Properties of hybridomas generated by spleen cells of IL-4- and IL-13-knockout mice.
Interleukin-4 (IL-4)- and IL-13-knockout mice were immunized with murine recombinant IL-4 or IL-13, and spleen cells were fused with P3X63-Ag8.653 myeloma cells. Selection of the antigen-positive hybridomas was fulfilled in the presence of IL-6 containing thymic stroma cell supernatant (TSS). All of the selected anti- IL-4- and anti-IL-13-specific hybridoma clones (eight and 10, respectively) required the presence of TSS (0.5-2.5%) for their cloning, stable growth in large-scale cultures, and production of monoclonal antibodies (MAbs). Several of the anti-IL-4-specific clones were adapted to growth without TSS. However, the loss of antibody-secreting capacity in the process of adaptation to TSS-free growth was detected. The data demonstrate that cytokine-deficient mice technology can be used for generation of MAbs to autologous cytokines.
Shichkin
Valentin P
VP
Division of Immunology, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA. shvpmail@yahoo.com
Spivak
Nikolai Y
NY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D003470:Culture Media; D005240:Feasibility Studies; D005260:Female; D006825:Hybridomas; D007114:Immunization; D018793:Interleukin-13 / Q000235:genetics* / Q000276:immunology*; D015847:Interleukin-4 / Q000235:genetics* / Q000276:immunology*; D015850:Interleukin-6 / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D011994:Recombinant Proteins / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D017154:Stromal Cells; D013950:Thymus Gland / Q000166:cytology
D000911:Antibodies, Monoclonal; D003470:Culture Media; D018793:Interleukin-13; D015850:Interleukin-6; D011994:Recombinant Proteins; D015847:Interleukin-4