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Correction of murine Rag1 deficiency by self-inactivating lentiviral vector-mediated gene transfer.
- Source :
- Leukemia (08876924); Sep2011, Vol. 25 Issue 9, p1471-1483, 13p, 1 Chart, 7 Graphs
- Publication Year :
- 2011
-
Abstract
- Severe combined immunodeficiency (SCID) patients with an inactivating mutation in recombination activation gene 1 (RAG1) lack B and T cells due to the inability to rearrange immunoglobulin (Ig) and T-cell receptor (TCR) genes. Gene therapy is a valid treatment option for RAG-SCID patients, especially for patients lacking a suitable bone marrow donor, but developing such therapy has proven challenging. As a preclinical model for RAG-SCID, we used Rag1-/- mice and lentiviral self-inactivating (SIN) vectors harboring different internal elements to deliver native or codon-optimized human RAG1 sequences. Treatment resulted in the appearance of B and T cells in peripheral blood and developing B and T cells were detected in central lymphoid organs. Serum Ig levels and Ig and TCR Vβ gene segment usage was comparable to wild-type (WT) controls, indicating that RAG-mediated rearrangement took place. Remarkably, relatively low frequencies of B cells produced WT levels of serum immunoglobulins. Upon stimulation of the TCR, corrected spleen cells proliferated and produced cytokines. In vivo challenge resulted in production of antigen-specific antibodies. No leukemia development as consequence of insertional mutagenesis was observed. The functional reconstitution of the B- as well as the T-cell compartment provides proof-of-principle for therapeutic RAG1 gene transfer in Rag1-/- mice using lentiviral SIN vectors. [ABSTRACT FROM AUTHOR]
- Subjects :
- SEVERE combined immunodeficiency
CYTOKINES
MUTAGENESIS
GENETIC transformation
GENE therapy
T cell receptors
BONE marrow transplantation
B cells
T cells
ANIMAL experimentation
BONE marrow
CELL physiology
CELL receptors
COMPARATIVE studies
ENZYME-linked immunosorbent assay
FLOW cytometry
GENES
GENETIC techniques
IMMUNOGLOBULINS
RESEARCH methodology
MEDICAL cooperation
MICE
POLYMERASE chain reaction
PROTEINS
RESEARCH
RESEARCH funding
RETROVIRUSES
RNA
SPLEEN
WESTERN immunoblotting
EVALUATION research
REVERSE transcriptase polymerase chain reaction
THERAPEUTICS
PHYSIOLOGY
Subjects
Details
- Language :
- English
- ISSN :
- 08876924
- Volume :
- 25
- Issue :
- 9
- Database :
- Complementary Index
- Journal :
- Leukemia (08876924)
- Publication Type :
- Academic Journal
- Accession number :
- 65221744
- Full Text :
- https://doi.org/10.1038/leu.2011.106