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  • Title: [Therapeutic effects of granulysin/interleukin-12 recombinant M. smegmatis in a murine model of M. tuberculosis infection].
    Author: Yang C, Wang YW, Yi ZJ, He YL, Li N, Xu L, Zhang L, Zhu DY.
    Journal: Zhonghua Jie He He Hu Xi Za Zhi; 2007 Feb; 30(2):133-7. PubMed ID: 17445478.
    Abstract:
    OBJECTIVE: To study the effects and mechanisms of recombinant Mycobacterium smegmatis (M. smegmatis) encoding human granulysin and murine interleukin-12 (IL-12) on murine M. tuberculosis infection. METHODS: Balb/c mice infected with M. tuberculosis were treated with normal saline, M. smegmatis, pZM03, recombinant M. smegmatis respectively. The numbers of viable bacteria in the lung and the spleen were counted. The levels of IL-12, IgG(2a) in serum and interferon-gamma (IFN-gamma) released from spleen lymphocytes stimulated with purified protein derivative (PPD) were detected with enzyme linked immunosorbent assay (ELISA). The expression of granulysin in tissue was detected with immunohistochemistry. Lungs and spleens were prepared for pathological analysis. RESULTS: The recombinant M. smegmatis group [(4.57 +/- 0.28) in lung, (3.47 +/- 0.25) in spleen] showed significantly reduced number of colony forming units (log CFU/g) compared with the control [(5.77 +/- 0.12) in lung, (4.66 +/- 0.18) in spleen], M. smegmatis [(5.62 +/- 0.14) in lung, (4.65 +/- 0.26) in spleen] and pZM03 [(5.04 +/- 0.22) in lung, (4.25 +/- 0.48) in spleen] group. The expression of granulysin in tissue, and increased level of IL-12 and IgG(2a) in serum were found in recombinant M. smegmatis group. IFN-gamma released from spleen lymphocytes stimulated with PPD in recombinant M. smegmatis group and pZM03 group was higher than that of other groups, but the difference between recombinant M. smegmatis group and pZM03 group was not significant. The pathological changes in lungs of the recombinant M. smegmatis group were localized, while those in the control group were extensive. Significant pathological changes were not found in the spleens of all groups. CONCLUSION: The recombinant M. smegmatis had immunotherapeutic effects, which were associated with a switch to Th1 response and the antibacterial activity of granulysin.
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