Quantitation of specific antibodies bound to feline leukemia virus in the plasma of pet cats.

A method is described for determining levels of circulating immune complexes (CIC) composed of feline leukemia virus (FeLV) antigens and corresponding antibodies in plasma of persistently-infected pet cats. The procedure is based on the ability of high-titered heterologous anti-FeLV serum to chase cat anti-FeLV IgG from dissociated CIC by successfully competing for binding of free antigen. The eluted cat antibody is then collected and quantitated. In a study of cats in the process of clearing persistent FeLV infections, measured levels of FeLV-specific CIC correlated well with fluctuating levels of free FeLV antigen and antibody. The Raji cell assay for CIC in those cats was of comparatively little value in following the clearance of the virus, presumably because that assay does not distinguish between CIC containing viral and those containing non-viral antigens. The method described can be adapted to studies of specific immune complexes associated with a variety of syndromes, provided that the antigen eliciting the immune response is known.

Selective removal of antigen-complexed IgG from cat plasma by adsorption onto a protein A-silica matrix.

The binding of normal cat IgG, heat-aggregated cat IgG and specific immune complexes (IC) containing cat IgG to a silica matrix containing covalently bound Staphylococcus aureus protein A was evaluated. The amounts of serum relative to protein A-silica, the flow rates and the perfusion times were representative of those existing when protein A-silica columns are used for therapeutic extracorporeal immunoadsorption of IgG and IC from humans and animals. When cat IgG was present in a large excess, approximately one molecule was bound to the matrix per molecule of solid-phase protein A with a KA of 1.5 X 10(6) 1/mol. Aggregated and immune complexed IgG bound to the matrix with relatively higher affinity. IC prepared in vitro between the purified envelope glycoprotein of the feline leukemia virus (FeLV gp70) and affinity-purified cat antibodies bound to the matrix even though normal IgG was present in greater than 10,000-fold excess. Once bound, IC were not eluted from columns upon further perfusion with normal serum. However, bound IgG was eluted from columns by further perfusion of normal serum or IC. IC were at least five-fold more efficient than normal IgG in exerting this effect. The results suggest that protein A-silica columns can be used for preferential removal of IC from plasma in a clinical or experimental setting.

Treatment of feline leukemia and reversal of FeLV by ex vivo removal of IgG: a preliminary report.

Cats that were spontaneously infected with feline leukemia virus (FeLv) were treated with a combination of low-dose irradiation and extracorporeal immunosorption using formalin and heat-fixed S. aureus as a non-specific immunosorbent to remove plasma IgG and immune complexes. The treatment resulted in reduction of circulating lymphoblasts within two weeks and clinical improvement of three of the five animals. A reversal of the FeLV status is reported in five of five cats. Two of the five cats remain FeLV negative and completely tumor free seven and eight months post-therapy at the time of writing (July 1979). A third cat returned to an FeLV positive state but remained tumor free for 24 weeks. Another cat responded to the therapy by reduction of lymphoblasts and became FeLV negative but died of a hemorrhage during an immunosorption. The last cat's status was FeLV positive, then FeLV negative, and finally FeLV positive again. He died 20 weeks after initiation of therapy. During the treatment there was a weight gain in the three cats responding by tumor regression. The results are discussed in terms of a removal of some type of immunoinhibiting factors such as antigen-antibody complexes or suppressor molecules.