Soybean protein fraction digested with neutral protease preparation, “Peptidase R”, produced by Rhizopus oryzae, stimulates innate cellular immune system in mouse

A soybean protein fraction was prepared from defatted soybean seed flour and digested with 29 kinds of commercially available protease originating from preparations of animals, plants, and microorganisms. Some digests, in particular, Ro-digest prepared using a Rhizopus oryzae neutral protease preparation (Peptidase R), displayed strong mitogenic activity toward C3H/HeN mouse spleen cells. The number of spleen CD11b+, CD49b+, interleukin (IL)-12+CD11b+, and interferon (IFN)-γ+CD49b+ cells significantly increased when cultured with Ro-digest. Similarly, the number of spleen IFN-γ+CD4+ cells significantly increased in the presence of Ro-digest while that of spleen IL-4+CD4+ cells was largely unchanged. Additionally, 5-week-old male C3H/HeN mice were given diets consisting of ovalbumin (OVA) alone (control diet) or a mixture of OVA and Ro-digest (Ro-digest-added diet) as a protein source for 5 weeks, and the immune properties of the mice were investigated. The number of IL-12+CD11b+ cells was greater in spleens from mice given the Ro-digest-added diet than in those given the control diet. The cytotoxic activity of spleen cells toward the human erythroleukemia cell line, K562, was significantly higher in mice given the Ro-digest-added diet than in those given the control diet. Furthermore, in a microarray analysis of mRNAs extracted from mice Peyer's patch cells, gene expression related to innate immune responses was increased in mice given the Ro-digest-added diet. These results indicate that the Ro-digest might stimulate cellular immune systems, in particular, an innate immunity in mice.