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ELISPOT assay in Transplantation

ELISPOT assay in studying immunosurveillance after transplantation ELISPOT and organ transpantation

In the field of organ transplantation, there is need for reliable in vitro assays to predict rejection and as guide for therapeutic interventions after transplantation. The T cell ELISPOT assay has proven to be a promising tool to generate these outcomes, since it can accurately quantify the frequency and cytokine profile of circulating donor-reactive T lymphocytes. Several studies have shown that the number of cytokine producing T cells correlates with post-transplantation outcomes1. An increased frequency of donor-reactive IL-21 producing cells before kidney transplantation correlates with risk of acute rejections and impaired post-transplantation function1,2. Additionally, the assay may have an important role for immune monitoring of alloreactivity after reduction or withdrawal of immunosuppressive therapy, and for identification of patients tolerant to their allograft. In addition, the immunosurveillance after transplantation can be monitored by T cell ELISPOT assay3,4,5,6. Frequent and reliable T cell monitoring is feasible by the T cell ELISPOT assay. Therefore, clinicians may use the ELISPOT assay in combination with diagnostic assays to optimize the immunosuppressive load with minimal side effects to prolong transplant and patient survival.

The ELISPOT assay can also be helpful in understating the functional properties of T cells that emerge around the onset of acute Graft-versus-Host Disease (GVHD) in allogeneic hematopoietic stem cell transplantation (HSCT). Acute GVHD is an immune cell-driven, potentially fatal complication of allogeneic HSCT that affects multiple organs, including the skin, liver and gastrointestinal tract. This study demonstrated that CXCR3+ T cells of donor origin prevalent in patients release IFN-γ and Granzyme B on short-term activation, indicating that these inflammation-promoting and tissue-destructive compounds may be co-drivers of acute GVHD7.

Both the T cell and B cell ELISPOT assay can be useful when studying T cell mediated help to B cells, by for example IL-21. This study shows that one month after vaccination, kidney transplant recipients have significantly lower levels of both SARS-CoV-2-specific IL-21 producing T cells and SARS-CoV-2-specific IgG producing B cells than healthy controls. This may be due to the use of specific immunosuppressive medications and may explain both the poor humoral and cellular responses to vaccination observed in these patients8.

 

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Examples of studies using our ELISPOT assay:

Click on the authors for the abstract of the below mentioned acticles or find them in our Reference Database.

 

  1. van Besouw et al. (2019) The Number of Donor-Specific IL-21 Producing Cells Before and After Transplantation Predicts Kidney Graft Rejection. Front Immunol 10: 748
  2. Mendoza R et al. (2021) Pre-transplant donor-reactive IL-21 producing T cells as a tool to identify an increased risk for acute rejection. Sci Rep 11(1): 12445
  3. Bererhi L et al. (2012) Clinical and immunological features of very long-term survivors with a single renal transplant. Transpl Int 25(5): 545-554
  4. Candon S et al. (2009) Humoral and cellular immune responses after influenza vaccination in kidney transplant recipients. Am J Transplant 9(10): 2346-2354
  5. Dedeoglu B et al. (2017) T-Cell Composition of the Lymph Node Is Associated with the Risk for Early Rejection after Renal Transplantation. Front Immunol 8: 1416
  6. Paul-Heng M et al. (2018) Direct recognition of hepatocyte-expressed MHC class I alloantigens is required for tolerance induction. JCI Insight 3(15): e97500
  7. van Halteren AGS et al. (2023) A unique immune signature in blood separates therapy-refractory from therapy-responsive acute graft-versus-host disease. Blood 141(11): 1277-1292
  8. Malahe SRK et al. (2023) The role of interleukin-21 in COVID-19 vaccine-induced B cell-mediated immune responses in patients with kidney disease and kidney transplant recipients. Am J Transplant 23(9): 1411-1424