Hybrid cells

Abstract

A rapid, simple-to-use method for preparing hybrid cells, applicable to fully differentiate, non-dividing cells, entails bringing at least two different cells into contact under conditions that promote cell fusion and then purifying the resultant hybrid without antibiotic or metabolic selection. This approach yields hybrid cells useful in a variety of applications, including clinical treatment regimens, as cellular modulators of the immune system.

1 .- 28 . (canceled)  
     
     
         29 . A composition comprising a hybrid cell preparation, wherein the hybrid cell preparation is prepared by a method comprising: 
 (a) staining a neoplastic cell population freshly isolated from a patient with a first marker,    (b) staining a dendritic cell population with a second marker, wherein the first marker is different from the second marker,    (c) contacting the neoplastic cell population and the dendritic cell population with one another under conditions that promote cell fusion,    (d) obtaining a resultant hybrid cell population stained with both the first and second markers, and    (e) purifying the resultant hybrid cell population;    wherein the method does not involve antibiotic or metabolic selection, the purifying is accomplished in less than about 24 to 48 hours after said contacting.    
     
     
         30 . The composition of  claim 29 , wherein the first marker is a fluorescent dye, the second marker is a fluorescent dye, and the resultant hybrid cell population is purified by fluorescence activated cell sorting.  
     
     
         31 . The composition of  claim 29 , wherein the resultant hybrid cell population contains less than 10% of its total population as reactant cells.  
     
     
         32 . The composition of  claim 29 , wherein the resultant hybrid cell population contains less than 5% of its total population as reactant cells.  
     
     
         33 . The composition of  claim 29 , further comprising a pharmaceutically acceptable vehicle.  
     
     
         34 . The composition of  claim 30 , wherein the first and second fluorescent dyes are endotoxin-free, pyrogen-free or both.  
     
     
         35 . The composition of  claim 29 , wherein the neoplastic cell is a tumor cell.  
     
     
         36 . The composition of  claim 35 , wherein the tumor cell is a primary tumor cell.  
     
     
         37 . A method of treating cancer, comprising: 
 (i) providing a hybrid cell preparation prepared by a method comprising: 
 (a) staining a neoplastic cell population freshly isolated from a patient with a first marker,  
 (b) staining a dendritic cell population with a second marker, wherein the first marker is different from the second marker,  
 (c) contacting said neoplastic cell population and the dendritic cell population with one another under conditions that promote cell fusion,  
 (d) obtaining a resultant hybrid cell population stained with both the first and second markers, and  
 (d) purifying the resultant hybrid cell population;  
 wherein the method does not involve antibiotic or metabolic selection, said purifying is accomplished in less than about 24 to 48 hours after the contacting;  
   (ii) resuspending the resultant hybrid cell preparation in a pharmaceutically acceptable vehicle, and    (iii) administering the hybrid cell preparation to a cancer patient.    
     
     
         38 . The method of  claim 37 , wherein the first marker is a fluorescent dye, the second marker is a fluorescent dye, and the resultant hybrid cell population is purified by fluorescence activated cell sorting.  
     
     
         39 . A cancer vaccine comprising a hybrid cell preparation and a pharmaceutically acceptable vehicle, wherein the hybrid cell preparation is prepared by a method comprising: 
 (a) staining a neoplastic cell population freshly isolated from a patient with a first marker,    (b) staining a dendritic cell population with a second marker, wherein the first marker is different from the second marker,    (c) contacting the neoplastic cell population and the dendritic cell population with one another under conditions that promote cell fusion,    (d) obtaining a resultant hybrid cell population stained with both the first and second markers,    (e) purifying the resultant hybrid cell population, and    (f) resuspending the resultant hybrid cell population in a pharmaceutically acceptable vehicle;    wherein the method does not involve antibiotic or metabolic selection, the purifying is accomplished in less than about 24 to 48 hours after the contacting.    
     
