IN VITRO PROPAGATION AND HOMING OF LIVER-DERIVED DENDRITIC... : Transplantation (original) (raw)

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IN VITRO PROPAGATION AND HOMING OF LIVER-DERIVED DENDRITIC CELL PROGENITORS TO LYMPHOID TISSUES OF ALLOGENEIC RECIPIENTS

Implications for the Establishment and Maintenance of Donor Cell Chimerism Following Liver Transplantation1,2

Thomson, Angus W.3,4,5; Lu, Lina3; Subbotin, Vladimir M.3; Li, Youping3; Qian, Shiguang3; Rao, Abdul S.3,6; Fung, John J.3; Starzl, Thomas E.3

Pittsburgh Transplantation Institute and Departments of Surgery, Pathology, and Molecular Genetics and Biochemistry, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania 15213

1 Presented at the 13th Annual Meeting of the American Society of Transplant Physicians, May 16-18, 1994, Chicago, IL.

2 This work was supported in part by National Institutes of Health Grant DK 29961-14.

3 Pittsburgh Transplantation Institute and Department of Surgery, University of Pittsburgh.

4 Department of Molecular Genetics and Biochemistry, University of Pittsburgh.

5 Address correspondence to Dr. Angus W. Thomson, Pittsburgh Transplantation Institute, University of Pittsburgh Medical Center, W1544 Biomedical Science Tower, 200 Lothrop St., Pittsburgh, PA 15213-2582.

6 Department of Pathology, University of Pittsburgh.

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Abstract

Dendritic cell (DC) progenitors were propagated in liquid culture from nonparenchymal cells resident in normal mouse (B10.BR; H-2k, I-E+) liver in response to granulocyte-macrophage colony stimulating factor (GM-CSF). The liver-derived DC progenitors were MHC class II−/dim and did not express counter receptors for CTLA-4, a structural homologue of the T cell activation molecule CD28. Following subcutaneous or intravenous injection, these liver-derived cells migrated to T cell-dependent areas of lymph nodes and spleen of unmodified, allogeneic (B10; H-2b; I-E−) recipients, where they were identified 1-5 days, and 1 and 2 months after injection by their strong surface expression of donor MHC class II (I-Ek) and their dendritic morphology. Maximal numbers of liver-derived DC in the spleen were recorded 5 days after injection. Both clusters of strongly donor MHC class II+ cells—and (more rarely) dividing cells—could also be identified, suggesting cell replication in situ. Using the same techniques employed to generate DC progenitors from normal liver, GM-CSF-stimulated cells were propagated for 10 days from the bone marrow and spleen of nonimmunosuppressed mice sacrificed 14 days after orthotopic liver transplantation (B10;H-2b → C3H;H-2k). Immunocytochemical staining for recipient and donor MHC class II phenotype revealed the growth both of host cells with DC characteristics, and of cells expressing donor alloantigens (I-Ab). These results are consistent with the growth, in response to GM-CSF, of donor-derived DC from progenitors seeded from the liver allograft to recipient lymphoid tissue. The functional activity of the progenitors of chimeric DC and the possible role of these cells in the establishment and maintenance of donor-specific tolerance following liver transplantation remain to be determined.