Neutrophil Inhibitory Factor Abrogates Neutrophil Adhesion by Blockade of CD11a and CD11b β2 Integrins
- Siu K. Lo1,
- Arshad Rahman1,
- Ning Xu,
- Ming Yuan Zhou,
- Pablito Nagpala,
- Howard A. Jaffe and
- Asrar B. Malik
Abstract
We studied the basis of inhibition of polymorphonuclear leukocyte (PMN) adhesion induced by neutrophil inhibitory factor (NIF), a 41-kDa CD11/CD18 β2 integrin-binding protein isolated from the canine hookworm (Ancylostoma caninum). NIF blocked PMN adhesion in a concentration-dependent manner with complete blockade occurring at ∼10 nM NIF. Because CD11a and CD11b β2integrins are functionally active on stimulated PMNs, and yet NIF is postulated to inhibit only CD11b integrin by binding to its I domain, we evaluated the contributions of CD11a and CD11b β2integrins in the mechanism of inhibition of PMN adhesion to endothelial cells. We observed an additive inhibitory effect (>90% inhibition) of PMN adhesion to endothelial cells when NIF was used in combination with anti-CD11b monoclonal antibodies, which alone at saturating concentrations reduced PMN adhesion by only 50%. NIF also prevented aggregation of phorbol ester-stimulated JY lymphoblastoid cells that expressed only the functionally active CD11a, suggesting that NIF also can inhibit CD11a-dependent response. We transduced the NIF cDNA into human dermal microvessel endothelial cells in which NIF synthesis and release prevented PMN adhesion to the transduced human dermal microvessel endothelial cells. These data indicated that the potent antiadhesive effect of NIF may be the result of inhibition of CD11a and CD11b β2 integrins on PMNs. Moreover, the strategy of NIF release from transduced endothelial cells suggests the feasibility of blocking the CD11a- and CD11b β2integrin-dependent PMN adhesion and PMN migration responses specifically at sites of endothelial cell activation.
Footnotes
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Send reprint requests to: Arshad Rahman, Department of Pharmacology, The University of Illinois College of Medicine, 835 South Wolcott Ave. (m/c 868), Room E403, Chicago, IL 60612. E-mail:ARahman{at}uic.edu
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↵1 S.K.L. and A.R. contributed equally to this work.
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This work supported by National Institutes of Health (Grants HL 27016, HL 46350, and HL 45638).
- Abbreviations:
- NIF
- neutrophil inhibitory factor
- PMN
- polymorphonuclear leukocyte
- ICAM-1
- intercellular adhesion molecule-1
- mAb
- monoclonal antibody
- CMV
- cytomegalovirus
- DMEM
- Dulbecco’s modified Eagle’s medium
- FBS
- fetal bovine serum
- HMEC
- human dermal microvascular endothelial cells
- HUVEC
- human umbilical vein endothelial cell
- HBSS
- Hanks’ balanced salt solution
- PMA
- phorbol-12-myristate-13-acetate
- TNF-α
- tumor necrosis factor alpha
- FACS
- fluorescence-activated cell-sorting analysis
- HLA
- human leukocyte antigen
- CM
- conditioned medium
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- Received March 1, 1999.
- Accepted July 28, 1999.
- The American Society for Pharmacology and Experimental Therapeutics
