Samuel C. Williams, Sandra Garcet, Hong Hur, Shunsuke Miura, Juana Gonzalez, Kristina Navrazhina, Mika Yamamura-Murai, Kazuhiko Yamamura, Xuan Li, John Frew, Vincent A. Fischetti, Uri Sela, James G. Krueger
{"title":"革兰氏阴性厌氧菌能引起强大的角质细胞免疫反应,有望揭示 HS 的发病机制","authors":"Samuel C. Williams, Sandra Garcet, Hong Hur, Shunsuke Miura, Juana Gonzalez, Kristina Navrazhina, Mika Yamamura-Murai, Kazuhiko Yamamura, Xuan Li, John Frew, Vincent A. Fischetti, Uri Sela, James G. Krueger","doi":"10.1111/exd.15087","DOIUrl":null,"url":null,"abstract":"<p>Hidradenitis Suppurativa (HS) is a chronic autoinflammatory skin disease with activated keratinocytes, tunnel formation and a complex immune infiltrate in tissue. The HS microbiome is polymicrobial with an abundance of commensal gram-positive facultative (GPs) <i>Staphylococcus</i> species and gram-negative anaerobic (GNA) bacteria like <i>Prevotella</i>, <i>Fusobacterium</i> and <i>Porphyromonas</i> with increasing predominance of GNAs with disease severity. We sought to define the keratinocyte response to bacteria commonly isolated from HS lesions to probe pathogenic relationships between HS and the microbiome. Type strains of <i>Prevotella nigrescens</i>, <i>Prevotella melaninogenica</i>, <i>Prevotella intermedia</i>, <i>Prevotella asaccharolytica</i>, <i>Fusobacterium nucleatum</i>, as well as <i>Staphylococcus aureus</i> and the normal skin commensal <i>Staphylococcus epidermidis</i> were heat-killed and co-incubated with normal human keratinocytes. RNA was collected and analysed using RNAseq and RT-qPCR. The supernatant was collected from cell culture for protein quantification. Transcriptomic profiles between HS clinical samples and stimulated keratinocytes were compared. Co-staining of patient HS frozen sections was used to localize bacteria in lesions. A mouse intradermal injection model was used to investigate early immune recruitment. TLR4 and JAK inhibitors were used to investigate mechanistic avenues of bacterial response inhibition. GNAs, especially <i>F. nucleatum</i>, stimulated vastly higher <i>CXCL8</i>, <i>IL17C</i>, <i>CCL20</i>, <i>IL6</i>, <i>TNF</i> and <i>IL36γ</i> transcription in normal skin keratinocytes than the GPs <i>S. epidermidis</i> and <i>S. aureus</i>. Using RNAseq, we found that <i>F. nucleatum</i> (and <i>Prevotella</i>) strongly induced the IL-17 pathway in keratinocytes and overlapped with transcriptome profiles of HS patient clinical samples. Bacteria were juxtaposed to activated keratinocytes in vivo, and <i>F. nucleatum</i> strongly recruited murine neutrophil and macrophage migration. Both the TLR4 and pan-JAK inhibitors reduced cytokine production. Detailed transcriptomic profiling of healthy skin keratinocytes exposed to GNAs prevalent in HS revealed a potent, extensive inflammatory response vastly stronger than GPs. GNAs stimulated HS-relevant genes, including many genes in the IL-17 response pathway, and were significantly associated with HS tissue transcriptomes. The close association of activated keratinocytes with bacteria in HS lesions and innate infiltration in murine skin cemented GNA pathogenic potential. These novel mechanistic insights could drive future targeted therapies.</p>","PeriodicalId":12243,"journal":{"name":"Experimental Dermatology","volume":null,"pages":null},"PeriodicalIF":3.5000,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/exd.15087","citationCount":"0","resultStr":"{\"title\":\"Gram-negative anaerobes elicit a robust keratinocytes immune response with potential insights into HS pathogenesis\",\"authors\":\"Samuel C. Williams, Sandra Garcet, Hong Hur, Shunsuke Miura, Juana Gonzalez, Kristina Navrazhina, Mika Yamamura-Murai, Kazuhiko Yamamura, Xuan Li, John Frew, Vincent A. Fischetti, Uri Sela, James G. Krueger\",\"doi\":\"10.1111/exd.15087\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Hidradenitis Suppurativa (HS) is a chronic autoinflammatory skin disease with activated keratinocytes, tunnel formation and a complex immune infiltrate in tissue. The HS microbiome is polymicrobial with an abundance of commensal gram-positive facultative (GPs) <i>Staphylococcus</i> species and gram-negative anaerobic (GNA) bacteria like <i>Prevotella</i>, <i>Fusobacterium</i> and <i>Porphyromonas</i> with increasing predominance of GNAs with disease severity. We sought to define the keratinocyte response to bacteria commonly isolated from HS lesions to probe pathogenic relationships between HS and the microbiome. Type strains of <i>Prevotella nigrescens</i>, <i>Prevotella melaninogenica</i>, <i>Prevotella intermedia</i>, <i>Prevotella asaccharolytica</i>, <i>Fusobacterium nucleatum</i>, as well as <i>Staphylococcus aureus</i> and the normal skin commensal <i>Staphylococcus epidermidis</i> were heat-killed and co-incubated with normal human keratinocytes. RNA was collected and analysed using RNAseq and RT-qPCR. The supernatant was collected from cell culture for protein quantification. Transcriptomic profiles between HS clinical samples and stimulated keratinocytes were compared. Co-staining of patient HS frozen sections was used to localize bacteria in lesions. A mouse intradermal injection model was used to investigate early immune recruitment. TLR4 and JAK inhibitors were used to