Pretransplant Donor-Specific Helper T Cell Reactivity as a Tool for Tailoring the Individual Need for Immunosuppression1

Bj, van der Mast; Nm, van Besouw; P, de Kuiper; Lm, Vaessen; Pj, Gregoor; Jn, Ijzermans; T, van Gelder; Fh, Claas; Weimar, Willem

doi:10.1097/00007890-200109150-00023

Cited by 16 publications

(2 citation statements)

References 29 publications

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“…Because the allo-reactive repertoire contains both naive and memory T cells [10], [47], [48], we judge that simultaneous quantification of both subsets is an advantage of the current technique compared to ELISPOT. Together, these differences may explain why published PF of donor-specific T cells in human organ transplant recipients detected by LDA [8], [9], [10], [13], [49], [50] or ELISPOT [22], [23], [51], [52], [53] are significantly lower compared to those observed in this study with the CFSE-MLR. Importantly, the use of CD40-B cells as stimulators to detect allogeneic T-cell responses [54], and CFSE-dilution as a technique to measure the proliferative response of T cells to allo-antigens [17], [24], [25], [46], [47] have both been described, but to our best knowledge these techniques have never been combined.…”

Section: Discussioncontrasting

confidence: 71%

Detailed Kinetics of the Direct Allo-Response in Human Liver Transplant Recipients: New Insights from an Optimized Assay

et al. 2010

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Conventional assays for quantification of allo-reactive T-cell precursor frequencies (PF) are relatively insensitive. We present a robust assay for quantification of PF of T-cells with direct donor-specificity, and establish the kinetics of circulating donor-specific T cells after liver transplantation (LTx). B cells from donor splenocytes were differentiated into professional antigen-presenting cells by CD40-engagement (CD40-B cells). CFSE-labelled PBMC from LTx-recipients obtained before and at several time points after LTx, were stimulated with donor-derived or 3rd party CD40-B cells. PF of donor-specific T cells were calculated from CFSE-dilution patterns, and intracellular IFN-γ was determined after re-stimulation with CD40-B cells. Compared to splenocytes, stimulations with CD40-B cells resulted in 3 to 5-fold higher responding T-cell PF. Memory and naïve T-cell subsets responded equally to allogeneic CD40-B cell stimulation. Donor-specific CD4+ and CD8+ T-cell PF ranged from 0.5 to 19% (median: 5.2%). One week after LTx, PF of circulating donor-specific CD4+ and CD8+ T cells increased significantly, while only a minor increase in numbers of T cells reacting to 3rd party allo-antigens was observed. One year after LTx numbers of CD4+ and CD8+ T cells reacting to donor antigens, as well as those reacting to 3rd party allo-antigens, were slightly lower compared to pre-transplant values. Moreover, CD4+ and CD8+ T cells responding to donor-derived, as well as those reacting to 3rd party CD40-B cells, produced less IFN-γ. In conclusion, our alternative approach enables detection of allo-reactive human T cells at high frequencies, and after application we conclude that donor-specific T-cell PF increase immediately after LTx. However, no evidence for a specific loss of circulating T-cells recognizing donor allo-antigens via the direct pathway up to 1 year after LTx was obtained, underscoring the relative insensitiveness of previous assays.

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Section: Discussioncontrasting

confidence: 71%

Detailed Kinetics of the Direct Allo-Response in Human Liver Transplant Recipients: New Insights from an Optimized Assay

et al. 2010

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“…Early studies have defined the relative donor-specific response rate either as hypo-responsiveness or as hyper-responsiveness in kidney transplant recipients and related these responses to good graft function or acute rejection within the first year after transplantation, respectively ( 61 , 62 ). However, limited predictive value of the MLR for acute rejection has been reported since ( 63 , 64 ).…”

Section: In Vitro Methods To Detect Alloreactive T and B Celmentioning

confidence: 99%

Pre-existing Alloreactive T and B Cells and Their Possible Relevance for Pre-transplant Risk Estimation in Kidney Transplant Recipients

2020

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In allogeneic transplantation, genetic disparities between patient and donor may lead to cellular and humoral immune responses mediated by both naïve and memory alloreactive cells of the adaptive immune system. This review will focus on alloreactive T and B cells with emphasis on the memory compartment, their role in relation to kidney rejection, and in vitro assays to detect these alloreactive cells. Finally, the potential additional value of utilizing donor-specific memory T and B cell assays supplementary to current routine pre-transplant risk assessment of kidney transplant recipients will be discussed.

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Assays for Alloreactive Responses by PCR

Stordeur

2007

Methods in Molecular Biology

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In organ transplantation, major efforts are currently developed to minimize or even to withdraw immunosuppressive drugs. Different protocols have therefore been proposed to induce graft tolerance, that is, the survival of an allograft in the absence of continuing immunosuppression. They generally involve pre-transplant conditioning followed by hematopoietic stem cell infusion. Follow-up of immune status is mandatory in this kind of protocol, in order to investigate tolerance and consequently to reduce or withdraw immunosuppression without graft rejection. However, assessment of the alloreactive response using currently available techniques is labor-intensive and requires significant volume of patient's blood. The implementation of such tolerance protocols is thus difficult in routine practice. In this chapter, we describe a novel real-time polymerase chain reaction method based on interferon-gamma and interleukin-2 mRNAs quantification that requires only 10 ml of blood. Moreover, results can be obtained within 48 h. A modified mixed lymphocyte reaction (MLR) using whole blood T lymphocytes as responsive cells and CD3-depleted peripheral blood mononuclear cells (PBMCs) as stimulators is described. An alternative, that is, the use of PBMC as responding cells instead of whole blood, is also depicted. We successfully applied the technique in the monitoring of anti-donor reactivity in living donor liver graft recipients. We suggest that this rapid MLR assay could be valuable for the monitoring of patients undergoing solid organ or hematopoietic stem cell transplantation.

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Pretransplant Donor-Specific Helper T Cell Reactivity as a Tool for Tailoring the Individual Need for Immunosuppression1

Cited by 16 publications

References 29 publications

Detailed Kinetics of the Direct Allo-Response in Human Liver Transplant Recipients: New Insights from an Optimized Assay

Detailed Kinetics of the Direct Allo-Response in Human Liver Transplant Recipients: New Insights from an Optimized Assay

Pre-existing Alloreactive T and B Cells and Their Possible Relevance for Pre-transplant Risk Estimation in Kidney Transplant Recipients

Assays for Alloreactive Responses by PCR

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