{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T13:31:53Z","timestamp":1768483913251,"version":"3.49.0"},"posted":{"date-parts":[[2019,1,9]]},"group-title":"Microbiology","reference-count":58,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2019,1,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  <jats:italic>Paracoccidioides brasiliensis<\/jats:italic>\n                  and\n                  <jats:italic>P. lutzii<\/jats:italic>\n                  , etiological agents of paracoccidioidomycosis (PCM), develop as mycelia at 25-30 \u00b0C and as yeast at 35-37 \u00b0C. Only a few\n                  <jats:italic>Paracoccidioides<\/jats:italic>\n                  spp. proteins are well characterized. Thus, we studied paracoccin (PCN) from\n                  <jats:italic>P. brasiliensis<\/jats:italic>\n                  , its role in the fungus biology, and its relationship with the host innate immune cells. Cloning and heterologous expression analysis revealed its lectin, enzymatic, and immunomodulatory properties. Recently, we employed a system based on\n                  <jats:italic>Agrobacterium tumefaciens<\/jats:italic>\n                  -mediated transformation to manipulate\n                  <jats:italic>P. brasiliensis<\/jats:italic>\n                  yeast genes to obtain clones knocked-down for PCN, which after all, are unable to transit from yeast to mycelium forms, causing a mild pulmonary disease. Herein, we generate\n                  <jats:italic>P. brasiliensis<\/jats:italic>\n                  overexpressing PCN (ov-PCN). To date, it was not explored the overexpressing of endogenous components in\n                  <jats:italic>Paracoccidioides<\/jats:italic>\n                  spp. Therefore, we investigate the role of PCN in fungal biology and pathogenesis. Augmented levels of PCN mRNA and protein, and N-acetylglucosaminidase activity confirmed PCN overexpression in ov-PCN of\n                  <jats:italic>P. brasiliensis<\/jats:italic>\n                  yeasts. Interestingly, PCN overexpression did not affect the yeasts\u2019 growth or viability and favored cell separation. The ov-PCN clones transitioned faster to the mycelium form than the wt-PCN yeasts. Concerning infection, while most of mice infected with the wt-yeasts (90%) survive at least until the 70\n                  <jats:sup>th<\/jats:sup>\n                  day, all mice infected with ov-PCN yeasts were already died at the 35\n                  <jats:sup>th<\/jats:sup>\n                  day post-infection. In vitro assays showed that ov-PCN were more susceptible to phagocytosis by macrophages. Finally, it was verified that the chitin particles isolated from the ov-PCN cells were smaller than those obtained from the wt-PCN yeasts. Macrophages stimulated with the chitin isolated from ov-PCN produce IL-10, whereas the particles with a wider size range harvested from wt-PCN yeasts induced TNF-\u03b1 and IL-1\u03b2 secretion. The anti-inflammatory microenvironment from macrophage stimulation with small chitin particles hampers the development of a protective immune response against the fungus. We postulated that the high grade of chitin cleavage, as the results of augmented PCN expression, favors pathogenesis following\n                  <jats:italic>P. brasiliensis<\/jats:italic>\n                  infection. Thus, PCN is a relevant virulence fungal factor.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>AUTHOR SUMMARY<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Paracoccidioides<\/jats:italic>\n                    spp. are pathogenic fungi that cause paracoccidioidomycosis (PCM) in humans, the main deep mycosis of Latin America. Recently, by knocking down the paracoccin gene, our group showed that this lectin is necessary for the morphological transition from yeast to hyphae, and that this decrease results in low\n                    <jats:italic>P. brasiliensis<\/jats:italic>\n                    virulence. Here, after overexpress PCN, we revealed the importance of the yeast chitin hydrolysis to the host response. Infection of mice with ov-PCN yeasts causes severe lung disease compared to moderate disease caused by wt-PCN yeasts. The release of smaller chitin particles was as a result of an accelerated chitin hydrolysis provided by ov-PCN yeasts. Interestingly, these smallest chitin particles are able to modulate host response by increasing IL-10 in the meantime that decrease TNF-\u03b1 secretion, thus hampering Th1 immune response that is crucial in the fight against this fungi. These findings represent a significant advance in the knowledge about the role of PCN chitinase in\n                    <jats:italic>P. brasiliensis<\/jats:italic>\n                    .\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/515056","type":"posted-content","created":{"date-parts":[[2019,1,9]],"date-time":"2019-01-09T16:45:12Z","timestamp":1547052312000},"source":"Crossref","is-referenced-by-count":1,"title":["Paracoccin overexpression in\n                  <i>Paracoccidioides brasiliensis<\/i>\n                  reveals the influence of chitin hydrolysis on fungal virulence and host immune response"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3037-3663","authenticated-orcid":false,"given":"Relber Aguiar","family":"Gon\u00e7ales","sequence":"first","affiliation":[]},{"given":"Vanessa Cristina","family":"Silva Vieira","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0020-6530","authenticated-orcid":false,"given":"Rafael","family":"Ricci-Azevedo","sequence":"additional","affiliation":[]},{"given":"Fabr\u00edcio Freitas","family":"Fernandes","sequence":"additional","affiliation":[]},{"given":"Sandra Maria de","family":"Oliveira Thomaz","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8935-8030","authenticated-orcid":false,"given":"Agostinho","family":"Carvalho","sequence":"additional","affiliation":[]},{"given":"Patr\u00edcia Ediv\u00e2nia","family":"Vendruscolo","sequence":"additional","affiliation":[]},{"given":"Cristina","family":"Cunha","sequence":"additional","affiliation":[]},{"given":"Maria Cristina","family":"Roque-Barreira","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8436-9398","authenticated-orcid":false,"given":"Fernando","family":"Rodrigues","sequence":"additional","affiliation":[]}],"member":"54368","reference":[{"key":"2021010418152399000_515056v1.1","doi-asserted-by":"publisher","DOI":"10.1080\/mmy.40.3.225.242"},{"key":"2021010418152399000_515056v1.2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.59.030804.121055"},{"issue":"6","key":"2021010418152399000_515056v1.3","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1111\/myc.12608","article-title":"Paracoccidioidomycosis infection in domestic and wild mammals by Paracoccidioides lutzii","volume":"60","year":"2017","journal-title":"Mycoses"},{"key":"2021010418152399000_515056v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.ympev.2009.04.005"},{"key":"2021010418152399000_515056v1.5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0041480"},{"issue":"1","key":"2021010418152399000_515056v1.6","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1128\/IAI.53.1.199-206.1986","article-title":"Exocellular components of Paracoccidioides brasiliensis: identification of a specific antigen","volume":"53","year":"1986","journal-title":"Infect Immun"},{"issue":"10","key":"2021010418152399000_515056v1.7","doi-asserted-by":"crossref","first-page":"2147","DOI":"10.1128\/JCM.26.10.2147-2151.1988","article-title":"Production of Paracoccidioides brasiliensis exoantigens for immunodiffusion tests","volume":"26","year":"1988","journal-title":"J Clin Microbiol"},{"issue":"2","key":"2021010418152399000_515056v1.8","doi-asserted-by":"crossref","first-page":"200","DOI":"10.4269\/ajtmh.1990.43.200","article-title":"Antibody response to the 43 kDa glycoprotein of Paracoccidioides brasiliensis as a marker for the evaluation of patients under treatment","volume":"43","year":"1990","journal-title":"Am J Trop Med Hyg"},{"issue":"3","key":"2021010418152399000_515056v1.9","first-page":"297","article-title":"Biochemistry and molecular biology of the main diagnostic antigen of Paracoccidioides brasiliensis","volume":"26","year":"1995","journal-title":"Arch Med Res"},{"key":"2021010418152399000_515056v1.10","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.8.4553"},{"key":"2021010418152399000_515056v1.11","doi-asserted-by":"publisher","DOI":"10.1080\/714031053"},{"key":"2021010418152399000_515056v1.12","doi-asserted-by":"publisher","DOI":"10.1128\/CDLI.9.2.374-377.2002"},{"key":"2021010418152399000_515056v1.13","doi-asserted-by":"crossref","unstructured":"Fernandes, F.F. , et al., Detrimental Effect of Fungal 60-kDa Heat Shock Protein on Experimental Paracoccidioides brasiliensis Infection. 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