Induction and specificity of syngeneic antiserum against rauscher murine leukemia virus-induced leukemia, RBL-5
✍ Scribed by Olusola Alaba; L. W. Law
- Publisher
- John Wiley and Sons
- Year
- 1980
- Tongue
- French
- Weight
- 753 KB
- Volume
- 26
- Category
- Article
- ISSN
- 0020-7136
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✦ Synopsis
Abstract
Plasma membranes purified from RBL‐5 leukemia cells and solubilized with Na deoxycholate were fractionated on an Ultrogel AcA34 column. Fractions containing most of the tumor‐rejection activity but low amounts of gp70 and p30 were consolidated and used to hyperimmunize semisyngeneic, CBF~1~, hybrid mice. Using the complement‐dependent cytotoxicity assay, we determined the antibody titers against RBL‐5 and several other leukemic cells. Only RBL‐5 and EL‐4 tumor cells were lysed. Similar results were obtained when the same tumor cells were used to absorb the syngeneic antiserum and the residual cytotoxic titer determined on RBL‐5 target cells. Coupling of the gamma‐globulins from this antiserum to Sepharose 4B facilitated the isolation of an antigen which inhibited the C'‐dependent lysis of RBL‐5 tumor cells and, when used for immunization, protected the recipient mice against a subcutaneous challenge of RBL‐5 leukemia cells. By means of discgel electrophoresis, in the presence of SDS and 2‐mer‐captoethanol, two polypeptide chains with molecular weights of 46,000 and 21,000 were resolved. The binding of the antigen to a lens culinaris lentil lectin column and its subsequent elution with α‐methyl mannoside suggests that the specific tumor antigen is a glycoprotein.