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Updated: Aug 14, 2026
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
Published on: March 22, 2016
Retroviral-mediated gene transfer of the leukocyte integrin CD18 subunit
1Medical Research Division, Seattle Veterans Administration Medical Center, WA 98108.
Insights
Gene therapy successfully restored CD18 expression in cells from children with leukocyte adherence deficiency (LAD). This demonstrates potential for treating LAD by correcting defects in CD18 gene function.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Leukocyte adherence deficiency (LAD) involves defects in the CD18 subunit of leukocyte integrins.
- These defects prevent the formation of functional CD11/CD18 adherence complexes, impairing immune cell function.
Purpose of the Study:
- To investigate the feasibility of gene therapy for LAD using a retroviral vector.
- To restore the surface expression of CD18 and functional CD11/CD18 complexes in LAD cells.
Main Methods:
- A retroviral vector (LCD18SN) carrying the CD18 cDNA was developed.
- The vector was used to transduce K562 human myeloid leukemia cells and Epstein-Barr virus (EBV)-transformed B-cells from a child with LAD.
Main Results:
- Transduction of K562 cells led to high levels of CD18 mRNA and intracellular protein.
- Gene transfer into LAD EBV B-cells resulted in measurable surface expression of the CD11a/CD18 complex.
Conclusions:
- Retroviral-mediated gene transfer of CD18 can restore surface expression of CD11a/CD18 complexes in LAD lymphocytes.
- Leukocyte adherence deficiency is a potential candidate disorder for gene therapy.
Abstract:
Children with leukocyte adherence deficiency (LAD) exhibit heterogeneous defects in the leukocyte integrin CD18 subunit that prevent surface expression of functional CD11/CD18 leukocyte integrin adherence complexes. We used a retroviral vector, designated LCD18SN, to transfer the CD18 cDNA into K562 human myeloid leukemia cells and into EBV B-cells from a child with LAD. Transfer of the LCD18SN retroviral construct, which expresses the CD18 cDNA from the Moloney Murine leukemia virus (MoMLV) long terminal repeat (LTR), into K562 cells resulted in relatively high levels of CD18 mRNA and intracellular protein. Retroviral-mediated gene transfer of CD18 into LAD EBV B-cells resulted in low, but readily measurable, levels of surface expression of the CD11a/CD18 complex in these previously deficient lymphocytes. The reconstitution of surface expression of the CD11a/CD18 complex by gene transfer of the CD18 cDNA into LAD EBV B-cells indicates that this syndrome represents a candidate disorder for gene therapy.
