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Rheumatology Advance Access published online on March 3, 2008
Rheumatology, doi:10.1093/rheumatology/ken073
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© The Author 2008. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Reduced folate carrier polymorphism determines methotrexate uptake by B cells and CD4+ T cells

B. Baslund1, J. Gregers2 and C. H. Nielsen3

1Department of Rheumatology, Rigshospitalet and the University of Copenhagen, 2Bonkolab and 3Institute for Inflammation Research, Rigshospitalet, The University Hospital of Copenhagen, Denmark.

Correspondence to: B. Baslund, Department of Rheumatology TA 4242, The University Hospital, Blegdamsvej 9, DK-2100 Copenhagen, Denmark. E-mail: bbaslund{at}gmail.com


  Abstract

Objective. To examine if polymorphism 80G -> A in the Reduced Folate Carrier (RFC) affects uptake of MTX in B- and CD4+ T-cells.

Methods. Mononuclear cells were isolated from peripheral blood of healthy persons. Real-time PCR was used to detect the RFC80 variants. FITC-labelled MTX was added to cells stimulated with Candida albicans or tetanus toxoid, and the uptake of MTX was measured by flow cytometry. A FITC-conjugated monoclonal antibody against RFC was used to detect the cellular RFC expression.

Results. Antigen-stimulated CD4+ T cells and B cells from individuals with the GG variant (n = 9) exhibited lower uptake of MTX than individuals expressing the AA variant (n = 8), or the GA variant (n = 8). No difference could be demonstrated between the three groups with respect to the expression of RFC by CD4+ T cells and B cells, and CD4+ T cells from individuals homozygous for the G allele exhibited lower uptake of MTX per receptor than CD4+ T cells from individuals homozygous for the A allele.

Conclusion. MTX is taken up more efficiently via the A allele than via the G allele. This difference between the variant forms of RFC suggests that genotyping could be relevant for determining the relevant dosage of MTX in the treatment of neoplastic and autoimmune disease.

KEY WORDS: Methotrexate, Genetic polymorphism, Reduced folate carrier

Submitted 12 November 2007; revised version accepted 30 January 2008.
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