Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Feb 2;132(4):585–593. doi: 10.1083/jcb.132.4.585

Different fates of phagocytosed particles after delivery into macrophage lysosomes

PMCID: PMC2199875  PMID: 8647890

Abstract

Phagocytosis in macrophages is often studied using inert polymer microspheres. An implicit assumption in these studies is that such particles contain little or no specific information in their structure that affects their intracellular fate. We tested that assumption by examining macrophage phagosomes containing different kinds of particles and found that although all particles progressed directly to lysosomes, their subsequent fates varied. Within 15 min of phagocytosis, >90% of phagosomes containing opsonized sheep erythrocytes, poly-e-caprolactone microspheres, polystyrene microspheres (PS), or polyethylene glycol- conjugated PS merged with the lysosomal compartment. After that point, however, the characteristics of phagolysosomes changed in several ways that indicated differing degrees of continued interaction with the lysosomal compartment. Sheep erythrocyte phagolysosomes merged together and degraded their contents quickly, poly-e-caprolactone phagolysosomes showed intermediate levels of interaction, and PS phagolysosomes became isolated within the cytoplasm. PS were relatively inaccessible to an endocytic tracer, Texas red dextran, added after phagocytosis. Moreover, immunofluorescent staining for the lysosomal protease cathepsin L decreased in PS phagolysosomes to 23% by 4 h after phagocytosis, indicating degradation of the enzyme without replacement. Finally, PS surface labeled with fluorescein-labeled albumin showed a markedly reduced rate of protein degradation in phagolysosomes, when compared to rates measured for proteins in or on other particles. Thus, particle chemistry affected both the degree of postlysosomal interactions with other organelles and, consequently, the intracellular half-life of particle-associated proteins. Such properties may affect the ability of particles to deliver macromolecules into the major histocompatibility complex class I and II antigen presentation pathways.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alonso M. J., Cohen S., Park T. G., Gupta R. K., Siber G. R., Langer R. Determinants of release rate of tetanus vaccine from polyester microspheres. Pharm Res. 1993 Jul;10(7):945–953. doi: 10.1023/a:1018942118148. [DOI] [PubMed] [Google Scholar]
  2. Amigorena S., Drake J. R., Webster P., Mellman I. Transient accumulation of new class II MHC molecules in a novel endocytic compartment in B lymphocytes. Nature. 1994 May 12;369(6476):113–120. doi: 10.1038/369113a0. [DOI] [PubMed] [Google Scholar]
  3. Armstrong J. A., Hart P. D. Response of cultured macrophages to Mycobacterium tuberculosis, with observations on fusion of lysosomes with phagosomes. J Exp Med. 1971 Sep 1;134(3 Pt 1):713–740. doi: 10.1084/jem.134.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berthiaume E. P., Medina C., Swanson J. A. Molecular size-fractionation during endocytosis in macrophages. J Cell Biol. 1995 May;129(4):989–998. doi: 10.1083/jcb.129.4.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berón W., Alvarez-Dominguez C., Mayorga L., Stahl P. D. Membrane trafficking along the phagocytic pathway. Trends Cell Biol. 1995 Mar;5(3):100–104. doi: 10.1016/s0962-8924(00)88958-8. [DOI] [PubMed] [Google Scholar]
  6. Chen J. W., Murphy T. L., Willingham M. C., Pastan I., August J. T. Identification of two lysosomal membrane glycoproteins. J Cell Biol. 1985 Jul;101(1):85–95. doi: 10.1083/jcb.101.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen J. W., Pan W., D'Souza M. P., August J. T. Lysosome-associated membrane proteins: characterization of LAMP-1 of macrophage P388 and mouse embryo 3T3 cultured cells. Arch Biochem Biophys. 1985 Jun;239(2):574–586. doi: 10.1016/0003-9861(85)90727-1. [DOI] [PubMed] [Google Scholar]
