RESUMEN
Weak D red cell phenotype (formerly Du) exhibits weaker serological reaction with anti-D antibodies. Weak D occurs in 0.2% to 1% of whites and is caused by qualitatively altered RhD proteins called partial D or normal, only weakly expressed RhD proteins that are called weak D. Partial D genes are hybrid alleles between RHD an RHCE genes. 23 partial RHD alleles are described. Weak D phenotypes with reduced expression are likely to posses the normal RHD gene, but the latest findings indicate that weak D alleles carry at least one point mutation. The aim of the present work was to answer an important question how to approach partial and weak D identification in diagnostic use and if it is possible to distinguish between partial D and weak D using commercially available anti-D reagents for routine use. We also wanted to evaluate D-screen kit for partial D identification. We compared phenotypes identified by serological testing and genotypes identified by RHD Multiplex PCR and DVII specific ASPA PCR. Our results showed that it is not possible to distinguish between partial and weak D using commercially available anti-D reagents for routine use. D-screen proved to be useful for DVI and DVII identification, whereas for partial DDFR identification we must look for another set of monoclonal antibodies or simply use genotyping methods. In 44 samples with not interpretable serological results out of 80 we found all RHD specific exons present and we classified the samples as weak D. Fourteen types of weak D with at least one point mutation were recently proposed. Designing of allele specific PCRs for identification of proposed types of weak D is in progress.
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Tumor necrosis factor α (TNF-α) and its receptors (TNFRI and TNFRII) which exist in soluble form as a product of cleavage of the extracellular domain of membrane integrated receptors, still rise debate about their importance. It was reported that TNF-α has numerous actions in diseases such as inflammation, autoimmunity, infectious diseases, septic shock and many types of cancer [1, 2]. Several authors have reported the significance of sTNFRI level in serum of cancer patients [3, 4]. This study was performed in collaboration with the Institute of Oncology of Slovenia.At least two different mouse monoclonal antibodies (MAbs) against human sTNFRI have been prepared to obtain a sensitive and reliable sandwich ELISA. It was compared with commercially available R&D and Endogen ELISAs for the determination of sTNFRI. Groups of patients with different stages of melanoma and epithelial ovarian carcinoma were tested and their clinical records were reexamined. Levels of sTNFRI were measured and compared with the normal serum levels of sTNFRI.
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Macrophage Migration Inhibitory Factor (MIF) is a crucial component of the immune system acting together with glucocorticosteroids to regulate immunity and inflammation. Understanding of its many putative functions and action mechanisms is still ambiguous. Due to the newest findings that a local MIF expression is up regulated in allograft rejection and in glomerulonephritis, an interest in MIF research is increasing and is focused on possibilities of anti-MIF treatment.In the present work new murine monoclonal antibodies (MAbs) directed against human recombinant MIF (hrMIF) are described. hrMIF protein used for the immunisation was tested for its biological activity and has evident macrophage migration inhibitory activity. The selected MAbs were purified and further characterised. They recognised MIF in a Western blot experiment after a native IEF. Anti-MIF MAb designated as Ml inhibited MIF activity in the test, which was performed in the 48 well Boyden chamber system. It is presumed that Ml MAb could be used as a potential therapeutic agent.
