In vitro assay for quantitating secreted antibodies in lymphocyte supernatant for evaluation of vaccine or antigen induced specific antibody secretion from ex vivo circulating antibody-secreting lymphocytes

Abstract

The present invention provides methods of quantitating recent secreted antigen specific antibodies from supernatant of antibody secreting cells (ASC) in vitro culture for evaluation of vaccine or antigen induced antigen specific antibody secretion without ex vivo antigen stimulation.

1 . A method for detecting the level or the presence of antigen-specific antibodies secreted from an mammal's antibody secreting cells, said method comprising the steps of (i) obtaining a sample comprising the supernatant of an in vitro culture of antibody secreting cells isolated from an mammal after excluding serum antibodies from the in vitro culture, and (ii) determining the level or the presence of said antigen-specific antibodies in said sample;
 wherein the antibody secreting cells are in vivo stimulated by an antigen in the mammal before isolating the antibody secreting cells from the mammal, and the antibody secreting cells or the culture are not in vitro stimulated by the antigen or a part thereof prior to the determining step.   
     
     
         2 . The method of  claim 1 , wherein said antibody secreting cells are peripheral blood mononuclear cells. 
     
     
         3 . The method of  claim 1 , wherein said antibodies comprises whole immunoglobulins or fragments thereof. 
     
     
         4 . The method of  claim 1 , wherein said antigen is derived from a pathogen. 
     
     
         5 . The method of  claim 1 , wherein said antigen is an antigenic part of a vaccine. 
     
     
         6 . The method of  claim 1 , wherein said antigen is derived from an antigenic substance, which is capable of eliciting an antibody immune response specific to said antigen in the said subject. 
     
     
         7 . The method of  claim 1 , wherein said subject is immunized with a vaccine comprising said antigen before the obtaining step. 
     
     
         8 . The method of  claim 1 , wherein said subject is infected with a pathogen from which said antigen is derived before the obtaining step. 
     
     
         9 . The method of  claim 1 , wherein said subject is in vivo stimulated with said antigenic substance before the obtaining step. 
     
     
         10 . The method of  claim 1 , wherein the level or the presence of said antigen-specific antibodies is determined by an enzyme-linked immunosorbent assay. 
     
     
         11 . The method of  claim 1 , wherein the level or the presence of said antigen-specific antibodies is determined by an assay that quantifies or determines the presence of said antigen-specific antibodies. 
     
     
         12 . The method of  claim 1 , wherein the level or the presence of said antigen-specific antibodies is determined by an immunoassay.