Method for prediction of clinical course of autoimmune thyroiditis

FIELD: medicine.

SUBSTANCE: peripheral venous blood of the patients with autoimmune thyroiditis is examined for the absolute transferring receptor (CD71+) T-lymphocyte count and the interleukin-2 (IL-2) concentration. If the CD71+ T-lymphocyte count exceeds 0.3×109 cell/l, and the IL-2 concentration is less than 25 pg/ml, a favourable clinical course of the disease is predicted, while the CD71+ T-lymphocyte count less than 0.3×109 cell/l and the IL-2 concentration exceeding 25 pg/ml shows an unfavourable clinical course of autoimmune thyroiditis.

EFFECT: method provides more accurate and objective prediction with using the material, reagents and research methods widely applied in common clinical-laboratory practise.

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The invention relates to immunology, endocrinology, laboratory diagnosis, and is intended for prediction of clinical course of autoimmune thyroiditis.

Autoimmune thyroiditis (AIT) suffer from 3-4% of the world's population, with age, the number of patients increases, while the prevalence in women is 3 to 6 % per year. Recently, clinicians in many countries the increasing number of patients with autoimmune thyroiditis among young people, especially among children and adolescents (thyroid Disease, Ed. of Bravermann LI - M.: Medicine, 2000). In General, the analysis of blood in patients with autoimmune thyroiditis possible lymphocytosis, monocytosis, leukopenia and increased ESR. Immunological blood test allows to detect the increase in the number and activity of T-helper and T-killer cells by reducing the number of T-suppressor cells, increased levels of immunoglobulins.

There is a method of evaluating the secretion of TSH, which includes sample with thyroliberin. The thyrotropin-releasing hormone stimulates the release of TSH thyroid-stimulating cells adenogipofiza, so normal after the introduction of thyroliberin TSH level increases. Sample with thyroliberin used to assess secretion of TSH reserve and the degree of suppression of TSH secretion (thyrotoxicosis). Response to thyrotropin-releasing hormone reduced or delayed amid suppressive therapy levotiroxina recent sample with thyroliberin rarely used, because of the new highly sensitive methods for the determination of TSH (Endocrinology edited Nlevine. - M., 1999).

There is a way to evaluate the effectiveness of treatment of AIT, including the definition in the serum of patients contents of interferon-gamma (Patent No. 2323443, Russia. Publ. 27.04.2008). After 1 month of treatment with the concentration of interferon-gamma less than 35 PG/ml appreciate the treatment as effective, and when the content is more than 35 PG/ml evaluate treatment as ineffective and adjust the dose of therapeutic drug or change the medication.

Also known is a method for predicting and diagnosing diseases of the thyroid gland (US 2010/0131286 A1, 27.05.2010), which investigated the level of expression of specific marker genes, indicating that the risk of the development and distribution of endocrine pathologies, including autoimmune thyroiditis. In the specified method is used, however, expensive equipment and reagents, which is not always economically feasible.

The prototype is to study the serum levels of cytokine BLyS, stimulating b-lymphocytes for the diagnosis, prognosis and screening the effectiveness of therapy of autoimmune diseases (WO 2009043848, 09.04.2009). The method includes comparing the concentration of BLyS with a reference value and the subsequent calculation of variances for the purpose of diagnosis, prognosis and screening. However, the content of titoki is and BLyS serum increases not only when AIT, systemic lupus erythematosus, rheumatoid arthritis and sjögren's syndrome, but also in patients with multiple myeloma, chronic b-cell leukemia that is not sufficiently objective criterion for predicting the nature of the flow AIT. We stated the invention improves the accuracy and objectivity of forecasting, using two criteria: the absolute content of T-lymphocytes with the transferrin receptor (CD71+) and the concentration of interleukin-2 (IL-2). When the content of lymphocytes CD71+ more than 0.3×109CL/l and the concentration of IL-2 less than 25 PG/ml predicts favorable course of the disease, and when the content of lymphocytes CD71+ less than 0.3×109CL/l and the concentration of IL-2 for more than 25 PG/ml predict unfavorable for AIT. Used in the way materials, reagents and methods are widely used in standard clinical and laboratory practice.

The claimed method is standard in the following way. Blood should be taken from the cubital vein in a minimal volume (1 ml) to 10 o'clock in the morning, on an empty stomach. The content of T-lymphocytes CD71+ were determined using indirect immunoperoxidase reactions with the use of monoclonal antibodies for the preparations of lymphocytes type "dried droplet" production RPC Medispectra Moscow. For the quantitative determination of IL-2 is used serum or PLA is mA blood analyzed by ELISA with the use of a diagnostic kit for ELISA. The results of the reaction are measured on a spectrophotometer with a wavelength of 450 nm.

In this way were examined persons rehabilitated in a medical company "Biochar" Arkhangelsk: 28 patients with autoimmune thyroiditis and 15 people have a course of hormone treatment of thyroid cancer and are in remission (control). Diagnosed by the doctors of the city endocrinology Advisory centre of Arkhangelsk, the survey was carried out in compliance with the basic norms of biomedical ethics. The results were processed using the software package Statistica 6" ("StatSoft, USA). Study type retrospective, random sample, simultaneously. General population - residents of the North of European Russia. The significance of differences between groups was evaluated using a parametric t-student test for independent samples and non-parametric criterion of Wilcoxon signed. Statistical significance was assigned at p<0,05.

It is established that under unfavorable course of the disease (increase in blood concentrations of antibodies to thyroperoxidase) patients the decrease of the absolute content of T-lymphocytes CD71+ from 0.47±0,05 to 0.26±0,02×l09to the/l and increased concentrations of IL-2 23,8±5.3 to 154,5±11,7 compared to controls (p< 0,05). Under favorable for AIT (decrease in the level of antibodies to thyroperoxidase) increases the content of lymphocytes CD71+ to 0.56±0,05×109CL/l and normalized index of IL-2 (less than 25 PG/ml).

There is information on the stimulating effect of thyroxine on the release of soluble alpha-receptors to IL-2 (Zwirska-Korczala K. Influence of thyroxine on serum soluble interleukin-2 receptor alpha levels in thyroid disorders / K. Zwirska-Korczala, A. Berdowska, J. Jochem et al. // J. Clin. Pharm.Therapeut. - 2004. - Vol. 29, N 2. - R. 151-156), the latter can neutralize the excess concentration of IL-2 and thereby reduce the level of autoimmune inflammation in the thyroid gland. It is known that IL-2 and transferrin, are powerful enough proliferative potential (Zinkernagel R. fundamentals of immunology: TRANS. with it. - M.: Mir, 2008. - 135 C.). At the same time, given that transferrin supports cell proliferation, and blockade of receptors for transferrin in transparentbikini cells induces in them apoptosis, reduction in the blood concentration of CD71+ cells can be considered as a compensatory mechanism in the regulation of proliferative process (Neckers L. Geldanamycin as a potential anti-cancer agent: its molecular target and biochemical activity/ L.Neckers, T.W.Schulte, E.Mimnaugh // Investigational New Drugs. - 1999. - V.17, No. 4. - S-373).

Examples of use of the method.

1. Patient P., aged 51 years. Diagnosis: autoimmune thyroiditis, confirmed high levels of antibodies to thyroperoxidase 56 IU/ml The patient received treatment with L-thyroxine in a dose of 100 mcg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes CD71+ 0,24×109CL/l, the level of IL-2 were 286 PG/ml Made a forecast of unfavorable course of the disease, which is confirmed by the further increase in the concentration of antibodies to thyroperoxidase.

2. Patient W., age 54 years. Diagnosis: autoimmune thyroiditis, confirmed high levels of antibodies to thyroperoxidase 176 IU/ml the Patient is assigned to treatment with L-thyroxine in a dose of 75 mcg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes CD71+ 0,22×109CL/l, the level of IL-2 was 89 PG/ml Made a forecast of unfavorable course of the disease, which is confirmed by the further increase in the concentration of antibodies to thyroperoxidase.

3. Patient B., aged 54 years. The diagnosis of autoimmune thyroiditis with a characteristic increase in the level of antibodies to thyroperoxidase 186 IU/ml the Patient is assigned to treatment with L-thyroxine in a dose of 50 mg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes with the transferrin receptor (CD71+) of 0.62×109CL/l, the level of IL-2 was 17.3 PG/ml the forecast has been Made favorable for the it, that is confirmed by further reducing the concentration of antibodies to thyroperoxidase.

A method for predicting the clinical course of autoimmune thyroiditis, characterized in that the peripheral venous blood of patients with autoimmune thyroiditis determine the absolute content of T-lymphocytes with the transferrin receptor (CD71+) and the concentration of interleukin-2 (IL-2); when the content of lymphocytes CD71+ more than 0.3×109CL/l and the concentration of IL-2 less than 25 PG/ml predicts favorable course of the disease, and when the content of lymphocytes CD71+ less than 0.3×109CL/l and the concentration of IL-2 for more than 25 PG/ml predict an unfavorable course of autoimmune thyroiditis.



 

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