Positive selection of hematopoietic CD34+ stem cells provides 'indirect purging' of CD34- lymphoid cells and the purging efficiency is increased by anti-CD2 and anti-CD30 immunotoxins

R. M. Lemoli, P. L. Tazzari, A. Fortuna, A. Bolognesi, S. C. Gulati, F. Stirpe, S. Tura

Research output: Contribution to journalArticlepeer-review

Abstract

Human CD34+ hematopoietic cells were purified using the avidin-biotin immunoabsorption technique. The selected population showed 78.6 ± 3% CD34+ cells and the overall recovery of CD34+ cells, CFU-GM and BFU-E from the starting population was 34 ± 5%, 71 ± 4% and 67 ± 2%, respectively. Hematopoietic progenitor cell purification also resulted in 3 log of normal T cell depletion from the bone marrow (BM) by immunofluorescence analysis. Moreover, when unseparated BM cells were mixed with the CD34- lymphoma cell lines D430B and Raji, the removal of greater than 3 log of tumor cells from the enriched CD34+ cell fraction was demonstrated. To increase the neoplastic cell purging, several immunotoxins (IT) containing the ribosome-inactivating protein (RIP) saporin and directed toward the lymphoid-associated antigens CD30 and CD2 were prepared. Our experiments showed only a minimal toxicity of the immunoconjugates on colony-forming cells (CFC) derived from purified CD34+ cells. Conversely, all the ITs were very effective in inhibiting protein synthesis and growth of normal and neoplastic lymphoid cells. Further experiments demonstrated that the sequential combination of CD34+ cells purification and IT treatment resulted in 5 or more log of tumor cell purging with no additional loss of BM progenitor cells.

Original languageEnglish
Pages (from-to)465-471
Number of pages7
JournalBone Marrow Transplantation
Volume13
Issue number4
Publication statusPublished - 1994

ASJC Scopus subject areas

  • Hematology
  • Transplantation

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