Patent
EX vivo effector cell activation for target cell killing
| العنوان: | EX vivo effector cell activation for target cell killing |
|---|---|
| Patent Number: | 4,844,893 |
| تاريخ النشر: | July 04, 1989 |
| Appl. No: | 06/916,173 |
| Application Filed: | October 07, 1986 |
| مستخلص: | A method and composition for killing target cells is disclosed. The method utilizes ex vivo IL-2 activation of leucocyte effector cells and arming the activated leucocyte effectors with monoclonal antibodies whose Fc portions bind to the IL-2-activated effectors and whose paratopic portions immunoreact with an epitope expressed on the surfaces of the target cells. The composition contains a cytolytic amount of the armed, IL-2-activated effector cells dispersed in an aqueous physiologically tolerable diluent medium. |
| Inventors: | Honsik, Cyril J. (La Jolla, CA); Reisfeld, Ralph A. (La Jolla, CA) |
| Assignees: | Scripps Clinic and Research Foundation (La Jolla, CA) |
| Claim: | We claim |
| Claim: | 1. A method of specifically killing of target cells consisting essentially of the steps of |
| Claim: | (a) activating a culture of leucocytes containing IgG1, IgG2a, IgG2b or IgG3 antibody Fc receptors ex vivo with an amount of interleukin-2 sufficient to enhance the natural killer activity of said cells to form IL-2-activated effector cells; |
| Claim: | (b) separating said effector cells from toxic amounts of interleukin-2; |
| Claim: | (c) binding the Fc receptors of said IL-2-activated effector cells with Fc-containing monoclonal antibodies of class IgG1, IgG2a, IgG2b or IgG3 whose paratopic portions immunoreact with an antigen expressed on the surface of target cells to form armed, IL-2-activated effector cells; |
| Claim: | (d) contacting target cells with a cytotoxic amount of said armed, IL-2-activated effector cells in the absence of exogenous IL-2; and |
| Claim: | (e) maintaining said contact for a time period sufficient for said armed, IL-2-activated effector cells to kill said target cells. |
| Claim: | 2. The method according to claim 1 wherein said target cells are tumor cells, and said antigen expressed on the surface of said target tumor cells is disialogangliosides GD2 or GD3. |
| Claim: | 3. The method according to claim 2 wherein said monoclonal antibodies are secreted by the hybridoma having ATCC accession number HB 8890. |
| Claim: | 4. The method according to claim 2 wherein said monoclonal antibodies are secreted by the hybridoma having ATCC accession number HB 9118. |
| Claim: | 5. The method of claim 1 further comprising contacting said target cells with a cytotoxic amount of bispecific hybrid monoclonal paratopic molecules that contain a first anti-T3 paratopic portion that immunoreacts with the T3 antigen expressed on the surfaces of T cells and a second paratopic portion that immunoreacts with a second epitope expressed on the surface of the target cells, said second epitope being different from the antigen with which said first-named Fc-containing monoclonal antibodies immunoreact. |
| Claim: | 6. The method of claim 5 wherein said anti-T3 paratopic portion that immunoreacts with the T3 antigen is secreted by the hybridoma having ATCC accession number CRL 8001. |
| Claim: | 7. The method of claim 6 wherein said target cells express both disialogangliosides GD2 and GD3, said first-named Fc-containing monoclonal antibodies immunoreact with one of said disialogangliosides, and said second paratopic portions of said hybrid molecule immunoreact with the other of said disialogangliosides. |
| Claim: | 8. The method of claim 5 wherein said bispecific hybrid molecules are contacted with said target cells substantially simultaneously with the contacting of said target cells with said armed, IL-2-activated effector cells. |
| Claim: | 9. The method of claim 8 wherein said bispecific hybrid molecules are coated on the surfaces of anti-T3-activated T cells that express the T3 antigen prior to said target cell contacting. |
| Claim: | 10. The method of claim 9 wherein said anti-T3-activated T cells are formed by the steps of |
| Claim: | (a) admixing and contacting a T cell population containing T3.sup.+ T cells with an activating amount of anti-T3 monoclonal antibodies; |
| Claim: | (b) maintaining said contact under biological conditions for a time period sufficient for the concentration of T3.sup.+ cells to diminish to about 75 percent of the concentration at the start of the contacting, while the number of cells in the population remains at least about constant to form anti-T3-activated T cells; |
| Claim: | (c) separating said anti-T3-activated T cells from said anti-T3 antibodies; and |
| Claim: | (d) coating said separated anti-T3-activated t cells with said anti-T3-containing bispecific hybrid molecules. |
| Claim: | 11. A method of enhancing ADCC of tumor cells consisting essentially of the steps of |
| Claim: | (a) activating a culture of peripheral blood mononuclear cells containing IgG1, IgG2a, IgG2b or IgG3 antibody Fc receptors ex vivo with an amount of interleukin-2 sufficient to enhance the natural killer activity of said cells to form IL-2-activated effector cells; |
