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

Bone Marrow Transplant. 1994 Apr;13(4):465-71.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD* / analysis
  • Antigens, CD34
  • Antigens, Differentiation, T-Lymphocyte / analysis
  • Antigens, Differentiation, T-Lymphocyte / immunology*
  • Bone Marrow Purging / methods*
  • CD2 Antigens
  • Cell Separation / methods*
  • Cytotoxicity, Immunologic
  • Hematopoietic Stem Cells* / chemistry
  • Humans
  • Immunosorbent Techniques
  • Immunotoxins / pharmacology*
  • Ki-1 Antigen / analysis
  • Ki-1 Antigen / immunology*
  • Lymphocyte Activation / drug effects
  • N-Glycosyl Hydrolases*
  • Plant Proteins / pharmacology*
  • Protein Biosynthesis
  • Receptors, Immunologic / analysis
  • Receptors, Immunologic / immunology*
  • Ribosome Inactivating Proteins, Type 1
  • Saporins

Substances

  • Antigens, CD
  • Antigens, CD34
  • Antigens, Differentiation, T-Lymphocyte
  • CD2 Antigens
  • Immunotoxins
  • Ki-1 Antigen
  • Plant Proteins
  • Receptors, Immunologic
  • Ribosome Inactivating Proteins, Type 1
  • N-Glycosyl Hydrolases
  • Saporins