RESUMEN
BACKGROUND: Since the declaration of the coronavirus 2019 (COVID-19) outbreak as pandemic, health workers have shown an incredible commitment to their patients, sometimes in apocalyptic conditions. We explored ways to deal with the coronavirus stressor and psychological outcomes among physicians and nurses. SUBJECTS AND METHODS: 124 healthcare workers in General Hospital Nasice (Croatia) were invited to participate in a study by performing within the period of March 26 to April 6 2020 questionnaire collected information on socio-demographic characteristics and living conditions that may be risk factors for covid-19 concern, Short form health survey-36, Depression Anxiety Stress Scales (DASS-21) and Ways of Coping Questionnaire (WOC; consisting of 8 subscales: Confrontive Coping, Distancing, Self-Controlling, Seeking Social Support, Accepting Responsibility, Escape-Avoidance, Planful Problem Solving, Positive Reappraisal). RESULTS: 11% healthworkers reports moderate to very-severe depression, 17% moderate to extremely-severe anxiety and 10% for moderate to extremely-severe stress. 67% of medical staff are worried. No statistically significant differences in the scales of depression, anxiety, and stress were found between nurses and physicians, but differences were found on Escape-Avoidance and Positive Reappraisal subscales. Nurses use significantly more avoiding coping style and positive reappraisal than doctors. Seeking social support is more pronounced in those over 40 years old, while those under 40 use more avoidable stress management techniques. CONCLUSIONS: Monitoring and ensuring the mental health of coronavirus care staff is crucial for global health. The education of medical staff in the field of stress management is a conditio sine qua non of the issue of an adequate relationship with the COVID-19 pandemic.
Asunto(s)
Adaptación Psicológica , Infecciones por Coronavirus/psicología , Encuestas Epidemiológicas , Salud Mental/estadística & datos numéricos , Enfermeras y Enfermeros/psicología , Médicos/psicología , Neumonía Viral/psicología , Estrés Psicológico , Adulto , COVID-19 , Niño , Infecciones por Coronavirus/epidemiología , Croacia/epidemiología , Humanos , Persona de Mediana Edad , Pandemias , Neumonía Viral/epidemiologíaRESUMEN
NK cells kill various cells using activating receptors, such as the natural cytotoxicity receptors (NCRs). NKp46 is a major NCR and is the only NCR expressed in mice (denoted Ncr1). Using Ncr1-deficient mice (Ncr1(gfp/pfp)) we demonstrated that Ncr1 controls various pathologies, and that in its absence Ncr1-related functions are impaired. In 2012, another Ncr1-related mouse was generated, named Noé, in which a random mutation, W32R, in position 32, impaired the Ncr1-Noé cell surface expression. Interestingly, in the Noé mice, Ncr1-dependent deficiencies were not observed. Additionally, the Noé-NK cells were hyperactivated, probably due to increased Helios expression, and the Noé mice demonstrate increased clearance of influenza and murine CMV. In contrast, in the Ncr1(gfp/pfp) mice infection with influenza was lethal and we show in the present study no difference in murine CMV infection between Ncr1(gfp/pfp) and wild-type (WT) mice. Because the foremost difference between the Noé and Ncr1(gfp/gfp) mice is the presence of a mutated Ncr1-Noé protein, we studied its properties. We show that Ncr1-Noé and various other Ncr1 mutants in position 32 can be expressed on the surface, albeit slowly and unstably, and that ligand recognition and function of the various Ncr1-Noé is similar to the WT Ncr1. We further show that the glycosylation pattern of Ncr1-Noé is aberrant, that the Ncr1-Noé proteins accumulate in the endoplasmic reticulum, and that the expression of Ncr1-Noé proteins, but not WT Ncr1, leads to increased Helios expression. Thus, we suggest that the NK hyperactivated phenotype observed in the Noé mice might result from the presence of the Ncr1-Noé protein.
