Monomeric and dimeric CXCL8 are both essential for in vivo neutrophil recruitment.
Rapid mobilization of neutrophils from vasculature to the site of bacterial/viral infections and tissue injury is a critical step in successful resolution of inflammation. The chemokine CXCL8 plays a central role in recruiting neutrophils. A characteristic feature of CXCL8 is its ability to reversibly exist as both monomers and dimers, but whether both forms exist in vivo, and if so, the relevance of each form for in vivo function is not known. In this study, using a 'trapped' non-associating monomer and a non-dissociating dimer, we show that (i) wild type (WT) CXCL8 exists as both monomers and dimers, (ii) the in vivo recruitment profiles of the monomer, dimer, and WT are distinctly different, and (iii) the dimer is essential for initial robust recruitment and the WT is most active for sustained recruitment. Using a microfluidic device, we also observe that recruitment is not only dependent on the total amount of CXCL8 but also on the steepness of the gradient, and the gradients created by different CXCL8 variants elicit different neutrophil migratory responses. CXCL8 mediates its function by binding to CXCR2 receptor on neutrophils and glycosaminoglycans (GAGs) on endothelial cells. On the basis of our data, we propose that dynamic equilibrium between CXCL8 monomers and dimers and their differential binding to CXCR2 and GAGs mediates and regulates in vivo neutrophil recruitment. Our finding that both CXCL8 monomer and dimer are functional in vivo is novel, and indicates that the CXCL8 monomer-dimer equilibrium and neutrophil recruitment are intimately linked in health and disease.
Das
Sandhya Thulasi
ST
Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, Texas, United States of America.
Rajagopalan
Lavanya
L
Guerrero-Plata
Antonieta
A
Sai
Jiqing
J
Richmond
Ann
A
Garofalo
Roberto P
RP
Rajarathnam
Krishna
K
D000818:Animals; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006025:Glycosaminoglycans / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D016209:Interleukin-8 / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D046210:Microfluidic Analytical Techniques; D008954:Models, Biological; D020556:Neutrophil Infiltration / Q000187:drug effects / Q000502:physiology*; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D023063:Receptors, Interleukin-8B / Q000378:metabolism
D006025:Glycosaminoglycans; D016209:Interleukin-8; D023063:Receptors, Interleukin-8B