Development Of Natural Killer Cells And Functional Natural Killer Cell Lines

Abstract

The invention relates to natural killer cells and methods for the development of immortalized natural killer cells and use of the natural killer cells. A growth and culture system is described that supports increased natural killer cell development, and provides for the establishment of continuous natural killer cell lines. Additionally, the disclosed method for generating natural killer cells may be used to produce large numbers of natural killer cells for therapeutic applications and for natural killer cell research.

1 .- 39 . (canceled) 
     
     
         40 . A method of screening potential cancer therapeutic, the method comprising:
 (a) culturing a first population of natural killer precursor and/or natural killer cell engineered to express Jagged2 for between 35 and 45 days in the presence of compound suspected of being a cancer therapeutic,   (b) culturing a second population of natural killer precursor and/or natural killer cell engineered to express Jagged2 in the absence of the compound suspected of being a cancer therapeutic;   (c) determining the rate of growth of said first population of natural killer precursor and/or natural killer cells cultured in the presence of the compound suspected of being a cancer therapeutic and determining the rate of growth said second population of natural killer precursor and/or natural killer cells cultured in the absence of the compound suspected of being a cancer therapeutic; and   (d) comparing the growth rate of said first and second populations of natural killer precursor and/or natural killer cells   wherein an increased rate of growth of said first population of natural killer precursor and/or natural killer cells is indicative of a cancer therapeutic.   
     
     
         41 . The method of  claim 40 , wherein Jagged2 is provided by a population of cells expressing Jagged2. 
     
     
         42 . The method of  claim 40 , wherein said population of cells is OP-9 stromal cells as deposited with ATCC, deposit number PTA-10785, and which have been subsequently engineered to express Jagged2. 
     
     
         43 . The method of  claim 40 , wherein the natural killer precursor cells are NK1.1 +  CD3 −  TCRab − , TCRdg − , CD4 − , CD8 − , CD19 − , CD25 + , CD43 + , CD45 + , CD49b − , CD51 + , CD94 + , NKG2D + , Mac-1 −/low , B220 − , c-kit + , perforin I + , granzyme B + , Notch-1 + , and cytotoxic. 
     
     
         44 . The method of  claim 40 , wherein the cancer therapeutic is useful in treating leukemia or lymphoma. 
     
     
         45 . A method of screening for a potential new drug to treat a disorder susceptible to natural killer precursor cells and/or natural killer cells, the method comprising:
 (a) providing a population of natural killer precursor and/or natural killer cells engineered to express Jagged2, wherein the population of natural killer precursor and/or natural killer cells engineered to express Jagged are produced by bringing a population of hematopoietic progenitor cells or light-density mononuclear cells or embryonic stem cells into contact with IL-7, Flt3L and Jagged2;   (b) culturing the population of natural killer precursor and/or natural killer cells engineered to express Jagged2 in the presence of potential new drug; and   (c) detecting the presence or absence of an effect of the potential new drug on the survival of the population of natural killer precursor and/or natural killer cells;   wherein survival of the population of natural killer precursor and/or natural killer cells is indicative of a drug that is capable of treating a disorder susceptible to natural killer precursor cells and/or natural killer cells.   
     
     
         46 . The method of  claim 45 , further comprising after step (b), detecting morphological, functional and/or physiological characteristics of the population of natural killer precursor and/or natural killer cells. 
     
     
         47 . The method of  claim 45 , further comprising culturing a second population of natural killer precursor and/or natural killer cells engineered to express Jagged2 in the absence of the potential new drug, wherein the second population of natural killer precursor and/or natural killer cells is produced using the same methods as used to produce the population of natural killer precursor and/or natural killer cell engineered to express Jagged2 in step (a). 
     
     
         48 . The method of  claim 47 , further comprising detecting the presence or absence of an effect of the potential new drug on the survival of the second population of natural killer precursor and/or natural killer cells. 
     
     
         49 . The method of  claim 47 , further comprising after step (b), detecting morphological, functional and/or physiological characteristics of the second population of natural killer precursor and/or natural killer cells. 
     
     
         50 . The method of  claim 48 , further comprising comparing the morphological, functional and/or physiological characteristics of the population of natural killer precursor and/or natural killer cells with the survival of the second population of natural killer precursor and/or natural killer cells. 
     
     
         51 . The method of  claim 49 , further comprising comparing the survival of the population of natural killer precursor and/or natural killer cells with the morphological, functional and/or physiological characteristics of the second population of natural killer precursor and/or natural killer cells.