Background: Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB),
especially the drug-resistant MTB, causes serious challenges for people healthcare
worldwide. Cytotoxic T lymphocytes (CTLs) play a vital role in immune defense
Objective: To identify novel CTL epitopes that could induce cellular immunity against
Methods: The HLA-A*0201 restricted CTL epitopes of the drug-resistant protein
InhA from MTB were predicted by online algorisms and synthesized by the Fmoc
solid phase method. The candidate peptides were used to induce CTLs from human
peripheral blood mononuclear cells (PBMCs) of HLA-A*0201 healthy donors and the
mice. IFN-γ productions of CTLs were detected by enzyme linked
immunospot assay (ELISPOT), flow cytometry and enzyme-linked immunosorbent
assay (ELISA), and cytotoxicity was analyzed by lactate dehydrogenase (LDH) assay.
Results: A group of 4 epitopes were screened out with high affinity to HLA-A*0201.
ELISPOT and flow cytometry analysis indicated these peptides significantly induced
IFN-γ release of CTLs from the HLA-A*0201+/PPD+ donors, as the mutant
analogues had more potent stimulation effects. LDH assay showed that CTLs from
PPD+ donors and the immunized mice exhibited significant cytotoxicity and low
cross-reactivity. ELISA analysis revealed comparative levels of IFN-γ were released
by CTLs isolated from the mice spleen.
Conclusion: Our study has identified 4 novel CTL epitopes of InhA that could elicited
potent CTL immunity, establishing foundation for development for multivalent
peptide vaccine against the drug-resistant MTB.
Keywords: Amyloid-β protein, endoplasmic reticulum stress, GRP78, oligomers, Rab protein, presenilin 1, toxic turn, Mycobacterium tuberculosis, drug-resistant, Cytotoxic T lymphocyte,
epitope, vaccine, IFN-γ
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