RESUMEN
Background: CD2v, a critical outer envelope glycoprotein of the African swine fever virus (ASFV), plays a central role in the hemadsorption phenomenon during ASFV infection and is recognized as an essential immunoprotective protein. Monoclonal antibodies (mAbs) targeting CD2v have demonstrated promise in both diagnosing and combating African swine fever (ASF). The objective of this study was to develop specific monoclonal antibodies against CD2v. Methods: In this investigation, Recombinant CD2v was expressed in eukaryotic cells, and murine mAbs were generated through meticulous screening and hybridoma cloning. Various techniques, including indirect enzyme-linked immunosorbent assay (ELISA), western blotting, immunofluorescence assay (IFA), and bio-layer interferometry (BLI), were employed to characterize the mAbs. Epitope mapping was conducted using truncation mutants and epitope peptide mapping. Results: An optimal antibody pair for a highly sensitive sandwich ELISA was identified, and the antigenic structures recognized by the mAbs were elucidated. Two linear epitopes highly conserved in ASFV genotype II strains, particularly in Chinese endemic strains, were identified, along with a unique glycosylated epitope. Three mAbs, 2B25, 3G25, and 8G1, effectively blocked CD2v-induced NF-κB activation. Conclusions: This study provides valuable insights into the antigenic structure of ASFV CD2v. The mAbs obtained in this study hold great potential for use in the development of ASF diagnostic strategies, and the identified epitopes may contribute to vaccine development against ASFV.
Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Anticuerpos Monoclonales , Mapeo Epitopo , FN-kappa B , Animales , Virus de la Fiebre Porcina Africana/inmunología , FN-kappa B/metabolismo , FN-kappa B/inmunología , Porcinos , Ratones , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/virología , Anticuerpos Monoclonales/inmunología , Proteínas del Envoltorio Viral/inmunología , Epítopos/inmunología , Anticuerpos Antivirales/inmunología , Ratones Endogámicos BALB CRESUMEN
Background: Fibromyalgia (FM) is a syndrome characterized by chronic and widespread musculoskeletal pain. A number of studies have implied a potential association between gut microbiota and FM. However, the casual association between gut microbiota and FM remains unknown. Method: Mendelian randomization (MR) study was conducted using the summary statistics of genetic variants from the genome-wide association study (GWAS). Inverse variance weighted (IVW), combined with MR-Egger and weighted median were used to investigate the causal association between 119 gut microbiota genera and FM. Sensitivity analyses were performed on the MR results, including heterogeneity test, leave-one-out test and pleiotropy test. Results: A total of 1,295 single nucleotide polymorphism (SNPs) were selected as instrumental variables (IVs), with no significant heterogeneity and pleiotropy according to the sensitivity analyses. Five gut microbiota genera were found to have significant casual association with FM. Coprococcus2 (OR = 2.317, p-value = 0.005, 95% CI: 1.289-4.167), Eggerthella (OR = 1.897, p-value = 0.001, 95% CI: 1.313-2.741) and Lactobacillus (OR = 1.576, p-value =0.020, 95% CI: 1.073-2.315) can increase the risk of FM. FamillyXIIIUCG001 (OR = 0.528, p-value = 0.038, 95% CI: 0.289-0.964) and Olsenella (OR = 0.747, p-value = 0.050, 95% CI: 0.557-1.000) can decrease the risk of FM. Conclusion: This MR study found that gut microbiota is casually associated with FM. New insights into the mechanisms of FM mediated by gut microbiota are provided.