Recombinant Mycobacterium smegmatis expressing Hsp65-hIL-2 fusion protein and its influence on lymphocyte function in mice

作     者：Xiao-Qing Guo Yan-Ming Wei Bo Yu 

作者机构：Animal Medical CollegeGansu Agricultural University Affiliated Hospital of Gansu College of Traditional Chinese Medicine 

出 版 物：《Asian Pacific Journal of Tropical Medicine》 (亚太热带医药杂志（英文版）)

年 卷 期：2012年第5卷第5期

页      面：347-351页

核心收录：

学科分类：1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学] 

基　　金：funded by the National Natural Science Eoundation of China(Grant ID:30972767) 

主　　题：Mycobacterium tuberculosis Mycobacterium smegmatis Hsp65 IL-2 Fusion protein Lymphocytes Mice 

摘      要：Objective:To coastruct a strain of recombinant Mycobacterium smegmatis expressing the heat shock protein 65(Hsp65) and human interleukin 2(IL-2) fusion protein(rMS-Hsp65/IL-2) and to explore the effect of this construct on lymphocyte function in ***:The fusion gene encoding Hsp65-hIL-2 was cloned into shuttle vector *** recombinant plasmid pSMT3- Hsp65-hIL-2 was transferred to Mycobacterium smegmatis by *** clones were selected by hygromycin and identified by *** expression of fusion protein Hsp65- hIL-2 was verified using indirect immunofluorescence *** were immunized for two times by subcutaneously injection with 1×10~6 CFU rMS-Hsp65/IL-2 at a three-week *** weeks after the second immunization,mice were sacrificed and the serum samples were collected for determination of anli-Hsp65 specific *** lymphocytes were isolated and treated with the rMS-Hsp65/IL-2 to determine lymphocytic proliferation activity by MTT ***-γand IL-2 in the medium of the treated cells were also determined by ***:Successful construction of rMS-Hsp65/IL-2 was verified by PCR and immunofluorescence *** to the splenic lymphocytes isolated from mice immunized with Bacille Calmette-Guerin or mice immunized with Mycobacterium smegmatis alone,the splenic lymphocytes isolated from mice immunized with rMS-Hsp65/IL-2 showed a marked increase in the proliferation of lymphocytes, together with an increased production of important cytokines such as IFN- 7 and IL-2. Conclusions:rMS-Hsp65/IL-2 markedly enhances lymphocyte ***,the fusion protein generated by rMS-Hsp65/lL-2 may be of potential value in generating an effective vaccine against tuberculosis.