Epidermal Langerhans cells promote skin allograft rejection in mice with NF-kappa B-impaired T cells.

T cells play a major role in the acute rejection of transplanted organs. Using mice transgenic for a T-cell-restricted NF-kappaB super-repressor (IkappaBalphaDeltaN-Tg mice), we have previously shown that T-cell-NF-kappaB is essential for the acute rejection of cardiac but not skin allografts. In this study, we investigated the mechanism by which skin grafts activate IkappaBalphaDeltaN-Tg T cells. Rejection was not due to residual T-cell-NF-kappaB activity as mice with p50/p52(-/-) T cells successfully rejected skin grafts. Rather, skin but not cardiac allografts effectively induced proliferation of graft-specific IkappaBalphaDeltaN-Tg T cells. Rejection of skin grafts by IkappaBalphaDeltaN-Tg mice was in part dependent on the presence of donor Langerhans cells (LC), a type of epidermal dendritic cells (DC), as lack of LC in donor skin grafts resulted in prolongation of skin allograft survival and injection of LC at the time of cardiac transplantation was sufficient to promote cardiac allograft rejection by IkappaBalphaDeltaN-Tg mice. Our results suggest that LC allow NF-kappaB-impaired T cells to reach an activation threshold sufficient for transplant rejection. The combined blockade of T-cell-NF-kappaB with that of alternative pathways allowing activation of NF-kappaB-impaired T cells may be an effective strategy for tolerance induction to highly immunogenic organs.

Codominant expression of the polymorphic MICA alloantigens encoded by genes in the HLA region.

The HLA-related, polymorphic MHC class I-related chain A (MICA) gene encodes a 383-amino acid polypeptide, with three extracellular domains (alpha1, alpha2 and alpha3), a transmembrane region and a cytoplasmic tail. We have previously shown that freshly isolated endothelial cells, fibroblasts, keratinocytes and monocytes express MICA, while peripheral blood CD4+, CD8+ or CD19+ lymphocytes do not. This polymorphic MICA molecule is a target for specific alloantibodies in sera from kidney, heart and lung transplant recipients, although its possible role during graft rejection remains to be demonstrated. In this study we investigated whether there is codominance in the expression of MICA. We isolated RNA from a heterozygous cell line (HCT116), previously shown by sequencing-based typing to be MICA*001/MICA*00902, as well as 12 clones derived from it. Thereafter, we retrotranscribed the RNA into cDNA, and performed a molecular typing using MICA-sequence specific oligonucleotides (SSOP). Using this approach, we detected the RNA encoding MICA*001 and MICA*00902 in all the clones and in the parental cell line, indicating that MICA is codominantly expressed. This codominant expression was further confirmed by cloning and sequencing plasmids encoding these two alleles produced from the same HCT116 RNA preparation. We also produced the two recombinant MICA proteins (MICA*001 and MICA*00902). They reacted with rabbit anti-MICA polyclonal antibodies by ELISA and Western blot, indicating that the plasmids carrying the cDNA inserts probably encode functional MICA proteins. This strongly suggests that, like the HLA class I and class II proteins, MICA is codominantly expressed. The codominant expression of the polymorphic, HLA-like MICA alloantigens may have implications for the immune response elicited by the allograft in organ transplantation.