Ivanka I. Kostadinova

Clinical and laboratory study of pro-inflammatory and anti-inflammatory cytokines in women with multiple sclerosis

Abstract Multiple sclerosis (MS) is an autoimmune demyelinating disorder of the central nervous system characterised with a complex system of interactions between proinflammatory and anti-inflammatory cytokines in its course. Aim: The aim of the present study was to investigate the serum levels of cytokines TNF-α, IFN- γ, IL- 4 and IL- 10 in female patients with MS and healthy individuals, the changes occurring in the relapse and remission phases of the disease and their correlation with the severity of the neurological deficit. Patients and methods: Thirty-five women with relapsing-remitting MS were examined. The patients’ age ranged between 18 and 50 years and MS was verified clinically and by magnetic resonance imaging according to the McDonald criteria. Thirteen of the patients were treated with interferon-β-1b. The serum concentrations of TNF-α, IFN- γ, IL- 4 and IL- 10 were determined twice - in relapse and in remission - using an enzyme-linked immunosorbent assay (EL ISA). The control group consisted of 35 age-matched healthy females. Results: The comparison of cytokine serum concentrations during the two phases of the disease showed significant elevation of the TNF-α serum levels in the relapse phase and of IL- 4 - in the remission phase. The comparison between the patients and the healthy control subjects demonstrated statistically significant lower concentrations of TNF-α in remission patients and higher concentrations of IL- 10 in relapse patients. The patients with interferon-β-1b treatment showed different profile of cytokine secretion from the patients without interferon-β-1b treatment. Interferon-β-1b-treated patients showed significantly lower serum levels of TNF-α and IFN- γ during the relapse phase and higher TNF-α and IL- 10 serum levels during the remission phase compared with the untreated patients. Conclusions: Serum levels of TNF-α and IL- 4 objectively reflect the immune response during relapse and remission of the disease. The severity of neurological deficit as estimated with the expanded disability status scale (EDSS ) does not depend on the serum levels of TNF-α, IL- 10 and IFN- γ in the two phases of MS. Резюме Введение: Множественный склероз (МС) представ- ляет собой аутоиммунное, демиелинизирующее за- болевание центральной нервной системы в ходе которого развивается сложная сеть взаимодействий между проинфляматорными и антиинфляматорными цитокинами. Цель: Исследовать сывороточные уровни TNF-α, IFN-γ, IL-4 и IL-10 у женщин с МС и у здоровых лиц, а также и изменения, наступающие во время приступа и во время ремиссии заболевания и их связь с тяжестью неврологического дефицита. Материал и методы: Обследовано 35 женщин в возрасте от 18 до 50 лет с приступно-ремитент- ным, клиническим и резонансно подтвержденным МС по критериям Мс Donald. 13 женщин лечены интерфероном β-1b. Сывороточные концентрации TNF-α, IFN-γ, IL-4 и IL-10 определены двукратно - во время приступа и во время ремиссии с помощью энзимосвязанного иммуносорбентного теста (ELISA). Контрольная группа включает 35 здоровых женщин того же возрастного интервала. Результаты: При сравнении сывороточных концен- траций цитокинов во время обеих фаз заболевания устанавливаются сигнификантно более высокие уровни TNF-α во время приступа и IL-4 - во время ремиссии. При сопоставлении результатов пациенток и здоровых женщин регистрированы статистически значимо более ниская концентрация TNF-α у больных во время ремиссии и более высокая концентрация IL-10 во время приступа. У пациенток с и без интерферона β-1b наблюдается различный профиль секреции цитокинов. Пациентки, леченные препара- том интерферон β-1b, имеют сигнификантно более ниские TNF-α и IFN-γ во время приступа и более высокие TNF-α и IL-10 в состоянии ремиссии по сравнению с нелеченными. Выводы: Сывороточные уровни TNF-α и IL-4 объ- ективно отражают активность иммунной реакции во время двух периодов клинических проявлений МС (приступ и ремиссия). Тяжесть неврологического дефицита, оцененная с помощью EDSS, не зависит от сывороточных уровней TNF-α, IL-10 и IFN-γ во время двух периодов клинических проявлений.

Experimental study on the role of 5-HT2 serotonin receptors in the mechanism of anti-inflammatory and antihyperalgesic action of antidepressant fluoxetine.

ABSTRACT INTRODUCTION: Fluoxetine is an antidepressant that has anti-inflammatory and antihyperalgesic effects in experimental models of pain and inflammation. The AIM of the present study was to determine the role of 5-HT2 receptors in the mechanism of anti-inflammatory and antihyperalgesic action of fluoxetine after single and repeated administration of the drug. MATERIALS AND METHODS: 40 male Wistar rats were randomly divided in five groups (n = 8) treated for 14 days with saline (control), diclofenac (positive control), fluoxetine, cyproheptadine (5-HT2 antagonist), and fluoxetine + cyproheptadine, respectively. We used the experimental model of inflammation induced by intraplantar injection of carrageenan and nociceptive test with mechanical pressure on the inflamed hind paw. RESULTS: Single and repeated administration of fluoxetine showed that it had significant anti-inflammatory and antihyperalgesic effects when compared with the control (p < 0.05). Cyproheptadine did not change significantly the anti-inflammatory effect of fluoxetine in the first 4 hours, after a single administration. At 24 hours the combination did not differ statistically when compared with the control. Cyproheptadin did not change significantly the anti-inflammatory effect of fluoxetine after repeated administration. After prolonged treatment the group that received fluoxetine + cyproheptadine showed a statistically significant increase in paw pressure to withdraw the hind paw compared with that treated with fluoxetine alone (p < 0.05). CONCLUSIONS: Fluoxetine has anti-inflammatory and antihyperalgesic effects in the carrageenan model of inflammation. 5-HT2 receptor mediated its anti-inflammatory effect in single dose treated animals. Spinal 5-HT2 receptors are involved in the antihyperalgesic effect of fluoxetine after repeated administration РЕЗЮМЕ ВВЕДЕНИЕ: Флуоксетин представляет собой антидепресант, обладающий противовоспалительным и антигиперальгезическим эффектами при экспериментальных моделях боли и воспаления. ЦЕЛ: Настоящее исследование ставит себе целью установить роль 5-НТ2 рецепторов в механизме противовоспалительного и антигиперальгезического действий флуоксетина при однократном и многократном применениях. МАТЕРИАЛ И МЕТОДЫ: На 40 мужских крыс породы „Вистар”, разделенных на 5 групп (n = 8), в течение 14 дней воздействовали физиологическим раствором (контрольная группа); диклофенаком (позитивная контрольная группа); флуоксетином, ципрохептадином (5-НТ2 антагонист) и флуоксетином + ципрохептадином. Использованы модель воспаления (интрапланетарноe введениe карагенина) и ноцицептивный тест (механическоe давлениe на воспаленную лапку). РЕЗУЛЬТАТЫ: При однократном и многократном применениях флуоксетина наблюдались сигнификантный противовоспалительныйи антигиперальгезический эффекты - сравнение с контрольной группой (р < 0.05). При однократном воздействии ципрохептадин достоверно не изменяет протививовоспалительный эффект флуоксетина в первые четыре часа, но на 24-ый час комбинация не показывает статистическую разницу при сравнении с контрольной группой. При многократном воздействии ципрохептадин достоверно не изменяет противовоспалительный эффект флуоксетина. Группа крыс, подвергнутая продолжительному воздействию флуоксетином и ципрохептадином, показала сигнификантное увеличение силы давления при отдергивании лапки по сравнению с группой, получившей только флуоксетин (р < 0.05). ВЫВОДЫ: Флуоксетин обладает противовоспалительным и антигиперальгезическим эффектами при карагениновой модели воспаления. 5-НТ2 рецепторы медиируют его противовоспалительный эффект при однократном воздействии. Спинальные 5- НТ2 рецепторы участвуют в реализации антигиперальгезического эффекта флуоксетина при его продолжительном применении

Female sex hormones and cytokine secretion in women with multiple sclerosis

Abstract Data from experimental and clinical research suggest that sex hormones may influence the autoimmune process in multiple sclerosis (MS). Studies on the hormonal profile of patients with MS and its relation to the disease activity provide heterogeneous results. Objectives: The aim of this study is to investigate the changes in serum levels of estradiol and progesterone and their correlations with the cytokine profile and the degree of disability in women with relapsing-remitting MS (RRMS). Methods: The serum concentrations of estradiol, progesterone, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukine-4 (IL-4) and interleukine-10 (IL-10) were measured and the degree of disability was determined in 35 women with RRMS, during relapse and remission. Serum levels of hormones were measured by micro-particle enzyme immunoassay and ELISA was used for the cytokines concentrations. The degree of disability was assessed by the Expanded Disability Status Scale and the Scripps Neurological Rating Scale. Results: Sixty per cent of patients had serum concentrations of estradiol and/or progesterone below the lower limit of normal in one or both phases of MS. Hormonal levels increased significantly during remission in these patients. Women with and without hormonal abnormalities differed in terms of cytokine profile during relapse and remission. Significantly higher TNF-alpha in both phases and IFN-gamma in remission was found for the patients with hormonal disturbances compared to these with normal hormonal status. Conclusions: Our study finds high frequency of hormonal disturbances among female patients with RRMS. Abnormally low concentrations of sex hormones are associated with higher serum levels of TNF-alpha and IFN-gamma, which could suggest suppressive effect of estradiol and progesterone on pro-inflammatory cytokine secretion.

EFFECT OF TESTOSTERONE PROPIONATE ON ERYTHROPOIESIS AFTER EXPERIMENTAL ORCHIECTOMY

ABSTRACT INTRODUCTION: Androgen deficiency anemia occurs most frequently in pharmacogenic suppression of androgen synthesis or with advancing age in men. Bilateral orchiectomy is a surgical modality used in the treatment of metastatic prostate carcinoma. It is accompanied by marked decrease in circulating serum levels of androgens. AIM: The aim of the experimental study was to determine the effect of substitution therapy with testosterone propionate (TP) on some haematological parameters of erythropoiesis in male rats after orchiectomy. MATERIAL AND METHODS: Eighty Wistar male rats with mean weight of 252.3 g were used in the study. The animals were allocated into 2 control orchidectomized groups, 2 shamoperated groups and 4 experimental orchidectomized groups. Testosterone propionate was administered intramuscularly, once a week at a dose of 4 mg and 8 mg per kilogram of body weight for 15 days and for 15 weeks. Erythrocyte count was performed and hemoglobin and hematocrit levels were measured. RESULTS: In the chronic experiment there was a significant decrease in red blood cells and hemoglobin, and a tendency of decrease in hematocrit after orchiectomy. The effect of TP on erythropoiesis in orchiectomised rats is dose-dependent. CONCLUSION: TP replacement therapy in doses of 4 mg/kg and 8 mg/kg has a stimulating effect on erythropoiesis only in chronic administration. РЕЗЮМЕ ВВЕДЕНИЕ: Андрон-дефицитная анемия наблюдается чаще всего при фармакогенном подавлении андроген- синтеза или с возрастом у мужчин. Билатеральная орхиэктомия представляет метод, применяемый в терапии метастатической карциномы предста- тельной железы, сопровождающейся выраженным понижением циркулирующих сывороточных уровней андрогенов. ЦЕЛЬ: Установить влияние заместительной терапии тестостерон - пропионатом - ТП- на некоторые гематологические показатели эритро- поэза у мужских крыс после орхиэктомии. МАТЕРИАЛ И МЕТОД: Использовано 80 мужских крыс породы Wistar, средняя масса тела - 252.3 гр. Животные разделены следующим образом: 2 контрольные орхиэктомированные, 2 симулятивно оперированные и 4 опытные орхиэтомированные группы. ТП применен мышечно раз в неделю в дозе 4 и 8 мг/кг массы тела; период применения - 15 дн. и 15 нед. Прослежены: число эритроцитов, уровень гемоглобина и гематокрита. РЕЗУЛЬТАТЫ: При хроническом опыте наблюдается сигнификантное снижение эритроцитов и гемогло- бина и тенденция к снижению гематокрита после орхиэктомии. Эффект ТП на эритропоэз у крыс с орхиэктомией дозозависима. ЗАКЛЮЧЕНИЕ: Заместительная терапия с помо- щью ТП в дозах 4 и 8 мг/кг оказывает стимули- рующий эффект на эритропоэз только в случаях продолжительного применения.

Effect of atorvastatin and rosuvastatin on learning and memory in rats with diazepam-induced amnesia.

ABSTRACT During the past decade, evidence has emerged that statins have neuroprotective effects. AIM: The aim of this study was to investigate the effects of atorvastatin and rosuvastatin on learning and memory in rats with diazepam-induced amnesia. MATERIAL AND METHODS: Experiments were carried out on 48 white male Wistar rats, divided into 6 groups, each of 8 rats. The experimental animals were treated per os for 14 days with atorvastatin and rosuvastatin in doses of 10 mg/kg and 20 mg/kg body weight, respectively. To induce amnesia diazepam was administered intraperitoneally in a dose of 2.5 mg/kg bw. Cognitive skills of the animals were examined after the induction of amnesia with active avoidance test using autonomic reflex conditioner (shuttle box) and passive avoidance tests (step-through and step down) (Ugo Basile, Italy). The following parameters were assessed: number of conditioned responses (avoidances), number of unconditioned responses (escapes) and number of intertrial crossings in the active avoidance test; latency of reactions was measured in the passive avoidance tests. RESULTS: We found a significant increase of conditioned responses in atorvastatin treated animals (in a dose of 10 mg/kg bw) in active avoidance training. In the animals treated with rosuvastatin in both doses there was a statistically significant increase of unconditioned responses. In the step-through passive avoidance test there was significant improvement of short-term and long-term memory following administration of atorvastatin (10 mg/kg bw). Rosuvastatin (10 mg/kg bw) preserves long-term memory. In the step-down passive avoidance test, atorvastatin (10 mg/kg bw) and rosuvastatin (10 mg/kg bw and 20 mg/kg bw) preserve long-term memory. CONCLUSIONS: Atorvastatin (10 mg/kg bw) and rosuvastatin (10 mg/kg and 20 mg/kg bw) improve cognitive functions in rats with diazepam-induced amnesia and preserve longterm memory. РЕЗЮМЕ ВВЕДЕНИЕ: В последнее десятилетие появились данные о нейропротективном действии статинов. ЦЕЛЬ: Изучить влияние Atorvastatin-а и Rosuvastatin- а на процессы обучения и памяти у крыс с Diazepam-индуцированной амнезией. МАТЕРИАЛ И МЕТОДЫ: Опыты поставлены на 46 белых мужских крысах породы Wistar, разделенных на 6 групп по 8 крысам. Подопытные животные в течение 14 дней перорально получали atorvastatin и rosuvastatin в дозах 10 mg/kg b.w и 20 mg/kg b.w. С целью причинить амнезию диазепам вводят интра- перитонеально (i.p.) в дозе 2,5 mg/kg b.w. Иссле- дованы когнитивные способности животных после индуцирования амнезии с помощью теста активного обучения посредством autonomic reflex conditioner (shuttle box) и пассивного обучения посредством passive avoidance controllers (step-through и step down) (Ugo Basile, Italy). Установлены следующие показатели: число условных ответов (avoidance), число безусловных ответов (escapes) и межтрени- ровочные переходы при тесте активного обучения; латентный период при тестах пассивного обучения. РЕЗУЛЬТАТЫ: При активном обучении наблюдается сигнификантное увеличение условных ответов у животных, получавших atorvastatin 10 mg/kg b.w. У животных, получавших rosuvastatin и при обеих дозах статистически значимо увеличиваются без- условные ответы. При тесте пассивного обучения step-trough step-trough регестрируется сигнификант- ное улучшение кратковременной и долговременной памяти, после приема atorvastatin 10 mg/kg b.w. Rosuvastatin 10 mg/kg b.w сохраняется долговре- менная память. При тесте пассивного обучения step-down atorvastatin 10 mg/kg b.w и rosuvastatin 10 mg/kg b.w и 20 mg/kg b.w сохраняется долгов- ременная память. ВЫВОДЫ: Применение atorvastatin-а 10 mg/kg b.w и rosuvastatin-а 10 и 20 mg/kg b.w улучшает когни- тивные функции у крыс с Diazepam-индуцированной амнезией и сохраняет долговременную память.

Vitamin D immunomodulatory potential in multiple sclerosis.

ABSTRACT Multiple sclerosis (MS) is an autoimmune disease of unknown etiology whose treatment is of limited efficiency and therefore has a high social burden. As it has been suggested that myelin destruction model, the clinical manifestation and the potential of therapeutic response in MS are correlated, it is quite justifiable that we study various factors (genetic, hormonal, environmental) that take part in the autoimmune process in order to improve the control over the disrupted immune regulation. Results from epidemiological and clinical studies clearly suggest that changes in vitamin D serum concentrations are correlated with the magnitude of the risk of developing MS, the phases of relapse and remittance and with gender differences in vitamin D metabolism. Experimental and clinical studies also have established that 25-hydroxy vitamin D (25(OH)D) and 1,25-dihydroxy vitamin D (1,25(OH)2D) exert an immunomodulatory effect in the central nervous system and peripheral organs of the immune system. The standard reference range of vitamin D concentration in serum is 50-80 nmol/l - it provides normal calcium metabolism. Issues that are discussed include the vitamin D serum concentration needed to suppress the aberrant immune response in MS patients; a subgroup of MS patients suitable for vitamin D treatment, the vitamin D being applied in optimally effective and safe dosage. MS prevalence rate in Bulgaria has increased two-fold in 17 years but this is a rather short interval to be able to assume that the gene pool of the population changes. Thus further studies on possible interactions between different environmental factors and these factors’ role in the disease pathogenesis are justified and necessary. РЕЗЮМЕ Множественный склероз - МС - представляет собой значимое иммунообуславленное заболевание неизвест- ной этиологии, лечение которого с ограниченной эффективностью. Концепция зависимости между моделью деструкции миелина, клиническими прояв- лениями и потенциалом терапевтического ответа является основанием изучить участие различных факторов в аутоиммунном процессе - генетических, гормональных, факторов окружающей среды - с целью улучшения контроля над расстроенной им- мунной регуляцией. Результаты эпидемиологических и клинических наблюдений показывают связь изме- нений сывороточных концентраций витамина Д со степенью риска развития МС, с фазами приступа и ремиссии, с половыми различиями в метаболизме витамина Д. Иммуномодулирующая активность 25- hydroxy vitamin D (25(OH)D) и 1,25-dihydroxy vitamin D (1,25(OH)2D) в центральной нервной системе и в периферических органах иммунной системы устанавливается при экспериментальных и клинических наблюдениях. Принятый стандарт референтных границ витамина Д в сыворотке 50-80 nmol/l обеспечивает нормальный кальциевый метаболизм. Дискуссионными остаются вопросы о: сывороточной концентрации витамина Д, необходи- мой для подавления аберантного иммунного ответа у пациентов с МС; субгруппе МС пациентов, под- ходящих для лечения витамином Д, примененным в оптимально эффективной и безопасной дозе. В нашей стране заболеваемость МС повыси- лась двукратно за последние 17 лет - короткий интервал, чтобы обсуждать вопрос: возможны ли изменения в генном фоне населения. С такой точки зрения исследования взаимодействия между факто- рами окружающей среды и их роли в патогенезе болезни обоснованы и необходимы.

Study on anti-inflammatory and immunomodulatory effects of fluoxetine in rat models of inflammation

The aim of the present study was to evaluate the anti-inflammatory effect of fluoxetine in carrageenan- and lipoplysaccharide-induced models of inflammation by investigating the changes in serum levels of pro-inflammatory cytokine TNF-α and anti-inflammatory cytokines IL-10 and TGF-β after single and repeated administration of the drug. To study the effect of a single and repeated dose fluoxetine on carrageenan-induced paw edema male Wistar rats were divided into five groups (n = 8): control group; positive control group; and three experimental groups treated with 5, 10, and 20 mg/kg bodyweight (bw) fluoxetine, respectively. To study the effect of a single and repeated dose of fluoxetine on serum cytokine levels, the animals were divided in four groups (n = 8): two control groups treated with saline and two experimental groups treated with fluoxetine 20 mg/kg bw. Carrageenan and LPS were injected immediately after fluoxetine or saline injection. Serum cytokine concentrations were tested by enzyme immunoassay. In single administration only the highest dose used inhibited carrageenan-induced inflammation. Edema inhibition was seen with 10 and 20 mg/kg bw fluoxetine after repeated administration. At 24 h a statistically significant effect on inhibition of carrageenan edema was found only in rats treated with 20 mg/kg bw fluoxetine In carrageenan-induced inflammation, fluoxetine significantly increased Il-10 and decreased TNF-α after repeated administration. Surprisingly, in single-dose treated animals an increase in TNF-α values upon fluoxetine administration was observed in this model of inflammation. In LPS-induced inflammation, fluoxetine significantly decreased TNF-α after single and repeated treatment. Fluoxetine has anti-inflammatory and immunomodulatory effect in the carrageenan-induced model of exudative inflammation. In LPS-induced inflammation it showed an immunomodulatory effect manifested with a decrease in pro-inflammatory cytokine TNF-α.

Antidepressant Effect and Recognition Memory Improvement of Two Novel Plant Extract Combinations - Antistress I and Anti-stress II on Rats Subjected to a Model of Mild Chronic Stress

Abstract Background: Chronic stress is one of the main factors which lead to depression – a psychiatric disorder affecting millions of people and predicted to be the second ranked cause of premature death in 2020. Depression is often associated with cognitive disturbances and memory deficit. Plant based therapy could be effective in the treatment of mild to moderate depression due to its low level of adverse reaction, its good tolerability and compliance. Materials and methods: 72 male Wistar rats, divided in 9 groups were given orally for 8 weeks two combinations of dry plant extracts – Antistress I and Antistress II and five individual dry extracts obtained from Serratula coronata, Hypericum perforatum, Valeriana officinalis, Crataegus monogyna and Melissa officinalis. The animals were exposed to a chronic unpredictable mild stress for 8 weeks. The depression-like symptoms were evaluated with Forced swim test while the assessment of the memory deficit was performed with Novel object recognition test. Results: Antistress II demonstrates antidepressant effect while Antistress I doesn’t improve the depressive-like symptoms. The individual extracts of Hypericum perforatum and Valeriana officinalis also possess antidepressant properties. Antistress II improves the cognition as well as the individual extracts of Hypericum perforatum, Valeriana officinalis and especially Serratula coronata. Dry extract from Serratula tend to have the best effect regarding the recognition memory. The effect of Antistress I on memory deficit is negligible. Conclusions: Antistress II possesses antidepressant effect and improves the recognition memory while Antistress I doesn’t demonstrate any of the above-described effects.

Vitamin D and environmental factors in multiple sclerosis

Multiple sclerosis (MS) is an autoimmune disease of unknown etiology whose treatment is of limited efficiency and therefore has a high social burden. As it has been suggested that myelin destruction model, the clinical manifestation and the potential of therapeutic response in MS are correlated, it is quite justifiable that we study various factors (genetic, hormonal, environmental) that take part in the autoimmune process in order to improve the control over the disrupted immune regulation. Results from epidemiological and clini¬cal studies clearly suggest that changes in vitamin D serum concentrations are correlated with the magnitude of the risk of developing MS, the phases of relapse and remittance and with gender differences in vitamin D metabolism. Experimental and clinical studies also have established that 25-hydroxy vitamin D (25(OH)D) and 1,25-dihydroxy vitamin D (1,25(OH)2D) exert an immunomodulatory effect in the central nervous system and peripheral organs of the immune system. The standard reference range of vitamin D con¬centration in serum is 50-80 nmol/l it provides normal calcium metabolism. Issues that are discussed include the vitamin D serum concentration needed to suppress the aberrant immune response in MS patients; a subgroup of MS patients suitable for vitamin D treat¬ment, the vitamin D being applied in optimally effective and safe dosage. MS prevalence rate in Bulgaria has increased two-fold in 17 years but this is a rather short interval to be able to assume that the gene pool of the population changes. Thus further studies on pos¬sible interactions between different environmental factors and these factors’ role in the disease pathogenesis are justified and necessary.

25 Hydroxyvitamin D and Cytokines in Multiple Sclerosis.

Abstract INTRODUCTION: Clinical trials of patients with multiple sclerosis (MS) have produced inconsistent results for the profile of cytokine secretion in serum and cerebrospinal fluid in patients with multiple sclerosis during periods of relapse and remission. Epidemiological and clinical observations data reveal an association of the changes in vitamin D serum concentration with the risk of developing MS. AIM: To evaluate changes in serum concentrations of 25(OH)D, IL17, IFN-gamma, TGFβ1, IL4, IL10 in relapse and remission and their correlation with the severity of disability. PATIENTS AND METHODS: Fifty-three persons (30 clinically healthy controls and 23 patients with relapsing-remitting multiple sclerosis) living between 41° and 42° northern latitude were registered during the astronomical winter period (October 2012- May 2013). -Patients were diagnosed according to Mc Donald 2010 criteria. The degree of neurological deficit was assessed by EDSS. Serum concentrations of 25(OH)D (nmol/l) and cytokines (pg/ml) were tested by ELISA - once for controls and twice for patients (during relapse and remission). RESULTS: In the studied population average levels of 25(OH)D were close to insufficiency, most pronounced in patients in relapse, as differences were not statistically significant. A reverse correlation was found between the levels of 25(OH)D and the deficit in relapse and remission. Concentrations of TGFβ1 significantly increased in remission compared with exacerbation and controls. Serum level of IL4 was significantly lower in relapse compared with controls. In remission there was a marked tendency of increase compared with exacerbation. During clinical improvement IL17 and IFN-gamma tended to decrease compared to the average levels in relapse. In both periods, the average concentrations of IFN-gamma in patients were significantly lower compared with controls. No statistically significant differences were found comparing cytokine changes with those of 25(OH)D and deficit. CONCLUSION: Persistent cytokine imbalance in patients compared with controls is a marker for Th1-mediated CNS demyelination. Anti-inflammatory TGFβ1, IL4 are indicators of immune response intensity. The deficit severity does not depend on changes of the tested cytokines, but correlates with 25(OH)D levels during periods of relapse and remission.