RESUMEN
Previous studies using ELISA and virus neutralization test (VNT) have proved the presence of Murine gammaherpesvirus 68 (MHV-68) serum antibodies in sera of laboratory staff working with MHV-68, as well as in the general population. In this study, we investigated the incidence of serum antibodies to MHV-68 and to human herpesviruses presumably antigenically similar to MHV-68, as Herpes simplex virus 1 (HSV-1), Human cytomegalovirus (HCMV), and Epstein-Barr virus (EBV), in general population using ELISA, VNT, and immunofluorescence assay (IFA). We also searched for the possible detection of false-positive reaction of MHV-68 antibodies due to cross-reactions between MHV-68 and the antibodies to the herpesviruses mentioned above. We found 16% of positive sera for MHV-68 antibodies by ELISA (titers of 1,600-102,400) and 4.5% by VNT and IFA (titers of 8-32). Tested human sera, either positive or negative for MHV-68 antibodies, were positive for antibodies to HSV-1, HCMV, and EBV (71/69%, 69/65%, and 66/49%, respectively). We concluded that the false-positivity of the sera for MHV-68 antibodies detected by ELISA was due to the nonspecific cross-reactions with antibodies to antigenically similar EBV.
Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Herpesviridae/inmunología , Rhadinovirus/inmunología , Infecciones Tumorales por Virus/inmunología , Reacciones Cruzadas , Citomegalovirus/inmunología , Ensayo de Inmunoadsorción Enzimática , Reacciones Falso Positivas , Técnica del Anticuerpo Fluorescente Indirecta , Herpesvirus Humano 1/inmunología , Herpesvirus Humano 4/inmunología , Humanos , Pruebas de NeutralizaciónRESUMEN
A tumor cell line, NB-78, was derived from a lymphoma from a BALB/c mouse infected with Murine gammaherpesvirus 78 (MHV-78). Cultures of the cells of this line underwent till now more than 100 passages, displaying an epitheloid transformed morphology and a diploid complement of 40 chromosomes. Viral antigen was detected in 12% of cells by immunofluorescence (IF) test. Reactivation of latent MHV-78 was proved by detecting infectious virus in culture medium only at passages 4345. The presence of viral M1, M2, M3, and M4 gene sequences in the genome of the cells was demonstrated by PCR. NB-78 is the first continuous cell line, which originates from a tumor of a MHV-78-infected host, harbors viral genome or at least its several genes, and produces infectious virus only rarely upon reactivation. It can be assumed that this cell line is primarily associated with MHV-78 and will serve as an invaluable tool for studying the MHV-78 latency.
Asunto(s)
Línea Celular Tumoral/virología , Transformación Celular Viral , Gammaherpesvirinae/genética , Animales , Antígenos Virales/biosíntesis , Diploidia , Femenino , Técnica del Anticuerpo Fluorescente , Gammaherpesvirinae/fisiología , Genes Virales , Genoma/genética , Inmunoquímica , Cariotipificación , Linfoma , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa , Activación Viral , Latencia del VirusRESUMEN
OBJECTIVE: Oxidative stress plays a crucial role in the development of complications in Diabetes mellitus (DM). Individual sensitivity against stress, however, varies among DM-patients and results, therefore, in differential severity of consequent complications. To allow more complex interpretation of a delicate antioxidant/free radicals balance and its effect on cellular functions in individual DM-patients, we analysed a correlation between total antioxidant status (TAS), antioxidant gap (AtxGap), level of free radicals (FR), routine clinical biochemical parameters in blood and differential gene expression in circulating leukocytes of DM-patients versus non-diabetic individuals. RESULTS AND CONCLUSIONS: Positive correlation was found between TAS and creatinine (p=0.05), AtxGap and iron (p=0.025), and between AtxGap and anti-streptolysin O (p=0.025). Whereas no correlation was found between FR and any of the routine clinical parameters tested, a negative correlation was observed between AtxGap and glucose content (p=0.025) and between AtxGap and gamma-glutamyltransferase (p=0.05). An increased content of FR was shown to influence significantly an expression of selected stress responsible genes in leukocytes. Transcription levels of NF-kappaB, XRCC1 and 90-kDa heat-shock protein A were increased in all DM-patients compared to non-diabetic individuals. In contrast, an expression of XIAP and cytochrome P450 reductase was up-regulated in patients with decreased levels of both FR and TAS and increased body mass index. This differential expression of the stress responsible genes might be further considered as a potential risk factor for diverse DM-complications helping also in reliable monitoring of supplemental antioxidant therapy and more complex interpretation of delicate antioxidant/free radicals balance.