     
         40 . The cancer vaccine of  claim 39 , wherein the first marker is a fluorescent dye, the second marker is a fluorescent dye, and the resultant hybrid cell population is purified by fluorescence activated cell sorting.  
     
     
         41 . The cancer vaccine of  claim 39 , wherein the pharmaceutically acceptable vehicle is saline.  
     
     
         42 . A tumor vaccine comprising a hybrid cell preparation and a pharmaceutically acceptable vehicle, wherein the hybrid cell preparation is prepared by a method comprising: 
 (a) staining a neoplastic cell population freshly isolated from a patient with a first marker,    (b) staining a dendritic cell population with a second marker, wherein the first marker is different from the second marker,    (c) contacting the neoplastic cell population and the dendritic cell population with one another under conditions that promote cell fusion,    (d) obtaining a resultant hybrid cell population stained with both the first and second markers,    (e) purifying the resultant hybrid cell population, and    (f) resuspending the resultant hybrid cell population in a pharmaceutically acceptable vehicle;    wherein the method does not involve antibiotic or metabolic selection, the purifying is accomplished in less than about 24 to 48 hours after the contacting.    
     
     
         43 . The tumor vaccine of  claim 42 , wherein the first marker is a fluorescent dye, the second marker is a fluorescent dye, and the resultant hybrid cell population is purified by fluorescence activated cell sorting.  
     
     
         44 . The tumor vaccine of  claim 42 , wherein the pharmaceutically acceptable vehicle is saline.  
     
     
         45 . A method for treating an autoimmune disease, comprising: 
 (i) preparing a hybrid cell preparation prepared by a method comprising 
 (a) contacting a population of target cells with a first marker,  
 (b) contacting a population of antigen presenting cells lacking an accessory molecule with a second marker,  
 (c) contacting said target cells and said antigen presenting cells deficient in an accessory interaction under conditions that promote cell fusion,  
 (d) obtaining a resultant hybrid cell population stained with both the first and second markers, and  
 (e) purifying the resultant hybrid cell population; and  
 (f) resuspending the resultant hybrid cell population in a pharmaceutically acceptable vehicle,  
 wherein said method does not involve antibiotic or metabolic selection;  
   (ii) resuspending the resultant hybrid cell preparation in a pharmaceutically acceptable vehicle, and    (iii) administering the hybrid cell preparation.    
     
     
         46 . A method of preparing a formulation for tolerizing an immune system against a target cell comprising: 
 (a) contacting a population of target cells with a first marker,    (b) contacting a population of antigen presenting cells lacking an accessory molecule with a second marker,    (c) contacting said target cells and said antigen presenting cells deficient in an accessory interaction with one another under conditions that promote cell fusion,    (d) obtaining a resultant hybrid cell population stained with both the first and second markers,    (e) purifying the resultant hybrid cell population by cell sorting, and    (f) resuspending the resultant hybrid cell population in a pharmaceutically acceptable buffer;    wherein said cell sorting does not involve antibiotic or metabolic selection.    
     
     
         47 . The method of  claim 46 , wherein the first marker is a fluorescent dye, the second marker is a fluorescent dye, and the resultant hybrid cell population is purified by fluorescence activated cell sorting.  
     
     
         48 . A kit comprising 
 (i) at least two dyes that are essentially endotoxin-free and    (ii) instructions for preparing hybrid cells from reactant cells by a method comprising: 
 (a) staining a neoplastic cell population freshly isolated from a patient with a first dye,  
 (b) staining a dendritic cell population with a second dye, wherein said first dye is different from said second dye,  
 (c) contacting said neoplastic cell population and said dendritic cell population with one another under conditions that promote cell fusion,  
 (d) obtaining a resultant hybrid cell population stained with both the first and second dyes, and  
 (e) purifying the resultant hybrid cell population;  
 wherein said method does not involve antibiotic or metabolic selection, said purifying is accomplished in less than about 24 to 48 hours after said contacting.