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AIM: The epidemic of bovine spongiform encephalopathy and subsequent emergence of a new variant of Creutzfeldt-Jakob disease have raised great public concern, initiating improved and prospective surveillance of human prion diseases in Europe and all over the world. This report briefly presents the epidemiology, clinical data, neuropathology, immunohistochemistry, biochemistry, and prion-protein gene analysis of Slovenian cases of Creutzfeldt-Jakob disease from January 1985 to the end of 2003. MATERIALS, METHODS AND RESULTS: During the 19-year period, 39 suspected cases of Creutzfeldt-Jakob disease were referred and 22 were confirmed. The prion-protein gene was analyzed in 12 of the confirmed cases and the protein glycosylation pattern in 11. There was a low average incidence of Creutzfeldt-Jakob disease (0.5/million) throughout the surveillance period, but a pronounced increase between January 2001 and December 2003 (to 1.9/million/year). A high female to male ratio (2.5/1) was noted. All of the confirmed cases were defined as sporadic Creutzfeldt-Jakob disease based on the clinical data, neuropathological findings, glycosylation pattern, and gene analysis. All tested cases had a type-2 glycosylation pattern; eleven of the twelve tested patients were homozygous at codon 129 of the prion-protein gene (1 VV and 10 MM) and one was heterozygous. CONCLUSION: The small number of Slovenian cases of sporadic Creutzfeldt-Jakob disease during the last 19 years has shown a pronounced increase in incidence, reflecting improved surveillance, and a high female to male ratio, where female cases are more than twice as numerous as male cases.
Asunto(s)
Síndrome de Creutzfeldt-Jakob/epidemiología , Síndrome de Creutzfeldt-Jakob/genética , Medición de Riesgo/métodos , Distribución por Edad , Anciano , Femenino , Predisposición Genética a la Enfermedad/epidemiología , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Proteínas PrPSc/genética , Prevalencia , Factores de Riesgo , Distribución por Sexo , Eslovenia/epidemiología , Análisis de SupervivenciaRESUMEN
A specific monoclonal antibody (mAb) V5B2 that discriminates between brain tissue of Creutzfeldt-Jakob disease patients and that from normal controls without proteinase K digestion has been prepared using a 13-residue synthetic peptide P1 from the primary structure of human PrP. In the light of the specific interaction between mAb V5B2 and the pathological isoform of PrP (PrP(Sc)), we investigated the solution behavior of antigen P1 and its interactions with mAb V5B2. Our results show that V5B2 recognizes epitope P1 in dimeric/oligomeric forms in solution and in the fibril-like aggregates, as well as in PrP(Sc) aggregates, and demonstrate that the specific epitope is present in all of these forms, but not in PrP(C).
Asunto(s)
Anticuerpos Monoclonales/inmunología , Síndrome de Creutzfeldt-Jakob/diagnóstico , Fragmentos de Péptidos/análisis , Fragmentos de Péptidos/inmunología , Proteínas PrPSc/análisis , Proteínas PrPSc/inmunología , Priones/análisis , Priones/inmunología , Dimerización , Humanos , Concentración de Iones de Hidrógeno , Epítopos Inmunodominantes/análisis , Epítopos Inmunodominantes/química , Epítopos Inmunodominantes/inmunología , Fragmentos de Péptidos/química , Proteínas PrPSc/química , Priones/química , Isoformas de Proteínas/análisis , Isoformas de Proteínas/química , Isoformas de Proteínas/inmunología , Soluciones/química , TemperaturaRESUMEN
Current methods for diagnosing transmissible spongiform encephalopathies rely on the degradation of the cellular prion protein (PrP(C)) and the subsequent detection of the protease-resistant remnant of the pathological prion isoform PrP(Sc) by antibodies that react with all forms of PrP. We report on a monoclonal antibody, V5B2, raised against a peptide from the C-terminal part of PrP, which recognizes an epitope specific to PrP(Sc). In cryostat sections from Creutzfeldt-Jacob's disease (CJD) patients' brains, V5B2 selectively labels various deposits of PrP(Sc) without any pretreatment for removal of PrP(C). V5B2 does not bind to non-CJD brain samples or to recombinant PrP, either in its native or denatured form. Specificity for PrP is confirmed by a sandwich enzyme-linked immunosorbent assay utilizing V5B2, which discriminates between CJD and normal samples without proteinase K treatment, and by immunoprecipitation from CJD brain homogenate. The PrP(Sc)-specific epitope is disrupted by denaturation. We conclude that the C-terminal part of PrP in disease-associated PrP(Sc) aggregates forms a structural epitope whose conformation is distinct from that of PrP(C).