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Impressum
Upregulation of TNF-alpha and IL-6 in keratocytes from patients with corneal ulcerations associated with rheumatoid arthritis
1Prada J., 2Noelle B., 1Pleyer U.
1Department of Ophthalmology, Charité Campus Virchow, Humboldt University, Berlin, Germany and 2Department of Ophthalmology, Christian Albrechts University, Kiel, Germany.
Objective: Sterile corneal ulceration is a well-recognized manifestation
of rheumatoid arthritis. The exact mechanism responsible for corneal melting
remains unknown, but is considered to be immune mediated leading to an
activation of metalloproteinases. A striking feature of rheumatoid corneal
ulceration is a limited infiltration of inflammatory cells, which further
leads to the hypothesis that keratocytes are actively involved in this
process. Therefore, we have analysed the gene expression of the proinflammatory
cytokines Tumor Necrosis Factor-alpha (TNF-alpha) and Interleukin-6 (IL-6)
in corneal samples from patients with corneal ulcerations and/or perforations
associated with rheumatoid arthritis.
Methods: Corneal samples from seven patients with rheumathoid arthritis
and corneal ulcerations and/or perforations were collected and stored
in 4% paraformaldehyde, previously to be paraffin-embedded in "RNAase-free"
conditions. Paraffin sections of 4-5 µm were fixed on silan-coated
slides and further analysed by systematic non-radioactive in situ hybridization,
using specific gene probes for TNF and IL-6 labelled with digoxigenin
(DIG). Labelling of gene probes with digoxigenin as well as the detection
of the resulting hybrids was carried out with the DIG labelling and detection
system from Boehringer Mannheim, Mannheim (Germany).
Results: An extended TNF-alpha gene expression could be clearly
observed in the keratocytes surrounding the corresponding corneal ulcerations
and/or perforations from 5 of the 7 analysed patients. In addition, all
7 patients presented clearly positive results for a widely extended IL-6
gene expression in keratocytes from the same analysed tissue samples.
Conclusion: Our observations indicate that corneal stroma cells
surrounding corneal ulcerations and/or perforations from patients with
rheumatoid arthritis often show an upregulation of the cytokines TNF-alpha
and IL-6. These findings support the hypothesis that proinflammatory cytokines
released by keratocytes may modify the production of metalloproteinases
in the corresponding cells, further resulting in collagenolytic corneal
damage.