Last year represented a transition in the major focus of my laboratory. Research has progressed from the purification, immunologic characterization, sequencing, and cloning of uromodulin and 85 KD immunorepressive glycoprotein derived from pregnancy urine to now focusing on the characterization of glycosylation patterns responsible for the biologic activity of uromodulin. Utilizing a combination of protease digestion and multiple chromatographic steps, we have purified to homogeneity and structurally characterized by 1H NMR two profoundly biologically active high mannose oligosacharides. Based on these structural observations, alternative sources for active oligosaccharides have been found and include egg white protein, soybean meal, and yeast mannan. These compounds inhibit and enhance the immune response both in vitro and in vivo. Utilizing defined oligosaccharides ranging from 7-11 sugar residues, we have performed a number of structure function studies. These compounds inhibit T cell responsiveness, enhance PGE2 synthesis, induce collagenase release, block hydrogen peroxide formation, activate the hexose monophosphate shunt, bind to IL-l, TNF, and IL-2, and regulate the expression of DR. We have applied for a patent covering the immunoregulatory activity of these structurally unique compounds.
