Human seminal fibronectin fragmentation patterns and their domain immunoreactivities in leucocytospermic patients

Abstract

The aim of the work was to analyse fibronectin (FN) domain immunoreactivities and profiles of FN fragmentation in seminal plasmas of fertile normozoospermic and infertile leucocytospermic male patients. ELISA with domain-specific monoclonal antibodies and immunoblotting were used in these measurements. Immunoblotting of normal and leucocytospermic seminal plasmas revealed the presence of twelve FN bands of ~70-196kDa with nearly identical FN profiles under reducing and non-reducing conditions. The epitopes of the cell-, fibrin-, collagen-binding FN domains and the extra domain A (EDA) FN segment retained the ability to bind their specific monoclonal antibodies, whereas the fibrin-heparin domain (N-terminal end) and the area around the disulfide bridges (C-terminal end) of the FN polypeptide did not show any reactivities with their respective specific antibodies. The mean values of cell- (338.4±138.4 and 398.3±310mgL(-1)), fibrin- (79.1±38.5 and 145.2±188.8mgL(-1)) and collagen-binding (19±19.8 and 50.9±73.4mgL(-1)) FN domain immunoreactivities and the relative amount of (EDA)FN did not show any significant differences between the normal and leucocytospermic groups. The high values of standard deviations for the FN domain immunoreactivities in the leucocytospermic group probably results from different aetiology of leucocytospermia. The profile of FN fragmentation and alterations of FN domain immunoreactivities in seminal plasma may influence their engagement in the fertilisation process. The analysis of seminal FN molecular status would be helpful for selecting the highest quality spermatozoa for use in assisted reproduction techniques.

Variability of serum IgG sialylation and galactosylation degree in women with advanced endometriosis

“…It should also be mentioned that in lectin-ELISA method, glycan-lectin reaction reflect the reactions that occur in living organisms, including the availability of sugar residues for lectins, which additionally allows to deepen the knowledge about mechanisms of these interactions. The profile and degree of IgG sialylation and galactosylation/agalactosylation was determined using modified solid phase enzyme-linked immunosorbent assay, lectin-ELISA described by us earlier [31][32][33][34] . The method was based on the reactivity of IgG glycan moieties with specific biotinylated lectins: Maackia amurensis agglutinin (MAA, recognizing sialic acid α2,3-linked) and Sambucus nigra agglutinin (SNA, which recognize sialic acid α2,6-linked), Ricinus communis agglutinin I (RCA-I) which detect the terminal galactose; Griffonia simplicifolia lectin II (GSL-II) detecting the terminal GlcNAc (Vector Laboratories Inc., Burlingame, CA, USA).…”

“…Another important aspect was the comparison of native serum IgG glycosylation with glycosylation of IgG isolated from serum, and evaluation the potential diagnostic suitability of glycan analysis when glycoprotein is not isolated earlier from biological fluid. The degree of IgG sialylation and galactosylation/agalactosylation was analysed using a modified solid phase enzyme-linked immunosorbent assay, lectin-ELISA [31][32][33][34] .…”