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.
Department of Genetic Medicine, Weill Medical College of Cornell University, New York, New York, USA.
Minoru
M
Tahara
Robert G
RG
Pergolizzi
Hiroyasu
H
Kobayashi
Anja
A
Krause
Karsta
K
Luettich
Martin L
ML
Lesser
Ronald G
RG
Crystal
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