Antibodies against human lymphokines: I. Methods for induction of antibodies capable of neutralizing stable (α) and unstable (β) lymphotoxins released in vitro by activated human lymphocytes

Abstract

Various methods were employed to induce antibodies in rabbits that were capable of neutralizing different families of lymphotoxins (LT). Both stable (α-LT) and unstable (β-LT) molecules, released by activated human lymphocytes in vitro, were neutralized. The different LT families were first separated into their respective groups by physical-chemical methods. Immunization with small quantities of antigen yielded a high percentage of responder animals. Techniques were developed for eliciting α-LT antibodies using as little as 2–3 ml of a cell-free supernatant. The situation was more difficult, however, when the unstable β-LT molecules were employed as antigens. We found that because of the low concentration and lability of β-LT in supernatants, the immunizing dose had to be: a) handled rapidly, b) larger than that used with the α-LT, and c) injected at closer intervals and over a longer immunization protocol. Physical-chemical studies supperted the concept that the LT neutralizing activity in the immune serum was immunoglobulin.