Skin explant model of human graft-versus-host disease: prediction of clinical outcome and correlation with biological risk factors.

A human skin explant model has been used to predict the clinical outcome and to study the immunopathology of human graft-versus-host disease (GVHD). Whether the model gives the same predictive effect for GVHD in different hematopoietic stem cell transplantation (HSCT) settings has not been assessed. It is also unknown whether the skin explant result reflects the known biological risk factors for clinical GVHD. In this study, the skin explant model was used to detect graft-versus-host reactions (GVHR) in vitro for 225 eligible patient/donor pairs. The predicted skin GVHR grade was correlated with the outcome of clinical GVHD, as well as HLA matching status, sex mismatches, and patient age. In sibling HSCT under either myeloablative or reduced-intensity conditioning, a significant correlation was observed between the predicted skin GVHR and clinical GVHD (P < .001 and P = .033, respectively). In HSCT using unrelated donors, the involvement of T-cell depletion led to a sharp increase in false-positive GVHR results, and no correlation was observed between the predicted skin GVHR and clinical GVHD. The skin GVHR grade correlated significantly with the HLA matching status (HLA-matched sibling pairs, HLA-matched unrelated pairs, and HLA-unmatched unrelated pairs). Furthermore, HLA-matched sibling pairs with a female-to-male sex mismatch had a significantly higher overall skin GVHR grade and a higher ratio of high- versus low-grade skin GVHR than the sibling pairs with all other sex combinations. Patient age was not reflected in the skin explant result. In conclusion, the predictive value of the skin explant model for aGVHD varies depending on the clinical transplant protocols, such as the type of GVHD prophylaxis used. Nevertheless, the skin explant model remains a unique in vitro system that provides an in situ histopathologic readout for studying alloreactivity and human GVHD. The model has also the potential to aid the development of novel prophylaxis and treatment for GVHD.

Ex vivo expansion of long-term culture initiating marrow cells by IL-10, SCF, and IL-3.

BACKGROUND: Ex vivo expansion of progentior cells may shorten hematopoietic regeneration after myeloablative chemoradiotherapy, increase target cells for gene therapy, and improve purging of progenitor cell components. STUDY DESIGN AND METHODS: Marrow cells were incubated for 1 week in suspension culture with and without IL-10, IL-3, and SCF. As long-term culture initiating cells (LTC-ICs) represent early hematopoietic progenitors in vitro, these cells were quantified at initiation and after a 1-week culture period in a limiting dilution assays. Additionally, immunophenotyping of cells before and after culture was performed. RESULTS: In six experiments, marrow cells cultured for 1 week with IL-10, IL-3, and SCF showed a significant increase (almost doubling) in LTC-ICs as compared with marrow cells before expansion. Additionally, an increased proliferative capacity of LTC-ICs was achieved with a sevenfold increase of committed colony-forming cells and a 10-fold proliferation of high proliferative potential colony-forming cells. Immunophenotyping revealed a sevenfold increase of CD34+ CD45 RA- cells in IL-10-, IL-3-, SCF-stimulated suspension cultures. In unstimulated cultures, no LTC-ICs were maintained after 1 week. CONCLUSION: Expansion of LTC-ICs by IL-10, IL-3, and SCF has not been shown so far. This in vitro model allows expansion of LTC-IC if compared with the input of progenitor cells without extensive progenitor cell manipulation. This should be an attractive model for in vitro purging, gene transfer, or expansion of progenitor cells to allow rapid engraftment after myeloablative chemotherapy.