Circulating human CD4 and CD8 T cells do not have large intracellular pools of CCR5.
CC Chemokine Receptor 5 (CCR5) is an important mediator of chemotaxis and the primary coreceptor for HIV-1. A recent report by other researchers suggested that primary T cells harbor pools of intracellular CCR5. With the use of a series of complementary techniques to measure CCR5 expression (antibody labeling, Western blot, quantitative reverse transcription polymerase chain reaction), we established that intracellular pools of CCR5 do not exist and that the results obtained by the other researchers were false-positives that arose because of the generation of irrelevant binding sites for anti-CCR5 antibodies during fixation and permeabilization of cells.
Department of Medicine, Division of Infectious Diseases, Case Western Reserve University/University Hospitals/Case Medical Center, Cleveland, OH, USA.
Heather A
HA
Pilch-Cooper
Scott F
SF
Sieg
Thomas J
TJ
Hope
Ann
A
Koons
Jean-Michel
JM
Escola
Robin
R
Offord
Ronald S
RS
Veazey
Donald E
DE
Mosier
Brian
B
Clagett
Kathy
K
Medvik
Julie K
JK
Jadlowsky
Mark R
MR
Chance
Janna G
JG
Kiselar
James A
JA
Hoxie
Ronald G
RG
Collman
Nadeene E
NE
Riddick
Valentina
V
Mercanti
Oliver
O
Hartley
Michael M
MM
Lederman
D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000737:chemistry / Q000378:metabolism*; D002469:Cell Separation; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D005189:False Positive Reactions; D005434:Flow Cytometry; D006801:Humans; D019713:Receptors, CCR5 / Q000032:analysis / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016707:Tissue Fixation
D019713:Receptors, CCR5