[Differentiation and function of human dendritic cells influenced by heat shock protein gp96 purified from K562 cells].
This study was to establish the method of purifying heat shock protein GP96 from K562 cells and explore the differentiation and function of human DC influenced by heat shock prolein (HSP). Using ammonium sulfate precipitation, conA-sepharose affinity chromatography and DEAE-sephacel anion exchange chromatography GP96 from K562 cells lysate was isolated and purified. The identification of the purified protein was controlled by Western blot with anti-human GP96 IgG. Human dendritic cell derived from peripheral blood mononuclear cell were cultured with purified GP96. The phenotype changes of DC were analyzed by flow cytometry and MLR was detected by MTT. The results showed that 60-80 microg GP96 was purified successfully from 1 x 10(10) K562 cells. DC stimulated with HSP-GP96 had higher expression rates of CD83, CD86, HLA-DR and lower expression rates of CD1a and had stronger ability to induce T cells proliferation. It is concluded that heat shock protein GP96 can be isolated and purified from K562 cells and could induce maturation of dendritic cell. HSP-DC vaccine show stronger ability to induce T cell proliferation than DC.
Chen
Yi-Juan
YJ
Department of Hematology, Fuxing Hospital, Capital University of Medical Sciences, Beijing 100038, China. chenyijuan@x263.net
Shi
Bin
B
Wang
Shen-Wu
SW
Cui
Jian-Ying
JY
D000951:Antigens, Neoplasm / Q000302:isolation & purification / Q000494:pharmacology*; D019496:Cancer Vaccines / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D020014:K562 Cells / Q000737:chemistry*
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; C052646:sarcoma glycoprotein gp96 rejection antigens