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Principal Investigator: Styliani Karanika
Organization: JOHNS HOPKINS UNIVERSITY
Fiscal Year: 2024
Award: $200,880
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
Host status plays a major role in tuberculosis (TB) disease outcomes, with many efforts to develop host-directed
therapies to boost immune response. It is unknown how these strategies will perform in people living with
HIV/AIDS (PLWHA) who are at increased risk of adverse TB treatment outcomes. Persistent Mycobacterium
tuberculosis (Mtb), which obligates prolonged TB treatment, highly expresses the stringent response protein
RelMtb. One attractive host-directed therapeutic strategy is boosting RelMtb-specific cellular responses. We
developed a novel therapeutic DNA TB vaccine that promotes immune responses to RelMtb, which are further
enhanced by fusion with a chemokine gene, Macrophage Inflammatory Protein-3 alpha (MIP-3α), that targets
relMtb to immature dendritic cells (DCs). Our data show that an intranasal (IN) or intramuscular (IM) DNA vaccine
expressing MIP-3α/relMtb (“fusion” vaccine) demonstrated greater adjunctive therapeutic efficacy when combined
with the first-line TB treatment in Mtb-infected mice as compared to IM vaccination with relMtb alone (“non-fusion”
vaccine). This proposal will dissect the mechanisms of this novel therapeutic strategy, specifically with IN
administration which had the highest efficacy. We will also test whether this strategy is likely to perform in
PLWHA, analyzing whether RelMtb-specific immune responses in PLWHA can predict therapeutic TB outcomes.
Aim 1 will investigate the molecular mechanism by which the IN administration potentiates anti-TB drugs, while
Aim 2 will focus on the role of DCs in the adjunctive therapeutic efficacy of the fusion vaccination strategy. Aim
3 will test the translational potential of the RelMtb-specific T-cell immunity in PLWHA. In Aim 1, we test if IN
vaccine administration enhances the host response to Mtb infection through increased IL-17A secretion. We will
perform single-cell transcriptomics on cells from murine lungs to confirm the upregulation of IL-17A response
pathways in the IN arm, followed by Mtb-infected IL-17A knockout murine experiments. In Aim 2, we hypothesize
that the IN fusion vaccine leads to more efficient systemic and local T-cell maturation, activation, and
differentiation through enhanced DC activation compared to the IN non-fusion vaccine. Aim 2a will assess
differences in DC activation by flow cytometry, DC and T-cell co-localization in murine tissues, and differences
in T-cell activation/differentiation using co-culture experiments with DCs derived from each vaccination group. In
Aim 2b, we will adoptively transfer DCs from mice immunized with either vaccine into Mtb-infected mice
concurrently with TB treatment to compare the adjunctive therapeutic efficacy. In Aim 3, we hypothesize that
RelMtb-specific Th1/Th17 cell responses in peripheral blood mononuclear cells derived from HIV-infected and
uninfected patients receiving treatment for pulmonary TB, correlate inversely with sputum culture conversion and
TB recurrence rates, independently of HIV status. The proposed studies will uncover critical details of the
mechanism of a novel therapeutic vaccination approach against TB with important implications in PLWHA and
help the PI to gain significant expertise in TB/HIV immunology.
Terms: <AIDS Virus><AIDS/HIV><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Adoptive Transfer><Affect><Antigens><Antitubercular Drugs><Blast Transformation><Blastogenesis><Body Tissues><CCL20><CCL20 gene><CD4 Cells><CD4 Lymphocyte Count><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ Cell Counts><CD4+ Counts><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CTLA-8><CTLA-8 Gene><CTLA8><CTLA8 Gene><Cell Body><Cell Maturation><Cells><Chemokine, CC Motif, Ligand 20><Chemotactic Cytokines><Clinical Management><Co-culture><Cocultivation><Coculture><Coculture Techniques><Collecting Cell><Coon's Technic><Coon's Technique><Cytotoxic T-Lymphocyte-Associated Antigen 8><Cytotoxic T-Lymphocyte-Associated Antigen 8 Gene><Cytotoxic T-Lymphocyte-Associated Serine Esterase 8><Cytotoxic T-Lymphocyte-Associated Serine Esterase 8 Gene><DNA><DNA Vaccines><DNA-based therapeutics><Data><Dendritic Cells><Dendritic cell activation><Deoxyribonucleic Acid><Disease Outcome><Enzyme Gene><Enzymes><Exhibits><Exodus 