Changes in the urinary protein pattern in SDS polyacrylamide gel electrophoresis in normal and hypertensive pregnancies.

The urinary protein pattern was determined in 46 healthy male and female subjects, 64 patients with an uncomplicated course of pregnancy and 88 hypertensive pregnant women by use of a special urine preparation and a modified SDS-polyacrylamide gel electrophoresis (SDS-PAGE). There were no differences in the protein electrophoresis pattern between the control subjects, independent of sex and age, and the pregnant women. The number of protein bands did not change in the course of pregnancy and in the post partum period. In both groups, an intensively stained protein band with an apparent molecular weight of 105 kD was detected. In 71 of the 88 hypertensive pregnant women we found a marked reduction in intensity or complete disappearance of the 105 kD protein band. Follow-up analysis in 30 of these pregnant women showed, that in 24 cases disappearance of the 105 kD band occurred simultaneously with and in 6 women before clinical manifestation of the disease. There were no differences in the protein electrophoresis pattern between patients with preexisting renal or hypertensive disease and hypertensive women without a complicated history. In 49 of the 52 hypertensive pregnant women complete reappearance of the 105 kD band was observed 2 to 14 days after delivery. By using silver staining and Western blot, the 105 kD band was identified as Tamm-Horsfall protein. Our findings may reflect a transitory tubular dysfunction in cases of preeclampsia and support the hypothesis of an immunological pathogenesis of the disease

IFN-γ Enhances T_H1 Polarisation of Monocyte-derived Dendritic Cells Matured with Clinical-greade Cytokines Using Serum-free Conditions

Using serum-free conditions, human monocyte-derived dendritic cells (MoDCs) tend to mature insufficiently in a TH1-polarizing direction under approved and standardized clinical conditions. However, for the initiation of an efficient tumour antigen-specific cytotoxic T-cell response, the induction of a distinct TH1 response is favourable. Therefore, to improve TH1 polarisation, the influence of interferon-γ (IFN-γ) on the maturation of MoDCs was investigated with clinical-grade cytokines or lipopolysaccharide (LPS) in serum-free medium focusing on the viability, phenotypic characteristics, cytokine profile and restimulating capacities. As in previous research, we confirmed that in respect of viability and phenotypic characteristics, cytokine cocktails consisting of tumour necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin (PG) E2, mature MoDCs most efficiently. However, these cytokine-matured MoDCs secreted relatively high levels of IL-10 and only low levels of IL-12p70. Remarkably, if IFN-γ was added, significantly lower levels of IL-10 concomitant with higher levels of IL-12p70 could be detected. Pretreatment with IFN-γ did not improve the phenotypic characteristics nor the TH1 polarisation of MoDCs. Nevertheless, MoDCs matured with clinical-grade cytokines and IFN-γ could be re-stimulated most effectively with IFN-γ. In conclusion, our work demonstrates that addition of INF-γ to clinical-grade cytokine cocktails readily matures MoDCs and enhances their TH1 polarisation efficiently under serum-free conditions