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Distribution of Radiolabeled Alloantibodies in Mice Bearing 3-Methylcholanthrene-induced Sarcomas¹

Department of Therapeutic Radiology [D. J. B., P. C. W.] and Division of Biometry [E. A. J.], University of Minnesota, Minneapolis, Minnesota 55455

The distribution of purified ¹²⁵I-labeled alloantibodies, prepared from the serum of DBA/2J mice obtained after immunization with C3H/HeJ spleen cells, was studied in immunosuppressed DBA/2J mice bearing either allogeneic C3H/HeJ 3-methylcholanthrene sarcomas growing s.c. or syngeneic SaD2 3-methylcholanthrene sarcomas. Once purified radiolabeled antibody was isolated from ¹²⁵I-labeled immune -globulin by a single adsorption onto C3H/HeJ RBC and elution from stroma prepared from these cells, by using 0.1 M glycine buffer (pH 3.0). Twice-purified alloantibody was then produced by Bio-Gel P-200 or Sephadex G-200 gel filtration chromatography or DEAE A-50 ion-exchange chromatography. In vitro, such purified antibodies bound specifically to C3H/HeJ RBC. In vivo, they localized to a significant extent following i.p. injection, preferentially in C3H/HeJ 3-methylcholanthrene sarcomas (4.4 to 8.9% of the injected dose per g of tumor equal to 1% of mouse weight), with mean tumor/blood ratios of 4.0 to 7.8, at 24 or 48 hr after injection. The percentage of injected dose localized in tumor and the tumor/blood ratio did not show significant differences with respect to time or method of antibody purification. Normal tissue/blood ratios in C3H/HeJ or SaD2 sarcoma-bearing mice were less than 0.9. The tumor/blood ratios in SaD2 sarcomas were approximately 0.6. Injection of ¹³¹I-labeled normal DBA/2J -globulin resulted in normal tissue/blood and tumor/blood ratios of less than 0.9 in C3H/HeJ tumor-bearing mice.

¹ This work was supported in part by an American Cancer Society Institutional research grant and by National Cancer Institute Grant CA 15548.

² To whom requests for reprints should be addressed, at Box 494, University Hospitals, 420 Delaware Street, S.E., Minneapolis, Minn. 55455.

Received 12/27/78. Accepted 5/24/79.