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- W2528540568 abstract "Graft versus host disease remains a major complication after allogeneic bone marrow transplantation, both in the HLA matched and unmatched settings. We have established a model system for the selective removal of alloreactive lymphocytes from blood mononuclear preparations. Initial work was done with unrelated donor-recipient pairs and we demonstrated the specific depletion of alloreactivity to the recipient while retaining third party responses. We have now extended our work to HLA-matched pairs Peripheral blood mononuclear cell preparations from HLA matched donorrecipient pairs were mixed in a modified primary mixed lymphocyte culture (MLC). This involved pre-treating the cells of the recipient either with phytohaemagglutin or with cytokines ( gamma-interferon or tumour necrosis factor-alpha). IL2 was also added to the MLC. A modification of the standard MLC was necessary as the latter is often unreactive between HLA matched pairs. The cells from the recipient were irradiated prior to mixing. Alloreactive donor lymphocytes were then identified by the expression of activation markers, specifically CD69. CD69 positive lymphocytes (alloreactive cells) were then removed by passing the cells through a magnetic cell sorter (MACS) using CD69-FITC and Anti-FITC microbeads. There was a median depletion of 67% in alloreactive cells and good recovery of lymphocytes of up to 77%. The depleted donor fraction was then retested with the original irradiated recipient cells in a secondary mixed lymphocyte culture. As a control, a third party irradiated stimulator was mixed with donor lymphocytes in a primary MLC and also with the CD69 depleted fraction post sorting. Both the primary and secondary MLC were pulsed with tritiated thymidine and harvested 18 hours later. A marked reduction ( up to 85%) in the proliferative response of the donor-recipient pair was noted after selective depletion. In contrast the proliferative response to third party control was unchanged. On a clinical level, this may translate into the abrogation of graft versus host disease while preserving graft versus leukaemia and anti-viral effect. The other advantage of this method is that magnetic sorting physically separates out the alloreactive lymphocytes without destroying them. This CD69 enriched alloreactive fraction may then be stored and used as donor lymphocyte infusions if leukaemia relapse occurs." @default.
- W2528540568 created "2016-10-14" @default.
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- W2528540568 date "1997-12-01" @default.
- W2528540568 modified "2023-09-26" @default.
- W2528540568 title "Selective depletion of alloreactive lymphoctyes in HLA matched pairs" @default.
- W2528540568 hasPublicationYear "1997" @default.
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