Asunto(s)
Antígenos Ly/inmunología , Regulación de la Expresión Génica/inmunología , Receptor 1 Gatillante de la Citotoxidad Natural/inmunología , Animales , Antígenos Ly/genética , Línea Celular Tumoral , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Glicosilación , Humanos , Factor de Transcripción Ikaros/genética , Factor de Transcripción Ikaros/inmunología , Ratones , Ratones Transgénicos , Mutación , Receptor 1 Gatillante de la Citotoxidad Natural/genética , Factores de Transcripción/genética , Factores de Transcripción/inmunologíaRESUMEN
Varicella-zoster virus (VZV) is the etiological agent of chickenpox and shingles. Due to the virus's restricted host and cell type tropism and the lack of tools for VZV proteomics, it is one of the least-characterized human herpesviruses. We generated 251 monoclonal antibodies (MAbs) against 59 of the 71 (83%) currently known unique VZV proteins to characterize VZV protein expression in vitro and in situ. Using this new set of MAbs, 44 viral proteins were detected by Western blotting (WB) and indirect immunofluorescence (IF); 13 were detected by WB only, and 2 were detected by IF only. A large proportion of viral proteins was analyzed for the first time in the context of virus infection. Our study revealed the subcellular localization of 46 proteins, 14 of which were analyzed in detail by confocal microscopy. Seven viral proteins were analyzed in time course experiments and showed a cascade-like temporal gene expression pattern similar to those of other herpesviruses. Furthermore, selected MAbs tested positive on human skin lesions by using immunohistochemistry, demonstrating the wide applicability of the MAb collection. Finally, a significant portion of the VZV-specific antibodies reacted with orthologs of simian varicella virus (SVV), thus enabling the systematic analysis of varicella in a nonhuman primate model system. In summary, this study provides insight into the potential function of numerous VZV proteins and novel tools to systematically study VZV and SVV pathogenesis.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Herpesvirus Humano 3/metabolismo , Proteínas Virales/inmunología , Proteínas Virales/metabolismo , Animales , Western Blotting , Línea Celular , Línea Celular Tumoral , Varicela/virología , Células Epiteliales/virología , Técnica del Anticuerpo Fluorescente Indirecta , Herpes Zóster/virología , Herpesvirus Humano 3/inmunología , Humanos , Ratones , Ratones Endogámicos BALB C , Proteómica , Piel/inmunología , Piel/virologíaRESUMEN
Members of the alpha- and beta-subfamily of herpesviridae encode glycoproteins that specifically bind to the Fc part of immunoglobulin (Ig)G. Plasma membrane resident herpesviral Fc receptors seem to prevent virus-specific IgG from activating antibody-dependent effector functions. We show that the mouse cytomegalovirus (MCMV) molecule fcr-1 promotes a rapid down-regulation of NKG2D ligands murine UL16-binding protein like transcript (MULT)-1 and H60 from the cell surface. Deletion of the m138/fcr-1 gene from the MCMV genome attenuates viral replication to natural killer (NK) cell response in an NKG2D-dependent manner in vivo. A distinct N-terminal module within the fcr-1 ectodomain in conjunction with the fcr-1 transmembrane domain was required to dispose MULT-1 to degradation in lysosomes. In contrast, down-modulation of H60 required the complete fcr-1 ectodomain, implying independent modes of fcr-1 interaction with the NKG2D ligands. The results establish a novel viral strategy for down-modulating NK cell responses and highlight the impressive diversity of Fc receptor functions.
