Purified complement C3b triggers phagocytosis and activation of human neutrophils via complement receptor 1.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
06 01 2023
06 01 2023
Historique:
received:
09
05
2022
accepted:
29
12
2022
entrez:
7
1
2023
pubmed:
8
1
2023
medline:
11
1
2023
Statut:
epublish
Résumé
The complement system provides vital immune protection against infectious agents by labeling them with complement fragments that enhance phagocytosis by immune cells. Many details of complement-mediated phagocytosis remain elusive, partly because it is difficult to study the role of individual complement proteins on target surfaces. Here, we employ serum-free methods to couple purified complement C3b onto E. coli bacteria and beads and then expose human neutrophils to these C3b-coated targets. We examine the neutrophil response using a combination of flow cytometry, confocal microscopy, luminometry, single-live-cell/single-target manipulation, and dynamic analysis of neutrophil spreading on opsonin-coated surfaces. We show that purified C3b can potently trigger phagocytosis and killing of bacterial cells via Complement receptor 1. Comparison of neutrophil phagocytosis of C3b- versus antibody-coated beads with single-bead/single-target analysis exposes a similar cell morphology during engulfment. However, bulk phagocytosis assays of C3b-beads combined with DNA-based quenching reveal that these are poorly internalized compared to their IgG1 counterparts. Similarly, neutrophils spread slower on C3b-coated compared to IgG-coated surfaces. These observations support the requirement of multiple stimulations for efficient C3b-mediated uptake. Together, our results establish the existence of a direct pathway of phagocytic uptake of C3b-coated targets and present methodologies to study this process.
Identifiants
pubmed: 36609665
doi: 10.1038/s41598-022-27279-4
pii: 10.1038/s41598-022-27279-4
pmc: PMC9822988
doi:
Substances chimiques
Complement C3b
80295-43-8
Receptors, Complement 3b
0
Complement System Proteins
9007-36-7
Immunoglobulin G
0
Receptors, Complement
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
274Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM098060
Pays : United States
Informations de copyright
© 2023. The Author(s).
Références
Proc Natl Acad Sci U S A. 2021 Feb 16;118(7):
pubmed: 33563762
Adv Immunol. 1989;46:183-219
pubmed: 2551147
J Exp Med. 1982 Oct 1;156(4):1149-64
pubmed: 7153708
Immunol Rev. 2001 Apr;180:112-22
pubmed: 11414353
Nat Cell Biol. 2019 Nov;21(11):1357-1369
pubmed: 31659275
PLoS One. 2015 Jun 12;10(6):e0129522
pubmed: 26070210
J Exp Med. 1977 Feb 1;145(2):357-71
pubmed: 833545
Cell Regul. 1991 Jan;2(1):41-55
pubmed: 1826092
Am J Blood Res. 2017 Jul 25;7(4):41-48
pubmed: 28804681
Mol Biol Cell. 2008 Nov;19(11):4628-39
pubmed: 18768756
Biochim Biophys Acta. 1986 Nov 4;853(1):65-89
pubmed: 3021215
J Exp Med. 2017 May 1;214(5):1509-1528
pubmed: 28389502
Clin Immunol Immunopathol. 1982 May;23(2):335-57
pubmed: 7049467
Trends Biotechnol. 2013 Apr;31(4):258-67
pubmed: 23490213
J Immunol. 1983 Oct;131(4):1930-5
pubmed: 6225800
J Biol Chem. 2021 Jan-Jun;296:100256
pubmed: 33839682
Immunol Rev. 2016 Nov;274(1):33-58
