CCR2-64I polymorphism is not associated with altered CCR5 expression or coreceptor function.
A polymorphism in the gene encoding CCR2 is associated with a delay in progression to AIDS in human immunodeficiency virus (HIV)-infected individuals. The polymorphism, CCR2-64I, changes valine 64 of CCR2 to isoleucine. However, it is not clear whether the effect on AIDS progression results from the amino acid change or whether the polymorphism marks a genetically linked, yet unidentified mutation that mediates the effect. Because the gene encoding CCR5, the major coreceptor for HIV type 1 primary isolates, lies 15 kb 3' to CCR2, linked mutations in the CCR5 promoter or other regulatory sequences could explain the association of CCR2-64I with slowed AIDS pathogenesis. Here, we show that CCR2-64I is efficiently expressed on the cell surface but does not have dominant negative activity on CCR5 coreceptor function. A panel of peripheral blood mononuclear cells (PBMC) from uninfected donors representing the various CCR5/CCR2 genotypes was assembled. Activated primary CD4(+) T cells of CCR2 64I/64I donors expressed cell surface CCR5 at levels comparable to those of CCR2 +/+ donors. A slight reduction in CCR5 expression was noted, although this was not statistically significant. CCR5 and CCR2 mRNA levels were nearly identical for each of the donor PBMC, regardless of genotype. Cell surface CCR5 and CCR2 levels were more variable than mRNA transcript levels, suggesting that an alternative mechanism may influence CCR5 cell surface levels. CCR2-64I is linked to the CCR5 promoter polymorphisms 208G, 303A, 627C, and 676A; however, in transfected promoter reporter constructs, these did not affect transcriptional activity. Taken together, these findings suggest that CCR2-64I does not act by influencing CCR5 transcription or mRNA levels.
Mariani
R
R
Aaron Diamond AIDS Research Center, The Rockefeller University, New York, New York 10016, USA.
Wong
S
S
Mulder
L C
LC
Wilkinson
D A
DA
Reinhart
A L
AL
LaRosa
G
G
Nibbs
R
R
O'Brien
T R
TR
Michael
N L
NL
Connor
R I
RI
Macdonald
M
M
Busch
M
M
Koup
R A
RA
Landau
N R
NR
D001782:Blood Donors; D002460:Cell Line; D018925:Chemokines / Q000096:biosynthesis; D005838:Genotype; D006678:HIV / Q000254:growth & development; D006801:Humans; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D054390:Receptors, CCR2; D019713:Receptors, CCR5 / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D019718:Receptors, CXCR4 / Q000032:analysis; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine / Q000032:analysis / Q000235:genetics*; D015728:Receptors, HIV / Q000502:physiology*; D013601:T-Lymphocytes / Q000821:virology