Genetically modified human natural killer cell lines
View Patent ↗The invention provides a natural killer cell, NK-92, modified to express an Fc receptor on the surface of the cell, such as CD16 (FcγRIII-A), or other Fcγ or Fc receptors. The modified NK-92 cell can be further modified to concurrently express an associated accessory signaling protein, such as FcεRI-γ, TCR-ζ, or to concurrently express interleukin-2 (IL-2) or other cytokines. Additional methods are disclosed for various assays, assessments, and therapeutic treatments with the modified NK-92 cells.
1. A method for assaying a candidate antibody for efficacy in killing a cancerous or infected cell, which method comprises:
(i) selecting a candidate antibody that specifically binds to an antigen that is expressed by a tumor or infected cell, and that also binds to an NK-92 cell modified to express an FyγRIII-A receptor on a surface of the NK-92 cell, wherein said NK-92 cell is available from American Type Culture Collection (ATCC) as Accession No. PTA-6670,
(ii) incubating in vitro a cancerous or infected cell with the candidate antibody to form a mixture under a condition that the antibody can bind to the cancerous or infected cell;
(iii) incubating in vitro the mixture with the modified NK-92 cell under conditions wherein the modified NK-92 cell is capable of killing said cancerous or infected cell; and
(iv) determining the efficacy of said antibody in killing said cancerous or infected cell.
2. The method of claim 1 , wherein the FcγRIII-A receptor has the amino acid sequence of SEQ ID NO:2.
3. The method of claim 1 , wherein the FcγRIII-A receptor consists of a polypeptide of SEQ ID NO:1.
4. The method of claim 1 , wherein a ratio of the modified NK-92 cell to the target cell is between 0.5:1 and about 100:1.
5. The method of claim 4 , wherein a ratio of the modified NK-92 cell to the target cell is between about 1:1 and about 20:1.
6. The method of claim 1 , wherein the target cell is SKOV-3 available from American Type Culture Collection as Deposit No. HTB-77.
7. The method of claim 1 , wherein the antibody is a monoclonal antibody or a polyclonal antibody.
8. The method of claim 1 , wherein the antibody is a chimeric antibody.
9. The method of claim 1 , wherein the antibody is a hybridoma supernate.
10. The method of claim 1 , wherein prior to incubating the mixture with the modified NK-92 cell, the modified NK-92 cell is cultured with a cytokine selected from the group consisting of IL-2, IL-12, IL-15, and IL-18.
11. The method of claim 1 , wherein prior to incubating the mixture with the modified NK-02 cell, the modified NK 92 cell is cultured with interleukin-2.