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Open Access Publications from the University of California

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

  • Author(s): Lewis, JE
  • Carmack, CE
  • Yamamoto, R
  • Granger, GA
  • et al.

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. © 1977.

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