A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers
ABSTRACT Haemophilus ducreyi causes the genital ulcer disease chancroid and cutaneous ulcers in children. To study its pathogenesis, we developed a human challenge model in which we infect the skin on the upper arm of human volunteers with H. ducreyi to the pustular stage of disease. The model has b...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2025-03-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.03885-24 |
| _version_ | 1826532115500498944 |
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| author | Julie A. Brothwell Yuhui Wei Jia Wang Tingbo Guo Chi Zhang Kate R. Fortney Rory Duplantier Li Chen Teresa A. Batteiger Mark H. Kaplan Stanley M. Spinola Sha Cao |
| author_facet | Julie A. Brothwell Yuhui Wei Jia Wang Tingbo Guo Chi Zhang Kate R. Fortney Rory Duplantier Li Chen Teresa A. Batteiger Mark H. Kaplan Stanley M. Spinola Sha Cao |
| author_sort | Julie A. Brothwell |
| collection | DOAJ |
| description | ABSTRACT Haemophilus ducreyi causes the genital ulcer disease chancroid and cutaneous ulcers in children. To study its pathogenesis, we developed a human challenge model in which we infect the skin on the upper arm of human volunteers with H. ducreyi to the pustular stage of disease. The model has been used to define lesional architecture, describe the immune infiltrate into the infected sites using flow cytometry, and explore the molecular basis of the immune response using bulk RNA-seq. Here, we used single cell RNA-seq (scRNA-seq) and spatial transcriptomics to simultaneously characterize multiple cell types within infected human skin and determine the cellular origin of differentially expressed transcripts that we had previously identified by bulk RNA-seq. We obtained paired biopsies of pustules and wounded (mock infected) sites from five volunteers for scRNA-seq. We identified 13 major cell types, including T- and NK-like cells, macrophages, dendritic cells, as well as other cell types typically found in the skin. Immune cell types were enriched in pustules, and some subtypes within the major cell types were exclusive to pustules. Sufficient tissue specimens for spatial transcriptomics were available from four of the volunteers. T- and NK-like cells were highly associated with multiple antigen presentation cell types. In pustules, type I interferon stimulation was high in areas that were high in antigen presentation—especially in macrophages near the abscess—compared to wounds. Together, our data provide a high-resolution view of the cellular immune response to the infection of the skin with a human pathogen.IMPORTANCEA high-resolution view of the immune infiltrate due to infection with an extracellular bacterial pathogen in human skin has not yet been defined. Here, we used the human skin pathogen Haemophilus ducreyi in a human challenge model to identify on a single cell level the types of cells that are present in volunteers who fail to spontaneously clear infection and form pustules. We identified 13 major cell types. Immune cells and immune-activated stromal cells were enriched in pustules compared to wounded (mock infected) sites. Pustules formed despite the expression of multiple pro-inflammatory cytokines, such as IL-1β and type I interferon. Interferon stimulation was most evident in macrophages, which were proximal to the abscess. The pro-inflammatory response within the pustule may be tempered by regulatory T cells and cells that express indoleamine 2,3-dioxygenase, leading to failure of the immune system to clear H. ducreyi. |
| first_indexed | 2025-03-14T01:46:06Z |
| format | Article |
| id | doaj.art-9a1f95bb88f545129fc2ad7ef914ff90 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2025-03-14T01:46:06Z |
| publishDate | 2025-03-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-9a1f95bb88f545129fc2ad7ef914ff902025-03-12T13:00:57ZengAmerican Society for MicrobiologymBio2150-75112025-03-0116310.1128/mbio.03885-24A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteersJulie A. Brothwell0Yuhui Wei1Jia Wang2Tingbo Guo3Chi Zhang4Kate R. Fortney5Rory Duplantier6Li Chen7Teresa A. Batteiger8Mark H. Kaplan9Stanley M. Spinola10Sha Cao11Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Medical and Molecular Genetics and Center for Computational Biology and Bioinformatics, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Computer Science, Indiana University, Bloomington, Indiana, USADepartment of Biomedical Engineering, Oregon Health and Science University, Portland, Oregon, USADepartment of Medical and Molecular Genetics and Center for Computational Biology and Bioinformatics, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Biostatistics, University of Florida, Gainesville, Florida, USADepartment of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USADepartment of Medical and Molecular Genetics and Center for Computational Biology and Bioinformatics, Indiana University School of Medicine, Indianapolis, Indiana, USAABSTRACT Haemophilus ducreyi causes the genital ulcer disease chancroid and cutaneous ulcers in children. To study its pathogenesis, we developed a human challenge model in which we infect the skin on the upper arm of human volunteers with H. ducreyi to the pustular stage of disease. The model has been used to define lesional architecture, describe the immune infiltrate into the infected sites using flow cytometry, and explore the molecular basis of the immune response using bulk RNA-seq. Here, we used single cell RNA-seq (scRNA-seq) and spatial transcriptomics to simultaneously characterize multiple cell types within infected human skin and determine the cellular origin of differentially expressed transcripts that we had previously identified by bulk RNA-seq. We obtained paired biopsies of pustules and wounded (mock infected) sites from five volunteers for scRNA-seq. We identified 13 major cell types, including T- and NK-like cells, macrophages, dendritic cells, as well as other cell types typically found in the skin. Immune cell types were enriched in pustules, and some subtypes within the major cell types were exclusive to pustules. Sufficient tissue specimens for spatial transcriptomics were available from four of the volunteers. T- and NK-like cells were highly associated with multiple antigen presentation cell types. In pustules, type I interferon stimulation was high in areas that were high in antigen presentation—especially in macrophages near the abscess—compared to wounds. Together, our data provide a high-resolution view of the cellular immune response to the infection of the skin with a human pathogen.IMPORTANCEA high-resolution view of the immune infiltrate due to infection with an extracellular bacterial pathogen in human skin has not yet been defined. Here, we used the human skin pathogen Haemophilus ducreyi in a human challenge model to identify on a single cell level the types of cells that are present in volunteers who fail to spontaneously clear infection and form pustules. We identified 13 major cell types. Immune cells and immune-activated stromal cells were enriched in pustules compared to wounded (mock infected) sites. Pustules formed despite the expression of multiple pro-inflammatory cytokines, such as IL-1β and type I interferon. Interferon stimulation was most evident in macrophages, which were proximal to the abscess. The pro-inflammatory response within the pustule may be tempered by regulatory T cells and cells that express indoleamine 2,3-dioxygenase, leading to failure of the immune system to clear H. ducreyi.https://journals.asm.org/doi/10.1128/mbio.03885-24Haemophilus ducreyihuman infectionscRNA-seqspatial transcriptomics |
| spellingShingle | Julie A. Brothwell Yuhui Wei Jia Wang Tingbo Guo Chi Zhang Kate R. Fortney Rory Duplantier Li Chen Teresa A. Batteiger Mark H. Kaplan Stanley M. Spinola Sha Cao A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers mBio Haemophilus ducreyi human infection scRNA-seq spatial transcriptomics |
| title | A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers |
| title_full | A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers |
| title_fullStr | A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers |
| title_full_unstemmed | A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers |
| title_short | A high-resolution view of the immune and stromal cell response to Haemophilus ducreyi infection in human volunteers |
| title_sort | high resolution view of the immune and stromal cell response to haemophilus ducreyi infection in human volunteers |
| topic | Haemophilus ducreyi human infection scRNA-seq spatial transcriptomics |
| url | https://journals.asm.org/doi/10.1128/mbio.03885-24 |
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