Kevin N Brown1, Simon M Barratt-Boyes. 1. Department of Infectious Diseases and Microbiology, Graduate School of Public Health, and Center for Vaccine Research, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Abstract
BACKGROUND: The study of dendritic cell (DC) biology in the rhesus macaque is becoming increasingly important but is limited by incomplete characterization and the lack of a rapid assay to quantify cells. METHODS: We characterized the surface phenotype of myeloid (mDC) and plasmacytoid DC (pDC) subsets in healthy rhesus macaque blood and developed a flow cytometry-based assay for absolute DC determinations. RESULTS: Rhesus CD11c(+) mDC were CD16(+) CD11b(+) CD56(lo) CD8(-) CD1c(-) whereas CD123(+) pDC lacked expression of these markers. Precise DC determinations were performed using a rapid two-step assay combining the analysis of whole blood and peripheral blood leukocytes (PBL). CONCLUSIONS: Antibodies to CD11b, CD56 and CD16 must be omitted from the lineage antibody cocktail to prevent inadvertent gating-out of DC when analyzing rhesus blood. The combined whole-blood/PBL quantification assay will be invaluable for the rapid and repeated monitoring of blood DC counts in this species.
BACKGROUND: The study of dendritic cell (DC) biology in the rhesus macaque is becoming increasingly important but is limited by incomplete characterization and the lack of a rapid assay to quantify cells. METHODS: We characterized the surface phenotype of myeloid (mDC) and plasmacytoid DC (pDC) subsets in healthy rhesus macaque blood and developed a flow cytometry-based assay for absolute DC determinations. RESULTS: Rhesus CD11c(+) mDC were CD16(+) CD11b(+) CD56(lo) CD8(-) CD1c(-) whereas CD123(+) pDC lacked expression of these markers. Precise DC determinations were performed using a rapid two-step assay combining the analysis of whole blood and peripheral blood leukocytes (PBL). CONCLUSIONS: Antibodies to CD11b, CD56 and CD16 must be omitted from the lineage antibody cocktail to prevent inadvertent gating-out of DC when analyzing rhesus blood. The combined whole-blood/PBL quantification assay will be invaluable for the rapid and repeated monitoring of blood DC counts in this species.
Authors: J Banchereau; F Briere; C Caux; J Davoust; S Lebecque; Y J Liu; B Pulendran; K Palucka Journal: Annu Rev Immunol Date: 2000 Impact factor: 28.527
Authors: S Pichyangkul; P Saengkrai; K Yongvanitchit; C Limsomwong; M Gettayacamin; D S Walsh; V A Stewart; W R Ballou; D G Heppner Journal: J Immunol Methods Date: 2001-06-01 Impact factor: 2.303
Authors: S Vuckovic; D Gardiner; K Field; G V Chapman; D Khalil; D Gill; P Marlton; K Taylor; S Wright; A Pinzon-Charry; C M Pyke; R Rodwell; R L Hockey; M Gleeson; S Tepes; D True; A Cotterill; D N J Hart Journal: J Immunol Methods Date: 2004-01 Impact factor: 2.303
Authors: Benoît Malleret; Ingrid Karlsson; Benjamin Manéglier; Patricia Brochard; Benoît Delache; Thibault Andrieu; Michaela Muller-Trutwin; Tim Beaumont; Joseph M McCune; Jacques Banchereau; Roger Le Grand; Bruno Vaslin Journal: Immunology Date: 2008-01-24 Impact factor: 7.397
Authors: Gerrit Koopman; Henk Niphuis; Anton G M Haaksma; Ann M Farese; Dan B Casey; Larry E Kahn; Dean Mann; Thomas J MacVittie; Susan L Woulfe; Jonathan L Heeney Journal: Hum Immunol Date: 2004-04 Impact factor: 2.850
Authors: P Toby H Coates; Simon M Barratt-Boyes; Linyou Zhang; Vera S Donnenberg; Peta J O'Connell; Alison J Logar; F Jason Duncan; Michael Murphey-Corb; Albert D Donnenberg; Adrian E Morelli; Charles R Maliszewski; Angus W Thomson Journal: Blood Date: 2003-06-26 Impact factor: 22.113
Authors: Viskam Wijewardana; Anthea L Bouwer; Kevin N Brown; Xiangdong Liu; Simon M Barratt-Boyes Journal: Immunology Date: 2014-10 Impact factor: 7.397
Authors: Chie Sugimoto; Atsuhiko Hasegawa; Yohei Saito; Yayoi Fukuyo; Kevin B Chiu; Yanhui Cai; Matthew W Breed; Kazuyasu Mori; Chad J Roy; Andrew A Lackner; Woong-Ki Kim; Elizabeth S Didier; Marcelo J Kuroda Journal: J Immunol Date: 2015-07-15 Impact factor: 5.422
Authors: Elizabeth R Wonderlich; Zachary D Swan; Stephanie J Bissel; Amy L Hartman; Jonathan P Carney; Katherine J O'Malley; Adebimpe O Obadan; Jefferson Santos; Reagan Walker; Timothy J Sturgeon; Lonnie J Frye; Pauline Maiello; Charles A Scanga; Jennifer D Bowling; Anthea L Bouwer; Parichat A Duangkhae; Clayton A Wiley; JoAnne L Flynn; Jieru Wang; Kelly S Cole; Daniel R Perez; Douglas S Reed; Simon M Barratt-Boyes Journal: J Immunol Date: 2017-01-06 Impact factor: 5.422
Authors: Elizabeth R Wonderlich; Amy L Caroline; Cynthia M McMillen; Aaron W Walters; Douglas S Reed; Simon M Barratt-Boyes; Amy L Hartman Journal: J Virol Date: 2018-01-17 Impact factor: 5.103