Use of allogeneic cell lines to load antigen-presenting cells to elicit or eliminate immune responses

Abstract

Novel antigen-presenting cells, including but not limited to dendritic cells, that are loaded with antigens from dead or dying cells including allogenic cell lines, and the methods for making such antigen-presenting cells are described. These loaded antigen-presenting cells induce therapeutic immune responses in humans. Such loaded antigen-presenting cells are useful in the management of cancer. Antigen-loaded dendritic cells prepared as described here can prime naïve T cells to differentiate into effector cells able to recognize multiple and/or shared tumor antigens that are expressed either on the tumor cells that are used to load the dendritic cells and/or on other tumor cells. The cytotoxic T cells generated by exposure to antigen-loaded dendritic cells prepared as described here can be used in adoptive therapy. This induction of responses against multiple antigens shared between different cells, for instance tumor cells, as described here is important as it leads to broad immune responses.

1 . A method of inducing tumor specific immune response in a tumor-bearing patient comprising:
 (a) isolating antigen-presenting cells from said patient;   (b) coincubating said antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells; and   (c) administering said loaded antigen-presenting cells to said patient, wherein said patient's cells upon contact with said loaded antigen-presenting cells mature to form cytotoxic cells expressing cytotoxic activity against said tumor cells.   
     
     
         2 . The method of  claim 1 , wherein said dead or dying cell portions are obtained from allogeneic cell lines. 
     
     
         3 . The method of  claim 1 , wherein said tumor specific immune response is directed against said tumor antigen, against multiple tumor antigens, against shared tumor antigens, or any combinations thereof. 
     
     
         4 . The method of  claim 1 , wherein said antigen-presenting cells are at various stages of differentiation and/or maturation and selected from the group consisting of dendritic cells, progenitors or precursors of dendritic cells from which dendritic cells are generated ex vivo, lymphoid dendritic cells, myleloid dendritic cells, interstitial dendritic cells, and Langerhans cells. 
     
     
         5 . The method of  claim 1 , wherein the cytotoxic cells comprise at least one of CD4 positive cells or CD8 positive cells. 
     
     
         6 . A method of inducing tumor specific responses in a tumor-bearing patient comprising:
 (a) isolating antigen-presenting cells and cytotoxic cell precursors from said patient;   (b) coincubating antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells;   (c) coculturing said loaded antigen-presenting cells with cytotoxic cell precursors isolated from said patient under conditions for their maturation to form cytotoxic cells; and   (d) administering said cytotoxic cells to said patient, wherein said cytotoxic cells expressing cytotoxic activity against said tumor cells.   
     
     
         7 . The method of  claim 6 , wherein said dead or dying cell portions are obtained from allogeneic cell lines. 
     
     
         8 . The method of  claim 6 , wherein said tumor specific immune response is directed against said tumor antigen, against multiple tumor antigens, against shared tumor antigens, or any combinations thereof. 
     
     
         9 . The method of  claim 6 , wherein said antigen-presenting cells are at various stages of differentiation and/or maturation and selected from the group consisting of dendritic cells, progenitors or precursors of dendritic cells from which dendritic cells are generated ex vivo, lymphoid dendritic cells, myleloid dendritic cells, interstitial dendritic cells, and Langerhans cells. 
     
     
         10 . The method of  claim 6 , wherein the cytotoxic cells comprise at least one of CD4 positive cells or CD8 positive cells. 
     
     
         11 . A method of inducing tumor specific responses in a tumor-bearing patient comprising:
 (a) coincubating antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells;   (b) coculturing said loaded antigen-presenting cells with naïve T cells under conditions for T cell maturation to form cytotoxic T cells; and   (c) administering said cytotoxic T cells to said patient, wherein said cytotoxic T cells expressing cytotoxic activity against said tumor cells.   
     
     
         12 . The method of  claim 11 , wherein said dead or dying cell portions are obtained from allogeneic cell lines. 
     
     
         13 . The method of  claim 11 , wherein said tumor specific immune response is directed against said tumor antigen, against multiple tumor antigens, against shared tumor antigens, or any combinations thereof. 
     
     
         14 . The method of  claim 11 , wherein said antigen-presenting cells are at various stages of differentiation and/or maturation and selected from the group consisting of dendritic cells, progenitors or precursors of dendritic cells from which dendritic cells are generated ex vivo, lymphoid dendritic cells, myleloid dendritic cells, interstitial dendritic cells, and Langerhans cells. 
     
     
         15 . The method of  claim 11 , wherein cytotoxic T cells comprise at least one of CD4 positive cells or CD8 positive cells. 
     
     
         16 . A method for mounting an immune response, eliciting tumor rejection, or both mediated by cytotoxic T cells comprising
 (a) coincubating antigen-presenting cells with dead or dying cell portions possessing at least one antigen to which the immune response is desired to form loaded antigen-presenting cells;   (b) coculturing said loaded antigen-presenting cells with naïve T cells under conditions for T cell maturation to form cytotoxic T cells; and   (c) contacting said cytotoxic T cells with target cells possessing said antigen, wherein said cytotoxic T cells express cytotoxic activity against said target cells possessing said antigen.   
     
