Optimizing Culture Conditions for the Generation of Human Regulatory Dendritic Cells

by Megan Kruckenberg* (BVU), Sabrina Scroggins B.S. and Annette Schlueter MD PhD. (Graduate Program in Immunology & Department of Pathology, University of Iowa)
Biochemistry
Faculty advisor: Dr. Brian Lenzmeier

Graft-versus-host disease (GVHD) is a problem that arises from allogeneic bone marrow transplants that are used to treat hematopoietic malignancies, inherited hematopoietic disorders, and aplastic anemia.  GVHD is the result of donor transplant tissue recognizing the recipient tissue as “non-self,” and therefore, mounting an immune response to attack recipient tissue. GVHD occurs in about 50% of allogeneic transplants and is directly fatal in about 30% of those patients, despite matching a donor to the recipient and prescribing immunosuppressive drugs1,2. It would be useful to find a treatment for GVHD that minimizes the side effects common with administration of immunosuppressive drugs. Mouse models have been created to look at the possibility of culturing regulatory dendritic cells (DCreg) from the recipient that could be injected after transplantation to regulate T cell activity. Such studies appear promising to prevent GVHD while allowing appropriate immune responses to occur. 

Our work aims to develop a technique for creating DCreg for human use. We cultured peripheral blood mononuclear cells (PBMNCs) isolated from leukoreduction chambers (LRS chambers) under DC reg conditions. Activated cultures were stimulated with TNF alpha, or cultured after freezing in different freeze medias. Our results show that DCreg can be cultured from both young and older donors from both fresh and frozen samples. We have found that samples from both age groups can be stimulated, and that purifying the samples prior to culture yields a better product.