Methods for treating hiv infected subjects

Abstract

Methods for inducing a population of T cells to proliferate by activating the population of T cells and stimulating an accessory molecule on the surface of the T cells with a ligand which binds the accessory molecule are described. T cell proliferation occurs in the absence of exogenous growth factors or accessory cells. T cell activation is accomplished by stimulating the T cell receptor (TCR)/CD3 complex or the CD2 surface protein. To induce proliferation of an activated population T cells, an accessory molecule on the surface of the T cells, such as CD28, is stimulated with a ligand which binds the accessory molecule. The T cell population expanded by the method of the invention can be genetically transduced and used for immunotherapy or can be used in methods of diagnosis.

1 . A method for selectively enriching a population of T cells in TH1 cells in vitro comprising:
 a) contacting a population of T cells with:
 (1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and 
 (2) an anti-CD28 antibody or a CD28-binding fragment thereof, thereby stimulating the activated T cells, such that the T cells are stimulated to proliferate and produce at least one TH1 cytokine, 
   thereby selectively enriching the population of T cells in TH1 cells.   
     
     
         2 . The method of  claim 1 , further comprising:
 b) determining the amount of at least one TH1 cytokine, wherein an increase in the amount of the at least one TH1 cytokine following activation and stimulation of the T cells compared to the amount of the at least one TH1 cytokine produced by T cells that have not been activated as in (1) and stimulated in (2), indicates enrichment of the population of T cells in TH1 cells.   
     
     
         3 . The method of  claim 1 , further comprising:
 c) monitoring proliferation of the T cells; and   d) reactivating and restimulating the T cells when the rate of T cell proliferation has decreased to induce further proliferation of the T cells, such that the population of T cells is enriched in TH1 cells.   
     
     
         4 . The method of  claim 1 , wherein the agent which provides a primary activation signal and the anti-CD28 antibody or CD28-binding fragment thereof are attached to a surface. 
     
     
         5 . The method of  claim 1 , wherein the agent which provides a primary activation signal and the anti-CD28 antibody or CD28-binding fragment thereof are attached to the same surface. 
     
     
         6 . The method of  claim 5 , wherein the surface is surface is a bead, a tissue culture flask, or a cell surface. 
     
     
         7 . The method of  claim 6 , wherein the bead is a magnetic bead or a biodegradable bead. 
     
     
         8 . The method of  claim 3 , wherein monitoring proliferation of the T cells is by examining cell size or determining the level of expression of a cell surface molecule selected from the group consisting of B7-1, B7-2, and combinations thereof, and the method further comprises reactivating and restimulating the T cells when T cell size has decreased or the level of the cell surface molecule has decreased to induce further proliferation of the T cells. 
     
     
         9 . A method for selectively enriching a population of T cells in TH2 cells in vitro comprising
 a) contacting a population of T cells with:
 (1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and 
 (2) a stimulatory form of a natural ligand of CD28 selected from the group consisting of B7-1, a CD28-binding fragment of B7-1, B7-2, and a CD28-binding fragment of B7-2, such that the T cells are stimulated to proliferate and produce at least one TH2 cytokine, 
   thereby enriching the population of T cells in TH2 cells.   
     
     
         10 . The method of  claim 9 , further comprising:
 b) determining the amount of at least one TH2 cytokine, wherein an increase of the at least one TH2 cytokine following the activation and stimulation of the T cells compared to the amount of the at least one TH2 cytokine produced by T cells that have not been activated as in (1) and stimulated in (2), indicates enrichment of the population of T cells in TH2 cells.   
     
     
         11 . The method of  claim 9 , further comprising:
 c) monitoring the proliferation of the T cells; and   d) reactivating and restimulating the T cells when the rate of T cell proliferation has decreased to induce further proliferation of the T cells, such that the population of T cells is enriched in TH2 cells.   
     
     
         12 . The method of  claim 9 , wherein the agent which provides a primary activation signal and the stimulatory form of a natural ligand of CD28 are attached to a surface. 
     
     
         13 . The method of  claim 9 , wherein the agent which provides a primary activation signal and the stimulatory form of a natural ligand of CD28 are attached to the same surface. 
     
     
         14 . The method of  claim 13 , wherein the surface is a bead, a tissue culture flask, or a cell surface. 
     
     
         15 . The method of  claim 13 , wherein the surface is a CHO cell surface. 
     
     
         16 . The method of  claim 11 , wherein monitoring proliferation of the T cells is by examining cell size or determining the level of expression of a cell surface molecule selected from the group consisting of B7-1, B7-2, and combinations thereof, and the method further comprises reactivating and restimulating the T cells when T cell size has decreased or the level of the cell surface molecule has decreased to induce further proliferation of the T cells. 
     
     
         17 . A method for increasing the proportion of TH1 versus TH2 cells in a population of T cells, comprising:
 a) contacting the population of T cells with:
 (1) an agent which provides a primary activation signal to the T cells thereby activating the T cells, wherein the agent is selected from the group consisting of an anti-CD3 antibody or a CD3-binding fragment thereof, an anti-CD2 antibody or a CD2-binding fragment thereof, and an antigen in a form suitable to trigger a primary activation signal in the T cell when complexed with the TCR/CD3 complex; and 
 (2) an anti-CD28 antibody or a CD28-binding fragment thereof, wherein the anti-CD28 antibody or a CD28-binding fragment is attached on a surface, 
   thereby increasing the proportion of TH1 versus TH2 cells in the population of T cells compared to the proportion of TH1 versus TH2 cells in a population of T cells not contacted with an agent which provides a primary activation signal to the T cells and an anti-CD28 antibody or a CD28-binding fragment thereof.   
     
     
         18 . The method of  claim 17 , wherein the agent which provides a primary activation signal is attached to a surface. 
     
     
         19 . The method of  claim 18 , wherein the agent which provides a primary activation signal and the anti-CD28 antibody or a CD28-binding fragment thereof are attached to the same surface. 
     
     
         20 . The method of  claim 19 , wherein the surface is a bead, a tissue culture flask, or a cell surface.