1><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Fluorescent Antibody Technic><Fluorescent Antibody Technique><Fluorescent Antinuclear Antibody Test><Genes><HIV><HIV/AIDS><HIV/Mtb><HIV/TB><HIV/mycobacterium tuberculosis><HIV/tuberculosis><Homologous Chemotactic Cytokines><Human><Human Immunodeficiency Viruses><IL-17><IL-17 Gene><IL-17A><IL-17A Gene><IL17><IL17 Protein><IL17 gene><IL17A><IL17A Gene><Image><Immune response><Immunity><Immunization><Immunize><Immunofluorescence Technic><Immunofluorescence Technique><Immunological response><Immunology><Intercrines><Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8)><Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8) Gene><Interleukin 17 Precursor><Interleukin 17 Precursor Gene><Interleukin-17><International><Intramuscular><Intranasal Administration><Intranasal Drug Administration><Knock-out><Knockout><LARC><LAV-HTLV-III><Lung><Lung Respiratory System><Lung TB><Lung Tuberculosis><Lymphadenopathy-Associated Virus><Lymphoblast Transformation><Lymphocyte Activation><Lymphocyte Stimulation><Lymphocyte Transformation><M tb><M tuberculosis><M tuberculosis infection><M. tb><M. tb infection><M. tuberculosis><M. tuberculosis infection><M. tuberculosis/HIV><M.tb infection><M.tuberculosis infection><MIP3A><MTB infection><MTB vaccine><Macrophage Inflammatory Protein 3-Alpha><Measures><Mentors><Mice><Mice Mammals><Modern Man><Molecular><Murine><Mus><Mycobacterium tuberculosis><Mycobacterium tuberculosis (MTB) infection><Mycobacterium tuberculosis infection><Naked DNA Vaccines><Outcome><PBMC><Pathway interactions><Patients><Peripheral Blood Mononuclear Cell><Persons><Physicians><Play><Proteins><Pulmonary TB><Pulmonary Tuberculosis><RNA vaccine><RNA-based vaccine><Receptor Protein><Recombinant DNA Vaccines><Recurrence><Recurrent><Reporting><Resistance><Risk><Role><Route><SCYA20><SIS cytokines><Scientist><Small Inducible Cytokine Subfamily A, Member 20><Spleen><Spleen Reticuloendothelial System><Sputum><Suspension substance><Suspensions><T cell response><T memory cell><T-Cell Activation><T-Cells><T-Lymphocyte><T4 Cells><T4 Lymphocyte Count><T4 Lymphocytes><TB drugs><TB infection><TB therapy><TB treatment><TB vaccine><Testing><Therapeutic><Tissues><Treatment Efficacy><Treatment Failure><Treatment outcome><Tuberculosis><Tuberculosis Vaccines><Up-Regulation><Upregulation><VAC-TX><Vaccinated><Vaccination><Vaccine Therapy><Vaccine for TB><Vaccine for Tuberculosis><Vaccines><Veiled Cells><Virus-HIV><Work><activate T cells><active method><active technique><active treatment><adverse drug reaction><anti-TB drugs><anti-TB vaccine><anti-tuberculosis drugs><arm><chemoattractant cytokine><chemokine><chronic infection><deliver vaccines><disseminated TB><disseminated tuberculosis><draining lymph node><experiment><experimental research><experimental study><experiments><flow cytophotometry><fluorescent antibody><high risk group><high risk individual><high risk people><high risk population><host response><imaging><imaging system><immune system response><immunization strategy><immunogen><immunoresponse><in vivo><infection due to Mycobacterium tuberculosis><intervention efficacy><mRNA vaccine><mRNA-based vaccine><memory T lymphocyte><mtb><mycobacterial><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutics><new therapy><new therapy approaches><new treatment approach><new treatment strategy><next generation therapeutics><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutics><novel therapy><novel therapy approach><pathway><persistent infection><pulmonary><receptor><regional lymph node><resistant><response><skills><social role><spectral image><spectral imagery><spectrograph><spectrum image><spectrum imagery><therapeutic DNA><therapeutic efficacy><therapeutic evaluation><therapeutic testing><therapeutic vaccination><therapeutic vaccine><therapy efficacy><therapy failure><thymus derived lymphocyte><transcriptomics><translational opportunities><translational potential><treat M. tuberculosis><treat Mtb><treat Mycobacterium tuberculosis><treat tb><treat tuberculosis><treatment vaccines><tuberculosis drugs><tuberculosis infection><tuberculosis therapy><tuberculosis treatment><tuberculous spondyloarthropathy><vaccination strategy><vaccine against M. tuberculosis><vaccine against Mtb><vaccine against Mycobacterium tuberculosis><vaccine against TB><vaccine against tuberculosis><vaccine candidates against tuberculosis><vaccine delivery><vaccine for the treatment><vaccine for treatment><vaccine platform><vaccinology>