Cells expressing Th1 characteristics and cytolytic properties
A novel cell type has been generated that has both Th1 characteristics and cytolytic activity. These Th1/killer cells are CD4+ cells purified from peripheral blood and manipulated to have Th1 characteristics such as production of IFN-gamma combined with cytolytic activity similar to cytotoxic T-cells (CTL). The CTL activity is targeted toward diseased cells, not normal cells. The cytolytic activity of the Th1/killer cells is mediated by Granzyme B-Perforin mechanism and results in apoptotic death of diseased cells. Methods of producing and using these Th1/killer cells include isolating CD4+ cells from peripheral blood, activating the CD4+ T-cells to form Th1/killer cells and administering these Th1/killer cells with the cytolytic activity to a patient wherein the Th1/killer cells are allogeneic to the patient.
1. A method for killing tumor cells comprising:
contacting the tumor cells from a tumor cell line with a composition comprising ex-vivo activated CD4+ Th1 cells without the presence of any other cell types, wherein the activated CD4+ Th1 cells have cytolytic activity mediated by Granzyme B and perforin, wherein the CD4+ Th1 cells are frozen after harvesting from cell culture, thawed reactivated by both binding of the CD3/CD28 cell surface molecules by agents and cross-linking of these agents in the composition by the presence of the cross-linking agents in the composition, washed and transferred to a non-nutritive buffer, wherein the CD4+ Th1 cells express IFN-gamma at greater than 1000 pg/10 6 cells and wherein the interaction of the CD4+ Th1 cells with the tumor cells leads to the killing of the tumor cells.
2. The method of claim 1 wherein the CD4+ Th1 cells lack expression of IL-4.
3. The method of claim 1 wherein the CD4+ Th1 cells are activated by cross-linked anti-CD3/anti-CD28 monoclonal antibodies.
4. The method of claim 1 wherein the tumor cell line is ARH77.
5. The method of claim 1 wherein the ratio of the CD4+ Th1 cells to cells of the tumor cell line is at least 1:9.
6. The method of claim 1 wherein the composition comprises at least about 10 7 cells/ml.