T Cell Immune Deficiency inzap70Mutant Zebrafish.
ZAP70[zeta-chain (TCR)-associated protein kinase,70-kDa], is required for T cell activation.ZAP70deficiencies in humans and null mutations in mice lead to severe combined immune deficiency. Here, we describe azap70loss-of-function mutation in zebrafish (zap70y442) that was created using transcription activator-like effector nucleases (TALENs). In contrast to what has been reported for morphant zebrafish,zap70y442homozygous mutant zebrafish displayed normal development of blood and lymphatic vasculature. Hematopoietic cell development was also largely unaffected in mutant larvae. However, mutant fish had reducedlck:GFP+thymic T cells by 5 days postfertilization that persisted into adult stages. Morphological analysis, RNA sequencing, and single-cell gene expression profiling of whole kidney marrow cells of adult fish revealed complete loss of mature T cells inzap70y442mutant animals. T cell immune deficiency was confirmed through transplantation of unmatched normal and malignant donor cells intozap70y442mutant zebrafish, with T cell loss being sufficient for robust allogeneic cell engraftment.zap70mutant zebrafish show remarkable conservation of immune cell dysfunction as found in mice and humans and will serve as a valuable model to studyzap70immune deficiency.
Molecular Pathology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.|Cancer Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
John C
JC
Moore
Laboratory of Molecular Genetics, NICHD, NIH, Bethesda, Maryland, USA.
Timothy S
TS
Mulligan
Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Harvard University, Cambridge, Massachusetts, USA.
Nora Torres
NT
Yordán
Laboratory of Molecular Genetics, NICHD, NIH, Bethesda, Maryland, USA.
Daniel
D
Castranova
Laboratory of Molecular Genetics, NICHD, NIH, Bethesda, Maryland, USA.
Van N
VN
Pham
Molecular Pathology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.|Cancer Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
Qin
Q
Tang
Molecular Pathology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.|Cancer Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
Riadh
R
Lobbardi
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Anthony
A
Anselmo
Izaak Walton Killam Health Center, Dalhousie University, Halifax, Nova Scotia, Canada.
Robert S
RS
Liwski
Izaak Walton Killam Health Center, Dalhousie University, Halifax, Nova Scotia, Canada.
Jason N
JN
Berman
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Ruslan I
RI
Sadreyev
Laboratory of Molecular Genetics, NICHD, NIH, Bethesda, Maryland, USA flyingfish2@nih.gov dlangenau@mgh.harvard.edu.
Brant M
BM
Weinstein
Molecular Pathology, Massachusetts General Hospital, Charlestown, Massachusetts, USA flyingfish2@nih.gov dlangenau@mgh.harvard.edu.|Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.|Cancer Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
David M
DM
Langenau