Models for Lymphoma

Abstract

This unit describes experimental procedures for development of therapeutic vaccines, particularly “second‐generation” recombinant vaccines. Specifically, a general procedure for handling and culturing lymphoma cell lines in vitro and their subsequent challenge into syngeneic mice is described. Several protocols describe the production of various idiotype (Id)‐based or cellular lymphoma vaccine formulations. In particular, the novel approach of rendering nonimmunogenic lymphoma‐derived scFv or Id immunogenic by fusing it with a chemokine moiety is described. As an alternative, a protocol for expression and purification of these chemokine‐fusion proteins from E. coli is included. A general procedure is included for cloning of cytokine genes, for example, murine GM‐CSF, in A20 lymphoma cells. Support protocols are presented for chemically conjugating intact Ig protein with KLH to produce a prototype protein vaccine and using the Helios Gene Gun System to immunize mice with recombinant DNA tumor vaccines.