD25 antibodies in combination with immunostimulation is an attractive modality especially in anti-tumour immunotherapy. However, CD25 is not expressed exclusively on Tregs but also on subpopulations of activated lymphocytes. Therefore, the modulatory effects of the specific anti-CD25 antibodies can also be partially attributed to their interactions with the effector cells. Here, the effector functions of iNKT cells were analysed in combination with anti-CD25 mAb PC61. Upon PC61 administration, α-galactosylceramide (α-GalCer)-mediated activation of iNKT cells resulted in decreased IFN-γ but not IL-4 production. In order to determine whether mutual interactions between Tregs and iNKT cells take place, we compared IFNγ production after α-GalCer administration in anti-CD25-treated and "depletion of regulatory T cell" (DEREG) mice. Since no profound effects on IFNγ induction were observed in DEREG mice, deficient in FoxP3(+) Tregs, our results indicate that the anti-CD25 antibody acts directly on CD25(+) effector cells. In vivo experiments demonstrated that although both α-GalCer and PC61 administration inhibited TC-1 tumour growth in mice, no additive/synergic effects were observed when these substances were used in combination therapy.
- MeSH
- antigeny CD1d imunologie metabolismus MeSH
- ELISA MeSH
- experimentální nádory farmakoterapie imunologie patologie MeSH
- exprese genu účinky léků imunologie MeSH
- forkhead transkripční faktory imunologie metabolismus MeSH
- galaktosylceramidy aplikace a dávkování imunologie farmakologie MeSH
- heparin-vázající růstový faktor podobný EGF MeSH
- interferon gama genetika imunologie metabolismus MeSH
- interleukin-4 genetika imunologie metabolismus MeSH
- Kaplanův-Meierův odhad MeSH
- mezibuněčné signální peptidy a proteiny genetika imunologie metabolismus MeSH
- monoklonální protilátky aplikace a dávkování imunologie farmakologie MeSH
- myši inbrední C57BL MeSH
- myši transgenní MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- NKT buňky účinky léků imunologie metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protokoly protinádorové kombinované chemoterapie terapeutické užití MeSH
- průtoková cytometrie MeSH
- receptor interleukinu-2 - alfa-podjednotka imunologie metabolismus MeSH
- regulační T-lymfocyty účinky léků imunologie metabolismus MeSH
- tumor burden účinky léků imunologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alpha-galactosylceramide MeSH Prohlížeč
- antigeny CD1d MeSH
- forkhead transkripční faktory MeSH
- Foxp3 protein, mouse MeSH Prohlížeč
- galaktosylceramidy MeSH
- Hbegf protein, mouse MeSH Prohlížeč
- heparin-vázající růstový faktor podobný EGF MeSH
- interferon gama MeSH
- interleukin-4 MeSH
- mezibuněčné signální peptidy a proteiny MeSH
- monoklonální protilátky MeSH
- PC61 monoclonal antibody MeSH Prohlížeč
- receptor interleukinu-2 - alfa-podjednotka MeSH
Invariant natural killer T (iNKT) cells play an important role in the immune response against various infectious agents. In this study we investigated their role in human defense against the varicella zoster virus. We observed decreased numbers of iNKT cells in patients who failed to control latent varicella zoster virus infection, e.g. underwent several reactivations of the virus. The residual population of iNKT cells expressed significantly higher levels of inhibitory receptor CD158a that was further up-regulated in the course of acute viral infection. Both of these abnormalities might contribute to impaired control of varicella zoster virus in human.
- MeSH
- aktivace lymfocytů imunologie MeSH
- buňky NK imunologie metabolismus patologie MeSH
- CD antigeny metabolismus MeSH
- cytokiny metabolismus MeSH
- herpes zoster imunologie patologie MeSH
- lektinové receptory NK-buněk - podrodina K metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- NKT buňky imunologie metabolismus patologie MeSH
- počet lymfocytů MeSH
- podskupiny lymfocytů imunologie metabolismus patologie MeSH
- receptory KIR2DL1 metabolismus MeSH
- T-lymfocyty imunologie metabolismus patologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CD antigeny MeSH
- cytokiny MeSH
- KIR2DL1 protein, human MeSH Prohlížeč
- lektinové receptory NK-buněk - podrodina K MeSH
- receptory KIR2DL1 MeSH
iNKT cells, CD1d dependent natural killer T cells are a unique population of T cells. The capacity of iNKT cells to produce regulatory cytokines first provided an indication of their regulatory potential. Later on, in experimental models as well as in patients afflicted with an auto-immune disease, such as Type 1 diabetes mellitus, multiple sclerosis, and systemic lupus erythematosus along with others, a deficit in iNKT cell number was observed, suggesting the role these cells may possibly have in the prevention of auto-immune diseases. More importantly, experimental strategies which focused on increasing the volume or stimulation of iNKT cells in laboratory animals, demonstrated an improved level of protection against the development of auto-immune diseases. This article reviews the mechanism of protection against autoimmunity by iNKT cells, discusses the obstacles against and indications for the potential use of iNKT cell manipulation in the treatment of human auto-immune diseases.
- MeSH
- antigeny CD1d imunologie metabolismus MeSH
- autoimunita imunologie MeSH
- autoimunitní nemoci imunologie MeSH
- diabetes mellitus 1. typu imunologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- NKT buňky imunologie metabolismus MeSH
- roztroušená skleróza imunologie MeSH
- systémový lupus erythematodes imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- antigeny CD1d MeSH
The aim of this study was to investigate differences in the relative distributions of subsets of natural killer (NK) cells, including immunoregulatory NK cells (CD56(+)CD16(-)), cytotoxic NK cells (CD56(+)CD16(+)), as well as total NK cells (CD56(+)CD3(-)), and NKT cells (CD56(+)CD3(+)) in peripheral blood and follicular fluid in subjects with successful or unsuccessful IVF treatment. The immunoregulatory NK cell population in follicular fluid of women who failed to achieve pregnancy after IVF treatment was significantly decreased compared to women who became pregnant after IVF. Conversely, the NKT cell population in the follicular fluid of women with unsuccessful treatment was significantly elevated compared with those with successful IVF. Understanding the changes in the distribution of NK and NKT cell populations in follicular fluid might serve as the basis for a more detailed study to determine whether NK cell parameters have prognostic value in guiding the selection of individual ova for use in IVF procedures.
- MeSH
- biologické markery metabolismus MeSH
- buňky NK imunologie metabolismus patologie MeSH
- CD antigeny biosyntéza MeSH
- dospělí MeSH
- fertilizace in vitro * MeSH
- folikulární tekutina cytologie imunologie metabolismus MeSH
- lidé MeSH
- NKT buňky imunologie metabolismus patologie MeSH
- počet buněk MeSH
- prognóza MeSH
- průtoková cytometrie MeSH
- separace buněk MeSH
- těhotenství MeSH
- výsledek terapie MeSH
- ženská infertilita krev diagnóza patologie patofyziologie terapie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- biologické markery MeSH
- CD antigeny MeSH
