Enzyme Linked Immunosorbent Spot (ELISPOT) Assay for Interferon‐Gamma Independently Predicts Renal Function in Kidney Transplant Recipients

Abstract

Post-transplant monitoring of cellular immunity might be useful in predicting long-term outcomes of kidney transplant recipients. We used an enzyme linked immunoabsorbent spot (ELISPOT) assay to serially measure the frequency of peripheral blood lymphocytes producing interferon-gamma in response to stimulator cells from donors or third parties in 55 primary kidney transplant recipients. Mean frequencies measured during the first 6 months after transplantation correlated significantly with the serum creatinine concentration at both 6 and 12 months following transplantation. The mean frequencies were higher in patients with acute rejection than in those without acute rejection. Multiple regression analyses indicated that the correlations between the early ELISPOT measurements of interferon-gamma and serum creatinine were independent of acute rejection, delayed graft function, or the presence of panel reactive antibodies before transplantation. Patients with low mean frequencies of interferon-producing cells in the early post-transplant period were generally free from acute rejection and exhibited excellent renal function at 6 and 12 months post-transplant. In conclusion, using the ELISPOT assay, we show an independent correlation between early cellular alloreactivity and long-term renal function. Increased levels of early alloreactivity measured with this assay may serve as a surrogate for chronic allograft dysfunction.

How Can We Measure Immunologic Tolerance in Humans?

“…In addition, the assay has been optimized for use of frozen cells that can be easily stored. Although no studies to date have reported the use of this assay to identify functionally tolerant humans after transplantation, the frequency of IFN-␥-producing cells has been reported to correlate with the incidence of acute and chronic rejection as well as allograft function after renal transplantation (45)(46)(47). Obtaining and storing sufficient numbers of donor cells to perform the assay repeatedly is a practical limitation, particularly in recipients of deceased-donor organs.…”

Achieving Donor-Specific Hyporesponsiveness Is Associated with FOXP3+ Regulatory T Cell Recruitment in Human Renal Allograft Infiltrates

“…Certainly, the donorspecific immune evaluation is feasible nowadays; on one hand, donorspecific alloantibodies can be precisely monitored by flow cytometric, cytotoxicity, or ELISA crossmatching techniques, and on the other, high frequency of donor-specific alloreactive T cells can be evaluated by the ELISPOT assay. Indeed, these procedures have been shown to be useful for predicting graft outcome (27)(28)(29)(30)(31)(32).…”

HSV infection and immunosuppression

“…Measurements of mitogen-stimulated ATP production by T-lymphocytes, and of immunoreactive Tcells by techniques such as the ELISPOT assay or flow cytometry, can better educate and inform the clinician confronting the unique treatment dilemma presented by this patient. 9,10 Rakesh Sindhi, MD, FACS Hillman Center for Pediatric Transplantation Children's Hospital of Pittsburgh Pittsburgh, PA…”