investigate mechanistic avenues of bacterial response inhibition. GNAs, especially <i>F. nucleatum</i>, stimulated vastly higher <i>CXCL8</i>, <i>IL17C</i>, <i>CCL20</i>, <i>IL6</i>, <i>TNF</i> and <i>IL36γ</i> transcription in normal skin keratinocytes than the GPs <i>S. epidermidis</i> and <i>S. aureus</i>. Using RNAseq, we found that <i>F. nucleatum</i> (and <i>Prevotella</i>) strongly induced the IL-17 pathway in keratinocytes and overlapped with transcriptome profiles of HS patient clinical samples. Bacteria were juxtaposed to activated keratinocytes in vivo, and <i>F. nucleatum</i> strongly recruited murine neutrophil and macrophage migration. Both the TLR4 and pan-JAK inhibitors reduced cytokine production. Detailed transcriptomic profiling of healthy skin keratinocytes exposed to GNAs prevalent in HS revealed a potent, extensive inflammatory response vastly stronger than GPs. GNAs stimulated HS-relevant genes, including many genes in the IL-17 response pathway, and were significantly associated with HS tissue transcriptomes. The close association of activated keratinocytes with bacteria in HS lesions and innate infiltration in murine skin cemented GNA pathogenic potential. These novel mechanistic insights could drive future targeted therapies.</p>\",\"PeriodicalId\":12243,\"journal\":{\"name\":\"Experimental Dermatology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2024-04-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/exd.15087\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Experimental Dermatology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/exd.15087\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"DERMATOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental Dermatology","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/exd.15087","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DERMATOLOGY","Score":null,"Total":0}
Gram-negative anaerobes elicit a robust keratinocytes immune response with potential insights into HS pathogenesis
Hidradenitis Suppurativa (HS) is a chronic autoinflammatory skin disease with activated keratinocytes, tunnel formation and a complex immune infiltrate in tissue. The HS microbiome is polymicrobial with an abundance of commensal gram-positive facultative (GPs) Staphylococcus species and gram-negative anaerobic (GNA) bacteria like Prevotella, Fusobacterium and Porphyromonas with increasing predominance of GNAs with disease severity. We sought to define the keratinocyte response to bacteria commonly isolated from HS lesions to probe pathogenic relationships between HS and the microbiome. Type strains of Prevotella nigrescens, Prevotella melaninogenica, Prevotella intermedia, Prevotella asaccharolytica, Fusobacterium nucleatum, as well as Staphylococcus aureus and the normal skin commensal Staphylococcus epidermidis were heat-killed and co-incubated with normal human keratinocytes. RNA was collected and analysed using RNAseq and RT-qPCR. The supernatant was collected from cell culture for protein quantification. Transcriptomic profiles between HS clinical samples and stimulated keratinocytes were compared. Co-staining of patient HS frozen sections was used to localize bacteria in lesions. A mouse intradermal injection model was used to investigate early immune recruitment. TLR4 and JAK inhibitors were used to investigate mechanistic avenues of bacterial response inhibition. GNAs, especially F. nucleatum, stimulated vastly higher CXCL8, IL17C, CCL20, IL6, TNF and IL36γ transcription in normal skin keratinocytes than the GPs S. epidermidis and S. aureus. Using RNAseq, we found that F. nucleatum (and Prevotella) strongly induced the IL-17 pathway in keratinocytes and overlapped with transcriptome profiles of HS patient clinical samples. Bacteria were juxtaposed to activated keratinocytes in vivo, and F. nucleatum strongly recruited murine neutrophil and macrophage migration. Both the TLR4 and pan-JAK inhibitors reduced cytokine production. Detailed transcriptomic profiling of healthy skin keratinocytes exposed to GNAs prevalent in HS revealed a potent, extensive inflammatory response vastly stronger than GPs. GNAs stimulated HS-relevant genes, including many genes in the IL-17 response pathway, and were significantly associated with HS tissue transcriptomes. The close association of activated keratinocytes with bacteria in HS lesions and innate infiltration in murine skin cemented GNA pathogenic potential. These novel mechanistic insights could drive future targeted therapies.
期刊介绍:
Experimental Dermatology provides a vehicle for the rapid publication of innovative and definitive reports, letters to the editor and review articles covering all aspects of experimental dermatology. Preference is given to papers of immediate importance to other investigators, either by virtue of their new methodology, experimental data or new ideas. The essential criteria for publication are clarity, experimental soundness and novelty. Letters to the editor related to published reports may also be accepted, provided that they are short and scientifically relevant to the reports mentioned, in order to provide a continuing forum for discussion. Review articles represent a state-of-the-art overview and are invited by the editors.