Restoration of human B-cell differentiation into NOD-SCID mice engrafted with gene-corrected CD34+ cells isolated from Artemis or RAG1-deficient patients

Mol Ther. 2008 Feb;16(2):396-403. doi: 10.1038/sj.mt.6300353. Epub 2007 Nov 27.

Abstract

Severe combined immunodeficiency (SCID) caused by mutation of the recombination-activating gene 1 (RAG1) or Artemis gene lead to the absence of B- and T-cell differentiation. The only curative treatment is allogeneic bone marrow (BM) transplantation, which displays a high survival rate when an HLA compatible donor is available but has a poorer prognosis when the donor is partially compatible. Consequently, gene therapy may be a promising alternative strategy for these diseases. Here, we report that lentiviral gene-corrected BM CD34(+) cells (isolated from Artemis- or RAG1-deficient patients) sustain human B-cell differentiation following injection into non-obese diabetic/SCID (NOD-SCID) mice previously infused with anti-interleukin-2 receptor beta chain monoclonal antibody. In most of the mice BM, engrafted with Artemis-transduced cells, human B-cell differentiation occurred until the mature stage. The B cells were functional as human immunoglobulin M (IgM) was present in the serum. Following injection with RAG1-transduced cells, human engraftment occurred in vivo but B-cell differentiation until the mature stage was less frequent. However, when it occurred, it was always associated with human IgM production. This overall approach represents a useful tool for evaluating gene transfer efficiency in human SCID forms affecting B-cell development (such as Artemis deficiency) and for testing new vectors for improving in vivo RAG1 complementation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antigens, CD34 / genetics
  • Antigens, CD34 / physiology*
  • B-Lymphocytes / cytology*
  • B-Lymphocytes / metabolism
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / metabolism
  • Bone Marrow Transplantation / methods*
  • Cell Differentiation / genetics
  • Cell Differentiation / physiology*
  • DNA-Binding Proteins
  • Endonucleases
  • Homeodomain Proteins / genetics
  • Humans
  • Immunoglobulin M / metabolism
  • Interleukin-2 Receptor beta Subunit / immunology
  • Lentivirus / genetics
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Nuclear Proteins / genetics

Substances

  • Antibodies, Monoclonal
  • Antigens, CD34
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • Immunoglobulin M
  • Interleukin-2 Receptor beta Subunit
  • Nuclear Proteins
  • RAG-1 protein
  • DCLRE1C protein, human
  • Endonucleases