Open AccessAntigen-specific purification of blocking factors from sera of mice with chemically induced tumors.
Author(s) -
Gerald T. Nepom,
Ingegerd Hellström,
Karl Erik Hellström
Publication year - 1977
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.74.10.4605
Subject(s) - immune system , antigen , concanavalin a , antibody , microbiology and biotechnology , affinity chromatography , glycoprotein , chemistry , blocking (statistics) , homologous chromosome , biology , immunology , biochemistry , in vitro , enzyme , statistics , mathematics , gene
Serum from mice with growing tumors can prevent ("block") the destruction of tumor cells by immune lymphocytes, as measured in a microcytotoxicity assay. Factors responsible for this blocking activity were purified by binding to immune adsorbents that had been prepared from antibodies obtained by immunizing BALB/c mice to the homologous tumors. Two transplantable BALB/c sarcoma lines with individually different tumor-specific transplantation antigens were studied in parallel. The original tumor-specific blocking activity was recovered by elution of the immune adsorbents; that is, (i) eluates blocked the reduction of surviving tumor cell targets by immune lymphocytes only if the tumor specificity, and (ii) immune adsorbent columns prepared from tumor-immune sera recognized the purified blocking fractions in a tumor-specific fashion, indicating that a portion of the humoral response in the immune mice was directed against a factor that was individually distinct for each tumor. Absorption of eluates with the homologous tumor cells removed their blocking activity, indicating that the blocking factors have antigen-binding properties. Blocking activity in the purified fractions resided in molecules presumptively identified as glycoproteins by affinity chromatography on concanavalin A-Sepharose.