In vitro immunocompetence of two compounds isolated from Polygala tenuifolia and development of resistance against grass carp reovirus (GCRV) and Dactylogyrus intermedius in respective host.

The present study was undertaken to isolate some compounds from methanol extract of Polygala tenuifolia and evaluate their immunostimulatory properties and antiviral activity using grass carp Ctenopharyngodon idella kidney (CIK) cells and GCRV. By applying insecticidal bioassay-guided, chromatography techniques and successive recrystallization, two purified compounds were obtained. The changes of expression of selected immune genes (Mx1, IL-1ß, TNFα, MyD88 and IgM) in C. idella kidney cell lines were evaluated after exposure to these isolated compounds. The results showed that compound 1 and 2 up-regulated to varying degrees of Mx1, IL-1ß, TNFα, and MyD88 in C. idella kidney cells. WST-8 kit assay verified the two compounds has no toxic effects on CIK cell, and furthermore, have in vitro antivirus activity. Especially, that there is keeping 79% cell viability when exposure to compound 2 (100 mg L(-1)). According to in vivo insecticidal assays against Dactylogyrus intermedius, compound 2 exhibited higher efficacy than compound 1, which was found to be 87.2% effective at the concentrations of 5 mg L(-1) and safe to goldfish (Carassius auratus). Besides, the purified compounds were identified by spectral data as: (1) 1,5-Anhydro-D-glucitol and (2) 3,4,5-trimethoxy cinnamic acid. Overall, the results indicate that bath administration of these compounds modulates the immune related genes in C. idella kidney cells and to some extent, eliminate the virus and parasitic infections.

Antigen-enhanced glucosamine incorporation by peritoneal macrophages in cell-mediated hypersensitivity. I. Studies on biology and mechanism.

The interaction between sensitized lymphocytes and specific antigen occurring in classic delayed hypersensitivity causes guinea pig peritoneal macrophages to incorporate increased amounts of glucosamine into TCA precipitable, membrane-associated, cell surface material. Antigen-induced stimulation of glucosamine also occurred in peritoneal exudate cells (PEC) isolated from animals primed for cutaneous basophil hypersensitivity with certain strong antigens (KLH, vaccinia virus) in incomplete Freund's adjuvant (IFA), and lymphocytes from such animals elaborated MIF when cultured with specific antigen. Thus, the use of complete Freund's adjuvant is not obligatory for the induction of sensitized lymphocytes capable of secreting MIF or stimulating macrophage glucosamine incorporation; however, the potency of the immunogen employed is a critical variable since lymphocytes from animals primed with weaker antigens (HSA, BGG) in IFA did not have these capabilities. Significantly enhanced incorporation of radioactive glucosamine by macrophages occurred when normal PEC were cultured in lymphokine-containing supernatants, but the magnitude of incorporation was smaller than that of sensitized PEC stimulated by antigen. The final 24 hr of macrophage culture was critically important because lymphokines were equally effective in promoting glucosamine incorporation when present for only this interval. The kinetics of this response are thus very similar to those reported for macrophage "activation". The mechanism by which sensitized lymphocytes and their products stimulate glucosamine incorporation is not established, but at least part of the increment may be attributed to enhanced transport of glucosamine across the macrophage plasma membrane. The plant lectins Con A and PHA stimulated unsensitized plastic-adherent cells to increased glucosamine in corporation and exerted a further additive stimulation on sensitized PEC when nonadherent sensitized lymphocytes were present. It is likely that these mitogens stimulate glucosamine incorporation by two distinct mechanisms, one involving sensitized nonadherent lymphocytes and a second involving only adherent cells (macrophages and/or plastic adherent lymphocytes.