Generation of Human Monoclonal Antibody Specific for Propionibacterium Acnes by In Vitro Immunization

摘要

Propionibacterium acnes (P. acnes) is known to be a main factor of pimple deterioration that occurs in hair follicle. P. acnes destroys skin structure and its resultant scars remain for a long time. Recently, bloods kept in blood banks were reported to be contaminated with P. acnes, which might cause serious posttransfusion infection in blood donors. The aim of this study is to generate human monoclonal antibody specific for P. acnes to treat skin problems caused by P. acnes and to apply it to examination for blood contamination. To develop human monoclonal antibody specific for P. acnes, we performed in vitro immunization (IVI) of human peripheral blood mononuclear cells (PBMCs). Sonicated or autoclaved P. acnes were used as sensitizing antigen inIVI. PBMC were treated with L-leucyl-L-leucine methyl ester (LLME) to remove lysosome-rich cells including monocytes and NK cells, and sensitized with antigen in the presence of IL-2 and IL-4. Cells were cultured for 7 days and supernatants were appliedto ELISA and ELISPOT analyses. To select antigen-specific phage antibodies, we performed phage display using solid-panning system. Results demonstrate that in vitro-immunized PBMC produce P. acnes-specific antibody, suggesting that IVI protocol is usefulfor inducing antibody production specific for pathogenic bacteria.