IgE receptors (FcR epsilon) modifications. II. Effect of regulator soluble factors of human IgE synthesis isolated by affinity chromatography.
Tabar AI, Sanz ML, Oehling A.
Departamento de Alergología, Facultad de Medicina, Universidad de Navarra, Pamplona, Spain
In a series of works done on humans patterned after animal experimental models, we created an in vitro model which allowed us to produce soluble IgE regulator factors. We observed its biological activity in IgE synthesis and constructed changes that protein fractions from affinity chromatography (fractions enriched in soluble regulator IgE factors) exercised on IgE receptor expression. We then decided to determine the molecular weight of each one of the active protein fractions.
We performed electrophoresis on the gel of polyacrylamide (Weber-Osborn technique). Protein bands were processed, with the aim of isolating the gel and quantifying the biological activity. The protein fractions obtained from the chromatographies with affinity to CON-A sepharose were subjected to electrophoresis in polyacrylamide gel, with the aim of obtaining molecular weights of proteins comprising it. Using rosette technique, there were no changes seen in Fc epsilon expression just as what happened in the experiment with material fractionated by chromatography. However, these changes are produced and are statistically significant using the immunofluorescence technique. The highest inhibition of rosettes or immunofluorescence is that of the protein band of 10,000 daltons extracted in NaCl and that of 20,000 daltons extracted in PBS. Both effects are similar.
The inhibition reached by the 20,000 dalton band in NaCl is less although there are no significant differences. The material extracted in PBS of the band between 6.9 and 7.2 mm of gel (G4) did not have any changes. None of the proteins added to the culture medium had any effects of receptor gamma expression. The average values of the receptor expression was similar to basal levels.
CITATION Allergol Immunopathol (Madr). 1990 Jan-Feb;18(1):13-7