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Principal Investigator: MICHAEL N STARNBACH
Organization: HARVARD MEDICAL SCHOOL
Fiscal Year: 2023
Award: $247,824
Funding agency: National Institute of Allergy and Infectious Diseases
Chlamydia trachomatis is an obligate intracellular bacterium and the most prevalent sexually transmitted
infection in the United States. If untreated, infection can lead to pelvic inflammatory disease, ectopic
pregnancy, and infertility. Although C. trachomatis infection is treatable with antibiotics, many cases are
asymptomatic, and repeat infections are common. Therefore, a vaccine is the best public health solution for
managing and preventing infection. To develop a truly effective vaccine, it is critical to understand how C.
trachomatis evades adaptive immunity to establish persistent infection during natural infection. CD8 T cells are
normally integral for controlling intracellular pathogen infections, but during C. trachomatis infection, the CD8 T
cell response is significantly impaired. We have shown that upregulation of the immunoinhibitory ligand PD-L1
during infection contributes to the diminution of the CD8 T cell response. Here we propose to identify C.
trachomatis virulence genes that are responsible for upregulating PD-L1, as we hypothesize that these
virulence genes are necessary to manipulate CD8 T cell immunity. In our first aim, we propose two
complimentary approaches to identify C. trachomatis genes responsible for PD-L1 upregulation during
infection. The first is to create and screen a lentiviral library containing individual C. trachomatis genes, testing
the ability of individual bacterial genes to upregulate PD-L1. The second approach is based on a fortuitous
phenotype in HeLa cells where only C. muridarum and not C. trachomatis is capable of upregulating PD-L1. By
using a C. trachomatis/C. muridarum chimera collection developed by collaborators, we will identify portions of
the C. muridarum chromosome that are sufficient to upregulate PD-L1. After generating a list of gene
candidates from these two approaches, we will create C. trachomatis strains with null mutations in genes
responsible for upregulating PD-L1 (CtNP), to validate their necessity for PD-L1 upregulation during infection in
vitro and in vivo. In aim 2, we will use the CtNP strains to test if bacterial genes responsible for PD-L1
upregulation are required to manipulate the CD8 T cell response and hinder CD8 T cell-mediated protection.
First, we will compare the proliferation and homing of C. trachomatis specific CD8 T cells during the course of
infection with CtNP strains or WT C. trachomatis. Secondly, we propose to evaluate the protective capacity of
memory CD8 T cells isolated from mice infected with CtNP strains or WT C. trachomatis by performing CD8 T
cell transfer experiments into naïve mice. This work will provide the foundation for future studies to interrogate
the mechanism by which C. trachomatis upregulates PD-L1. By understanding how C. trachomatis impairs the
T cell response, we can develop vaccines that stimulate superior immunity compared to natural infection.
Terms: <Antibiotic Agents><Antibiotic Drugs><Antibiotics><B7-H1><B7H1><Bacteria><Bacterial Genes><C muridarum><C trachomatis><C. muridarum><C. trachomatis><CD274><CD8><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CD8B><CD8B1><CD8B1 gene><Cancers><Candidate Disease Gene><Candidate Gene><Cannot achieve a pregnancy><Cell Body><Cell Communication and Signaling><Cell Function><Cell Isolation><Cell Process><Cell Segregation><Cell Separation><Cell Separation Technology><Cell Signaling><Cell physiology><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chimera><Chimera organism><Chlamydia Infections><Chlamydia muridarum><Chlamydia trachomatis><Chlamydial Infection><Chromosomes><Chronic><Chronic depressive disorder><Collection><Communities><Difficulty conceiving><Ectopic Pregnancy><Epidemic><Epithelial Cells><Foundations><Future><Genes><Genetic><Genome Library><Genomic Library><Goals><HeLa><Hela Cells><Homing><Immune><Immune Evasion><Immune memory><Immune response><Immunes><Immunity><Immunologic Memory><Immunological Memory><Immunological response><Immunomodulation><Impairment><In Vitro><Individual><Infection><Infection prevention><Infertility><Intracellular Communication and Signaling><LYT3><Left><Lentiviral Vector><Lentivirus Vector><Libraries><Ligands><Malignant Neoplasms><Malignant Tumor><Mediating><Memory><Mice><Mice Mammals><Miscellaneous Antibiotic><Murine><Mus><PD 1><PD-1><PD-L1><PD1><PDL-1><PDL1><Pathogenicity Factors><Pathway interactions><Pelvic Inflammatory Disease><Phenotype><Prevent infection><Programmed Cell Death 1 Ligand 1><Programmed Death Ligand 1><Proliferating><Proteins><Public Health><Regulation><Reporting><Rickettsia trachomae><Sampling><Secondary to><Sexually Transmitted Diseases><Sexually Transmitted Disorder><Sexually Transmitted Infection><Signal Transduction><Signal Transduction Systems><Signaling><Subcellular Process><T cell response><T memory cell><T-Cells><T-Lymphocyte><T8 Cells><T8 Lymphocytes><Testing><Transcript><United States><Up-Regulation><Upregulation><Uterus><Vaccines><Venereal Diseases><Venereal Disorders><Venereal Infections><Virulence Factors><Virus><Work><adaptive immunity><anamnestic reaction><bacteria pathogen><bacterial pathogen><biological signal transduction><cell sorting><chimeras><chlamydial disease><chronic depression><chronic infection><develop a vaccine><develop vaccines><development of a vaccine><experiment><experimental research><experimental study><experiments><extrauterine pregnancy><fertility cessation><fertility loss><gain of function><gene null><genitourinary tract><host response><immune evasive><immune modulation><immune regulation><immune system response><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><in vivo><infertile><mRNA Expression><malignancy><memory T lymphocyte><mutant><neoplasm/cancer><null mutation><pathogen><pathogenic bacteria><pathogenicity gene><pathway><pelvic inflammation disease><pelvic inflammation disorder><pelvic inflammatory disorder><pelvic inflammatory syndrome><persistent infection><prevent><preventing><programmed cell death 1><programmed cell death ligand 1><programmed cell death protein 1><programmed cell death protein ligand 1><programmed death 1><protein death-ligand 1><reproductive><response><screening><screenings><secondary immune response><sle2><systemic lupus erythematosus susceptibility 2><thymus derived lymphocyte><tool><urogenital tract><vaccine development><virulence gene><virulent gene><womb>