Expression of human CD18 in murine granulocytes and improved efficiency for infection of deficient human lymphoblasts.
The CD18 gene encodes the beta 2-subunit of leukocyte integrins, and mutations in this gene cause extreme host susceptibility to bacterial and fungal infection. Because expression of CD18 is restricted to bone marrow-derived cells, this disorder is considered an excellent candidate for somatic gene therapy utilizing ex vivo infection of bone marrow stem cells. We have constructed a retroviral vector expressing CD18 with the Moloney murine leukemia virus (Mo-MLV) long terminal repeat (LTR) as the promoter, and high-titer ecotropic and amphotropic producer cell lines were isolated using the GP+E-86 and GP+envAM12 safe packaging cell lines. Infection of CD18-deficient lymphoblasts resulted both in expression of immunodetectable CD18 at 35-40% of normal levels on 55-60% of cells and in functional restoration of CD18-dependent aggregation. All of 16 mice transplanted with syngeneic bone marrow infected with the CD18 retrovirus expressed human CD18 on 17-36% of granulocytes at 2 weeks after transplantation, and expression was appropriately up-regulated in response to stimulation with zymosan-activated serum. This recombinant retrovirus should prove useful for further studies of somatic gene therapy for CD18 deficiency.
Wilson
R W
RW
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Yorifuji
T
T
Lorenzo
I
I
Smith
W
W
Anderson
D C
DC
Belmont
J W
JW
Beaudet
A L
AL
D016475:3T3 Cells; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D016026:Bone Marrow Transplantation; D018821:CD18 Antigens; D002448:Cell Adhesion; D002478:Cells, Cultured; D004247:DNA; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors; D006098:Granulocytes / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000628:therapy; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016028:Receptors, Leukocyte-Adhesion / Q000096:biosynthesis* / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D014162:Transfection
D015703:Antigens, CD; D018821:CD18 Antigens; D016028:Receptors, Leukocyte-Adhesion; D004247:DNA