High titre anticytokine antibodies obtained by intralymphnode immunization with low amounts of antigen

Intralymphnode immunization was performed on rabbits to obtain anticytokine antibodies using low or very low amounts of the following purified cytokines: CSF-1 (or M-CSF: 10, 2 or 0.2 microgram/injection), GM-CSF (10 micrograms/injection), IL-2 (10 micrograms/injection) and HILDA/LIF (10 micrograms for the first injection and 5 micrograms/injection for boosts). This technique is easily performed by dissection of the popliteal lymphnode. Specific high titre antibodies were obtained after the first or second boost for antigen doses between 10 (for all cytokines tested) and 0.2 microgram (for CSF-1) per injection. In most cases, these antibodies could be used for immunoprecipitation, competition assays, dot immunoblotting, neutralization of biological activity and receptor binding inhibition. Some applications show that these tools are useful for cytokine research projects. For newly identified cytokines available in limited amounts, this method of obtaining specific polyclonal antibodies is an interesting alternative to the expensive, time-consuming and technically more demanding monoclonal antibody method.