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Infection and Immunity, April 2000, p. 2286-2293, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Intracellular Trafficking and Killing of Streptococcus pneumoniae by Human Alveolar Macrophages Are Influenced by Opsonins

Stephen B. Gordon,dagger Glen R. B. Irving, Roderick A. Lawson, Margaret E. Lee, and Robert C. Read*

Division of Molecular and Genetic Medicine, University of Sheffield Medical School, Sheffield S10 2RX, United Kingdom

Received 30 September 1999/Returned for modification 16 November 1999/Accepted 20 December 1999

Human alveolar macrophages (HAM) are the major resident phagocytic cells of the gas-exchanging areas of the lung. Following contact with macrophages, bacteria enter phagosomes, which gradually acquire the characteristics of terminal phagolysosomes, with incorporation of lysosome-associated membrane protein (LAMP). We measured the binding of type 1 Streptococcus pneumoniae to the surface of HAM and then measured subsequent internalization and phagosomal incorporation of LAMP-1 under various opsonic conditions. Opsonization with serum containing immunoglobulin resulted in significantly greater binding of pneumococci to HAM compared with opsonization with immunoglobulin G (IgG)-depleted serum containing complement, which in turn resulted in marginally increased binding over that observed in the absence of opsonization. Internalization of opsonized S. pneumoniae gradually increased to a maximum of 20% of bound bacteria by 120 min of warm incubation, with 20% of internalized pneumococci being localized within LAMP-containing compartments by 80 min. Internalization of opsonized S. pneumoniae by HAM correlated with a reduction of bacterial viability. When inocula were adjusted so that pneumococcal binding under different conditions was equalized, subsequent internalization, trafficking to LAMP-containing compartments, and reduction of bacterial viability were less efficient in the absence of opsonization than that observed following opsonization with adsorbed or IgG-replete adsorbed serum. Once bound to the surface of HAM, pneumococci opsonized with adsorbed serum with or without IgG were internalized, processed, and killed equally well. In conclusion, binding, intracellular trafficking, and killing of S. pneumoniae by HAM are each significantly increased by opsonization with serum containing immunogloblin and/or complement.

* Corresponding author. Mailing address: Division of Molecular & Genetic Medicine, Sheffield University Medical School, Sheffield, United Kingdom. Phone: 44 114 272 4072. Fax: 44 114 273 9926. E-mail: r.c.read{at}shef.ac.uk.

dagger Present address: The Wellcome Trust Research Laboratories, Universities of Malawi and Liverpool, College of Medicine, Blantyre, Malawi.

Infection and Immunity, April 2000, p. 2286-2293, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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