[Purification by FPLC and characterization of Mycobacterium tuberculosis heat-resistant peptide antigen stimulating proliferation of human gammadelta+ T cells].

Aim: To purify Mycobacterium tuberculosis heat-resistant peptide antigen(Mtb-Ag) that can stimulate gammadelta+ T cells.

Methods: Mtb-Ag was first separated by fast performance liquid chromatography (FPLC) with S-100 column, and then the components including low molecular weight peptide antigens (Mtb-LW-Ag) peaks were purified by FPLC with Mono Q column. Activity of the purified Mtb-LW-Ag that stimulates human gammadelta+ T cell proliferation was examined by flow cytometry.

Results: Mtb-Ag was separated into four peaks(A, B, C and D) by FPLC S-100 column and the peak B, C and D contained Mtb-LW-Ag. The peak B was further separated into six main peaks(B-I-VI), peak C only one main peak(C-main), and peak D eight main peaks(D-I-VIII) by FPLC Mono Q column. Furthermore, peak B, C, B-III and C-main peptide could significantly stimulate human gammadelta+ T cell proliferation in a dose of 0.1 mg/L.

Conclusion: Varied Mtb-LW-Ag peptides are purified from Mtb-Ag by FPLC, and the B-III and C-main peptides may be the main components of stimulating human gammadelta+ T cell proliferation.