It has been previously reported that circulating anti-heat-shock-protein (Hsp) antibody levels are elevated in cardiovascular disorders. concentration by nephelometry. Plasma malondialdehyde levels were measured by the thiobarbituric-acid-based colorimetric assay. For statistical analyses, nonparametric methods were applied. Anti-Hsp60, anti-Hsp65, and anti-Hsp70 antibodies were detected in all of our serum samples. There were no significant differences in serum anti-Hsp60, anti-Hsp65, and anti-Hsp70 antibody levels between the control and preeclamptic groups. Serum levels of Hsp70 and CRP, as well as plasma levels of VWF antigen, fibronectin, and malondialdehyde, were significantly higher in preeclamptic patients than in normotensive healthy pregnant women. Serum anti-Hsp60 antibody levels showed significant correlations with serum anti-Hsp65 antibody levels both in the control and the preeclamptic groups (Spearman for 10?min. The aliquots of serum and plasma were stored at ?80C until the analyses were performed. Laboratory methods Anti-Hsp60 and anti-Hsp65 immunoglobulin G (IgG) levels were measured by enzyme-linked immunosorbent assay (ELISA), as explained previously (Prohaszka et al. 1999, 2001). In brief, plates were coated with 0.1?g per well human Hsp60 (recombinant human Hsp60, StressGen, SPP-740) or Hsp65 (recombinant Hsp65, Braunschweig, Germany). After washing and blocking (phosphate-buffered saline (PBS), 0.5% gelatine), the wells were incubated with 100?l of serum samples diluted 1:500 (PBS, 0.5% gelatine, 0.05% Tween 20). Bound anti-Hsp60/65 antibodies were detected by antihuman IgG peroxidase-labeled antibodies (Sigma, St. Louis, MO, USA) and test. As the continuous variables were not normally distributed, nonparametric statistical methods were used. To compare continuous variables between two groups, the MannCWhitney test was applied. The Fisher exact and Pearson test (categorical variables). Serum anti-Hsp60 antibody levels showed significant correlations with serum anti-Hsp65 antibody levels both in the control and the preeclamptic groups (Spearman R?=?0.55 and 0.59; p?PXD101 PXD101 parameters of normotensive healthy pregnant women and serum anti-Hsp antibody levels Table?4 Correlation coefficients between clinical characteristics and laboratory parameters of preeclamptic patients and serum anti-Hsp antibody levels Discussion In this study, we reported the presence of anti-Hsp60, anti-Hsp65, and anti-Hsp70 antibodies in the peripheral blood circulation of healthy pregnant women. However, neither were serum levels of anti-heat-shock-protein antibodies increased nor were these antibodies related to systemic inflammation, oxidative stress, and endothelial activation/injury in preeclampsia. Our findings that anti-Hsp60 and anti-Hsp70 antibodies were present in all of our serum samples are in agreement with the role of these antibodies as naturally occurring autoantibodies. Such antibodies are important for initial defense against invading pathogens (Lutz and Miescher 2008). Indeed, anti-Hsp70 antibody was detected in midtrimester amniotic fluid and its level correlated with intra-amniotic concentrations of antimicrobial immune mediators (Gelber et al. 2007). The lack of correlation between serum Rabbit Polyclonal to p70 S6 Kinase beta. Hsp70 and anti-Hsp70 antibody levels is consistent with earlier observations in nonpregnant women (Pockley et al. 1998; Rea et al. 2001). The strong positive correlation between anti-Hsp60 and anti-Hsp65 levels found in our study groups might reflect the presence of cross-reactive epitopes on the target molecules. Given the ubiquitous nature and the high degree of sequence homology between microbial and mammalian forms of warmth shock proteins, these molecules could act as harmful autoantigens and may provide a link between contamination and autoimmunity through molecular mimicry (Lamb et al. 1989). Most of the known risk factors of atherosclerosis (e.g., contamination, hemodynamic stress (hypertension), oxidative stress) are known to induce warmth shock protein expression in and/or release from your vessel wall. Cross-reactive anti-heat-shock-protein antibodies and T cells can damage vascular tissues overexpressing warmth shock proteins, contributing to the development of atherosclerosis (Mandal et al. 2004). Additionally, immune sensitization to human Hsp60, possibly developed as a consequence of contamination, may adversely impact pregnancy end result (Witkin et al. 1994, 1996). Furthermore, the presence of anti-Hsp60 and anti-Hsp70 antibodies in the serum and formation of Hsp60- and Hsp70-immune complexes in the placenta were associated with preterm birth (Ziegert et al. 1999). Recently, serum anti-Hsp70 levels were found to be significantly elevated at 16? weeks of gestation in women who later gave birth to babies with birth defects, suggesting.