A novel whole-bacterial enzyme linked-immunosorbant assay to quantify Chlamydia trachomatis specific antibodies reveals distinct differences between systemic and genital compartments

Autor: Alison J. Quayle, Rebecca A. Lillis, Pamela A. Kozlowski, Hannah L. Albritton, Chris L. McGowin, Stephanie N. Taylor, Julia Siren, Li Shen, Joyce A. Ibana, Lyndsey R. Buckner
Rok vydání: 2017
Předmět:
0301 basic medicine
Immunoglobulin A
Serotype
Physiology
Antibody Response
lcsh:Medicine
Chlamydia trachomatis
Cervix Uteri
medicine.disease_cause
Biochemistry
Epithelium
Immunoglobulin G
Chlamydia Infection
Mice
Animal Cells
Immune Physiology
Medicine and Health Sciences
Enzyme-Linked Immunoassays
lcsh:Science
Immune Response
Vaccines
Immune System Proteins
Multidisciplinary
biology
Antibodies
Bacterial
Infectious Diseases
medicine.anatomical_structure
Vagina
Female
Cellular Types
Anatomy
Antibody
Research Article
Adult
Infectious Disease Control
Immunology
030106 microbiology
Sexually Transmitted Diseases
Enzyme-Linked Immunosorbent Assay
Research and Analysis Methods
Antibodies
Cell Line
Young Adult
03 medical and health sciences
Antigen
Immunity
medicine
Animals
Humans
Immunoassays
Secretion
business.industry
lcsh:R
Biology and Life Sciences
Proteins
Epithelial Cells
Cell Biology
Chlamydia Infections
Biological Tissue
030104 developmental biology
Immunologic Techniques
biology.protein
lcsh:Q
Physiological Processes
business
Zdroj: PLoS ONE, Vol 12, Iss 8, p e0183101 (2017)
PLoS ONE
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0183101
Popis: Chlamydia trachomatis (CT) is the leading sexually transmitted bacterial infection. The continued global burden of CT infection strongly predicates the need for a vaccine to supplement current chlamydial control programs. The correlates of protection against CT are currently unknown, but they must be carefully defined to guide vaccine design. The localized nature of chlamydial infection in columnar epithelial cells of the genital tract necessitates investigation of immunity at the site of infection. The purpose of this study was to develop a sensitive whole bacterial enzyme-linked immunosorbent assay (ELISA) to quantify and compare CT-specific IgG and IgA in sera and genital secretions from CT-infected women. To achieve this, elementary bodies (EBs) from two of the most common genital serovars (D and E) were attached to poly-L-lysine-coated microtiter plates with glutaraldehyde. EB attachment and integrity were verified by the presence of outer membrane antigens and the absence of bacterial cytoplasmic antigens. EB-specific IgG and IgA standards were developed by pooling sera with high titers of CT-specific antibodies from infected women. Serum, endocervical and vaginal secretions, and endocervical cytobrush specimens from CT-infected women were used to quantify CT-specific IgG and IgA which were then normalized to total IgG and IgA, respectively. Analyses of paired serum and genital samples revealed significantly higher proportions of EB-specific antibodies in genital secretions compared to sera. Cervical and vaginal secretions and cytobrush specimens had similar proportions of EB-specific antibodies, suggesting any one of these genital sampling techniques could be used to quantify CT-specific antibodies when appropriate normalization methodologies are implemented. Overall, these results illustrate the need to investigate genital tract CT antibody responses, and our assay provides a useful quantitative tool to assess natural immunity in defined clinical groups and CT vaccine trials.
Databáze: OpenAIRE
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