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. 2005 Apr 7;64(11):1542–1549. doi: 10.1136/ard.2004.032599

Involvement of neurotrophins and their receptors in spondyloarthritis synovitis: relation to inflammation and response to treatment

M Rihl 1, E Kruithof 1, C Barthel 1, F De Keyser 1, E Veys 1, H Zeidler 1, D Yu 1, J Kuipers 1, D Baeten 1
PMCID: PMC1755273  PMID: 15817657

Abstract

Objective: To investigate whether expression of the four members of the neurotrophin (NT) family and their four corresponding receptors is related to synovial inflammation in patients with spondyloarthritis (SpA).

Material and Methods: Synovial fluid (SF) and serum NTs and their receptors were measured by ELISA. Immunohistochemistry was used for synovial tissue biopsy specimens from patients with SpA, rheumatoid arthritis, and osteoarthritis (OA). In SpA synovium, immunoreactivity of the receptors trkA and NGFRp75 was also assessed before and after 12 weeks of treatment with the monoclonal anti-tumour necrosis factor α antibody, infliximab.

Results: mRNA transcripts of all NTs and receptors were expressed in the inflamed synovium. At the protein level, brain derived neurotrophic factor and NT-3 were significantly higher in the SF of patients with SpA than in those with OA. In contrast, ELISA of serum samples showed that the highest member in SpA was NT-4. Immunohistochemistry demonstrated that the NT receptors trkA and NGFRp75 were highly expressed in the inflamed synovium of patients with SpA, correlating with vascularity and lymphoid aggregates, respectively. Additionally, immunoreactivity of both receptors was significantly decreased after infliximab treatment.

Conclusions: NTs and their receptors are expressed in inflamed peripheral joints of patients with SpA. Their expression is not constitutive but related to inflammation and they may be involved in the local disease processes.

Full Text

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Figure 1.

Figure 1

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 Neurotrophins were measured by quantitative immunoassay in the SF of 15 patients with SpA, 15 with RA, and 10 with OA (A-D) and in the serum of these patients and 10 healthy controls (HC) (E–H). The scatter plots of all four neurotrophins indicate the individual values. Bold horizontal lines represent the median. Detectable concentrations (median, range; always given for the total group of patients) of SF BDNF were measured in 11/15 (73%) patients with SpA (4.5 pg/ml, 0–90), in 8/15 (53%) patients with RA (1.7 pg/ml, 0–218), and in 2/10 (20%) patients with OA (0 pg/ml, 0–24). SF BDNF concentrations were significantly higher in the SpA group than in patients with OA (A), *p = 0.025. SF NT-3 was detected in 5/15 (33%) patients with SpA (0 pg/ml, 0–200) and in 4/15 (27%) patients with RA (0 pg/ml, 0–17). SF NT-3 levels were also significantly higher in SpA than in OA (C), *p = 0.047). Serum BDNF (E) was detectable in all 50 subjects showing significantly higher levels in HC than all the other groups (SpA, *p<0.0001; RA, *p<0.0001; and OA, *p = 0.001), whereas serum NT-4 (H) was significantly higher in SpA (*p<0.001), RA (*p = 0.001), and OA (*p = 0.008) than in HC.

Figure 2.

Figure 2

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 Immunoreactivity of trkA (A–C) and NGFRp75 (D–F) in the lining layer, sublining layer, and endothelium of synovial tissue biopsy specimens obtained from clinically affected knee joints of 24 patients with SpA, 15 with RA, and 10 with OA. The results represent the individual scores on a semiquantitative 0 (no expression) to 3 (high expression) scale, with bars representing the median score. As indicated (*), the immunoreactivity of trkA was significantly higher in SpA lining than in RA lining (p = 0.047) and OA lining (p = 0.034). Similarly, trkA was more highly expressed in SpA and RA sublining than in OA (p = 0.001 and p = 0.003, respectively). NGFRp75 was almost absent in the lining layer and was equally expressed across disease groups in the sublining layer. However, some patients with SpA showed high endothelial expression, resulting in a significant difference in comparison with RA (p = 0.012) and OA (p = 0.041).

Figure 3.

Figure 3

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 Microscopic pictures of ST sections from 12 different patients are depicted. (A–F) immunostaining of synovial tissue samples of two different patients with SpA, RA, and OA each at low (A, B, C, x160) and high magnification (D, E, F, x320). In patients with SpA, intensive trkA staining predominantly in the lining layer (A) and less intense staining in the sublining layer and the endothelium (D) is seen. In patients with RA, a similar pattern for trkA staining is seen (B, x160 and E, x320) and correspondingly for patients with OA (C, F). (G-L) NGFRp75 immunoreactivity showing an intense staining in the sublining layer (G, J) of patients with SpA. However, immunoreactivity for NGFRp75 is less intense in RA (H, K) and OA (I, L) than in SpA. In all three groups, NGFRp75 staining is not seen in the lining layer, but it is found in the endothelium of patients with SpA.

Figure 4.

Figure 4

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 Significant correlations as determined by Spearman's test in patients with SpA between NT expression and measures of synovial inflammation were found for (A) NT-3 in SF and the degree of vascularisation; (B) immunoreactivity of trkA in the lining layer and the degree of vascularisation; and (C) immunoreactivity of NGFRp75 in the endothelium and the number of lymphoid aggregates.

Figure 5.

Figure 5

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 Immunohistochemistry on ST biopsy samples of selected patients with SpA before and after 12 weeks of infliximab treatment (original magnification is x160 in A, B and x320 in C, D). (A, C) show trkA staining at baseline (week 0) with a predominance of immunoreactivity in the lining and sublining layer. (B, D) Staining of synovial samples obtained in the same patients after 12 weeks of treatment with infliximab. (E–G) indicate the semiquantitative scores (0–3) for trkA expression in the lining and sublining layer and in the endothelium of all nine patients with SpA who were analysed before and 12 weeks after TNFα blocking treatment with infliximab. (E) The trkA reduction in the lining layer was significant at p = 0.03 (*).

Figure 6.

Figure 6

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 Immunohistochemistry on ST biopsy specimens of selected patients with SpA before and after 12 weeks of infliximab treatment (original magnification is x160 in A, B and x320 in C, D). (A, C) NGFRp75 staining at baseline, with a predominance in the sublining layer and the endothelium. (B, D) Synovial sections of the same patients stained after 12 weeks of anti-TNFα treatment with infliximab, showing clearly a less intense or even absent staining in both the lining and the sublining layer as well as in the endothelium. (E–G) indicate the semiquantitative scores (0–3) of NGFRp75 expression in the lining and sublining layer and in the endothelium of the nine patients with SpA before and 12 weeks after TNFα blocking treatment with infliximab. (F, G) NGFRp75 is significantly reduced in the sublining layer and in the endothelium (both at *p = 0.004).

Selected References

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