title: Double-staining artefact observed in certain individuals during dual-colour immunophenotyping of lymphocytes by flow cytometry.
abstract: An unusual double-staining artefact has been repeatedly observed in certain individuals (both HIV-positive and healthy laboratory controls) during 2-colour immunophenotyping of lymphocytes by flow cytometry after red cell lysis. This artefact can falsely suggest co-expression of CD4 and CD8, as well as some of the other monoclonal antibody pairs commonly used in the typing of lymphocytes. It is caused by a serum factor associated with the ammonium-sulphate-precipitated and gamma-globulin fractions. It appears to be fairly common in the population (being found in 6 out of 105 of our healthy laboratory controls and in 42 out of 247 of our HIV-1 seropositive subjects); it may be genetically determined, but may be acquired in some. It is a potential source of error in lymphocyte immunophenotyping, which can be avoided by simple adjustments to the recommended techniques.
authors.affiliation: Department of Immunology, St Mary's Hospital Medical School, Paddington, London, United Kingdom.
authors.forename: T
authors.initials: T
authors.lastname: Ekong
authors.forename: M
authors.initials: M
authors.lastname: Gompels
authors.forename: C
authors.initials: C
authors.lastname: Clark
authors.forename: J
authors.initials: J
authors.lastname: Parkin
authors.forename: A
authors.initials: A
authors.lastname: Pinching
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D016477:Artifacts; D015153:Blotting, Western; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006679:HIV Seropositivity; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D016131:Lymphocyte Subsets / Q000166:cytology; D008297:Male; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D016827:CD8 Antigens