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Infect Immun ; 88(9)2020 08 19.
Article in English | MEDLINE | ID: mdl-32601108

ABSTRACT

Chlamydia trachomatis infection of the human fallopian tubes can lead to damaging inflammation and scarring, ultimately resulting in infertility. To study the human cellular responses to chlamydial infection, researchers have frequently used transformed cell lines that can have limited translational relevance. We developed a primary human fallopian tube epithelial cell model based on a method previously established for culture of primary human bronchial epithelial cells. After protease digestion and physical dissociation of excised fallopian tubes, epithelial cell precursors were expanded in growth factor-containing medium. Expanded cells were cryopreserved to generate a biobank of cells from multiple donors and cultured at an air-liquid interface. Culture conditions stimulated cellular differentiation into polarized mucin-secreting and multiciliated cells, recapitulating the architecture of human fallopian tube epithelium. The polarized and differentiated cells were infected with a clinical isolate of C. trachomatis, and inclusions containing chlamydial developmental forms were visualized by fluorescence and electron microscopy. Apical secretions from infected cells contained increased amounts of proteins associated with chlamydial growth and replication, including transferrin receptor protein 1, the amino acid transporters SLC3A2 and SLC1A5, and the T-cell chemoattractants CXCL10, CXCL11, and RANTES. Flow cytometry revealed that chlamydial infection induced cell surface expression of T-cell homing and activation proteins, including ICAM-1, VCAM-1, HLA class I and II, and interferon gamma receptor. This human fallopian tube epithelial cell culture model is an important tool with translational potential for studying cellular responses to Chlamydia and other sexually transmitted pathogens.


Subject(s)
Epithelial Cells/immunology , Gene Expression Regulation/immunology , Host Microbial Interactions/immunology , T-Lymphocytes/immunology , Adult , Amino Acid Transport System ASC/genetics , Amino Acid Transport System ASC/immunology , Antigens, CD/genetics , Antigens, CD/immunology , Biomarkers/metabolism , Chemokine CCL5/genetics , Chemokine CCL5/immunology , Chemokine CXCL10/genetics , Chemokine CXCL10/immunology , Chemokine CXCL11/genetics , Chemokine CXCL11/immunology , Chlamydia Infections/genetics , Chlamydia Infections/immunology , Chlamydia Infections/microbiology , Chlamydia trachomatis/growth & development , Chlamydia trachomatis/immunology , Epithelial Cells/microbiology , Fallopian Tubes/cytology , Fallopian Tubes/surgery , Female , Fusion Regulatory Protein 1, Heavy Chain/genetics , Fusion Regulatory Protein 1, Heavy Chain/immunology , Histocompatibility Antigens Class I/genetics , Histocompatibility Antigens Class I/immunology , Histocompatibility Antigens Class II/genetics , Histocompatibility Antigens Class II/immunology , Host Microbial Interactions/genetics , Humans , Intercellular Adhesion Molecule-1/genetics , Intercellular Adhesion Molecule-1/immunology , Minor Histocompatibility Antigens/genetics , Minor Histocompatibility Antigens/immunology , Models, Biological , Primary Cell Culture , Receptors, Interferon/genetics , Receptors, Interferon/immunology , Receptors, Transferrin/genetics , Receptors, Transferrin/immunology , Salpingectomy , T-Lymphocytes/microbiology , Vascular Cell Adhesion Molecule-1/genetics , Vascular Cell Adhesion Molecule-1/immunology , Interferon gamma Receptor
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