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. 2004 Mar;63(3):310–317. doi: 10.1136/ard.2003.006148

The dinucleotide repeat polymorphism in the 3'UTR of the CD154 gene has a functional role on protein expression and is associated with systemic lupus erythematosus

M Citores 1, I Rua-Figueroa 1, C Rodriguez-Gallego 1, A Durantez 1, M Garcia-Laorden 1, C Rodriguez-Lozano 1, J Rodriguez-Perez 1, J Vargas 1, P Perez-Aciego 1
PMCID: PMC1754911  PMID: 14962968

Abstract

Objective: To investigate the association of the (CA)n dinucleotide repeat in the 3' untranslated region (3'UTR) of the CD154 gene with systemic lupus erythematosus (SLE), and its functional role in protein expression.

Methods: The allelic and genotypic distributions of the polymorphism were compared in 80 patients with SLE and 80 controls. A complete clinical and analytical database was recorded in each patient in order to correlate the clinical manifestations in SLE with different alleles. To investigate the functional role of the polymorphism, the CD154 protein expression on activated lymphocytes from healthy homozygous controls was evaluated by flow cytometry.

Results: The 24 CA allele was the most represented in controls (p = 0.029), whereas the alleles containing >24 CA repeats were found in patients (p = 0.0043). Furthermore, when only homozygous women were considered, most controls carried two 24 CA alleles (p = 0.041), whereas most patients carried two alleles containing >24 CA repeats (p = 0.032). Also, patients carrying at least one 24 CA allele had less neurological involvement (p = 0.034), and carriers of at least one allele with fewer than 24 CA repeats presented more livedo reticularis (p = 0.006) and anti-Sm (p = 0.01) and anti-RNP (p = 0.038) autoantibodies. CD154 maximum expression in activated lymphocytes from all controls was reached after 54 hours, but it was more prolonged in controls carrying two alleles with >24 CA repeats (p = 0.0068).

Conclusion: The CD154 3'UTR microsatellite is associated with SLE, and the most represented alleles in patients were accompanied by a more prolonged protein expression in activated lymphocytes from controls.

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Figure 1 .

Figure 1

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Frequency distribution of the number of (CA)n repeats in patients with SLE and controls.

Figure 2 .

Figure 2

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CD154 kinetic expression on PBMC from controls stimulated with PHA and anti-CD28. PBMC from three homozygote controls carrying two 24 CA alleles were stimulated with PHA and anti-CD28 antibody and were harvested after 24, 48, 54, 57, and 72 hours. CD154 expression was measured by flow cytometry on CD4+ T lymphocytes (continuous line) and B cells (dotted lines).

Figure 3 .

Figure 3

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Flow cytometry plots showing the CD154 expression on T lymphocytes from representative controls. PBMC from controls with different genotypes were activated with PHA and anti-CD28 for 54 (first column) or 72 hours (last column). Percentages of CD4+CD154- and CD4+CD154+ are indicated (left and right of each plot, respectively), and the percentage of the CD4+CD154+ cells from the total of CD4+ cells is shown in parenthesis. All the CD4+ cells were CD14-, all of them being CD4+ T lymphocytes.

Selected References

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