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Correlation of tumor specific delayed type hypersensitivity reaction and tumor protection to SV40-induced mKSA fibrosarcoma.

Mice immunized by excision of a primary, subcutaneously growing SV40-induced mKSA solid tumor which resisted challenge of homologous tumor cells administered at a contralateral site, were found to develop a specific DTH response to SV40 tumor associated transplantation antigens (TATA). In a two-way criss-cross experiment, this DTH response (assessed by direct challenge) was found to be one-way SV40 specific in that chemically induced, non SV40, MCA tumor failed to elicit a DTH response in mice primed by excision of mKSA tumor. These mice also showed a corresponding one-way specific protection against challenge with live homologous mKSA sarcoma cells. In contrast immunization and challenge of MCA-excised mice with either MCA or mKSA tumor cells, exhibited cross-reactivity in both DTH response and protection against either tumor. Unlike this cross-immunity by the direct challenge method, transfer of "immune" spleen cells from mKSA or MCA excision-primed mice demonstrated a specific DTH response and protection to the original immunizing, homologous but not heterologous tumor. Tumor resistant, DTH-primed mice remained DTH reactive to the primary tumor cells over a period of 4 weeks. Characterization of the splenic T-DTH cells in mice primed by excision of mKSA tumor, indicated a Lyt 1+2+ phenotype of cells conferring both the DTH response and the immune protection against mKSA sarcoma in a local (Winn) adoptive transfer assay, thus reinforcing the correlation between the DTH response and the antitumor protection.

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Comparison of excision versus cryosurgery of an HSV-2-induced fibrosarcoma. I. Survival, extent of metastatic disease and host immunocompetence following surgery.

Cryosurgery and excision were used to treat primary tumours of HSV-2-transformed hamster tumour sublines, and post-operative survival and the extent of metastatic disease were compared in the two groups. An inferior prognosis was observed following cryosurgery although the extent of metastatic disease was similar in both groups. Using this model it would appear that cryosurgery enhances the development of micrometastases rather than affecting the number of cells shed from the primary tumour during surgery. To investigate the underlying causes of the decrease in survival following cryosurgery, in vitro assays were used to monitor host immunocompetence following surgery. The results showed that whilst natural killer cell cytotoxicity was only marginally depressed, mitogen responsiveness and lymphocyte participation in a mixed lymphocyte reaction were severely reduced 3-7 days post-cryosurgery. In parallel with immunosuppression, extensive cell proliferation in the spleen of cryosurgically treated tumour-bearing animals was observed. Histological examination of the spleen demonstrated the presence of large numbers of transformed cells which correlated with the loss of mitogen responsiveness and the ability to participate in a mixed lymphocyte reaction. Further studies (manuscript submitted for publication) have demonstrated that spleen cells from animals whose tumour is treated by cryosurgery are capable of suppressing immunocompetence in vitro, implying they have a role in the uncontrolled growth of micrometastases in vivo.

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Selection of liver-colonizing tumor cells from a murine fibrosarcoma induced by methylcholanthrene.

An experimental tumor model was developed to study the organ preference of malignant tumors. The primary tumor ER 15-P was induced by inoculation of 1 mg methylcholanthrene in 0.1 ml sesame oil into the left femoral muscle of a female C57/Bl6J mouse. The tumor was palpable 100 days after induction. Spontaneous lung metastases were found at autopsy on day 128. Serial IM transplantation of tumor cells from the primary ER 15-P resulted in pulmonary metastases in all male and female mice. After IV injection of tumor cells from ER 15-P to male mice, colonies were found in lungs, thoracic cavity, liver, kidneys and occasionally also adrenals; female mice sometimes had ovarian metastases in addition, but no hepatic metastases. Liver-colonizing tumor cells were selected in male mice as follows: (a) IV injection of tumor cells from primary ER 15-P; (b) removal of tumor cells from liver tumor nodules, reinjection into mesenteric vein; (c) preparation of resulting tumors in the liver, reinjection of these cells through the portal system in one group of mice, and IV administration into tail vein in another group: (d) IM inoculation of tumor cells of the mesenteric passage in the left gastrocnemius muscle of mice prior to IV injection via tail vein in another group. Steps c and d were repeated three times. The procedure resulted in a highly significant decrease of tumor cell colonization to lungs and other organs, and a preferential increase of liver colonization. The liver preference of cell lines thus selected was obvious. Possible mechanisms for the organ preference of malignant tumors are discussed.

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Initiation of a transplantable fibrosarcoma by the synergism of two non-initiators, alpha-tocopherol and soya oil.

When, in the course of an ageing study, alpha-tocopherol (vitamin E), dissolved in soya oil, was given to 22 Balb/c mice once a week subcutaneously for 10 months, it caused the development of vigorously growing fibrosarcomata at the site of the injections in 17 (77.3%) of the animals. The tumors produced in this manner proved eminently transplantable into syngeneic Balb/c hosts, and have been serially transplanted every 3-4 weeks for over 3 years in such recipients, having reached their 44th transplantation cycle at the present time; upon transplantation, they now exhibit a 100% "take" incidence and proliferate extremely rapidly, growing from pin-head size to up to half the weight of a whole recipient mouse within 3 weeks. All fibrosarcomata showed marked mitotic activity, invasion of adjacent tissues and extensive necrotic areas, and they became more undifferentiated after the third transplantation cycle. Neither pure alpha-tocopherol alone nor soya oil alone produced any tumors when given subcutaneously once a week, for 10 months to groups of 22 Balb/c mice each. It is concluded that the two agents alpha-tocopherol and soya oil which proved non-carcinogenic when injected alone, developed a powerful carcinogenic effect when they acted on subcutaneous connective tissue simultaneously. The possible mechanisms of this phenomenon are discussed.

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