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A Renewable Source of Donor Cells for Repetitive Monitoring of T- and B-Cell Alloreactivity

2005; Elsevier BV; Volume: 5; Issue: 1 Linguagem: Inglês

10.1111/j.1600-6143.2003.00637.x

1600-6143

Martin S. Zand, Anirban Bose, Thuong Vo, Myra Coppage, Tina Pellegrin, Lois J. Arend, F. Eun-Hyung Lee, Adel Bozorgzadeh, Nufatt Leong,

Immune Cell Function and Interaction

A major impediment to repetitive monitoring of alloreactivity or tolerance is the limited supply of donor cells available for assays of host-versus-graft T- and B-cell reactivity. In this paper, we describe the use of CD40L stimulated CD19+ B cells as targets or stimulators in flow cytometric crossmatching (FXM), mixed lymphocyte reactivity and IFN-γ ELISPOT assays. Stimulated B cells (sBc) express high levels of MHC class I and II, as well as the costimulatory molecules CD80 and CD86. They can be polyclonally expanded and frozen for later use. We describe the use of sBc in ELISPOT, mixed lymphocyte cultures and FXM. CD4+ T cells exposed to sBc express a similar cytokine profile as those stimulated with unfractionated PBMC. We further analyzed T- and B-cell responses in 14 patients on the renal transplant waiting list, finding that those with an elevated panel reactive antibody (PRA) (>60%) had higher alloreactive T-cell precursor frequencies as measured by CDFSE MLR and IFN-γ ELISPOT. We conclude that sBc are a renewable source of donor-specific target/stimulator cells for use in repetitive and coordinate assays of B- and T-cell alloreactivity.