ELISPOT in Organ Transplantation

ELISPOT and organ transpantationIn the organ transplantation field, there is need for reliable in vitro assays to predict outcome and as guide for therapeutic interventions after transplantation. The ELISPOT assay has proven to be a promising tool to generate these markers, since it can accurately quantify the frequency and cytokine profile of circulating donor-reactive T lymphocytes. Different studies have shown that the number of IFN-γ producing T cells correlated with posttransplantation outcomes. Increased frequency of donor-specific IFN-γ producing cells before transplantation was related with risk of acute rejections and impaired posttransplant function. Additionally, the assay has high value in immune monitoring of alloreactivity after reduction or withdrawal of toxic immunosuppressants and identifying patients tolerant to their allograft. Frequent and reliable T-cell monitoring is feasible by the ELISPOT assay. Therefore, clinicians may use the ELISPOT assay in conjunction with additional tests as a routine basis to optimize immunosuppressive therapy with minimal side effects to prolong transplant and patient survival.

For example, Van Besouw et al. (2005) measured the number of IFN-γ producing cells reactive to donor cells in peripheral blood mononuclear cells (PBMC) of heart transplant patients with the human IFN-γ ELISPOT before, during and after a period of acute rejection (AR). The recognition of allo-major histocompatibility complex (MHC) antigens by T cells is the central event that initiates AR.
The authors showed that the T cell response directed against donor antigens was always readily detectable (Figure 1). This response increased significantly during an episode of AR and after successful treatment the response decreased (p<0.05). Van Besouw et al. showed in their study that the ELISPOT assay is a useful tool to determine T cell alloreactivity.

Human IFN-gamma ELISPOT and heart transplantation

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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.

van Besouw NM,  Zuijderwijk JM, Vaessen LMB, Balk AHMM, Maat APWM, van der Meide PH, and Weimar W.
The direct and indirect allogeneic presentation pathway during acute rejection after human cardiac transplantation.
Clin Exp Immunol 141: 1534-40 (2005).
U-CyTech products used in this study:
Human IFN-γ ELISPOT kit

Cunningham E.C., Tay S.S., Wang C., Rtshiladze M., Wang Z.Z., McGuffog C., Cubitt J., McCaughan G.W., Alexander I.E., Bertolino P., Sharland A.F., Bowen D.G. and Bishop A.G.
Gene therapy for tolerance: high-level expression of donor major histocompatibility complex in the liver overcomes naive and memory alloresponses to skin grafts.
Transplantation 95: 70-7 (2013)
U-CyTech products used in this study:
Mouse IFN-γ ELISPOT kit

Gerrits JH, van de Wetering J, Drabbels JJ, Claas FH, Weimar W, and van Besouw NM.
Donor-reactive cytokine profiles after HLA-identical living-related kidney transplantation.
Nephrol Dial Transplant 23:2016-23 (2008).
U-CyTech products used in this study:
Human IFN-γ ELISPOT kit
Human IL-10 ELISPOT kit
Human IL-13 ELISPOT kit
Human Granzyme B ELISPOT kit

Hochmeister S, Zeitelhofer M, Bauer J, Nicolussi EM, Fischer MT, Heinke B, Selzer E, Lassmann H, and Bradl M.
After injection into the striatum, in vitro-differentiated microglia- and bone marrow-derived dendritic cells can leave the central nervous system via the blood stream.
Am J Pathol 173:1669-81 (2008).
U-CyTech products used in this study:

Kloosterboer FM, van Luxemburg-Heijs SA, Willemze R, and Falkenburg JH.
Similar potential to become activated and proliferate but differential kinetics and profiles of cytokine production of umbilical cord blood T cells and adult blood naive and memory T cells.
Hum. Immunol. 67: 874-83 (2006).
U-CyTech products used in this study:
Human IFN-γ ELISPOT kit
Human IL-2 ELISPOT kit
Human IL-4 ELISPOT kit
Human IL-10 ELISPOT kit

Lv M, Li Y, Yu M, Sun Y, Lin Z, Qiao C, Luo Q, Gu X, Huang Y, Feng J, and Shen B.
Structured to reduce the mitogenicity of anti-CD3 antibody based on computer-guided molecular design.
Int J Biochem Cell Biol 39: 1142-55 (2007).
U-CyTech products used in this study:
Human IFN-γ ELISPOT kit
Human IL-2 ELISPOT kit

van der Meide PH, Joosten AM, Hermans P, Kloosterman TC, Olsson T, and de Labie MC.
Assessment of the inhibitory effect of immunosuppressive agents on rat T cell interferon-gamma production using an ELISPOT assay.
J Immunol Methods 144: 203-13 (1991).
U-CyTech products used in this study:
Rat IFN-γ ELISPOT antibody pair

Zanone MM, Favaro E, Quadri R, Miceli I, Giaretta F, Romagnoli R, David E, Perin PC, Salizzoni M, and Camussi G.
Association of cytomegalovirus infections with recurrence of humoral and cellular autoimmunity to islet autoantigens and of type 1 diabetes in a pancreas transplanted patient.
Transpl Int 23:333-7 (2010).
U-CyTech products used in this study:
Human IFN-γ ELISPOT kit