  8. Desjardins M. Biogenesis of phagolysosomes: the 'kiss and run' hypothesis. Trends Cell Biol. 1995 May;5(5):183–186. doi: 10.1016/s0962-8924(00)88989-8. [DOI] [PubMed] [Google Scholar]
  9. Desjardins M., Celis J. E., van Meer G., Dieplinger H., Jahraus A., Griffiths G., Huber L. A. Molecular characterization of phagosomes. J Biol Chem. 1994 Dec 23;269(51):32194–32200. [PubMed] [Google Scholar]
  10. Desjardins M., Huber L. A., Parton R. G., Griffiths G. Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus. J Cell Biol. 1994 Mar;124(5):677–688. doi: 10.1083/jcb.124.5.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dong J. M., Prence E. M., Sahagian G. G. Mechanism for selective secretion of a lysosomal protease by transformed mouse fibroblasts. J Biol Chem. 1989 May 5;264(13):7377–7383. [PubMed] [Google Scholar]
  12. Ferris A. L., Brown J. C., Park R. D., Storrie B. Chinese hamster ovary cell lysosomes rapidly exchange contents. J Cell Biol. 1987 Dec;105(6 Pt 1):2703–2712. doi: 10.1083/jcb.105.6.2703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Garcia-del Portillo F., Finlay B. B. Targeting of Salmonella typhimurium to vesicles containing lysosomal membrane glycoproteins bypasses compartments with mannose 6-phosphate receptors. J Cell Biol. 1995 Apr;129(1):81–97. doi: 10.1083/jcb.129.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Germain R. N. MHC-dependent antigen processing and peptide presentation: providing ligands for T lymphocyte activation. Cell. 1994 Jan 28;76(2):287–299. doi: 10.1016/0092-8674(94)90336-0. [DOI] [PubMed] [Google Scholar]
  15. Goren M. B., D'Arcy Hart P., Young M. R., Armstrong J. A. Prevention of phagosome-lysosome fusion in cultured macrophages by sulfatides of Mycobacterium tuberculosis. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2510–2514. doi: 10.1073/pnas.73.7.2510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harding C. V. Phagocytic processing of antigens for presentation by MHC molecules. Trends Cell Biol. 1995 Mar;5(3):105–109. doi: 10.1016/s0962-8924(00)88959-x. [DOI] [PubMed] [Google Scholar]
  17. Hart P. D., Young M. R. Interference with normal phagosome-lysosome fusion in macrophages, using ingested yeast cells and suramin. Nature. 1975 Jul 3;256(5512):47–49. doi: 10.1038/256047a0. [DOI] [PubMed] [Google Scholar]
  18. Joiner K. A., Fuhrman S. A., Miettinen H. M., Kasper L. H., Mellman I. Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts. Science. 1990 Aug 10;249(4969):641–646. doi: 10.1126/science.2200126. [DOI] [PubMed] [Google Scholar]
  19. Kirschke H., Langner J., Wiederanders B., Ansorge S., Bohley P. Cathepsin L. A new proteinase from rat-liver lysosomes. Eur J Biochem. 1977 Apr 1;74(2):293–301. doi: 10.1111/j.1432-1033.1977.tb11393.x. [DOI] [PubMed] [Google Scholar]
  20. Knapp P. E., Swanson J. A. Plasticity of the tubular lysosomal compartment in macrophages. J Cell Sci. 1990 Mar;95(Pt 3):433–439. doi: 10.1242/jcs.95.3.433. [DOI] [PubMed] [Google Scholar]
  21. Kornfeld S., Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol. 1989;5:483–525. doi: 10.1146/annurev.cb.05.110189.002411. [DOI] [PubMed] [Google Scholar]
  22. Kovacsovics-Bankowski M., Rock K. L. A phagosome-to-cytosol pathway for exogenous antigens presented on MHC class I molecules. Science. 1995 Jan 13;267(5195):243–246. doi: 10.1126/science.7809629. [DOI] [PubMed] [Google Scholar]
