Immunostimulation effects of proteose-peptone component 3 fragment on human hybridomas and peripheral blood lymphocytes.
Fat-free bovine milk fermented by 12 kinds of lactic acid bacteria and yeast enhanced monoclonal antibody production of human hybridoma HB4C5 cells 2.8-fold in serum-free medium. Immunoglobulin production of human peripheral blood lymphocytes (PBL) was also stimulated in vitro. IgM and IgG production of human PBL was accelerated up to 2.8-fold and 5.4-fold, respectively. Interferon-gamma production of human PBL was also accelerated 6.0-fold by 50 microg/ml of the fermented milk. However, interleukin-4 production of PBL was not affected, and tumor necrosis factor-alpha production was suppressed. The activity was enhanced 2.5-fold by the thermal treatment for 30 min at 65 degrees C and was completely lost by trypsin digestion. The findings suggested that the active substance in the fermented milk was heat stable protein. Gel-filtration and the SDS-PAGE analysis revealed that the molecular weight of the active substance was estimated as 19.0 kDa, which was not detected in fat-free bovine milk before fermentation. N-terminal amino acid sequence of the 19.0 kDa protein was highly homologous to proteose-peptone component 3 (PP3). Since molecular weight of PP3 is 28 kDa, it is suggested that the 19.0 kDa protein is derived from degradation of PP3 during fermentation of fat-free milk. Moreover, PP3 purified from fat-free milk also enhanced IgM production of HB4C5 cells.
Sugahara
Takuya
T
Faculty of Agriculture, Ehime University, 3-5-7, Tarumi, Matsuyama, Ehime 790-8566, Japan. mars95@agr.ehime-u.ac.jp
Onda
Hiroyuki
H
Shinohara
Yusuke
Y
Horii
Mayumi
M
Akiyama
Koichi
K
Nakamoto
Katsunori
K
Hara
Kazushi
K
D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D002364:Caseins / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology*; D006801:Humans; D006825:Hybridomas / Q000187:drug effects* / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology*; D008214:Lymphocytes / Q000187:drug effects* / Q000276:immunology*; D008894:Milk Proteins / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology*
D000276:Adjuvants, Immunologic; D002364:Caseins; D006023:Glycoproteins; D007075:Immunoglobulin M; D008894:Milk Proteins; C085522:PP3 protein, Bos taurus; D010446:Peptide Fragments