Human cytokine production from engineered stromal cells influences lineage development from CO-transplanted human CD 34' progenitors

Ma Dao, L. L. Wiltshire, K. A. Pepper, Jan Nolta

Research output: Contribution to journalArticle

Abstract

We developed a model system for the sustained growth of adult human bone marrow cells m immune deficient (bnx) mice, following retroviral-mediated gene transfer. Human hematopoiesis from CD34progenitors is supported by co-injection of human bone marrow stromal celts engineered to secrete human IL-3 (hIL-3) The species specificity of IL-3 allows support of human hemalopoietic cells in the mice without perturbing murine hematopoiesis. The /wu/hu cotransplantation system was used to demonstrate transduction of human hemalopoietic cells capable of giving rise to progeny of both T lymphoid and myeloid lineages, by clonal integration analysis (Noltt et al, PNAS 93, in prêts, 1996). The major limitation of our model is a total absence of human B cell development in the mice, which could be due to supcaphysiological levels of hIL-3 in the circulation. To try to obtain human B lymphoid, as well as T lymphoid and myeloid cell development m the mice, we co-injected CD34cclls with human marrow stromal cells engineered to secrete human EL-2, IL-7, Stem Cell Factor (SCF) or FLT3 ligand (Ft), +/- IL-3. No single factor other than IL-3 supported sustained human hematopoiesis in the mice. The use of IL-2 was discontinued due to adverse effects on the murine hematopoietic system. Co-expression of SCF or FL with IL-3 had no oven effect on human hematopoiesis. Production of both human IL-3 and IL 7 m the mice supported extrathymic development of human T lymphocytes for 7-8 months, but no B cells, myeloid cells, or colony-forming progenitors were detected. Human B cells have not yet been produced from CD34" cells in the bnx mice under any condition. Another limitation to the jb/w/hu system is a lack of maturation of human red blood cells, although BFU-E are maintained. Mature red celi development in the mice could provide a useful model to study gene therapy for globm disorders in vivo. We co-transplanted stromal cells secreting human erythropoietin (Epo) and hIL-3 into mice. An increase in hematocrit from 40-45% to 80-85% resulted, with production of human and murine red cells. All mice (N=6) displayed paralysis and died within one month. We are currently attempting lo refine this system by reducing the level of human Epo expression. The bnxfha cotransplantaiion model provides an interesting system in which to study human hematopoietic cell differentiation under the influence of various cytokines.

Original languageEnglish (US)
Pages (from-to)1117
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
StatePublished - 1996
Externally publishedYes

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

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