pubmed: 27782325
J Exp Med. 1996 Aug 1;184(2):627-37
pubmed: 8760816
Nat Commun. 2022 Apr 12;13(1):1955
pubmed: 35413960
Blood Cells Mol Dis. 2009 May-Jun;42(3):177-84
pubmed: 19246218
Microbes Infect. 2000 Mar;2(3):289-94
pubmed: 10758405
Microbiol Spectr. 2016 Nov;4(6):
pubmed: 27809953
J Cell Biol. 1994 Jun;125(6):1407-16
pubmed: 7515890
Cell. 2016 Jan 14;164(1-2):128-140
pubmed: 26771488
J Immunol. 1993 Jul 1;151(1):330-8
pubmed: 8326130
Plasmid. 2016 Sep - Nov;87-88:65-71
pubmed: 27693407
J Immunol. 1979 Oct;123(4):1839-46
pubmed: 573303
J Biol Chem. 2003 Nov 14;278(46):45720-9
pubmed: 12941957
Biomed Res Int. 2017;2017:9042851
pubmed: 28691037
J Exp Med. 1985 Jun 1;161(6):1414-31
pubmed: 2409200
Immunology. 1987 May;61(1):15-20
pubmed: 2953672
PLoS One. 2013;8(1):e54735
pubmed: 23349959
Mol Immunol. 2004 Nov;41(11):1089-98
pubmed: 15476920
Clin Exp Immunol. 1998 Apr;112(1):27-33
pubmed: 9566786
J Immunol. 1994 Jul 15;153(2):804-10
pubmed: 8021514
Nat Rev Immunol. 2019 Aug;19(8):503-516
pubmed: 31048789
J Immunol. 1999 Feb 15;162(4):2266-74
pubmed: 9973503
Biochemistry. 2015 Feb 17;54(6):1408-20
pubmed: 25613106
J Cell Biol. 1993 Feb;120(4):1031-43
pubmed: 7679388
BMC Biol. 2015 Nov 09;13:93
pubmed: 26552476
Scand J Immunol. 1977;6(8):797-807
pubmed: 561436
Cell Mol Bioeng. 2010 Jun 1;3(2):106-116
pubmed: 21532911
J Leukoc Biol. 1996 Jun;59(6):883-90
pubmed: 8691074
Cytometry. 1991;12(7):677-86
pubmed: 1838328
Nat Struct Mol Biol. 2017 Aug;24(8):643-651
pubmed: 28671664
Clin Exp Immunol. 1992 Jul;89(1):8-17
pubmed: 1385769
Int J Med Microbiol. 2015 Jan;305(1):55-64
pubmed: 25466204
PLoS Comput Biol. 2011 Jan 27;7(1):e1001068
pubmed: 21298079
J Exp Med. 1983 Jun 1;157(6):1844-54
pubmed: 6854210
Proc Natl Acad Sci U S A. 2013 Oct 8;110(41):16426-31
pubmed: 24065820
Complement. 1987;4(2):75-86
pubmed: 3040333
Proc Natl Acad Sci U S A. 2017 Mar 28;114(13):3403-3408
pubmed: 28292891
Clin Immunol Immunopathol. 1997 Jan;82(1):49-59
pubmed: 9000042
Complement. 1984;1(4):213-27
pubmed: 6242391
Blood. 2015 Apr 9;125(15):2359-69
pubmed: 25608561
Proc Natl Acad Sci U S A. 2021 Jun 29;118(26):
pubmed: 34155115
Cytokine. 1990 Nov;2(6):447-55
pubmed: 1716488
Langmuir. 2005 Mar 1;21(5):1962-71
pubmed: 15723496
J Leukoc Biol. 1985 Aug;38(2):267-78
pubmed: 2993460
J Clin Invest. 1989 Jul;84(1):107-12
pubmed: 2544621
J Cell Sci. 2011 Apr 1;124(Pt 7):1106-14
pubmed: 21385838
ACS Chem Biol. 2006 Nov 21;1(10):644-8
pubmed: 17175580
Biophys J. 2015 Aug 4;109(3):469-76
pubmed: 26244729
Immunity. 2006 Mar;24(3):305-16
pubmed: 16546099
J Immunol. 2002 Dec 1;169(11):6417-26
pubmed: 12444150
J Immunol. 1989 Apr 1;142(7):2352-8
pubmed: 2538507
J Cell Sci. 2006 May 1;119(Pt 9):1903-13
pubmed: 16636075
mBio. 2013 Apr 16;4(2):e00065
pubmed: 23592259
J Immunol. 1984 Jun;132(6):3028-33
pubmed: 6233366
J Vis Exp. 2013 Jul 17;(77):e50788
pubmed: 23893048
Microbiol Spectr. 2016 Jun;4(3):
pubmed: 27337463
Scand J Immunol. 1990 Dec;32(6):669-77
pubmed: 2148644
Sci Signal. 2018 Mar 13;11(521):
pubmed: 29535263
J Cell Sci. 2005 Nov 15;118(Pt 22):5205-20
pubmed: 16249234
Immunology. 1987 Sep;62(1):45-51
pubmed: 3653927
Blood. 2001 Apr 15;97(8):2478-86
pubmed: 11290613