  23. MacDonald R. G., Tepper M. A., Clairmont K. B., Perregaux S. B., Czech M. P. Serum form of the rat insulin-like growth factor II/mannose 6-phosphate receptor is truncated in the carboxyl-terminal domain. J Biol Chem. 1989 Feb 25;264(6):3256–3261. [PubMed] [Google Scholar]
  24. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  25. Morris W., Steinhoff M. C., Russell P. K. Potential of polymer microencapsulation technology for vaccine innovation. Vaccine. 1994 Jan;12(1):5–11. doi: 10.1016/0264-410x(94)90003-5. [DOI] [PubMed] [Google Scholar]
  26. Pitt A., Mayorga L. S., Stahl P. D., Schwartz A. L. Alterations in the protein composition of maturing phagosomes. J Clin Invest. 1992 Nov;90(5):1978–1983. doi: 10.1172/JCI116077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Portnoy D. A., Erickson A. H., Kochan J., Ravetch J. V., Unkeless J. C. Cloning and characterization of a mouse cysteine proteinase. J Biol Chem. 1986 Nov 5;261(31):14697–14703. [PubMed] [Google Scholar]
  28. Prence E. M., Dong J. M., Sahagian G. G. Modulation of the transport of a lysosomal enzyme by PDGF. J Cell Biol. 1990 Feb;110(2):319–326. doi: 10.1083/jcb.110.2.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rabinowitz S., Horstmann H., Gordon S., Griffiths G. Immunocytochemical characterization of the endocytic and phagolysosomal compartments in peritoneal macrophages. J Cell Biol. 1992 Jan;116(1):95–112. doi: 10.1083/jcb.116.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Racoosin E. L., Swanson J. A. Macrophage colony-stimulating factor (rM-CSF) stimulates pinocytosis in bone marrow-derived macrophages. J Exp Med. 1989 Nov 1;170(5):1635–1648. doi: 10.1084/jem.170.5.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Racoosin E. L., Swanson J. A. Macropinosome maturation and fusion with tubular lysosomes in macrophages. J Cell Biol. 1993 Jun;121(5):1011–1020. doi: 10.1083/jcb.121.5.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sturgill-Koszycki S., Schlesinger P. H., Chakraborty P., Haddix P. L., Collins H. L., Fok A. K., Allen R. D., Gluck S. L., Heuser J., Russell D. G. Lack of acidification in Mycobacterium phagosomes produced by exclusion of the vesicular proton-ATPase. Science. 1994 Feb 4;263(5147):678–681. doi: 10.1126/science.8303277. [DOI] [PubMed] [Google Scholar]
  33. Tulp A., Verwoerd D., Dobberstein B., Ploegh H. L., Pieters J. Isolation and characterization of the intracellular MHC class II compartment. Nature. 1994 May 12;369(6476):120–126. doi: 10.1038/369120a0. [DOI] [PubMed] [Google Scholar]
  34. Twining S. S. Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes. Anal Biochem. 1984 Nov 15;143(1):30–34. doi: 10.1016/0003-2697(84)90553-0. [DOI] [PubMed] [Google Scholar]
  35. Veras P. S., de Chastellier C., Moreau M. F., Villiers V., Thibon M., Mattei D., Rabinovitch M. Fusion between large phagocytic vesicles: targeting of yeast and other particulates to phagolysosomes that shelter the bacterium Coxiella burnetii or the protozoan Leishmania amazonensis in Chinese hamster ovary cells. J Cell Sci. 1994 Nov;107(Pt 11):3065–3076. doi: 10.1242/jcs.107.11.3065. [DOI] [PubMed] [Google Scholar]
  36. Veras P. S., de Chastellier C., Rabinovitch M. Transfer of zymosan (yeast cell walls) to the parasitophorous vacuoles of macrophages infected with Leishmania amazonensis. J Exp Med. 1992 Sep 1;176(3):639–646. doi: 10.1084/jem.176.3.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wang Y. L., Goren M. B. Differential and sequential delivery of fluorescent lysosomal probes into phagosomes in mouse peritoneal macrophages. J Cell Biol. 1987 Jun;104(6):1749–1754. doi: 10.1083/jcb.104.6.1749. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES