title: Efficiency of human HLA-mismatched CD34+ cells from unrelated donors in establishing in vitro hematopoiesis in allogeneic long-term marrow cultures.
abstract: We have examined the capacity of highly purified human CD34+ marrow cell isolates from unrelated, HLA-mismatched donors to establish in vitro hematopoiesis on recipient marrow stromal cells in 2-stage hematopoietic long-term marrow cultures (H-LTMC). HLA-typing of both peripheral blood mononuclear cells and CD34+ marrow cells was performed for both HLA class I and HLA class II antigens for eight healthy individuals. Significant antigenic mismatches for these molecules ranged from three to six antigens for each recipient-donor pair. Comparison of MHC antigen expression by peripheral blood cells and CD34+ marrow cell isolates confirmed the presence of identical HLA-A, -B, and -C, and -DR specificities on the surface of these cells. Typing of -DQ specificities, however, was not consistently reactive on CD34+ cells. The > or = 20% plating efficiency of purified CD34+ cells for BFU-E, CFU-GM, and CFU-MIX allowed us to use inoculum doses of 10(3), 10(4), and 10(5) cells to determine the efficiency of allogeneic CD34+ cells in achieving in vitro engraftment and the establishment of hematopoiesis in H-LTMC. Engraftment of adherent BFU-E, CFU-GM, and CFU-MIX was equally efficient for autologous and allogeneic CD34+ cells. In vitro hematopoiesis reflected by the cumulative recoveries of progenitor cells over time was also equivalent for allogeneic and autologous CD34+ cells. These results demonstrate that highly purified, HLA-mismatched CD34+ marrow cells proliferate and establish in vitro hematopoiesis as efficiently as autologous cells in marrow derived stromal cell cultures and confirm that interactions between stromal cells and highly purified CD34+, DR-, and CD34+, DR+ marrow cell isolates are not MHC-restricted.
authors.affiliation: Department of Hematology and Vascular Biology, Walter Reed Army Institute of Research, Washington, DC, USA.
authors.forename: V F
authors.initials: VF
authors.lastname: La Russa
authors.forename: S W
authors.initials: SW
authors.lastname: Kessler
authors.forename: V A
authors.initials: VA
authors.lastname: Henson
authors.forename: M
authors.initials: M
authors.lastname: Cutting
authors.forename: T
authors.initials: T
authors.lastname: Polsinelli
authors.forename: R D
authors.initials: RD
authors.lastname: Knight
authors.forename: D G
authors.initials: DG
authors.lastname: Wright
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002450:Cell Communication; D002478:Cells, Cultured; D006650:Histocompatibility Testing; D006801:Humans; D013234:Stem Cells / Q000166:cytology / Q000276:immunology*; D017154:Stromal Cells / Q000166:cytology / Q000276:immunology*; D014019:Tissue Donors
chemical_list: D018952:Antigens, CD34