{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:58:48Z","timestamp":1790611128055,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,24]]},"group-title":"Microbiology","reference-count":71,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Coxiella burnetii<\/jats:italic>\n                  (Cb), the causative agent of Q fever, replicates within host macrophages by modulating innate immune responses through its type IVB secretion system (T4SS). Host cells sense pathogens via Pattern recognition receptors (PRRs), including Toll-like receptors (TLRs), to recognize pathogen-associated molecular patterns (PAMPs) and initiate signaling pathways that drive pro-inflammatory cytokine and interferon responses. Toll-like receptor 3 (TLR3) triggers the nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-\u03baB) and Interferon regulatory factors (IRFs) resulting in pro-inflammatory cytokine production and type I interferon (IFN-I) induction. Here we demonstrate that Cb requires T4SS to suppress TLR3-induced NF-\u03baB and IRF transcriptional responses. Furthermore, RNA purified from both virulent and avirulent Cb is sufficient to activate TLR3, identifying pathogen RNA as a relevant PAMP in this context. Using a pulmonary infection model with virulent Cb, we found that TLR3\/ Toll-Interleukin-1 Receptor Domain-Containing Adapter Protein Inducing Interferon Beta (TRIF)-dependent innate immune pathway inhibits cachexia. In contrast, signaling through the interferon-\u03b1\/\u03b2 receptor (IFNAR) restricts bacterial dissemination, indicating that these pathways play distinct but complementary roles in host defense. Mechanistically, Cb suppresses TLR3\/TRIF signaling in a T4SS-dependent manner by preventing host TLR3 recruitment to the Coxiella-containing vacuole (CCV) and disrupting TRIF\u2013TNF Receptor-Associated Factor 6 (TRAF6) interactions, thereby selectively inhibiting impairing NF-\u03baB activation. Consistent with this, we identify five T4SS effector proteins that attenuate TLR3-induced NF-\u03baB signaling, including CBU1292, which suppresses both NF-\u03baB and IRF-dependent responses downstream of TLR3.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Author summary<\/jats:title>\n                  <jats:p>\n                    Pathogenic bacteria have evolved mechanisms to evade or manipulate host immune defenses.\n                    <jats:italic>Coxiella burnetii<\/jats:italic>\n                    (Cb), the causative agent of Q fever, replicates within immune cells and interferes with innate immune signaling pathways. We found that its type IVB secretion system (T4SS) express effectors that inhibit immune responses triggered by Toll-like receptor 3 (TLR3), a sensor that detects microbial RNA. Specifically, Cb suppresses signaling through the adaptor protein TRIF, thereby reducing activation of key immune regulators, NF-\u03baB and interferon regulatory factors (IRFs), which are required for inflammatory and antiviral responses. Using a mouse model of lung infection, we found that disruption of TLR3\/TRIF signaling and type I interferon receptor (IFNAR) signaling leads to different disease outcomes, indicating that these pathways play distinct roles in controlling infection and disease severity. We also identified several Cb-secreted effectors that interfere with TLR3-mediated immune activation, including one that inhibits both NF-\u03baB and interferon responses. Together, our findings reveal how Cb evades immune detection by targeting a critical RNA-sensing pathway and highlight distinct host defense mechanisms that limit bacterial infection and disease.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.23.753975","type":"posted-content","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:05:36Z","timestamp":1790294736000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Coxiella burnetii<\/i>\n                  type IVB secretion system modulates TLR3\/TRIF-dependent NF-\u03baB and IRF responses during infection"],"prefix":"10.64898","author":[{"given":"Saugata","family":"Mahapatra","sequence":"first","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"},{"name":"Image Analysis Laboratory, Texas A&M School of Veterinary Medicine & Biomedical Sciences, College Station, Texas 77843 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aryashree","family":"Arunima","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Busbee","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer E.","family":"Dumaine","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"},{"name":"Department of Chemistry and Life Science, United States Military Academy, West Point, New York 10996 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren","family":"Stranahan","sequence":"additional","affiliation":[{"name":"Department of Veterinary Pathobiology, Texas A&M School of Veterinary Medicine & Biomedical Sciences, College Station, Texas 77843 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ricardo A.","family":"Zeledon","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bobby","family":"Dow","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha","family":"Butler","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Clark","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2884-6895","authenticated-orcid":false,"given":"A. Phillip","family":"West","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"},{"name":"The Jackson Laboratory, Bar Harbor, Maine 04609 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2536-0857","authenticated-orcid":false,"given":"Erin J.","family":"van Schaik","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3896-8581","authenticated-orcid":false,"given":"James E.","family":"Samuel","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, Bryan, Texas 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"8","key":"2026092808351056000_2026.09.23.753975v1.1","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1038\/nrmicro3049","article-title":"Molecular pathogenesis of the obligate intracellular bacterium Coxiella burnetii","volume":"11","year":"2013","journal-title":"Nat Rev Micro"},{"key":"2026092808351056000_2026.09.23.753975v1.2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.024778-0"},{"key":"2026092808351056000_2026.09.23.753975v1.3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.5.3240"},{"key":"2026092808351056000_2026.09.23.753975v1.4","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2007.00901.x"},{"key":"2026092808351056000_2026.09.23.753975v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.11.005"},{"issue":"7","key":"2026092808351056000_2026.09.23.753975v1.6","doi-asserted-by":"crossref","first-page":"919","DOI":"10.2217\/fmb-2016-0044","article-title":"Right on Q: Genetics begin to Unravel Coxiella burnetii host cell Interactions","volume":"11","year":"2016","journal-title":"Fut Microbiol"},{"key":"2026092808351056000_2026.09.23.753975v1.7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007855"},{"key":"2026092808351056000_2026.09.23.753975v1.8","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-007-4315-1_4"},{"key":"2026092808351056000_2026.09.23.753975v1.9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006922"},{"key":"2026092808351056000_2026.09.23.753975v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.55.5.1144-1150.1987"},{"key":"2026092808351056000_2026.09.23.753975v1.11","first-page":"25","article-title":"The SCID Mouse Model for Identifying Virulence Determinants in Coxiella burnetii","volume":"7","year":"2017","journal-title":"Front Cell Infect Microbiol"},{"key":"2026092808351056000_2026.09.23.753975v1.12","doi-asserted-by":"crossref","unstructured":"Mahapatra S , Gallaher B , Smith SC , Graham JG , Voth DE , Shaw EI . 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