RESUMEN
Ebola hemorrhagic fever is a fatal disease caused by the negative-strand RNA of the Ebola virus. A high-intensity outbreak of this fever was reported in West Africa last year; however, there is currently no definitive treatment strategy available for this disease. In this study, we analyzed the molecular evolutionary history and attempted to determine the positive selection sites in the Ebola genes using multiple-genomic sequences of the various Ebola virus subtypes, in order to gain greater clarity into the evolution of the virus and its various subtypes. Only the glycoprotein (GP) gene was positively selected among the 8 Ebola genes, with the other genes remaining in the purification stage. The positive selection sites in the GP gene were identified by a random-site model; these sites were found to be located in the mucin-like region, which is associated with transmembrane protein binding. Additionally, different branches of the phylogenetic tree displayed different positive sites, which in turn was responsible for differences in the cell adhesion ability of the virus. In conclusion, the pattern of positive sites in the GP gene is associated with the epidemiology and prevalence of Ebola in different areas.
Asunto(s)
Ebolavirus/genética , Ebolavirus/patogenicidad , Glicoproteínas/metabolismo , Fiebre Hemorrágica Ebola/virología , Brotes de Enfermedades , Ebolavirus/clasificación , Evolución Molecular , Glicoproteínas/genética , Fiebre Hemorrágica Ebola/epidemiología , Fiebre Hemorrágica Ebola/metabolismo , Humanos , Filogenia , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/metabolismoRESUMEN
CD4(+) regulatory T lymphocytes (Treg cells) play a crucial role in maintaining the normal immune homeostasis. Foxp3, as a key marker for Treg cells, is widely used to identify Treg cells, not only in humans but also in other species, like mouse, porcine, ovine, and bovine. To detect reproducible Treg cells is important for evaluating the state of the immune system, and thus, it is necessary to optimize Foxp3 staining. Here, we present a comparative study of MF23 and FJK-16s clones of anti-mouse Foxp3 antibodies, used in combination with two different fixation/permeabilization buffers. For Foxp3 staining, the fixation/permeabilization buffer and Foxp3 antibody FJK-16s clone from eBioscience were better than those from BD Pharmingen, with the best fluorochrome PE. Moreover, when using the best combination, there was a highly significant positive correlation between CD25(+) T cells and CD25(+)Foxp3(+) T cells. Therefore, the CD25 marker can be used as an alternative to the Foxp3 antibody. As FJK-16s is also applicable for detecting bovine, porcine, canine, ovine, and equine Foxp3 antibodies, these results will be helpful not only in quantifying the frequencies of mouse Treg cells, but also in accurately detecting Treg cells of the other species mentioned above by multicolor flow cytometry.