Micronized and Heat-Treated LM1004 Stimulates Host Immune Responses Via the TLR-2/MAPK/NF-κB Signalling Pathway In Vitro and In Vivo.

Although nanometric dead has emerged as a potentially important modulator of immune responses, its underlying mechanism of action has not been fully understood. This study aimed to identify the detailed biochemical mechanism of immune modulation by micronized and heat-treated LM1004 (MHT-LM1004, <1 μm in size). MHT-LM1004 was prepared from LM1004 via culture in a specifically designed membrane bioreactor and heat treatment. MHT-LM1004 was shown to effectively induce the secretion of TNF-α and IL-6 and the mRNA expression of inducible nitric oxide synthase (iNOS). MHT-LM1004 enhanced the expression of TLR-2, phosphorylation of MAPKs (ERK), and nuclear translocation of NF-κB in a dose-dependent manner. Oral administration of MHT-LM1004 (4 × 10 or 4 × 10 cells/kg mouse body weight) increased the splenocyte proliferation and serum cytokine levels. These results suggested that MHT-LM1004 effectively enhances early innate immunity by activating macrophages via the TLR-2/MAPK/NF-κB signalling pathway and that this pathway is one of the major routes in immune modulation by the species.