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Determination of C3b Receptors: Title and subTitle BreakOn Normal and Patient Polymorphonuclear Neutrophils With C3b-Coated Fluorescent Microspheres FREE

James D. Ogle, PhD; Cora K. Ogle, PhD; J. Greg Noel, MS; Joseph S. Solomkin, MD; J. Wesley Alexander, MD, ScD
[+] Author Affiliations

Accepted for publication Sept 12, 1984.

Read before the Fourth Annual Meeting of the Surgical Infection Society, Montreal, May 1, 1984.

Reprint requests to Department of Biological Chemistry, University of Cincinnati Medical Center, 231 Bethesda Ave, Cincinnati, OH 45267-0522 (Dr J. D. Ogle).


Arch Surg. 1985;120(1):104-109. doi:10.1001/archsurg.1985.01390250092015
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• A method, devised in the authors' laboratories, for the determination of C3b receptors on normal and patient neutrophils using C3b-coated fluorescent microspheres, was applied to the quantitation of C3b receptors on the neutrophils of several patients suffering from burns and trauma and a patient with pancreatitis. From three to 11 days in the clinical course the relative number of C3b receptors was, or rose to, two to ten times the number of receptors present at later times in the clinical course and, in most of the cases studied, the increase in C3b receptor number coincided with enhanced neutrophil bactericidal function. The rise in C3b receptor number was ascribed to up-regulation by C5a and C5a des Arg from complement activation and also, in the cases where sepsis occurred, to the presence of bacterial chemotactic peptides. Preliminary experiments with zymosan-activated serum and the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine confirmed this explanation.

(Arch Surg 1985;120:104-109)

REFERENCES

Ross GD:  Analysis of different types of leukocyte membrane complement receptors and their interaction with the complement system . J Immunol Methods 1980;;37:197-211.
Dierich MP, Landen B:  Complement bridges between cells: Analysis of a possible cell-cell interaction mechanism . J Exp Med 1977;;146:1484-1489.
Lambris JD, Ross GD:  Assay of membrane complement receptors: (CR, and CR2) with C3b- and C3d-coated fluorescent microspheres . J Immunol 1982;;128:186-189.
Ogle JD, Ogle CK, Noel JG, et al: Studies on the binding of C3b-coated microspheres to normal and patient neutrophils. J Immunol Methods, 1984, in press.
Ogle JD, Ogle CK, Alexander JW:  Inhibition of neutrophil function by fluid phase C3b of complement . Infect Immun 1983;;40:967-976.
Alexander JW, Wenthurst DB, Good RA:  Improved tests for the evaluation of neutrophil function in human disease . J Lab Clin Med 1968;;72:136-148.
Ferrante A, Thong YH:  A rapid one-step procedure for purification of mononuclear and polymorphonuclear leukocytes from human blood using a modification of the Hypaque-Ficoll technique . J Immunol Methods 1978;;24: 389-393.
Fearon DT, Collins LA:  Increased expression of C3b receptors on polymorphonuclear leukocytes induced by chemotactic factors and by purification procedures . J Immunol 1983;;130:370-375.
Kirchanski SJ, Price BJ, Davidovits G, et al:  Flow cytometry measurements of membrane C3b receptors modulation during phagocytosis . Biotechniques 1983;; (June) /July:66-70.

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Ross GD:  Analysis of different types of leukocyte membrane complement receptors and their interaction with the complement system . J Immunol Methods 1980;;37:197-211.
Dierich MP, Landen B:  Complement bridges between cells: Analysis of a possible cell-cell interaction mechanism . J Exp Med 1977;;146:1484-1489.
Lambris JD, Ross GD:  Assay of membrane complement receptors: (CR, and CR2) with C3b- and C3d-coated fluorescent microspheres . J Immunol 1982;;128:186-189.
Ogle JD, Ogle CK, Noel JG, et al: Studies on the binding of C3b-coated microspheres to normal and patient neutrophils. J Immunol Methods, 1984, in press.
Ogle JD, Ogle CK, Alexander JW:  Inhibition of neutrophil function by fluid phase C3b of complement . Infect Immun 1983;;40:967-976.
Alexander JW, Wenthurst DB, Good RA:  Improved tests for the evaluation of neutrophil function in human disease . J Lab Clin Med 1968;;72:136-148.
Ferrante A, Thong YH:  A rapid one-step procedure for purification of mononuclear and polymorphonuclear leukocytes from human blood using a modification of the Hypaque-Ficoll technique . J Immunol Methods 1978;;24: 389-393.
Fearon DT, Collins LA:  Increased expression of C3b receptors on polymorphonuclear leukocytes induced by chemotactic factors and by purification procedures . J Immunol 1983;;130:370-375.
Kirchanski SJ, Price BJ, Davidovits G, et al:  Flow cytometry measurements of membrane C3b receptors modulation during phagocytosis . Biotechniques 1983;; (June) /July:66-70.

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