AUTHOR=Santos-Junior Manoel N. , Rezende Izadora S. , Souza Clarissa L. S. , Barbosa Maysa S. , Campos Guilherme B. , Brito Laís F. , Queiroz Éllunny C. , Barbosa Elaine N. , Teixeira Mariana M. , Da Silva Letícia O. , Silva Lucas S. C. , Nascimento Flávia S. , Da Silva Tassyo L. , Martens Adam A. , Siqueira Adriano F. P. , Assumpção Mayra E. O. D’Avila , Machado-Santelli Glaucia M. , Bastos Bruno L. , Guimarães Ana M. S. , Timenetsky Jorge , Marques Lucas M. 

TITLE=Ureaplasma diversum and Its Membrane-Associated Lipoproteins Activate Inflammatory Genes Through the NF-κB Pathway via Toll-Like Receptor 4

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.01538

DOI=10.3389/fmicb.2018.01538

ISSN=1664-302X

ABSTRACT=<p><bold>Objectives:</bold><italic>Ureaplasma diversum</italic> is a pathogen of cows that may cause intense inflammatory responses in the reproductive tract and interfere with bovine reproduction. The aims of this study were to evaluate the immune response of bovine blastocysts and macrophages to <italic>U. diversum</italic> infection and to evaluate the invasion capacity of this microorganism in bovine blastocysts.</p><p><bold>Methods:</bold> Viable and heat-inactivated <italic>U. diversum</italic> strains ATCC 49782 and CI-GOTA and their extracted membrane lipoproteins were inoculated in macrophages in the presence or absence of signaling blockers of Toll-Like Receptor (TLR) 4, TLR2/4, and Nuclear Factor KB (NF-κB). In addition, the same viable <italic>U. diversum</italic> strains were used to infect bovine blastocysts. RNA was extracted from infected and lipoprotein-exposed macrophages and infected blastocysts and assayed by qPCR to evaluate the expression of Interleukin 1 beta (IL-1β), Tumor Necrosis Factor Alpha (TNF-α), TLR2 and TLR4 genes. <italic>U. diversum</italic> internalization in blastocysts was followed by confocal microscopy.</p><p><bold>Results:</bold> Both <italic>Ureaplasma</italic> strains and different concentrations of extracted lipoproteins induced a higher gene expression of IL-1β, TNF-α, TLR2, and TLR4 in macrophages (<italic>p</italic> &lt; 0.05) when compared to non-infected cells. The used blockers inhibited the expression of IL-1β and TNF-α in all treatments. Moreover, <italic>U. diversum</italic> was able to internalize within blastocysts and induce a higher gene expression of IL-1b and TNF- α when compared to non-infected blastocysts (<italic>p</italic> &lt; 0.05).</p><p><bold>Conclusion:</bold> The obtained results strongly suggest that <italic>U. diversum</italic> and its lipoproteins interact with TLR4 in a signaling pathway acting via NF-kB signaling to stimulate the inflammatory response. This is the first study to evaluate the <italic>in vitro</italic> immunological response of macrophages and bovine blastocysts against <italic>U. diversum</italic>. These results may contribute to a better understanding of the immunomodulatory activity and pathogenicity of this infectious agent.</p>