Selective expression of folate receptor beta and its possible role in methotrexate transport in synovial macrophages from patients with rheumatoid arthritis.
OBJECTIVE
To investigate the expression of folate receptors (FR) and reduced folate carrier (RFC) and determine their relevance to methotrexate (MTX) transport in synovial mononuclear cells (SMC) from patients with rheumatoid arthritis (RA).


METHODS
Levels of FR and RFC messenger RNA (mRNA) were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) in SMC from RA patients and peripheral blood mononuclear cells from healthy donors. Expression of FR-beta mRNA and protein was determined by Northern blot and Western blot analyses in RA SMC and monocyte/macrophage-lineage cells. FR-beta expression and folic acid binding capacity on the cell surface were examined by flow cytometric analysis and 3H-folic acid binding analysis. Studies of the inhibition of 3H-MTX uptake in the presence of unlabeled folic acid were performed to investigate the uptake of MTX through FR in RA SMC.


RESULTS
RT-PCR, Northern blot, and Western blot analyses showed that FR-beta mRNA and protein were expressed selectively in activated monocytes and CD14+ RA SMC. These cells exhibited folic acid binding capacity. Furthermore, the FR-beta protein was shown to have folic acid binding capacity. Uptake of 3H-MTX through RA SMC was significantly inhibited in the presence of unlabeled folic acid.


CONCLUSIONS
These results demonstrate that FR-beta expression is selectively elevated in RA synovial macrophages and suggest that MTX is transported through FR-beta in RA synovial macrophages. The findings suggest that folate antagonists with higher affinity for FR-beta would be useful in the treatment of RA.
Nakashima-Matsushita
N
N
Kagoshima University School of Medicine, Japan.
Homma
T
T
Yu
S
S
Matsuda
T
T
Sunahara
N
N
Nakamura
T
T
Tsukano
M
M
Ratnam
M
M
Matsuyama
T
T
D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000473:pathology*; D001692:Biological Transport; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology*; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008727:Methotrexate / Q000493:pharmacokinetics*; D009000:Monocytes / Q000737:chemistry / Q000276:immunology; D012313:RNA / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis; D013583:Synovial Membrane / Q000473:pathology*
D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D018950:Lipopolysaccharide Receptors; D011956:Receptors, Cell Surface; D012313:RNA; D005492:Folic Acid; D008727:Methotrexate