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Enhanced immunogenicity of the tuberculosis subunit Rv0572c vaccine delivered in DMT liposome adjuvant as a BCG-booster 1 1 Introduction 1 2 Materials and methods 2 2.1 Cloning and purification 2 2.2 Patient participants 2 2.3 Whole blood IFN-γ assay 2 2.4 Experimental animals 3 2.5 Preparation of adjuvants and vaccines 3 2.6 Immunization protocol 3 2.7 Antibody titers 3 2.8 Th1 cytokines secreted by splenocytes 3 2.9 Flow cytometry 3 2.10 Cytokine mRNA expression in lung tissues 3 2.11 Statistical analysis 4 3 Results 4 3.1 Purification and identification of recombinant protein 4 3.2 Rv0572c-specific IFN-γ levels in human whole blood detected by WBIA 4 3.3 Antibody responses induced by DMT-adjuvanted rRv0572c 5 3.4 Rv0572c-specific Th1 cytokines induced by splenocytes 5 3.5 Functional T cell subsets response to PPD and rRv0572c 5 3.6 Rv0572c-specific cytokines mRNA expressed in lung tissues 5 4 Discussion 6 5 Conclusion 10 Ethic approval 10 Informed consent statement 10 Data availability statement 10 CRediT authorship contribution statement 10 Declaration of competing interest 10 Acknowledgements 10 Appendix A Supplementary data 10 References 10

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UND

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nonfiction