Purification of a 92 kDa human immunostimulating glycoprotein obtained from the Tamm-Horsfall glycoprotein

Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

“…To exclude contaminating molecules other than LPS, the standard THP preparation obtained by repeated NaCl precipitation from urine was further purified by 2 sequential procedures taking advantage of the fact that THP forms a gel when monovalent ions above 60 mM are present. After induction of gel formation by 70 mM sodium phosphate, THP was purified by ultracentrifugation (23). The pellet fraction containing THP was fully active on C57BL/6 splenocytes for TNF-α and IL-6 production ( Figure 2D and data not shown).…”

Tamm-Horsfall glycoprotein links innate immune cell activation with adaptive immunity via a Toll-like receptor-4–dependent mechanism

“…After induction of gel formation by 70 mM sodium phosphate, THP was purified by ultracentrifugation (23). The pellet fraction containing THP was fully active on C57BL/6 splenocytes for TNF-α and IL-6 production ( Figure 2D and data not shown).…”

“…Mouse THP was purified from sterile urine from female C57BL/6 mice by precipitation in 0.58 M NaCl as described (46) or by dialysis against PBS using a 50-kDa Spectra/Por Biotech RC dialysis membrane from Spectrum Laboratories Inc. To further purify THP, the THP precipitation was diluted in sodium phosphate buffer, pH 6.3, to a concentration of 70 mM to induce THP gel formation and was incubated for 1 hour at room temperature. Afterward, the sample was ultracentrifuged at 109,000 g for 30 minutes at 10°C, and the pellet was dissolved in H2O and dialyzed against water (23). The pellet fraction contained the majority of THP as assessed by SDS-PAGE.…”

Tamm–Horsfall protein isolated from urine of pregnant and non‐pregnant women has similar oligosaccharides

“…Recently, many studies have demonstrated that THP/UM plays a central role in the regulation of circulating levels and biological activities of cytokines including IL-1 [5] and TNF [6]. However, it has also been reported that THP/UM does not react with cytokines under physiological conditions [18], and Fontan et al [19] claimed that the immunoregulatory properties of THP were a consequence of its contamination by a 92-kD immunostimulating glycoprotein that is probably a part of THP. There is also evidence that THP is involved in a number of pathological conditions [20] (for references see also Ref.…”