Search

Introduction. In visceral leishmaniasis (VL), in old world caused by Leishmania donovani, elongation factor-1α (EF-1α) is an antigen-activator of SHP-1 phosphatase that inhibits the leishmanicidal activity of macrophages. However, the relevance of EF-1α from L. infantum in VL of the new world is still unknown. Objective. Obtain the recombinant protein of EF-1α from L. infantum (rEFLi). Material and methods. The EFLi gene was cloned, sequenced, and expressed as recombinant protein in a prokaryotic system. rEFLi was purified and its immunogenicity was verified. Results. The EFLi gene has 1350 pb, with 25 nucleotide differences, 10 amino acid differences and without structural differences (3D) compared to EFLd. rEFLi (50 kDa), was recognized by sera from mice immunized with its homologous genetic construction. Conclusion. Our evidence suggests that rEFLi can be evaluated as a biotechnological vaccine alone or in combination, for the prevention of VL. [ABSTRACT FROM AUTHOR]

Introduction: Chronic hyperglycemia affects neutrophil functions, leading to reduced pathogen killing and increased morbidity. This impairment has been directly linked to increased glycemia, however, how this specifically affects neutrophils metabolism and their differentiation in the bone marrow is unclear and difficult to study., Research Design and Methods: We used high-resolution respirometry to investigate the metabolism of resting and activated donor neutrophils, and flow cytometry to measure surface CD15 and CD11b expression. We then used HL-60 cells differentiated towards neutrophil-like cells in standard media and investigated the effect of doubling glucose concentration on differentiation metabolism. We measured the oxygen consumption rate (OCR), and the enzymatic activity of carnitine palmitoyl transferase 1 (CPT1) and citrate synthase during neutrophil-like differentiation. We compared the surface phenotype, functions, and OCR of neutrophil-like cells differentiated under both glucose concentrations., Results: Donor neutrophils showed significant instability of CD11b and OCR after phorbol 12-myristate 13-acetate stimulation at 3 hours post-enrichment. During HL-60 neutrophil-like cell differentiation, there was a significant increase in surface CD15 and CD11b expression together with the loss of mitochondrial mass. Differentiated neutrophil-like cells also exhibited higher CD11b expression and were significantly more phagocytic. In higher glucose media, we measured a decrease in citrate synthase and CPT1 activities during neutrophil-like differentiation., Conclusions: HL-60 neutrophil-like differentiation recapitulated known molecular and metabolic features of human neutrophil differentiation. Increased glucose concentrations correlated with features described in hyperglycemic donor neutrophils including increased CD11b and phagocytosis. We used this model to describe metabolic features of neutrophil-like cell differentiation in hyperglycemia and show for the first time the downregulation of CPT1 and citrate synthase activity, independently of mitochondrial mass., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)