Trans-splicing repair of CD40 ligand deficiency results in naturally regulated correction of a mouse model of hyper-IgM X-linked immunodeficiency.
X-linked immunodeficiency with hyper-IgM (HIGM1), characterized by failure of immunoglobulin isotype switching, is caused by mutations of the CD40 ligand (CD40L), which is normally expressed on activated CD4(+) T cells. As constitutive expression of CD40L induces lymphomas, we corrected the mutation while preserving the natural regulation of CD40L using pre-mRNA trans-splicing. Bone marrow from mice lacking CD40L was modified with a lentivirus trans-splicer encoding the normal CD40L exons 2-5 and was administered to syngenic CD40L-knockout mice. Recipient mice had corrected CD40L mRNA, antigen-specific IgG1 responses to keyhole limpet hemocyanin immunization, regulated CD4(+) T-cell CD40L expression after CD3 stimulation in primary and secondary transplanted mice, attenuation of Pneumocystis carinii pneumonia, and no evidence of lymphoproliferative disease over 1 year. Thus, HIGM1 can be corrected by CD40L trans-splicing, leading to functional correction of the genetic defect without the adverse consequences of unregulated expression of the CD40L gene.
Tahara
Minoru
M
Department of Genetic Medicine, Weill Medical College of Cornell University, New York, New York, USA.
Pergolizzi
Robert G
RG
Kobayashi
Hiroyasu
H
Krause
Anja
A
Luettich
Karsta
K
Lesser
Martin L
ML
Crystal
Ronald G
RG
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D023201:CD40 Ligand / Q000235:genetics*; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D040181:Genetic Diseases, X-Linked / Q000235:genetics / Q000628:therapy*; D005818:Genetic Engineering / Q000379:methods*; D006433:Hemocyanins / Q000276:immunology; D006942:Hypergammaglobulinemia / Q000235:genetics / Q000628:therapy*; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M; D016086:Lentivirus; D008168:Lung / Q000382:microbiology / Q000473:pathology; D008232:Lymphoproliferative Disorders / Q000175:diagnosis; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D045363:Pneumocystis carinii / Q000235:genetics / Q000276:immunology; D011020:Pneumonia, Pneumocystis / Q000628:therapy; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric
D017931:DNA Primers; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D012333:RNA, Messenger; D023201:CD40 Ligand; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin