Identification and characterization of VEGF-A-responsive neutrophils expressing CD49d, VEGFR1, and CXCR4 in mice and humans.
Vascular endothelial growth factor A (VEGF-A) is upregulated during hypoxia and is the major regulator of angiogenesis. VEGF-A expression has also been found to recruit myeloid cells to ischemic tissues where they contribute to angiogenesis. This study investigates the mechanisms underlying neutrophil recruitment to VEGF-A as well as the characteristics of these neutrophils. A previously undefined circulating subset of neutrophils shown to be CD49d(+)VEGFR1(high)CXCR4(high) was identified in mice and humans. By using chimeric mice with impaired VEGF receptor 1 (VEGFR1) or VEGFR2 signaling (Flt-1tk(-/-), tsad(-/-)), we found that parallel activation of VEGFR1 on neutrophils and VEGFR2 on endothelial cells was required for VEGF-A-induced recruitment of circulating neutrophils to tissue. Intravital microscopy of mouse microcirculation revealed that neutrophil recruitment by VEGF-A versus by the chemokine macrophage inflammatory protein 2 (MIP-2 [CXCL2]) involved the same steps of the recruitment cascade but that an additional neutrophil integrin (eg, VLA-4 [CD49d/CD29]) played a crucial role in neutrophil crawling and emigration to VEGF-A. Isolated CD49d(+) neutrophils featured increased chemokinesis but not chemotaxis compared with CD49d(-) neutrophils in the presence of VEGF-A. Finally, by targeting the integrin α4 subunit (CD49d) in a transplantation-based angiogenesis model that used avascular pancreatic islets transplanted to striated muscle, we demonstrated that inhibiting the recruitment of circulating proangiogenic neutrophils to hypoxic tissue impairs vessel neoformation. Thus, angiogenesis can be modulated by targeting cell-surface receptors specifically involved in VEGF-A-dependent recruitment of proangiogenic neutrophils without compromising recruitment of the neutrophil population involved in the immune response to pathogens.
Department of Medical Cell Biology and.
Sara
S
Massena
Department of Medical Cell Biology and.
Gustaf
G
Christoffersson
Department of Medical Cell Biology and.
Evelina
E
Vågesjö
Department of Medical Cell Biology and.
Cédric
C
Seignez
Department of Medical Cell Biology and.
Karin
K
Gustafsson
Department of Medical Cell Biology and.
François
F
Binet
Department of Medical Cell Biology and.
Carmen
C
Herrera Hidalgo
Department of Medical Cell Biology and.
Antoine
A
Giraud
Department of Medical Cell Biology and.
Jalal
J
Lomei
Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden; and.
Simone
S
Weström
Gakubunkan Institute of Physiology and Medicine, Jobu University, Gunma, Japan.
Masabumi
M
Shibuya
Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden; and.
Lena
L
Claesson-Welsh
Department of Medical Cell Biology and.
Pär
P
Gerwins
Department of Medical Cell Biology and.
Michael
M
Welsh
Department of Medical Cell Biology and.
Johan
J
Kreuger
Department of Medical Cell Biology and.
Mia
M
Phillipson
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D039441:Integrin alpha4 / Q000235:genetics / Q000378:metabolism*; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018715:Microscopy, Video; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000502:physiology*
D012333:RNA, Messenger; D019718:Receptors, CXCR4; D042461:Vascular Endothelial Growth Factor A; D039441:Integrin alpha4; D040281:Vascular Endothelial Growth Factor Receptor-1