RESUMEN
MICA and MICB are stress-induced ligands recognized by the activating receptor NKG2D. A microRNA encoded by human cytomegalovirus downregulates MICB expression by targeting a specific site in the MICB 3' untranslated region. As this site is conserved among different MICB alleles and a similar site exists in the MICA 3' untranslated region, we speculated that these sites are targeted by cellular microRNAs. Here we identified microRNAs that bound to these MICA and MICB 3' untranslated region sequences and obtained data suggesting that these microRNAs maintain expression of MICA and MICB protein under a certain threshold and facilitate acute upregulation of MICA and MICB during cellular stress. These microRNAs were overexpressed in various tumors and we demonstrate here that they aided tumor avoidance of immune recognition.
Asunto(s)
Antígenos de Histocompatibilidad Clase I/metabolismo , MicroARNs/fisiología , Receptores Inmunológicos/metabolismo , Estrés Fisiológico/inmunología , Antígenos de Superficie/metabolismo , Línea Celular , Vectores Genéticos , Antígenos de Histocompatibilidad Clase I/química , Humanos , MicroARNs/inmunología , Subfamilia K de Receptores Similares a Lectina de Células NK , Receptores de Células Asesinas Naturales , Estrés Fisiológico/genética , Estrés Fisiológico/metabolismoRESUMEN
The activity of natural killer (NK) cells is controlled by a balance of signals derived from inhibitory and activating receptors. TIGIT is a novel inhibitory receptor, recently shown in humans to interact with two ligands: PVR and Nectin2 and to inhibit human NK-cell cytotoxicity. Whether mouse TIGIT (mTIGIT) inhibits mouse NK-cell cytotoxicity is unknown. Here we show that mTIGIT is expressed by mouse NK cells and interacts with mouse PVR. Using mouse and human Ig fusion proteins we show that while the human TIGIT (hTIGIT) cross-reacts with mouse PVR (mPVR), the binding of mTIGIT is restricted to mPVR. We further demonstrate using surface plasmon resonance (SPR) and staining with Ig fusion proteins that mTIGIT binds to mPVR with higher affinity than the co-stimulatory PVR-binding receptor mouse DNAM1 (mDNAM1). Functionally, we show that triggering of mTIGIT leads to the inhibition of NK-cell cytotoxicity, that IFN-γ secretion is enhanced when mTIGIT is blocked and that the TIGIT-mediated inhibition is dominant over the signals delivered by the PVR-binding co-stimulatory receptors. Additionally, we identify the inhibitory motif responsible for mTIGIT inhibition. In conclusion, we show that TIGIT is a powerful inhibitory receptor for mouse NK cells.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Citotoxicidad Inmunológica , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Receptores Inmunológicos/metabolismo , Receptores Virales/metabolismo , Secuencia de Aminoácidos , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos T/metabolismo , Antígenos de Histocompatibilidad Clase I/inmunología , Interferón gamma/biosíntesis , Ratones , Ratones Endogámicos C57BL , Nectinas , Receptores Inmunológicos/química , Proteínas Recombinantes de Fusión/metabolismo , Alineación de Secuencia , Transducción de Señal , Resonancia por Plasmón de SuperficieRESUMEN
NK cells employ a variety of activating receptors to kill virally infected and tumor cells. Prominent among these receptors are the natural cytotoxicity receptors (NCRs) (NKp30, NKp44, and NKp46), of which only NKp46 has a mouse ortholog (NCR1). The tumor ligand(s) of NKp46/NCR1 is still unknown, but it was shown that the human NKp46 and the mouse NCR1 are involved in tumor eradication both in vitro and in vivo. Whether any of the NK activating receptors is involved in the prevention of tumor metastasis is unknown. To address this question, we studied the activity of the NK cell receptor NKp46/NCR1 in two spontaneous metastasis models, the B16F10.9 melanoma (B16) and the Lewis lung carcinoma (D122) in the NCR1 knockout mouse that was generated by our group, in various in vitro and in vivo assays. We demonstrated that all B16 and D122 tumors, including those generated in vivo, express an unknown ligand(s) for NKp46/NCR1. We have characterized the properties of the NKp46/NCR1 ligand(s) and demonstrated that NKp46/NCR1 is directly involved in the killing of B16 and D122 cells. Importantly, we showed in vivo that NKp46/NCR1 plays an important role in controlling B16 and D122 metastasis. Thus, to our knowledge, in this study we provide the first evidence for the direct involvement of a specific NK killer receptor in preventing tumor metastasis.
Asunto(s)
Antígenos Ly/inmunología , Receptor 1 Gatillante de la Citotoxidad Natural/inmunología , Animales , Antígenos Ly/genética , Antígenos Ly/metabolismo , Carcinoma Pulmonar de Lewis , Separación Celular , Citotoxicidad Inmunológica/inmunología , Citometría de Flujo , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptor 1 Gatillante de la Citotoxidad Natural/genética , Receptor 1 Gatillante de la Citotoxidad Natural/metabolismo , Invasividad Neoplásica/genética , Metástasis de la Neoplasia/prevención & controlRESUMEN
NK cell cytotoxicity is controlled by numerous NK inhibitory and activating receptors. Most of the inhibitory receptors bind MHC class I proteins and are expressed in a variegated fashion. It was recently shown that TIGIT, a new protein expressed by T and NK cells binds to PVR and PVR-like receptors and inhibits T cell activity indirectly through the manipulation of DC activity. Here, we show that TIGIT is expressed by all human NK cells, that it binds PVR and PVRL2 but not PVRL3 and that it inhibits NK cytotoxicity directly through its ITIM. Finally, we show that TIGIT counter inhibits the NK-mediated killing of tumor cells and protects normal cells from NK-mediated cytotoxicity thus providing an "alternative self" mechanism for MHC class I inhibition.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Citotoxicidad Inmunológica , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Receptores Inmunológicos/metabolismo , Receptores Virales/metabolismo , Animales , Moléculas de Adhesión Celular/química , Línea Celular , Antígenos de Histocompatibilidad Clase I/metabolismo , Humanos , Técnicas In Vitro , Subunidad beta del Receptor de Interleucina-2/metabolismo , Ligandos , Subgrupos Linfocitarios/inmunología , Subgrupos Linfocitarios/metabolismo , Ratones , Nectinas , Unión Proteica , Dominios y Motivos de Interacción de Proteínas , Receptores Inmunológicos/química , Receptores Inmunológicos/genética , Receptores Virales/química , Receptores Virales/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Human natural killer (NK) cells possess an arsenal of receptors programmed to regulate the NK cell functions, once encountering a target cell. In general, the activating receptors mediate cytotoxicity when engaged by their tumor specific, stress induced, virally encoded, or rarely, self ligands. Whereas, the inhibitory receptors bind self molecules, mostly MHC class I, presented on all normal and healthy nucleated cells. However, NK cells also possess numerous, highly homologous, pairs of receptors that sometimes even share the same ligands but display divergent functions. In this review we describe the NK cell repertoire of paired receptors and discuss questions regarding their function and mode of action. We focus primarily on the three PVR-binding receptors; the co-stimulatory DNAM1 and CD96 and the inhibitory TIGIT.