Method of testing a drug for anti-cancer activity

Abstract

A method for separating tumor cells with lymphotropic metastatic potential from those without lymphotropic metastatic potential in a human carcinoma. Cells of the carcinoma are transplanted in each of a plurality of fresh athymic mice. At least one of the athymic mice which does not develop a palpable tumor at the transplant site is treated to suppress the T-cell independent innate anti-tumor activity of natural killer cells therein. Tumor-forming cells at the transplant site of the treated athymic animal are harvested to obtain a cell line of cells with lymphotropic metastatic potential, which is also tested for the expression of T-lymphocyte associated molecules. Such cells are intimately associated with low or diminished angiogenicity and immunogenicity. The traditional scientific criteria for human cancer cells is re-defined, and therapeutic targets for human cancer cells is re-focused.

1 . A method of testing a drug for anti-cancer activity, the method comprising the steps of:
 a) obtaining a cell line of cells with lymphotropic metastatic potential;   b) injecting a quantity of cells of the cell line into a SCID animal; and   c) after lymph node metastasis occurs in the SCID animal, testing the drug on the SCID animal for anti-cancer activity.   
     
     
         2 . A method according to  claim 1  further comprising testing cells of the cell line for the presence of one or more lymphoid gene products to confirm that the cells of the cell line have lymphotropic metastatic potential. 
     
     
         3 . A method according to  claim 1  further comprising selecting the SCID animal to be a SCID mouse. 
     
     
         4 . A method according to  claim 1  wherein the step of obtaining a cell line of cells with lymphotropic metastatic potential comprises deriving the cell line from a patient's carcinoma to test the suitability of the drug for treatment of the patient's carcinoma. 
     
     
         5 . A method according to  claim 1  wherein the step of obtaining a cell line of cells with lymphotropic metastatic potential comprises the sub-steps of:
 a) transplanting cells of a carcinoma at a transplant site in each of a plurality of T-lymphocyte deficient environments which are adapted to suppress tumor growth of NK-sensitive cells; 
 b) waiting a period of time after the cells are transplanted to determine whether a palpable tumor develops at the transplant site of at least one of the environments; 
 c) after waiting the period of time, treating with antibodies to NK cells at least one of the environments which does not develop a palpable tumor at the transplant site to overcome tumor growth suppression of NK-sensitive tumor cells in the treated environment; and 
 d) harvesting tumor-forming cells at the transplant site of the treated environment to obtain the cell line of cells with lymphotropic metastatic potential. 
 
     
     
         6 . A method according to  claim 5  further comprising selecting the SCID animal to be a SCID mouse. 
     
     
         7 . A method according to  claim 5  further comprising selecting the environments to be athymic animals. 
     
     
         8 . A method according to  claim 5  wherein the step of transplanting cells includes limiting the number of cells being transplanted in each of the environments to prevent overwhelming of the NK cells in the environments respectively. 
     
     
         9 . A method according to  claim 5  further comprising selecting the environments to be athymic mice, and wherein the step of transplanting cells includes transplanting about 2 million of the cells in each of the athymic mice. 
     
     
         10 . A method according to  claim 5  further comprising testing cells of the cell line for the presence of one or more lymphoid gene products to confirm that the cells of the cell line have lymphotropic metastatic potential. 
     
     
         11 . A method according to  claim 5  further comprising cultivating cells of the cell line in vitro, and repeating sub-steps a to d to obtain a more pure cell line of cells with lymphotropic metastatic potential. 
     
     
         12 . A method according to  claim 5  wherein the step of obtaining a cell line of cells with lymphotropic metastatic potential comprises deriving the cell line from a patient's carcinoma to test the suitability of the drug for treatment of the patient's carcinoma. 
     
     
         13 . A drug which is tested in accordance with the method of  claim 1  and which is determined by the testing to have anti-cancer activity. 
     
     
         14 . A method of treatment of a patient who has a carcinoma, the method comprising the steps of:
 a) extracting cells from the carcinoma;   b) obtaining from the extracted cells a cell line of cells with lymphotropic metastatic potential;   c) injecting a quantity of cells of the cell line into a SCID animal;   d) after lymph node metastasis occurs in the SCID animal, testing drugs on the SCID animal for anti-cancer activity until at least one of the drugs is determined to have anti-cancer activity for the carcinoma; and   e) administering to the patient the at least one drug determined to have anti-cancer activity for the carcinoma.   
     
     
         15 . A method according to  claim 14  further comprising selecting the SCID animal to be a SCID mouse. 
     
     
         16 . A method according to  claim 14  wherein the step of obtaining a cell line of cells with lymphotropic metastatic potential comprises the sub-steps of:
 a) transplanting cells of the carcinoma at a transplant site in each of a plurality of T-lymphocyte deficient environments which are adapted to suppress tumor growth of NK-sensitive cells; 
 b) waiting a period of time after the cells are transplanted to determine whether a palpable tumor develops at the transplant site of at least one of the environments; 
 c) after waiting the period of time, treating with antibodies to NK cells at least one of the environments which does not develop a palpable tumor at the transplant site to overcome tumor growth suppression of NK-sensitive tumor cells in the treated environment; and 
 d) harvesting tumor-forming cells at the transplant site of the treated environment to obtain the cell line of cells with lymphotropic metastatic potential. 
 
     
     
         17 . A method according to  claim 16  further comprising selecting the environments to be athymic animals. 
     
     
         18 . A method according to  claim 16  wherein the step of transplanting cells includes limiting the number of cells being transplanted in each of the environments to prevent overwhelming of the NK cells in the environments respectively. 
     
     
         19 . A method according to  claim 16  further comprising selecting the environments to be athymic mice, and wherein the step of transplanting cells includes transplanting about 2 million of the cells in each of the athymic mice. 
     
     
         20 . A method according to  claim 16  further comprising cultivating cells of the cell line in vitro, and repeating sub-steps a to d to obtain a more pure cell line of cells with lymphotropic metastatic potential, and the method further comprising testing cells of the cell line for the presence of one or more lymphoid gene products to confirm that the cells of the cell line have lymphotropic metastatic potential.