Effective blocking of natural cytotoxicity of young rabbit serum on murine thymocytes by high concentration of glucose in complement-dependent cytotoxicity method.
By an accidental observation, the natural cytotoxicity of 21-day-old rabbit sera to murine thymocytes was found to be almost completely inhibitable by 4.5 mg/ml glucose. This enabled us to use unabsorbed young rabbit serum as a source of complement for antibody-mediated cytotoxic treatment of murine thymocytes. This glucose-sensitive natural cytotoxic activity was recovered in the immunoglobulin fraction purified on an affinity column, and was dependent on complement activity. The unabsorbed 21-day-old rabbit sera had much higher complement activity than those from 14-day-old rabbits and commercially available low background complement. Indeed, complete depletion of CD4+ cells from murine thymocytes and splenocytes was achieved using unabsorbed 21-day rabbit serum as a source of complement and a monoclonal antibody GK1.5 which has been claimed to be inefficient in complement-dependent killing. Owing to the increase in complement titer and circulating blood volume 14-21 days after birth, the use of 21-day-old rabbit sera assures a much more abundant supply of complement for the treatment of murine lymphocytes.
Park
S Y
SY
Laboratory of Cellular Immunology, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Kojima
M
M
Suzuki
H
H
Shinohara
N
N
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000187:drug effects*; D002846:Chromatography, Affinity; D003165:Complement System Proteins / Q000502:physiology*; D006224:Cricetinae; D003601:Cytotoxicity Tests, Immunologic; D004306:Dose-Response Relationship, Immunologic; D005947:Glucose / Q000494:pharmacology*; D007106:Immune Sera / Q000276:immunology*; D007518:Isoantibodies / Q000276:immunology; D008212:Lymphocyte Depletion; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D009005:Monosaccharides / Q000494:pharmacology; D011817:Rabbits / Q000276:immunology; D051381:Rats
D007106:Immune Sera; D007518:Isoantibodies; D009005:Monosaccharides; C035678:anti-Thy antibody; D003165:Complement System Proteins; D005947:Glucose