Asunto(s)
Proteínas Portadoras/metabolismo , Regulación hacia Abajo , Antígenos de Histocompatibilidad Clase I/metabolismo , Glicoproteínas de Membrana/metabolismo , Antígenos de Histocompatibilidad Menor/metabolismo , Muromegalovirus/metabolismo , Receptores Fc/metabolismo , Receptores Inmunológicos/metabolismo , Proteínas Virales/metabolismo , Animales , Proteínas Portadoras/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Inmunoglobulinas/inmunología , Ligandos , Lisosomas/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos BALB C , Antígenos de Histocompatibilidad Menor/inmunología , Muromegalovirus/fisiología , Células 3T3 NIH , Subfamilia K de Receptores Similares a Lectina de Células NK , Procesamiento Proteico-Postraduccional , Estructura Terciaria de Proteína , Receptores Inmunológicos/inmunología , Receptores de Células Asesinas Naturales , Replicación Viral/fisiologíaRESUMEN
The killing activity of NK cells is regulated by signals derived from inhibitory and activating NK cell receptors, including the CD300 family of proteins. CD300a was reported to be expressed on all NK cells and to deliver an inhibitory signal upon binding to a yet unknown ligand/s. The CD300a protein contains four ITIMs and is highly homologous to CD300c. Little is known about the function and distribution of these two receptors and the identity of their ligand/s. In this article, we show that CD300a is indeed an inhibitory receptor expressed by human NK cells, but surprisingly, we show that not all NK clones are inhibited in a CD300a-dependent manner. We demonstrate, using a panel of 13 new anti-CD300a and CD300c Abs that we generated, that CD300a and CD300c are indistinguishable on the surface of NK cells. Using mutational-analysis survey, we show that tyrosine 267 located in the third ITIM motif of the CD300a protein is important for the inhibitory function of CD300a.
Asunto(s)
Antígenos CD/biosíntesis , Antígenos CD/fisiología , Regulación de la Expresión Génica/inmunología , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Receptores Inmunológicos/biosíntesis , Receptores Inmunológicos/fisiología , Alanina/metabolismo , Secuencias de Aminoácidos/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/metabolismo , Antígenos CD/genética , Sitios de Unión de Anticuerpos , Línea Celular Tumoral , Células Clonales , Técnicas de Cocultivo , Reactivos de Enlaces Cruzados/metabolismo , Inhibidores de Crecimiento/antagonistas & inhibidores , Inhibidores de Crecimiento/biosíntesis , Inhibidores de Crecimiento/genética , Inhibidores de Crecimiento/fisiología , Humanos , Inmunofenotipificación , Ratones , Datos de Secuencia Molecular , Receptores Inmunológicos/antagonistas & inhibidores , Receptores Inmunológicos/genética , Tirosina/metabolismoRESUMEN
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
BACKGROUND: The aim of this study was to examine the impact of education, coronavirus disease 2019 (COVID-19), and risk factors on the quality of life in patients with type 2 diabetes. METHODS: A prospective study was conducted in three phases: before education, after education, and in the period of pandemic coronavirus disease 2019 (COVID-19). The subjects were diabetics on oral therapy. To determine the quality of life index, a standardized Ferrans and Powers survey questionnaire was used. RESULTS: A total of 205 participants took part in the study, of which 111 (54.1%) were men and 94 (46%) women. Participants were enrolled in the study between January 2019 and September 2020. Glycated hemoglobin values were significantly higher before education compared to post-education and at the time of COVID-19 (Friedman test, p = 0.002), and body mass index was significantly lower after education compared to values before education (Friedman test, p = 0.008). The quality of life was significantly lower in all domains in the COVID-19 period (Friedman test, p < 0.001). CONCLUSIONS: A significant predictor of worse assessment of overall quality of life was male gender and rural place of residence. Disease duration of up to 5 years was a significant predictor of worse assessment in the psychological/spiritual domain, while being married was a predictor of better assessment of the quality of life in the family domain. The education of diabetics brought an increase in the health and quality of life while the coronavirus disease pandemic had negative consequences on the same parameters. We consider it necessary to systematically educate diabetics about the comorbidity of COVID-19.
