Heat-killed cells of Lactobacillus plantarum L-137 are potent inducers of IL-12 in vitro as well as in vivo and have been shown to have antiallergic, antitumor, and antiviral effects through this induction, which leads to a Th1 type immune response. To determine why L-137 cells induce much greater IL-12 production than the type strain Lactobacillus plantarum JCM1149, we examined the differences in their CW components. The L-137 CW was found to have a higher alanine content and IL-12p40 induction was significantly greater in comparison with JCM1149 CW, whereas peptidoglycans isolated from both strains did not cause IL-12p40 induction. Because in purified CW preparations from gram-positive bacteria, the presence of LTA, the major proinflammatory structure on these bacteria, has been known to have high alanine content, we investigated the responsiveness of both strains to anti-LTA antibody by flow cytometry. L-137 cells reacted more with anti-LTA antibody than did JCM1149 cells. Furthermore, derivative strains of L-137, cured of a specific plasmid pLTK11 of the 15 endogenous plasmids in wildtype L-137, had poor responsiveness to anti-LTA antibody and showed lower IL-12p40 inducing activity than the wild-type L-137 with pLTK11. Our results suggest that LTA expression on the cell surface causes IL-12p40 induction, and that the above internal plasmid of L-137 influences LTA synthesis and expression on the cell surface. Cell wall, Lactobacillus plantarum, lipoteichoic acid, plasmid. LAB are the most common type of probiotic microbes. Reports of the beneficial roles of these bacteria in humans and animals have included effects on the immune system (1, 2). In this laboratory, HK-LP, a strain isolated from fermented food, has previously been shown to be a potent inducer of IL-12 in mice in vitro as well as in vivo production against a natural antigen in a mouse model of food allergy, but also inhibited tumor growth in mice transplanted with syngeneic tumor cells (3, 4). Furthermore, it was demonstrated that, in healthy subjects, daily intake of HK-LP enhanced acquired immunity, especially enhancing Th 1-related immune functions, in addition to producing subsequent improvements in the health-related Key words
This study reports the effect of heat treating Lactobacillus plantarum L-137 on its in vitro cytokine-inducing activity, on the stability of this activity in simulated digestive juices, and on its in vivo immunomodulatory properties. L-137 cells were harvested at the stationary phase with or without the subsequent heat treatment and then lyophilized. Heat-killed L-137 cells stimulated mouse spleen cells to produce more interleukin-12p40 than unheated L-137. The interleukin-12p40-inducing activity of unheated L-137 was significantly lower when incubated with simulated intestinal juice, but the activity of heat-killed L-137 cells was maintained. Furthermore, heat-killed L-137 was more protective than unheated L-137 in a mouse model of dextran sulfate sodium-induced colitis. A heat treatment may therefore be effective for enhancing the immunomodulatory activity of L-137 cells.
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