Objective · To evaluate the immune effect of major antigen components of Mycobacterium on NKT cells in C57BL/6 mouse livers. Methods · Mycobacterium bovis BCG cells, bacterial whole cell lysate (WCL), total lipid antigens (TLIP), culture filtrate protein B (CFP-B), and lipoglycan were used respectively to immunize C57BL/6 mice via tail vein injection. The liver cells were isolated 3, 6, and 12 days after immunization. Changes in NKT cell contents were measured with flow cytometry. Mice without antigen immunization were used as controls. Results · Flow cytometry showed that percentages of NKT cells in the control group, the lipoglycan group, the CFP-B group, the TLIP group, the WCL group, and the BCG group were (0.040 0±0.020 62)%, (0.027 8±0.039 93)%, (0.035 6±0.047 46)%, (0.411 1±0.245 38)%, (0.118 9±0.054 19)%, and (0.078 9±0.045 95)%, respectively. Statistical analysis showed that the content of NKT cells was significantly higher in the TLIP and WCL groups than in the control group (P<0.01) and was higher in the TLIP group than in the WCL group. The differences between other antigen groups and the control group were not statistically significant (P>0.05). Conclusion · Of major antigen components of Mycobacterium, TLIP can significantly increase the number of NKT cells in C57BL/6 mouse livers. TLIP is worthy of further investigation for its potential anti-tuberculosis immune function.