Asunto(s)
COVID-19 , Diabetes Mellitus Tipo 2 , Educación del Paciente como Asunto , Calidad de Vida , Comorbilidad , Femenino , Humanos , Masculino , Estudios Prospectivos , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
Many viruses encode proteins that inhibit the induction of programmed cell death at the mitochondrial checkpoint. Murine cytomegalovirus (MCMV) encodes the m38.5 protein, which localizes to mitochondria and protects human HeLa cells and fibroblasts from apoptosis triggered by proteasome inhibitors but not from Fas-induced apoptosis. However, the ability of this protein to suppress the apoptosis of murine cells and its role during MCMV infection have not been investigated previously. Here we show that m38.5 is expressed at early time points during MCMV infection. Cells infected with MCMVs lacking m38.5 showed increased sensitivity to cell death induced by staurosporine, MG132, or the viral infection itself compared to the sensitivity of cells infected with wild-type MCMV. This defect was eliminated when an m38.5 or Bcl-X(L) gene was inserted into the genome of a deletion mutant. Using fibroblasts deficient in the proapoptotic Bcl-2 family proteins Bak and/or Bax, we further demonstrated that m38.5 protected from Bax- but not Bak-mediated apoptosis and interacted with Bax in infected cells. These results consolidate the role of m38.5 as a viral mitochondrion-localized inhibitor of apoptosis and its functional similarity to the human cytomegalovirus UL37x1 gene product. Although the m38.5 gene is not homologous to the UL37x1 gene at the sequence level, m38.5 is conserved among rodent cytomegaloviruses. Moreover, the fact that MCMV-infected cells are protected from both Bak- and Bax-mediated cell death suggests that MCMV possesses an additional, as-yet-unidentified mechanism to block Bak-mediated apoptosis.
Asunto(s)
Muerte Celular , Muromegalovirus/inmunología , Proteínas Virales/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Animales , Línea Celular , Inhibidores de Cisteína Proteinasa , Fragmentación del ADN , ADN Viral/genética , Fibroblastos/virología , Eliminación de Gen , Prueba de Complementación Genética , Leupeptinas/farmacología , Ratones , Datos de Secuencia Molecular , Muromegalovirus/genética , Muromegalovirus/fisiología , Unión Proteica , Análisis de Secuencia de ADN , Estaurosporina/farmacología , Proteínas Virales/genética , Proteína X Asociada a bcl-2/antagonistas & inhibidoresRESUMEN
AIM: The aim of this study was to evaluate the presence of the cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in human periapical lesions. SUBJECTS AND METHODS: Samples were obtained from three groups of teeth: symptomatic teeth, asymptomatic lesions, and uninflamed periradicular tissues as a control. RESULTS: TNF-alpha levels were significantly increased in symptomatic lesions compared to control. Group with asymptomatic lesions had significantly higher concentrations compared to control. There were no significant differences in TNF-alpha levels between symptomatic and asymptomatic lesions. In group with symptomatic lesions, IL-6 levels were significantly higher than in group with asymptomatic lesions. The IL-6 levels in symptomatic group also showed significantly higher concentration in comparison with control group. In asymptomatic group, the IL-6 level had significantly higher concentrations compared to control. CONCLUSION: These results indicate that symptomatic lesions represent an immunologically active stage of disease, and asymptomatic lesions are the point from which the process advances toward healing.
Asunto(s)
Interleucina-6/metabolismo , Periodontitis Periapical/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Ensayo de Inmunoadsorción Enzimática , Humanos , Periodontitis Periapical/patologíaRESUMEN
Mouse cytomegalovirus (MCMV), a beta-herpesvirus that establishes latent and persistent infections in mice, is a valuable model for studying complex virus-host interactions. MCMV encodes the m145 family of putative immunoevasins with predicted major histocompatibility complex, class I (MHC-I) structure. Functions attributed to some family members include down-regulation of host MHC-I (m152) and NKG2D ligands (m145, m152, and m155) and interaction with inhibitory or activating NK receptors (m157). We present the cellular, biochemical, and structural characterization of m153, which is a heavily glycosylated homodimer, that does not require beta2m or peptide and is expressed at the surface of MCMV-infected cells. Its 2.4-A crystal structure confirms that this compact molecule preserves an MHC-I-like fold and reveals a novel mode of dimerization, confirmed by site-directed mutagenesis, and a distinctive disulfide-stabilized extended N terminus. The structure provides a useful framework for comparative analysis of the divergent members of the m145 family.