Detection of immune complexes by their complement-dependent binding to bovine conglutinin: Technical aspects of a solid-phase immunoenzyme microassay and its application to normal and pathological sera

Abstract

A micromethod for the solid-phase conglutinin binding assay (Con BA) for the detection of circulating immune complexes (CIC) is described, in which the use of microplates, glucose oxidase-coupled anti-human immunoglobulins and automatic OD recorder contributed to the speed and low cost of this reproducible and sensitive test. The Con BA values of a large population of healthy blood donors had a wide distribution which in our series of 189 appeared to be trimodal. The Con BA values clearly reflected the levels of the main Ig classes (M, G, and A). This was confirmed by follow up of 6 individuals with high initial Con BA values. For 4 of them, a parallel decrease in Con BA value and IgG concentration was observed. In the 2 others, the sustained high level of Con BA remained unexplained. Complement components and activity of these sera were within normal limits. Because of fluctuation in the aggregated human gamma-globulins (AHG) reference curve, expression of results was based upon the standard deviation of 6 normal sera. In view of the above results, these sera were selected from those with the lowest Con BA values. On the basis that Con BA and C1q BA may detect CIC differing in their ag/ab ratio, sera from rheumatoid arthritis and chronic active B hepatitis patients with or without conventional rheumatoid factors (RF) were analyzed by both Con BA and C1q BA. RF-containing sera, likely to contain CIC in antigen excess, contributed exclusively to the highest C1q BA and lowest Con BA values. In contrast C1q BA-Con BA+ sera were preferentially RF negative. It is proposed that the complexes thus detected may be of idiotype-anti-idiotype nature.