Phagocytosis: Techniques

Abstract

Phagocytosis is a complex cellular process in which particulate material is engulfed and eliminated. Cells involved in this process are called phagocytes, and their phagocytic abilities are essential to both innate and adaptive immunity. Many techniques have been developed to quantify phagocytic process and the subsequent microbicidal processes.

Key Concepts

  • Phagocytic process.
  • Methods for quantification of phagocytosis.
  • Bactericidal mechanisms.
  • Methods for quantification of bactericidal mechanisms.

Keywords: phagocytosis; antigen uptake; respiratory burst; bacterial killing; hydrogen peroxide; lysosomal fusion

Figure 1. Summary of opsonophagocytosis quantification methods. Opsonophagocytosis is the most specific method for quantifying engulfment because it is dependent on opsonisation (i.e. IgG or complement) and the subsequent internalisation by a corresponding receptor. Most of the techniques also allow for quenching of noninternalised particles (i.e. RBC lysis solution, trypan blue quenching fluorescein and antibiotics inhibiting bacterial proliferation), thereby providing less confounded results.
Figure 2. Quantification of bactericidal mechanisms. Efficacy of phagolysosomal microbial killing can be determined by a variety of techniques targeting key mechanisms such as NADPH oxidase and oxidative burst, H2O2 production as well as phagolysosomal fusion.
close

References

Araki N (2006) Role of microtubules and myosins in Fc gamma receptor‐mediated phagocytosis. Frontiers in Bioscience 11: 1479–1490.

Babior BM , Kipes RS and Curnette JT (1973) Biological defense mechanisms. The production by leukocytes of superoxide, a potential bactericidal agent. Journal of Clinical Investigation 52: 741–744.

Campbell DE and Douglas SD (1997) Phagocytic cell functions. I. Oxidation and chemotaxis. In: Rose RE , de Macario EC , Folds JD , Lane HC and Nakamura RM (eds) Manual of Clinical Laboratory Immunology, pp. 320–328. Washington, DC: ASM Press.

Chan ED , Chan J and Schluger NW (2001) What is the role of nitric oxide in murine and human host defense against tuberculosis? Current knowledge. American Journal of Respiratory Cell and Molecular Biology 25: 606–612.

Gallin JI (1992) Disorders of phagocytic cells. In: Gallin JI , Goldstein IM and Snyderman R (eds) Inflammation – Basic Principles and Clinical Correlates, pp. 859–872. New York: Raven Press.

O'Gorman MG (1997) Phagocytic cell functions. II. Adhesion and phagocytosis. In: Rose NR , de Macario EC , Folds JD , Lane HC and Nakamura RM (eds) Manual of Clinical Laboratory Immunology, pp. 329–333. Washington, DC: ASM Press.

Raad H , Paclet M‐H , Boussetta T , et al. (2009) Regulation of the phagocyte NADPH oxidase activity: phosphorylation of gp91phox/NOX2 by protein kinase C enhances its diaphorase activity and binding to Rac2, p67phox, and p47phox. The FASEB Journal 23 (4): 1011–1022.

Ravetch JV (1997) Fc receptors. Current Opinion in Immunology 9: 121–125.

Roy S , Ninkovic J , Banerjee S , et al. (2011) Opioid drug abuse and modulation of immune function: consequences in the susceptibility to opportunistic infections. Journal of Neuroimmune Pharmacology 6 (4): 442–465.

Shah B , Li Y , Stechschulte DJ and Dileepan KN (1995) Phagocytosis of mast cell granules results in decreased macrophage superoxide production. Mediators of Inflammation 4: 406–412.

Simpson DW , Roth R and Loose LD (1979) A rapid, inexpensive and easily quantified assay for phagocytosis and microbicidal activity of macrophages and neutrophils. Journal of Immunological Methods 29 (3): 221–226.

Szolnoky G , Bata‐Csörgö Z , Kenderessy AS , et al. (2001) A mannose‐binding receptor is expressed on human keratinocytes and mediates killing of Candida albicans . Journal of Investigative Dermatology 117 (2): 205–213.

Underhill DM and Ozinsky A (2002) Phagocytosis of microbes: complexity in action. Annual Review of Immunology 20: 825–852.

Yamada A , Dileepan KN , Stechschulte DJ and Suzuki T (1989) Regulation of FC 2a receptor‐mediated phagocytosis by a murine macrophage‐like cell line, P388D1. Involvement of casein kinase activity associated with FC 2a receptor. Journal of Molecular and Cellular Immunology 4: 191–201.

Zurier RB , Hoffstein S and Weissmann G (1973) Mechanisms of lysosomal enzyme release from human leukocytes. I. Effect of cyclic nucleotides and colchicine. Journal of Cell Biology 58 (1): 27–41.

Further Reading

Coligan JE , Kruisbeek AM , Margulies DH , Shevach EM and Strober W (eds) (1992) Current Protocols in Immunology. New York: John Wiley & Sons, Inc..

Klempner MS , Styrt B and Ho J (eds) (1989) Phagocytosis and Disease. Hingham, MA: Kluwer Academic Publishers.

Pauletti R , Notario A and Ricevutti G (eds) (1997) Phagocytes: Biology, Physiology, Pathology and Pharmacotherapeutics, vol. 832 (Annals of the New York Academy of Sciences). New York: The New York Academy of Sciences.

Stuart LM and Ezekowitz RA (2005) Phagocytosis: elegant complexity. Immunity 22 (5): 539–550.

Yeo JC , Wall AA , Stow JL and Hamilton NA (2013) High‐throughput quantification of early stages of phagocytosis. Biotechniques 55 (3): 115–124.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Ninkovic, Jana, and Roy, Sabita(Mar 2018) Phagocytosis: Techniques. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0001443.pub2]