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


CONCLUSIONS
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.
Chen
Yong
Y
Department of Immunology, Bengbu Medical College, Bengbu 233003, China.
Lu
He-zuo
HZ
Hu
Jian-guo
JG
Hou
Yan-qiang
YQ
He
Hai-hui
HH
Zhang
Hai-feng
HF
Li
Bai-qing
BQ
D000942:Antigens, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D049109:Cell Proliferation; D002851:Chromatography, High Pressure Liquid; D006358:Hot Temperature; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
D000942:Antigens, Bacterial; D016692:Receptors, Antigen, T-Cell, gamma-delta