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R. A m a r o w i c z , M . K a r a m a c ' , B. R u d n i c k a a n d E . C i s k a *Glucopyranosyl sinapate was obtained from rapeseeds using column chromatography on Sephadex LH-20 and RP-18 semipreparative HPLC. TLC analysis of this compound, sinapic acid and phenolic compounds fractions from column chromatography were conducted on plates coated with silica gel (standard and HPTLC), with cellulose and on RP-TLC plates. Eleven various solvent systems were applied to develop chromatograms. HPTL plates were found more efficient than those coated with standard silica gel. Compared to the latter cellulose ensured worse results of TLC analysis. The best developing system for plates coated with silica gel was benzene-methanol-acetic acid (90:16:8). Separation of the compounds analysed using RP-TLC depended on octadodecylsilanization degree of silica gel on TLC plate.

Advanced glycation endproducts (AGEs) are created during the spontaneous reaction that occurs in the body and during protein processing. AGEs cause chronic hyperglycemia, contribute to vascular and renal dysfunction, and are potential antigens. Our recent work showed the mucosal immune system responses to naive, hydrolysed and glycated ovalbumin (OVA‐H, ‐G) gavages to Balb/C mice. There were no differences between groups in anti‐OVA IgG, IgA serum and fecal titers. Cultured B lymphocytes did not show differences in proliferation during stimulation with OVA; OVA‐G/H. B cell ELISpot showed high expression of specific IgA in inductive tissues from OVA‐G and ‐H group. FACS analysis showed significantly expressed CD4+ in mesenteric LN and PP in OVA‐H. Significant levels of CD4+TGFβ were found in OVA‐G group in PP, MLN, iLP, and in OVA‐H group in SPL, PP and iLP. Sorted CD4+ populations were cultured with OVA, OVA‐H, OVA‐G. It was found significant level of TGF‐β and IL‐4 in culture supernatant. Adding OVA‐G to the culture significantly affected the levels of IL‐17 in culture supernatant. This expression suggests that AGEs could induce T cell population Th17, which is recognized as a mediator in autoimmune pathology. The presence of strong TGF‐β responses, together with significant levels of Il4, may indicate of expression Th3 cells, which provides help for IgA and has suppressive properties for some immune cells. All groups showed positive DTH test responses after 48h. The presented protein modifications modulated mucosal immune response for OVA. Using low dose antigen immunization and modification could be the convenient way to modulate immune responses to protect against food allergies and other diseases. This work was supported by MSHE in Poland, Grant N312 32/0159.