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PMID:12496435

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Citation

Long, KH, Gomez, FJ, Morris, RE and Newman, SL (2003) Identification of heat shock protein 60 as the ligand on Histoplasma capsulatum that mediates binding to CD18 receptors on human macrophages. J. Immunol. 170:487-94

Abstract

Histoplasma capsulatum (Hc), is a facultative intracellular fungus that binds to CD11/CD18 receptors on macrophages (Mphi). To identify the ligand(s) on Hc yeasts that is recognized by Mphi, purified human complement receptor type 3 (CR3, CD11b/CD18) was used to probe a Far Western blot of a detergent extract of Hc cell wall and cell membrane. CR3 recognized a single 60-kDa protein, which was identified as heat shock protein 60 (hsp60). Biotinylation of viable yeasts, followed by precipitation with streptavidin-coated beads, and Western blotting with anti-hsp60 demonstrated that hsp60 was on the surface of Hc yeasts. Electron and confocal microscopy revealed that hsp60 resided on the yeast cell wall in discrete clusters. Recombinant hsp60 (rhsp60) inhibited attachment of Hc yeasts to Mphi. Recombinant hsp60 and Abs to CD11b and CD18 inhibited binding of yeasts to Chinese hamster ovary cells transfected with CR3 (CHO3). Polystyrene beads coated with rhsp60 bound to Mphi, and attachment was inhibited by Abs to CD11 and CD18. Freeze/thaw extract (F/TE), a preparation of Hc yeast surface proteins that contained hsp60, inhibited the attachment of Hc yeasts to Mphi. Depletion of hsp60 from F/TE removed the capacity of F/TE to block binding of Hc to Mphi. Interestingly, rhsp60 did not inhibit binding of Hc yeasts to dendritic cells (DC), which recognize Hc via very late Ag 5. Moreover, F/TE inhibited attachment of Hc to DC even when depleted of hsp60. Thus, Hc hsp60 appears to be a major ligand that mediates attachment of Hc to Mphi CD11/CD18, whereas DC recognize Hc via a different ligand(s).

Links

PubMed

Keywords

Animals; Antigens, CD11/metabolism; Antigens, CD18/metabolism; Binding, Competitive/immunology; CHO Cells; Cell Adhesion/immunology; Cells, Cultured; Chaperonin 60/analysis; Chaperonin 60/metabolism; Chaperonin 60/physiology; Cricetinae; Dendritic Cells/immunology; Dendritic Cells/metabolism; Dendritic Cells/microbiology; Fungal Proteins/metabolism; Histoplasma/immunology; Histoplasma/metabolism; Histoplasma/physiology; Humans; Macrophage-1 Antigen/metabolism; Macrophages/immunology; Macrophages/metabolism; Macrophages/microbiology; Membrane Proteins/analysis; Membrane Proteins/biosynthesis; Membrane Proteins/metabolism; Protein Binding/immunology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:ITB2

enables

GO:0031072: heat shock protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P50142

F

Seeded From UniProt

complete

HUMAN:ITAM

enables

GO:0031072: heat shock protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P50142

F

Seeded From UniProt

complete

AJECA:HSP60

enables

GO:0005178: integrin binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P05107
UniProtKB:P11215

F

Seeded From UniProt

complete

AJECA:HSP60

located_in

GO:0005618: cell wall

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

AJECA:HSP60

enables

GO:0046812: host cell surface binding

ECO:0000314: direct assay evidence used in manual assertion

F

  • occurs_in:(CL:0000235)

Seeded From UniProt

complete

AJECA:HSP60

involved_in

GO:0044650: adhesion of symbiont to host cell

ECO:0000314: direct assay evidence used in manual assertion

P

  • occurs_in:(CL:0000235)

Seeded From UniProt

complete

HUMAN:CH60

GO:0046789: host cell surface receptor binding

ECO:0000314:

F

2D gel electrophoresis, flow cytometric analysis and other methods were used and analyzed to determine and identify the ligand(s) on the surface of Hc yeasts that binds to human macrophage CD11/CD18.

complete
CACAO 9339

See also

References

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