     
         17 . The method of  claim 16 , wherein the method is adapted to identify one or more novel tumor rejection antigens. 
     
     
         18 . A method of administering loaded antigen-presenting cells to a patient comprising:
 (a) isolating said antigen-presenting cells from said patient;   (b) coincubating said antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells; and   (c) administering said loaded antigen-presenting cells to said patient.   
     
     
         19 . A method of administering tumor specific cytotoxic cells to a patient comprising:
 (a) isolating antigen-presenting cells and cytotoxic cell precursors from said patient;   (b) coincubating said antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells;   (c) coculturing said loaded antigen-presenting cells with said cytotoxic cell precursors isolated from said patient under conditions for their maturation to form cytotoxic cells; and   (d) administering said cytotoxic cells to said patient, wherein said cytotoxic cells expressing cytotoxic activity against said tumor cells.   
     
     
         20 . A method of simultaneously administering loaded antigen-presenting cells and tumor specific cytotoxic cells to a patient comprising:
 (a) isolating said antigen-presenting cells and cytotoxic cell precursors from said patient;   (b) coincubating said antigen-presenting cells with dead or dying cell portions possessing at least one tumor antigen to form loaded antigen-presenting cells;   (c) coculturing said loaded antigen-presenting cells with said cytotoxic cell precursors isolated from said patient under conditions for their maturation to form cytotoxic cells; and   (d) simultaneously administering said antigen-presenting cells and said cytotoxic cells to said patient.   
     
     
         21 . A method of making Cytotoxic T cells, the method comprising the steps of:
 (a) coincubating antigen-presenting cells with dead or dying cell portions possessing at least one antigen to which the immune response is desired to form loaded antigen-presenting cells;   (b) coculturing said loaded antigen-presenting cells with naïve T cells under conditions for T cell maturation to form said cytotoxic T cells.   
     
     
         22 . The method of  claim 21 , wherein said cytotoxic T cells comprise at least one of CD4 positive cells or CD8 positive cells. 
     
     
         23 . A method for mounting an immune response mediated by cytotoxic cells comprising
 (a) coincubating antigen-presenting cells with dead or dying cell portions possessing at least one antigen to which the immune response is desired to form loaded antigen-presenting cells, wherein said dead or dying cell portions are obtained from a first source;   (b) coculturing said loaded antigen-presenting cells with cytotoxic cell precursors under conditions for their maturation to form cytotoxic cells; and   (c) contacting said cytotoxic cells with target cells possessing said antigen, wherein said target cells are obtained from a second source, wherein said cytotoxic cells express cytotoxic activity against said target cells possessing said antigen.   
     
     
         24 . The method of  claim 23 , wherein said target cells are contacted in vitro or in vivo. 
     
     
         25 . The method of  claim 23 , wherein said first source is at least one cell line cultured in vitro or one heterologous patient. 
     
     
         26 . A method for reducing graft rejection, graft versus host disease, or both of a transplant of donor tissue into a host comprising
 (a) coincubating antigen-presenting cells with dead or dying cell portions from said donor to form loaded antigen-presenting cells, said dead or dying cell portions possessing antigens to which the reduction of an immune response is desired;   (b) administering said loaded antigen-presenting cells into said host at an effective concentration and under conditions for cytotoxic cell precursor maturation into activated cytotoxic cells; and   (c) treating said host with a composition to reduce said activated cytotoxic cells prior to transplantation of donor tissue into said host.   
     
     
         27 . A method for reducing graft versus host disease of a transplant of donor tissue into a host comprising
 (a) coincubating antigen-presenting cells with dead or dying cell portions from said host to form loaded antigen-presenting cells, said dead or dying cell portions possessing antigens to which the reduction of an immune response is desired;   (b) contacting said donor tissue with said loaded antigen-presenting cells,   (c) allowing cytotoxic cell precursors in said donor tissue mature to form cytotoxic cells; and   (d) treating said donor tissue with a composition to eliminate said activated cytotoxic cells prior to transplantation of said donor tissue into said host.   
     
     
         28 . A method for inducing antigen-specific tolerance in vivo comprising administering antigen presenting cells loaded with killed cells expressing a desired antigen and treated so as to prevent expression of co-stimulatory molecules but insure antigen presentation. 
     
     
         29 . An in vitro assay to determine the degree of cytotoxic activity of cytotoxic cells against tumor cells from a patient comprising
 (a) coincubating of antigen-presenting cells with dead or dying cell portions, said dead or dying cell portions possessing antigens to which the immune response is desired;   (b) coculturing said loaded antigen-presenting cells with cytotoxic precursor cells under conditions for their maturation into cytotoxic cells;   (c) isolating said cytotoxic cells;   (d) incubating said cytotoxic cells with said tumor cells; and   (e) monitoring the death of said tumor cells, wherein the degree of tumor cell death is proportional to the cytotoxic activity of said cytotoxic cells.