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Volume 3 Supplement 2

21st European Workshop for Rheumatology Research

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Functional and biologic evaluation of immunoregulatory cytokines in the immunopathologic lesion of Sjogren's syndrome

Autoimmune diseases are commonly considered to involve a Th1/Th2 imbalance in favor of Th1. Studies in mice suggest that the Th1 cytokines overexpression may be implicated in the pathogenesis of autoimmunity. Recent results, though, in humans challenge this notion.

Conflicting data have been published on the cytokine profile in patients with Sjogren's syndrome (S.s.). These data were mainly obtained using PCR based techniques and in-situ hybridization. Some of them indicate that a Th1 response is prevalent. To clarify this issue, small salivary glands (SGL) from patients with S.s. with different grades of infiltration as well as patients without S.s. but with sicca manifestations, were cultured in RPMI plus 40 IU/ml of recombinant IL2. IL4, IL13 and IFNgamma production in the culture supernatants (SN) was evaluated with a sensitive ELISA and, in parallel, mRNA levels for the same cytokines were evaluated with a quantative RT-PCR.

Within a period of a week, lymphocytes were evident in the cultures in numbers depending on the infiltration of the gland. The phenotype of these lymphocytes, as determined by flow cytometry, was similar to that published previously from SGL immunochemistry, suggesting that these cells derive from the salivary gland. SN were collected on various time intervals and mRNA was extracted from the lymphocytes on day 12 of the culture. Interestingly, IL13 mRNA was detected in almost all (17/19) of the samples, and both IFNgamma and IL4 on the majority (16/19 and 14/19, respectively). The cytokine production in the culture SN was examined in a set of 8 biopsies. IL 4 couldn't be detected (<4 pg/ml), may be due to its binding on the IL4R. Both IFNgamma and IL13 were present and the ratio of IFNgamma/IL13 was related to the grade of infiltration (p < 0.05), indicating that the balance of Th1 vs Th2 changes in favor of the former along with the severity of the disease. Finally, it was demonstrated that the Th2 cytokines IL4 and IL13 are functional since IgE was present in 12 out of 28 SN of cultured biopsies. IgE was present in greater quantities in those biopsies that had low grade infiltration (0-0.3 according to Chisholm's score) compared to those with medium grade (0.3-0.6), while in biopsies with infiltration >0.6 it was undetectable.

These results show that in Sjogren's syndrome the distinction between Th1 and Th2 doesn't apply as both play a role to its pathogenesis. Moreover it seems that Th2 response prevails at the early stages whereas Th1 gradually augments as the disease progresses.

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Mitsias, D., Tzioufas, A., Veiopoulou, C. et al. Functional and biologic evaluation of immunoregulatory cytokines in the immunopathologic lesion of Sjogren's syndrome. Arthritis Res Ther 3 (Suppl 2), P017 (2001).

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