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Effect of Inoculation with Neuraminidase-treated Tumor Cells on Macrophage Cytotoxicity in Vitro¹

Department of Anatomy and MCV-VCU Cancer Center, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298

Adherent peritoneal exudate cells (PM) and macrophages isolated from tumor tissue from C57BL/6J x DBA/2J F₁ mice previously inoculated with tumor cells treated with Vibrio cholerae neuraminidase and mitomycin C were studied for their cytotoxic effect on established tumor cell monolayers by time-lapse cinematography. The two murine pulmonary tumors studied were the spontaneous Lewis lung tumor and a 3-methylcholanthrene-induced squamous cell carcinoma. Tumor cells incubated alone, with PM from untreated mice, and with macrophages isolated from tumor tissue from untreated or treated mice grew to confluence by 24 hr of coincubation and displayed no observable detrimental change by 96 hr. Cytotoxic PM were isolated from tumor-bearing mice that had received a s.c. inoculation of tumor cells treated with both V. cholerae neuraminidase and mitomycin C and from tumor-free animals (a) inoculated i.p. with tumor cells treated simultaneously with V. cholerae neuraminidase and mitomycin C or (b) administered tumor cells treated with mitomycin C alone. PM isolated from mice within 3 days of inoculation with treated tumor cells killed cultures of the identical tumor by 36 hr, but they did not kill cultures of another tumor until 48 hr of coincubation. If PM were isolated 18 days following the final inoculation of treated tumor cells, they killed cultures of the same tumor by 36 hr, but they had no cytotoxic effect upon those of another tumor. PM from mice treated with Bacillus Calmette-Guérin or B. Calmette-Guérin cell walls killed tumor cells in culture by 48 hr. It is concluded that specifically cytotoxic macrophages kill tumor cells by 36 hr, whereas nonspecifically cytotoxic macrophages do not kill until 48 hr of coincubation.

¹ Aided by Grant IN-105C from the American Cancer Society.

² To whom requests for reprints should be addressed. This work was submitted in partial fulfillment of requirements for the Ph.D. degree in the School of Basic Sciences and Graduate Studies, Medical College of Virginia-Virginia Commonweath University, Richmond, Va.

Received 1/16/78. Accepted 5/ 8/78.