| Summary: | Heparin-binding hemagglutinin (HBHA) and <i>M. tuberculosis</i> pili (MTP) are important antigens on the surface of <i>Mycobacterium tuberculosis</i>. To display these antigens effectively, the fusion protein HBHA-MTP with a molecular weight of 20 kD (L20) was inserted into the receptor-binding hemagglutinin (HA) fragment of influenza virus and was expressed along with matrix protein M1 in Sf9 insect cells to generate influenza virus-like particles (LV20 in short). The results showed that the insertion of L20 into the envelope of the influenza virus did not affect the self-assembly and morphology of LV20 VLPs. The expression of L20 was successfully verified by transmission electron microscopy. Importantly, it did not interfere with the immunogenicity reactivity of LV20 VLPs. We demonstrated that LV20 combined with the adjuvant composed of DDA and Poly I: C (DP) elicited significantly higher antigen-specific antibodies and CD4<sup>+</sup>/CD8<sup>+</sup> T cell responses than PBS and BCG vaccination in mice, and reduced the bacterial load in the lungs of mice infected with <em>M. tuberculosis</em> H37Ra. It suggests that the insect cell expression system is an excellent protein production system, and LV20 VLPs could be a novel tuberculosis vaccine candidate for further evaluation.
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