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 composition comprising Th1/killer cells wherein the Th1/killer cells have Th1 characteristics and cytolytic activity.
2 . The composition of claim 1 wherein the cytolytic activity comprises NK characteristics.
3 . The composition of claim 1 wherein the Th1/killer cells express granzyme B and perforin.
4 . The composition of claim 1 wherein the Th1/killer cells express IFN-gamma.
5 . The composition of claim 1 wherein the Th1/killer cells have substantially reduced or no expression of IL-4.
6 . The composition of claim 1 wherein the Th1/killer cells are CD4+ cells.
7 . The composition of claim 1 wherein the Th1/killer cells are formulated with cross-linking agent for CD3 and CD28
8 . The composition of claim 1 wherein the Th1/killer cells are derived from normal donor peripheral blood.
9 . The composition of claim 1 wherein the composition comprises clinically-relevant number of the Th1/killer cells.
10 . The composition of claim 1 wherein the composition comprises at least about 1×10 7 cells.
11 . The composition of claim 1 wherein the composition comprises at least about 1×10 8 cells.
12 . The composition of claim 1 wherein the cytolytic activity of the Th1/killer cells specifically inactivates diseased cells and not normal cells.
13 . The composition of claim 12 wherein the diseased cells comprise cancerous cells, infected cells or combinations thereof.
14 . A composition comprising Th1/killer cells wherein the Th1/killer cells are CD4+ cells having cytolytic activity against a tumor cell line.
15 . The composition of claim 14 wherein the Th1/killer cells comprise Th1 characteristics.
16 . The composition of claim 14 wherein the Th1/killer cells express granzyme B and perforin.
17 . The composition of claim 14 wherein the Th1/killer cells express EN-gamma.
18 . The composition of claim 14 wherein the tumor cell line is ARH77.
19 . The composition of claim 14 wherein the Th1/killer cells do not inactivate normal cells.
20 . A method for destroying diseased cells comprising contacting the diseased cells with a composition comprising allogeneic Th1/killer cells wherein the interaction of the diseased cells with the Th1/killer cells leads to destruction of the diseased cells.
21 . The method of claim 20 wherein the Th1/killer cells are CD4+ cells.
22 . The method of claim 20 wherein the Th1/killer cells express Th1 characteristics and cytolytic activity.
23 . The method of claim 22 wherein the cytolytic activity of the Th1/killer cells comprises expression granzyme B-perforin.
24 . The method of claim 22 wherein the Th1 characteristics comprise expression of IFN-gamma.
25 . The method of claim 22 wherein the Th1/killer cells lack expression of IL-4.
26 . The method of claim 20 wherein the Th1/killer cells are obtained by activation of CD4+ T-cells with cross-linked anti-CD3/anti-CD28 monoclonal antibodies.
27 . The method of claim 20 wherein the diseased cells comprise cancerous cells, infected cells or a combination thereof.
28 . A method of treating a patient comprising administering a composition comprising a clinically-relevant number of Th1/killer cells.
29 . The method of claim 28 wherein the Th1/killer cells are CD4+ cells.
30 . The method of claim 28 wherein the Th1/killer cells express Th1 characteristics and cytolytic activity.
31 . The method of claim 30 wherein the Th1/killer cells are activated with beads attached to anti-CD3/anti-CD28 monoclonal antibodies and crosslinked.
32 . The method of claim 28 wherein the Th1/killer cells are obtained from the peripheral blood of a normal donor.
33 . The method of claim 28 wherein the Th1/killer cells are allogeneic to the patient.
34 . The method of claim 28 wherein the clinically-relevant number of cells is at least 1×10 8 .