Media elaborated with newly synthesized antibodies (mensa) and uses thereof

Abstract

Disclosed are methods and kits for early detection of antigen exposure through the presence or absence of antigen-specific antibodies.

1 - 11 . (canceled) 
     
     
         12 . A method of assessing the efficacy of a vaccine against a disease or toxin in a subject comprising obtaining PBMC or whole blood from the subject between 3 and 45 days following administration of a vaccine; separating plasma from the whole blood or PBMC to produce separated cells; isolating the newly proliferated ASC from the separated cells; washing the newly proliferated ASC; culturing the newly proliferated ASC in media conducive to antibody production and expression (MENSA), measuring the number of vaccine specific antibody secreting cells (ASC) or antibodies in the MENSA, and comparing the number of ASC or antibodies to a standard or control, wherein the washed ASC comprise at least a 10 6  or 10 7 -fold reduction of contaminating pre-existing plasma antibodies relative to whole blood, plasma, or PBMC, and wherein an increase of vaccine specific ASC or antibodies in the MENSA relative to a standard or control indicates an efficacious vaccine. 
     
     
         13 . The method of  claim 12 , wherein the control is the number of pre-existing vaccine specific antibodies and/or pre-existing ASC. 
     
     
         14 - 17 . (canceled) 
     
     
         18 . The method of  claim 12 , wherein the disease is a viral disease selected from the group of viruses consisting of Herpes Simplex virus-1, Herpes Simplex virus-2, Varicella-Zoster virus, Epstein-Barr virus, Cytomegalovirus, Human Herpes virus-6, Variola virus, Vesicular stomatitis virus, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Hepatitis D virus, Hepatitis E virus, Rhinovirus, Coronavirus, Influenza virus A, Influenza virus B, Measles virus, Polyomavirus, Human Papilomavirus, Respiratory syncytial virus, Adenovirus, Coxsackie virus, Dengue virus, Mumps virus, Poliovirus, Rabies virus, Rous sarcoma virus, Reovirus, Yellow fever virus, Ebola virus, Marburg virus, Lassa fever virus, Eastern Equine Encephalitis virus, Japanese Encephalitis virus, St. Louis Encephalitis virus, Murray Valley fever virus, West Nile virus, Rift Valley fever virus, Rotavirus A, Rotavirus B, Rotavirus C, Sindbis virus, Simian Immunodeficiency virus, Human T-cell Leukemia virus type-1, Hantavirus, Rubella virus, Simian Immunodeficiency virus, Human Immunodeficiency virus type-1, and Human Immunodeficiency virus type-2. 
     
     
         19 . The method of  claim 12 , wherein the disease is a bacterial disease selected from the group of bacterium consisting of  M. tuberculosis, M. bovis, M. bovis  strain BCG, BCG substrains,  M. avium, M. intracellulare, M. africanum, M. kansasii, M. marinum, M. ulcerans, M. avium  subspecies paratuberculosis,  Nocardia asteroides , other  Nocardia  species,  Legionella pneumophila , other  Legionella  species,  Salmonella typhi , other  Salmonella  species,  Shigella  species,  Yersinia pestis, Pasteurella haemolytica, Pasteurella multocida , other  Pasteurella  species,  Actinobacillus pleuropneumoniae, Listeria monocytogenes, Listeria ivanovii, Brucella abortus , other  Brucella  species,  Cowdria ruminantium, Chlamydia pneumoniae, Chlamydia trachomatis, Chlamydia psittaci, Coxiella burnetti , other  Rickettsial  species,  Ehrlichia  species,  Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Bacillus anthracis, Escherichia coli, Vibrio cholerae, Campylobacter  species,  Neiserria meningitidis, Neiserria gonorrhea, Pseudomonas aeruginosa , other  Pseudomonas  species,  Haemophilus influenzae, Haemophilus ducreyi , other  Hemophilus  species,  Clostridium tetani, Clostridium difficile , other  Clostridium  species,  Yersinia enterolitica , and other  Yersinia  species. 
     
     
         20 - 24 . (canceled) 
     
     
         25 . The method of  claim 12 , wherein the toxin is selected from the group consisting of Abrin, Conotoxins Diacetoxyscirpenol Bovine spongiform encephalopathy agent, Ricin, Saxitoxin, Tetrodotoxin, epsilon toxin, Botulinum neurotoxins, Shigatoxin, Staphylococcal enterotoxins, T-2 toxin, Diphtheria toxin, Tetanus toxoid, and pertussis toxin. 
     
     
         26 . The method of  claim 12 , wherein the newly proliferated ASC are separated and isolated from the plasma via ficoll gradient separation or elutriation. 
     
     
         27 . The method of  claim 12 , wherein the newly proliferated ASC are separated and isolated from the separated cells via magnetic bead sorting or fluorescence acquired cell sorting (FACS). 
     
     
         28 . The method of  claim 12 , wherein newly proliferated ASC are separated and isolated from the separated cells through use of antibodies specific for one or more cell surface markers comprising CD38, CD27, CD19, CD138, IgD, and Ki67. 
     
     
         29 . The method of  claim 12 , wherein the ASC are measured by immunoassay. 
     
     
         30 . The method of  claim 29 , wherein the immunoassay is an ELISPOT assay.