| Claim: | (b) separating said IL-2-activated effector cells from toxic amounts of interleukin-2; |
| Claim: | (c) binding the Fc receptors of said IL-2-activated effector cells with Fc-containing monoclonal antibodies of class IgG1, IgG2a, IgG2b or IgG3 whose paratopic portions immunoreact with disialogangliosides GD2 or GD3 to form armed, IL-2-activated effector cells; |
| Claim: | (d) contacting a cytotoxic amount of said armed, IL-2-activated effector cells with tumor cells that express disialoganglioside selected from the group consisting cf GD2 and GD3 in the absence of exogenous IL-2; and |
| Claim: | (e) maintaining said contact for a time period sufficient for said armed, IL-2-activated effector cells to lyse said tumor cells. |
| Claim: | 12. The method of claim 11 wherein said contacting takes place in vitro. |
| Claim: | 13. The method of claim 11 wherein said contacting takes place in vivo. |
| Claim: | 14. The method of claim 11 wherein said target cells express both GD2 and GD3, and comprising the additional step of contacting said target cells with a cytotoxic amount of bispecific hybrid monoclonal paratopic molecules that contain first anti-T3 paratopic portions that immunoreact with the T3 antigen expressed on the surfaces of T cells and second paratopic portions that immunoreact with disialoganglioside GD2 or GD3, the ganglioside with which said second paratopic portions react being the disialoganglioside with which said Fc-containing monoclonal antibodies do not immunoreact. |
| Claim: | 15. The method of claim 14 wherein said anti-T3 paratopic portions of said bispecific hybrid molecules are secreted by the hybridoma having ATCC accession number CRL 8001, said Fc-containing monoclonal antibodies are secreted by a hybridoma selected from the group consisting of those having ATCC accession numbers HB 8890 and HB 9118, and the second paratopic portions of said bispecific hybrids are secreted by a hybridoma selected from the group consisting of those having ATCC accession numbers HB 8890, HB 9118 and HB 8568. |
| Claim: | 16. A target cell killing composition consisting essentially of an aqueous physiologically tolerable diluent medium free of exogenous IL-2, containing dispersed therein an amount of armed, IL-2-activated effector cells from a leucocyte population effective to kill target cells, said IL-2-activated effector cells being armed with monoclonal antibodies of class IgGl1 IgG2a, IgG2b or IgG3 bound to the Fc receptors of said cells, the paratopic portions of said monoclonal antibodies immunoreacting with an antigen expressed on the surfaces of said target cells. |
| Claim: | 17. The composition of claim 16 wherein said target cells express disialogangliosides GD2 or GD3 on their cell surfaces. |
| Claim: | 18. The composition of claim 17 wherein said monoclonal antibodies are secreted by a hybridoma having the ATCC accession number HB 8890 or HB 9118. |
| Claim: | 19. The composition of claim 16 further including a cytotoxic amount of bispecific hybrid paratopic molecules that contain first anti-T3 paratopic portions that immunoreact with the T3 antigen expressed on the surfaces of T cells and second paratopic portions that immunoreact with a second epitope expressed on the surfaces of said target cells, said second target cell epitope being different from the antigen with which said first-named monoclonal antibodies immunoreact. |
| Claim: | 20. The composition of claim 19 wherein said bispecific hybrid paratopic molecules of said composition are present coated on the surfaces of anti-T3-activated T cells as armed, anti-T3-activated T cells along with said armed, IL-2-activated effector cells. |
| Current U.S. Class: | 424/858; 424/93; 4352/402; 530/387; 530/828; 530/388; 530/808 |
| Current International Class: | A61K 39395 |
| Patent References Cited: | 4444878 April 1984 Paulus 4507391 March 1985 Pukel et al. 4675287 June 1987 Reisfeld |
| Other References: | Honsik, C. J. et al., Proc. Natl. Acad. Sci., U.S.A., 83:7893-7897, (10-1986). Cheresh, D. A. et al., Proc. Natl. Acad. Sci., U.S.A., 82:5155-5159, (8-1985). Park, M. M. et al., Cellular Immunology, 84:94-103, (1984). Mule, J. J. et al., Science, 225:1487-1489, (9-1984). Rosenberg, S. et al., New Engl. J. Medicine, 313(23):1485-1492, (12-1985). Perez, P. et al., J. Exper. Med., 163:166-178, (1-1986). Jung, G. et al., Proc. Natl. Acad. Sci., U.S.A., 83:4479-4483, (6-1986). Staerz, U. D. et al., Nature, 314:628-631, (4-1985). Cancer Research, (1984), 44:806-809, Dippold et al. Natural Immunity and Biological Response, (1985), 4:253, Honsik et al. Eur. J. Immunol., (1986), 16:263-270, Staerz et al. Proc. Nat'l Acad. Sci. U.S.A., (1986), 83:1453-1457, Staerz et al. Proc. Nat'l Acad. Sci. U.S.A., (1985), 82:1242-1246, Houghton et al. |
| Primary Examiner: | Moskowitz, Margaret |
| Assistant Examiner: | Kushan, Jeff. P. |
| Attorney, Agent or Firm: | Dressler, Goldsmith, Shore, Sutker & Milnamow, Ltd. |
| رقم الانضمام: | edspgr.04844893 |
| قاعدة البيانات: | USPTO Patent Grants |
| الوصف غير متاح. |