ABSTRACT
OBJECTIVE: To determine gene expression of the interleukin-17 (IL-17) family members (IL-17A-F) in rheumatoid subcutaneous nodules, and to assess the cytokines involved in regulating IL-17A expression. METHODS: Total RNA was isolated from 19 nodules obtained from 16 different patients with rheumatoid arthritis (RA). Reverse transcription-polymerase chain reaction (PCR) was used to screen for gene expression of the IL-17 subtypes (IL-17A-F) in all nodules. Quantitative real-time PCR was used to measure the expression of interferon-gamma (IFN gamma), IL-6, IL-23, IL-12, and transforming growth factor beta (TGFbeta), relative to GAPDH as control, in a subset of 10 nodules. RESULTS: IL-17A gene expression was present in only 1 of 19 nodules, IL-17B in 17 of 19 nodules, IL-17C in 18 of 19 nodules, IL-17D in 16 of 19 nodules, and IL-17E in 3 of 19 nodules. IL-17F was absent in all samples. Cytokines that stimulate IL-17A production (IL-6, IL-23) as well as those that inhibit IL-17A production (IL-12, IFN gamma, TGFbeta) were present in the majority of nodules. Quantitative real-time PCR showed a similar pattern of gene expression for the individual cytokines between the different nodules. The mean +/- SD expression of IL-6 relative to GAPDH was 2.28 +/- 2.2 ng, and that of TGFbeta was 2.96 +/- 1.14 ng. There was a lower relative expression of IL-23 (0.05 +/- 0.05 ng), while the expression of IFN gamma was 0.67 +/- 0.68 ng and that of IL-12 was 0.48 +/- 0.23 ng. CONCLUSION: IL-17 family members are varyingly expressed in rheumatoid nodules. The paucity of IL-17A in nodules suggests an important difference from that observed in the synovium. The expression of IL-23 below a critical threshold level seems the most likely explanation for the virtual absence of IL-17A. The presence of tissue destruction within the nodule despite the absence of IL-17A suggests that IL-17A may be an important amplifier rather than an absolute requirement for inflammation in RA.
Subject(s)
Interleukin-17/genetics , Rheumatic Nodule/genetics , Rheumatoid Nodule/genetics , Synovial Membrane/immunology , T-Lymphocyte Subsets/immunology , Aged , Cell Differentiation/immunology , Cohort Studies , Cytokines/genetics , Female , Gene Expression Profiling , Gene Expression Regulation , Humans , Male , Middle Aged , RNA/metabolism , Rheumatoid Nodule/immunologyABSTRACT
OBJECTIVE: To investigate the distribution of the A2756G polymorphism of the methionine synthase reductase (MTR) gene in patients with rheumatoid arthritis (RA) treated with methotrexate (MTX) compared with a healthy control group; and to examine the relationships among the A2756G polymorphism, plasma total homocysteine (tHcy), serum folate and vitamin B12 levels, disease activity, and MTX toxicity in patients with RA. METHODS: A cross-sectional study was performed on 86 MTX-treated RA patients, consisting of a clinical interview and physical examination to determine disease activity and MTX-related adverse reactions. Genotype analysis of the MTR gene was performed. Fasting plasma tHcy, serum folate, and vitamin B12 levels were measured. Allele and genotype distributions were compared to a healthy control group. RESULTS: The frequency of the 2756GG genotype (16.3%) in the RA study group was higher than that expected in the general population (3.6%; p < 0.000001). This genotype was associated with MTX-induced accelerated rheumatoid nodulosis (MIARN). No association of disease activity variables or plasma homocysteine with MTR A2756G polymorphisms was observed. The MTR 2756GG genotype, low plasma vitamin B12 levels, and the presence of rheumatoid nodules predicted MIARN. No association of nodulosis with any other indicator of disease activity or medical treatment was found. CONCLUSION: In our population of MTX-treated RA patients the 2756GG genotype of the MTR gene was more common than expected and was associated with MIARN.
Subject(s)
Antirheumatic Agents/adverse effects , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/genetics , Ferredoxin-NADP Reductase/genetics , Methotrexate/adverse effects , Polymorphism, Single Nucleotide/genetics , Rheumatic Nodule/chemically induced , Adult , Aged , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/blood , Case-Control Studies , Cross-Sectional Studies , Female , Genotype , Homocysteine/blood , Humans , Male , Methotrexate/therapeutic use , Middle Aged , Pteroylpolyglutamic Acids/blood , Rheumatic Nodule/genetics , Severity of Illness Index , Vitamin B 12/bloodABSTRACT
Two patients are described with longstanding, multiple, subcutaneous nodules with the histopathological features of rheumatoid nodules. Neither patient had any clinical evidence of rheumatic disease. One patient had a family history of smiliar nodules transmitted as an autosomal dominant trait. Leukocyte function studies failed to reveal any defect to account for the nodule formation.
