Interferon‐γ‐based mixed lymphocyte culture as a selection tool for allogeneic bone marrow donors other than identical siblings

Abstract

Selection procedures in bone marrow transplantation (BMT) would benefit from the development of easy-to-perform cellular assays with high discriminative power. We tested a cytokine-based mixed lymphocyte culture (MLC) and compared its outcome to the routinely used MLC, helper T-lymphocyte precursor (HTLp)-f and cytotoxic T-lymphocyte precursor (CTLp)-f assays. Interferon (IFN)gamma was selected as a marker cytokine for (deleterious) T-helper 1 like responses and 36 (potential) BMT donor-recipient pairs were analysed. The IFNgamma-MLC appeared sensitive to HLA class II (subtype) differences, but not to isolated class I differences, or to mismatches other than HLA (identical siblings). The test enabled a distinction between combinations with positive MLC (proliferation) and HTLp-f, exemplified by the fact that although high IFNgamma levels were observed in the class II mismatched group, certain DRB3, DQB1-subtype and DRB1-subtype mismatches did not give rise to IFNgamma production. This might be of relevance for the detection of so-called permissible mismatches. With regard to prediction of acute graft-versus-host disease (aGVHD) in unrelated BMT, the data indicated that high levels of IFNgamma coincided with severe aGVHD, whereas low levels were largely associated with grades 0-I. However, in the case of isolated class I mismatches the test had no predictive value. The cell-saving IFNgamma-MLC provides an alternative for the assays currently in use, but should be employed along with an assay that is sensitive to class I differences to correct for false negatives. Consequently, a combination of IFNgamma-MLC and CTLp-f assays seems most promising for donor selection, other than identical siblings.

CD57+ T cells augment IFN-γproduction in a one-way mixed lymphocyte reaction and their expansion after stem cell transplantation in paediatric patients

“…IFN‐ γ is also one of the important cytokines in controlling and amplifying the immune response against alloantigens or initiating acute GVHD [29]. On the other hand, MLR is a useful model of the immune response between donors and recipients [18] and IFN‐ γ is reported to play a key role in MLR [19].…”

“…We have shown that the IFN‐ γ production in MLR increased constantly with the responders’ age and also correlated with the proportion of CD57 + T cells in PBMC, regardless of the responders’ age. Because the levels of the IFN‐ γ produced in MLR can be used to predict the severity of acute GVHD [19], these findings may explain why an increasing donor's age for HSCT is associated with an increased risk of developing acute GVHD [32]. Several clinical studies have shown that acute GVHD can be prevented by removing T cells from the donor marrow.…”

“…The one‐way mixed lymphocyte reaction (MLR) is a useful predictor in vitro of acute GVHD that occurs after allogeneic HSCT [18]. The IFN‐ γ levels in MLR between donors and recipients have been reported recently to correlate with the severity of acute GVHD [19].…”

Immunologic monitoring

“…A comprehensive study of the kinetics of T helper (Th) 1 and Th2 cytokines produced in alloresponses has recently been published (39). The clinical utility of anti‐donor cytokine HTLp frequency measurement has been extensively demonstrated for IL‐2 (40, 41) and less extensively for IFN‐γ (42, 43). Our laboratory has data to support the assertion that the frequency of IFN‐γ‐secreting CD8 + T cells is of value in measuring the alloresponses to HLA class I disparities in unrelated donor hematopoietic cell transplantation (Brookes and Lechler, unpublished observations).…”

Zinc Modulates the Priming of T Helper 1, T Helper 17, and T Regulatory Cells in Allogeneic and Autologous in vitro Models

“…23,29,30 Graft versus host disease is a major challenge, and cytokine levels, such as IFN-γ, are sensitive indicators of graft rejection in MLC, hence correlating with the severity of acute GVHD. [31][32][33] For that reason, zinc can suppress allogeneic immune responses because of its ability to stabilize induced Tregs. 24 In our study, pro-inflammatory cytokine IFNγ production was significantly reduced in both zinc-treated restimulated T cells with autologous PBMC (p < 0.05) and allogeneic PBMC (p < 0.01) compared with untreated T cells.…”