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. Author manuscript; available in PMC: 2011 Jun 1.
Published in final edited form as: Hum Immunol. 2010 Mar 11;71(6):566–576. doi: 10.1016/j.humimm.2010.02.008

Figure 3. Enzyme-Linked ImmunoSpot (ELISPOT) assay for the enumeration of interferon-gamma and granzyme-B secreting cells in LRD -Kidney transplant recipients.

Figure 3

Figure 3

Two serial dilutions of recipient PMBC responder cells (3×105 & 3×104 per well) were stimulated with 3×105 per well T cell depleted donor or third party cells in capture antibody coated ELISPOT plates. After 36 hrs, the plates were washed and analyzed for the respective cytokines using biotinylated antibodies, followed by signal development and detection with ELISPOT reader. Each symbol represents data from individual recipients. The frequency of 75 cytokine secreting cells per million PMBC was considered the cut-off value for a positive response represented by the dashed line and the percentage of recipients demonstrating donor specific unresponsiveness are indicated below data symbols. The frequencies of IFN-γ (A) and Granzyme-B (B) secreting cells in the three groups of patients are shown.