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Heterologous Expression, Purification, and Immunomodulatory Effects of Recombinant Lipoprotein GUDIV-103 Isolated from Ureaplasma diversum.
Santos-Junior, Manoel Neres; Neves, Wanderson Souza; Santos, Ronaldo Silva; Almeida, Palloma Porto; Fernandes, Janaina Marinho; Guimarães, Bruna Carolina de Brito; Barbosa, Maysa Santos; da Silva, Lucas Santana Coelho; Gomes, Camila Pacheco; Sampaio, Beatriz Almeida; Rezende, Izadora de Souza; Correia, Thiago Macedo Lopes; Neres, Nayara Silva de Macedo; Campos, Guilherme Barreto; Bastos, Bruno Lopes; Timenetsky, Jorge; Marques, Lucas Miranda.
Afiliación
  • Santos-Junior MN; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Neves WS; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Santos RS; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Almeida PP; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Fernandes JM; Bioinformatics and Computational Biology Lab, Division of Experimental and Translational Research, Brazilian National Cancer Institute (INCA), Rio de Janeiro 20231-050, Brazil.
  • Guimarães BCB; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Barbosa MS; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • da Silva LSC; Department of Microbiology, Institute of Biomedical Science, University of São Paulo, São Paulo 05508-000, Brazil.
  • Gomes CP; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Sampaio BA; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Rezende IS; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Correia TML; Department of Microbiology, Institute of Biomedical Science, University of São Paulo, São Paulo 05508-000, Brazil.
  • Neres NSM; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Campos GB; Department of Biointeraction, Multidisciplinary Institute of Health, Federal University of Bahia, Vitória da Conquista 40170-110, Brazil.
  • Bastos BL; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Timenetsky J; Department of Biology, and Biotechnology of Microorganisms, State University of Santa Cruz (UESC), Ilhéus 45662-900, Brazil.
  • Marques LM; Department of Microbiology, Institute of Biomedical Science, University of São Paulo, São Paulo 05508-000, Brazil.
Microorganisms ; 10(5)2022 May 16.
Article en En | MEDLINE | ID: mdl-35630474
Ureaplasma diversum is a bacterial pathogen that infects cattle and can cause severe inflammation of the genital and reproductive systems. Lipid-associated membrane proteins (LAMPs), including GUDIV-103, are the main virulence factors in this bacterium. In this study, we heterologously expressed recombinant GUDIV-103 (rGUDIV-103) in Escherichia coli, purified it, and evaluated its immunological reactivity and immunomodulatory effects in bovine peripheral blood mononuclear cells (PBMCs). Samples from rabbits inoculated with purified rGUDIV-103 were analysed using indirect enzyme-linked immunosorbent assay and dot blotting to confirm polyclonal antibody production and assess kinetics, respectively. The expression of this lipoprotein in field isolates was confirmed via Western blotting with anti-rGUDIV-103 serum and hydrophobic or hydrophilic proteins from 42 U. diversum strains. Moreover, the antibodies produced against the U. diversum ATCC 49783 strain recognised rGUDIV-103. The mitogenic potential of rGUDIV-103 was evaluated using a lymphoproliferation assay in 5(6)-carboxyfluorescein diacetate succinimidyl ester−labelled bovine PBMCs, where it induced lymphocyte proliferation. Quantitative polymerase chain reaction analysis revealed that the expression of interleukin-1ß, toll-like receptor (TLR)-α, TLR2, TLR4, inducible nitric oxide synthase, and caspase-3−encoding genes increased more in rGUDIV-103−treated PBMCs than in untreated cells (p < 0.05). Treating PBMCs with rGUDIV-103 increased nitric oxide and hydrogen peroxide levels. The antigenic and immunogenic properties of rGUDIV-103 suggested its suitability for immunobiological application.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Suiza
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Microorganisms Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Suiza