ELISPOT History

The ELISPOT Assay is competing with other methods measuring single cell cytokine production, e.g. intracellular cytokine by FACS or Miltenyl cytokine secretion assay. Other types of lymphocyte frequency and function assays include limiting dilution assay (LDA), cytotoxic T cell assay (CTL), tetramer staining. Compared with respect to sensitivity the ELISPOT Assay is outranking other methods to define frequency of antigen-specific lymphocytes. ELISPOT Assays are more convenient, cheaper and faster than most of the other methods and may be implemented in a daily routine of immune diagnostics.

Milestones and major advances in the development and the applications of the ELISPOT Assay

  • 1983 Czerkinsky's first description of a method to enumerate frequency of B hybridoma cells
         secreting an antigen specific immunoglobulin.
  • 1988 Again, Czerkinsky is the first to develop an ELISA Spot Assay measuring the frequency of T
         lymphocytes secreting a specific lymphokine.
  • 1988 First description of a Dual-Color ELISPOT (guess who...:-)
  • 1993 First trials to use computer imaging for analysing and counting spots
  • First membrane bottomed plate to perform ELISPOT Assays
  • Several papers validating the ELISPOT Assay and comparing its results with related functional
         immune cell assays
  • ELISPOT as a basis of worldwide WHO vaccination studies
  • ELISPOT as an integral step to optimize cell based antigen-specific anti tumour therapy
  • Future development: e.g. Fluorescent multi-color ELISPOT and high throughput immune
         toxicological studies in drug development. 

    Historical references about ELISPOT Assays

    Czerkinsky, C., Nilsson, L.angstrom., Nygren, H., Ouchterlony, O., and Tarkowski, A. 1983. 
    A solid-phase enzyme-linked immuno-spot (ELISPOT) assay for enumeration of specific 
    antibody-secreting cells. J. Immunol. Methods 65:109-121.

    Sedgwick, J.D. and Holt, P.G. 1983. A solid-phase immunoenzymatic technique for the 
    enumeration of specific antibody-secreting cells. J. Immunol. Methods 57:301-309.

    Franci, C., Ingles, J., Castro, R., and Vidal, J. 1986. Further studies on the ELISA-spot technique. 
    Its application to particulate antigens and potential improvement in sensitivity. 
    J. Immunol. Methods 88:225-232.



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