Several commercially available anti-CCR5 monoclonal antibodies lack specificity and should be used with caution.
CCR5 (CD195) is a receptor for the chemokines RANTES, MIP-1α, and MIP-1β and is used by HIV-1 as a co-receptor for entry into macrophages and CD4+ T cells. CCR5 exists in multiple conformations in the membrane and is present at low levels on human macrophages, making it difficult to detect. Nine commercially available anti-CCR5 monoclonal antibodies were evaluated for their specificity and their recognition of CCR5 expressed by macrophages. Unexpectedly, we found that three of the nine clones tested displayed substantial background binding to CCR5 negative cells, suggesting that these antibodies may give unreliable results. We recommend the use of clones CTC8, 45531 (PE conjugated), and 45523 and advise that data obtained using CTC5, 45531 (unconjugated), and 45549 anti-CCR5 antibody clones should be interpreted with care.
Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom. aura.bernstone@nhs.net
Laura
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Bernstone
Bonnie
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van Wilgenburg
William
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James
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D002454:Cell Differentiation; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects / Q000276:immunology; D019713:Receptors, CCR5 / Q000276:immunology*
D000911:Antibodies, Monoclonal; D019713:Receptors, CCR5; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor