Fourier-transform infrared spectroscopy of human IgG Fc fragments, which are a promising drug for the treatment of autoimmune diseases

Abstract

IgG Fc fragments that expose regulatory rheumatoid factor epitopes (regRF epitopes) have emerged as a promising immunosuppressive drug. Immunization of rats with such Fc fragments reduced symptoms of experimental autoimmune diseases. The immunosuppressive effect of Fc fragments is based on stimulating the production of regRF, which kills activated CD4 T lymphocytes. The formation of regRF epitopes on Fc fragments was previously shown to be associated with a reduction in disulfide bonds in the fragments’ hinge region. However, the structure of Fc fragments that bear regRF epitopes remained largely unclear. Infrared spectra were compared for lyophilized Fc fragments displaying regRF epitopes and Fc fragments without such epitopes. FTIR spectroscopy found no differences in the amide I, amide II, and amide III bands, indicating that there are no distinctive features in the secondary structure of Fc fragments bearing regRF epitopes. The distinctive feature of Fc fragments bearing regRF epitopes, irrespective of whether the free SH groups in the hinge were preserved or lost after lyophilization, is the presence of a band or a fine structure in the region containing the bending vibrations of the SH groups. Furthermore, the Fc fragments with regRF epitopes differ from those without in that they have a band in the absorption region of aromatic amino acid rings. Taken together, these facts suggest that the appearance of regRF epitopes results from changes in the tertiary structure of the hinge and the domains that occur when the hinge is reduced, and they also indicate that these conformational changes are resistant to subsequent changes in the state of cysteine residues in the hinge. Bands in the regions of aromatic amino acids and the bending vibrations of SH groups are markers of the presence of regRF epitopes on IgG Fc fragments. FTIR spectroscopy can be used to detect these epitopes.