A Study On The Immune Receptors For Polysaccharopeptide Through TLR4-TIRAP/MAL-MyD88Signaling Pathway In Breast Cancer Patient PBMC

Objective: The present study was undertaken to identify whether PSPaffect TLR4, as well as to examine the TLR4-TIRAP/MAL-MyD88signaling pathway in activation of breast cancer patient’s peripheral bloodmononuclear cells (PBMCs).Methods: After isolation of PBMC from breast cancer patient orhealthy individual, cells were divided into control group and PSP group.Cellular immunofluorescence staining was applied with monoclonalanti-TLR4antibody. As to blocking, breast cancer patient’s PBMC weredivided into negative control group, TLR4antibody group, PSP group andPSP+TLR4antibody group, and then the relative mRNA (IL-12, IL-6andTNF-α) expression of each group was detected by quantitative real-timePCR (Q-PCR). Meanwhile, Q-PCR and Western blot were used to detectthe related gene and protein expression of TLR4pathway regulated by PSP. Results: Enhanced labeling is visualized in control group of healthyindividual’s PBMC, compared with PSP group. What’s more, it is moreobvious with breast cancer patient’s PBMC. Compared with negativecontrol group, genes besides IL-6in TLR4antibody group weredown-regulated significantly, while genes in PSP group were up-regulatedsignificantly (P <0.05). Genes in TLR4antibody group and PSP groupwere down-regulated significantly in the comparison with PSP+TLR4antibody group respectively (P <0.05). Genes in TLR/MyD88pathway wasup-regulated significantly by PSP: TLR4, TLR5, TLR6, IRAK4, TRAF6,IRF5, TAK1, IKKα, NF-κB, ERK, P38, JNK, AP-1, IL-1β and IL-8(P <0.01). Meanwhile, the terminal effect proteins (NF-κB, AP-1and IRF5)were up-regulated significantly by PSP.Conclusions: TLR4may be one of the immune receptors of PSP. PSPfunction its immunoregulatory effect through the regulation ofTLR4-TIRAP/MAL-MyD88signaling pathway.