Models for lymphoma.
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.
Biragyn
Arya
A
National Cancer Institute, Frederick, Maryland.
Kwak
Larry W
LW
National Cancer Institute, Frederick, Maryland.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007167:Immunotherapy; D008223:Lymphoma / Q000517:prevention & control*; D051379:Mice; D009368:Neoplasm Transplantation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000276:immunology; D019444:Vaccines, DNA
D000888:Antibodies, Anti-Idiotypic; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin