Резюме
Зєльоний І.І. Вплив лікопіду на продукцію інтерферону мононуклеарами периферійної крові хворих на рецидивуючу бешиху.
Вивчений інтерфероновий статус (ІФС) хворих з рецидивуючою бешихою (РБ). Встановлено, що у хворих на РБ мало місце пригнічення спонтанної та індукованої продукції  α- та γ-інтерферону мононуклеарами периферійної крові хворих  на РБ. При проведенні імунореабілітації з використанням імуноактивного засобу  лікопіду спостерігається нормалізація показників ІФС.
Ключові слова: рецидивуюча бешиха, інтерфероновий статус, продукція, лікопід, імунореабілітація.

Резюме
Зеленый И.И. Влияние ликопида на продукцию интерферона мононуклеарами переферической крови больных рецидивирующей рожей.
Изучен интерфероновый статус (ИФС) больных рецидивирующей рожей (РР). Установлено, что у больных РР наблюдалось угнетение спонтанной и индуцированной продукции α- и γ-интерферона мононуклеарами периферической крови больных  РР. При проведении иммунореабилитации с использованием иммуноактивного препарата ликопида наблюдается нормализация показателей ИФС.
Ключевые слова: рецидивирующая рожа, интерфероновый статус, продукция, ликопид, иммунореабилитация.

Summary
Zelenyi I.I. Influence of licopid at interferon production by mononuclears of peripheral blood in patients with recurrent erysipelas.
 Interferon status (IFS) of the patients with recurrent erysipelas (RE) was studied. It was set that at inspected patients with RE have decrease  spontaneous and induction production of α- and γ-interferon peripheral blood mononuclears in patients with RE. The immunorehabilitation by immunoactive preparation licopid provide IFS normalization.
 Key words: recurrent erysipelas, interferon status, production, licopid, immunorehabilitation. 

УДК 616.5.22-002:579.862.1

ДЗ «Луганський державний медичний університет»

ГЗ «Луганский государственный медицинский университет»

91045, кв. 50-летия Обороны Луганска, 1 г, Луганск, Украина

State Establishment "Lugansk State Medical University"

91045,50 Rokyv Oborony Luganska Block, 1G, Lugansk,Ukraine

igor_zelenyi@mail.ru

Introduction. According to modern statistical studies it is known, that nowadays the incidence of infections of the streptococcal etymology, including erysipelas, in Ukraine and other CIS countries is extremely high [2,5]. According to scientific literature the rate of recurrent among all officially recorded cases of erysipelas is from 32% to 48% [3]. Clinico-epidemiological, medical and statistical analysis showed that, despite the widespread appliance of modern antibiotics for the treatment of this disease, the number of recurrent forms of erysipelas has no tendency towards reduction [1, 13]. Immunopathological features of recurrent erysipelas are still almost unstudied, that’s why there haven’t been developed any rational preventive and immunocorrective approaches [5]. Over the past few years the attention of researchers and practitioners has been attracted by the appliance of immunoactive drugs in order to improve the immune status of  the patients, suffering from erysipelas and as a result, to reduce the chance of erysipelas’ recurrent in the future [6].

In this context, our attention was attracted by the possibility of applying a new immunoactive drug of natural origin – likopid [7]. Active substance of the drug is glucoseminylmuramildipeptidum, a biologically active substance, which stimulates functional activity of phagocytes (neutrophils, macrophages), enhances proliferation of T-and B-lymphocytes, and increases synthesis of specific antibodies. Pharmacological action is ensured by the enhancing of the production of interleukins (IL-1, IL-6, IL-12), TNFα, gamma interferon, colony stimulating factors. It also increases the activity of natural killer cells [7]. Likopid indications are the following: pathological conditions involving secondary immunodeficiency, such as pyoinflammatory diseases of the skin and soft tissues, including purulent-septic postoperative complications, herpetic infections of any location, chronic viral hepatitis B and C [7].

The connection of the article with academic programs, plans, themes. The article is written according to the basic plan of scientific works of Lugansk State Medical University. This article is a fragment of the topic: " The immunopathogenesis of complicated and recurrent forms of erysipelas, immuno correction and immunorehabilitation» state registration № 0110U002396).

The aim of the article was to study the influence of likopid on the production of interferon by the mononuclear cells of peripheral blood of the patients, suffering from recurrent erysipelas.

Materials and methods. During the experiment two groups of the patients, aged from 35 to 59 years, were examined. All the patients were divided into two groups: the basic group (48patients) and the group of comparison (44 patients). Both groups were randomized according to the patient’s age, sex, severity of disease, localization of local inflammation of the skin, the frequency and total number of previous recurrences of erysipelas. The patients in both groups during acuity received conventional therapy, which included antibacterial, antihistamines, anti-inflammatory drugs (amizon or mefenamic acid) ascorbic acid or askorutin. If it was necessary for detoxification, salt solutions or single dose of Reamberin were prescribed. At the end of treatment of the patients, suffering from erysipelas, in case of the necessity of medical rehabilitation (MR) and in case of the signs of immunodeficiency state [4], the preventive measures with benzathine benzylpenicillin, according to the recommendations were held [1,11]. Moreover, patients from the basic group during MR additionally received immunoactive drug Likopid 10mg. per os. (1 pill once a day on an empty stomach 30 minutes before meal, during 10 days). The prescription of Likopid was based on its previous testing in vitro with the help of E-PYK method and adheres test to explore the sensitivity of immune competent cells of the patient’s blood - general populations E-PYK (T-lymphocytes) and A-cells, i.e., cells of the mononuclear- monocytic kind [4].

All the patients, suffering from erysipelas underwent clinico-laboratorial examination. In order to determine the ability of mononuclear cells to produce interferon, after the separation in the ordinary ficoll-verografin density gradient, they were washed in 199 medium and resuspended in RPMI-1640 medium. In order to get supernatants with the spontaneous production of INF mononuclear cells in an amount of 1.5 x 106 cells / ml, the cells were incubated without adding any evocators for 24 h in a CO2 incubator at 37 ° C; to get supernatants with the induced activity, the incubation was carried out with mitogen (PHA) to assess the secretion in full accordance with the recommendations of professor G. M.  Drannik and prof. V. E. Driyanskoya [10, 12]. After the incubation, the cells were pelleted and then supernatants were collected, freezed and kept for further testing at -20 º C. The studying of immunological parameters was performed twice: before treatment,including immunorehabilitation with Likopid and at the of the therapy. The results were compared with the results, obtained after the examination of 32 healthy individuals. Statistical analysis of the results was carried out using PC AMD Athlon 3600 + with the help of multivariate dispersive analysis of the licensed software (Microsoft Windows professional, Microsoft Office 2007 Stadia 6.1/prof. and statistic),  basic principles of the appliance of statistical methods during the clinical testing of medicines were also taken into account [8, 9].

Results and discussion.

Clinical study before medical rehabilitation showed that all the patients had some abnormalities, concerning interferon status (table 1). As it can be seen from the 1^st table, before medical rehabilitation, the spontaneous production of IFN α-mononuclear cells in the basic group was on average (12,1 ± 0,2) pg / ml, which was in 1.53 times lower than normal state (P <0.01); in the group of comparison , the spontaneous production of IFN α-mononuclear cells was (12,5 ± 0,22) pg / ml, which was in 1.48 times lower than normal state (P <0.01). The production of IFN α-mononuclear cells induced by mitogen in the basic group was (25,1 ± 0,8) pg / ml, which was in 2.13 times lower than normal state (P < 0.001 ); in the group of comparison - ​​(25,4 ± 0,9) pg / ml, which was in 2.1 times lower than normal state (P < 0.001). The spontaneous production of γ-IFN in the basic group was (13,7 ± 0,5) pg / ml, which was in 1.9 times lower than normal state (P < 0.001 ); in the group of comparison - (14,3 ± 0,4) pg / ml, which was in 1.83 times lower than normal rate (P < 0.001). Induced production of γ-IFN was on average (28,7 ± 2,2) pg / ml, which was in 2.4 times lower than normal state (P < 0.001). In the group of comparison this index was (29,3 ± 2,3) pg / ml, which was in 2.3 times lower than normal state (P <0.001). Thus, as it can be seen from the received data, before treatment there was the inhibition of the spontaneous and induced production of α-IFN and γ-IFN.

Table 1

Note: in the table. 1-2: The probability of the difference with respect to the norm * - at P <0.05,

** - P <0.01, *** - P <0.001; Column P - the probability of the difference between the

basic group and the comparison group.

At the end of the medical rehabilitation the following results were obtained (table 2):

As it can be seen from the 2^nd table, studied indices of the patients from the basic group, who received Likopid in the complex of rehabilitation, at the end of rehabilitation reached the lower limit of normal state (P> 0.05), while the same indices of the patients from the group of comparison, despite positive dynamics, still were significantly below normal state (P < 0.05 -0.01). Thus, during medical rehabilitation the spontaneous production of α-IFN in the basic group was (18, 2 ± 0, 5) pg / ml, which was equal to the normal state (P> 0.05). In the group of comparison the spontaneous production of α-IFN was (14,5 ± 0,3) pg / ml, which was in 1.28 times lower than normal state (P < 0.05 ) and in 1.26 times lower than the same index in the basic group (P < 0.05). Stimulated production of α-IFN in the basic group was (51,6 ± 1,5) pg / ml, reaching the lower limit of the normal state (P> 0.05). At the same time in the group of comparison this index only (35,7 ± 1,6) pg / ml, which was in 1.5 times lower than normal state (P < 0.01) and in1.44 times lower than in the basic group ( P < 0.01). The spontaneous production of γ-IFN in the basic group was (25,8 ± 0,9) pg / ml, with the normal state of (26,2 ± 1,3) pg / ml (P> 0.05). In the group of comparison the spontaneous production of γ- IFN was (16,7 ± 0,9) pg /ml, which was on average in 1.6 times lower than normal state (P < 0.05 ) and in 1.54 times lower than in the basic group (P < 0.05). The induced production of γ-IFN in the basic group increased to (65,4 ± 1,9) pg / ml, which was equal to the normal state (P> 0.05). In the group of comparison this index was (39,7 ± 1,5) pg / ml, which was in 1.7 times lower than normal state (P < 0.01) and in 1.64 times lower than in the basic group ( P < 0.01). Thus, the inclusion of Likopid to the complex of medical rehabilitation of the patients, suffering from erysipelas, can help to renew the capacity of peripheral blood mononuclear cells to produce interferon. The received data shows that the appliance of Likopid in order to optimize the medical rehabilitation of the patients, suffering from erysipelas, is reasonable.

Conclusions.

1. Before medical rehabilitation all patients in the basic group had significant inhibition of the indices of interferon status – spontaneous production of α-IFN was reduced in 1.53 times; induced production of α-IFN - in 2.13 times; spontaneous production of γ-IFN - in 1.9 times and induced production of γ-IFN – in 2.4 times;
2. The inclusion of Likopid promoted the normalization of studied indices;
3. In case of conventional ways of treatment in the group of comparison, despite positive dynamics, there was no complete normalization of studied indices;
4. The received data shows that the inclusion of modern immunoactive drug Likopid into the complex of MR for the immunocorection can be considered pathogenetic reasonable and clinically perspective;
5. Prospects for further researches can be seen in the continuation of investigation of the effectiveness of the drug in the complex of rehabilitation of the patients, suffering from erysipelas , including its potential impact on the cytokine profile.

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