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H.B. Mateiko, I.I. Pyliuk Dynamics of Cytokines (IL-2, IL-4, Interferon-γ) and β2-Microglobulin Content in Cases of Pneumonia in Children Who Often Suffer from Acute Respiratory Disease Ivano-Frankivsk National Medical University, Ivano-Frankivsk, Ukraine Abstract The objective of the research was to study indices of cytokines (IL-2, IL-4, interferon- γ) and β2-microglobulin content in blood serum of children at the age of 3 to 8 who often suffered from acute respiratory diseases. Their dynamics before and after protocol pneumonia treatment was analyzed. Materials and methods of research. 80 children with pneumonia at the age of 3 to 8 were examined. They included 40 children who often suffered from acute respiratory diseases (the number of cases during the year was 6 and more) and 40 children who were sick not very frequently. Children of both groups received protocol therapy for pneumonia. Results of the research. More significant changes in the correlation of pro- and anti-inflammatory cytokines by reducing the levels of IL-2 and interferon-γ on the background of increased IL-4content were observed in children who often suffered from acute respiratory diseases. This indicated switching from a cellular immune response to humoral one. Reduction of β2-microglobulin as a marker of immunosuppression was observed. Conclusions. Protocol treatment for pneumonia in children who often suffered from acute respiratory infections contributed to normalization of above mentioned cytokines and β2-microglobulin indices indicating adequate immune response to inflammation. Keywords: children who often suffer from acute respiratory infections; pneumonia; treatment. Problem statement and analysis of the recent research The problem bronchopulmonary pathology continues to be the focus of pediatric science and practice and recently has ranked first in the incidence among children. Pneumonia plays the leading role in nosological structure of lower respiratory tract diseases. The urgency of the problem is caused by the high risk of complications, disability and mortality as well as significant economic costs associated with the treatment [2,3,5]. Particular attention should be paid to the development of pneumonia in children who often suffer from acute respiratory diseases (ARD) as ARD in these children may result in immunosuppression, antioxidant defense reduction, chronic pathology not only of respiratory, butother body systems too. On the one hand, frequent ARD help to create child’s immunity, on the other hand, they induce temporary immunosuppression leading to endless circle “infectionimmunosuppression-infection”. Therefore, a direct connection between the seasonal increase in the incidence of ARD and incidence of pneumonia was noted [1, 4, 6]. Lack of immune system spare capacity is found in children who often suffer from ARD. This is the result of long and mass antigenic effect on the body and manifests in the predominance of immune response of T-helper (Th) 2 over Th1 type. However, induced production of proinflammatory cytokines is insufficient for an adequate immune response [7,9]. Such condition of the immune system is a factor in the development of bacterial ARD complications, pneumonia in particular [10]. Major histocompatibility complex (MHC) class I cells are involved in the immune response with β2microglobulin (β2-MG) as a light part of their molecules. β2-microglobulin takes part in the monitoring of immunity control over body antigenic homeostasis which forms the basis of “self-recognition” phenomenon and acts as immunosuppression marker [8]. Materials and methods of the research 80 children with pneumonia at the age of 3 to 8 were examined. Main group consisted of 40 children who often suffered from ARD (the number of cases during the year was 6 and more). Experimental group included 40 children who were sick not very frequently. Children of both groups received protocol therapy for pneumonia (order of the Ministry of Heath of Ukraine №18 from 13.01.2005). Control group consisted of 20 practically healthy children of the same age. With the used of ELISA method the levels of IL-2 using test systems of “Orgenium” (Finland), IL-4, IFN-γ using “Vector Best” (Russia), β2-MG using “Orgentec” (Germany) were determined in all examined patients. Examination was conducted on the 1st -2nd and 12th-14th days of hospital treatment in a centralized laboratory for diagnosis of HIV infection,

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toxoplasmosis, veneral diseases and viral hepatitis at Ivano-Frankovsk Regional HIV/AIDS Prevention and Control Center. Results of the research and their discussion Reduction of IL-2 level compared with that of healthy children was observed in blood serum of children of the main group and the comparison group before the treatment during the study of cytokines content. The indices constituted 189.77±23.77 pg/ml and 330.50±39.25 pg/ml versus 512.15±67.26 pg/ml (р<0.001), (р<0.05). However, the level of IL-2 was 1.7 times lower in the main group of children than in the comparison group, (р<0.01). The normalization of IL-2 level in the main group of children was not observed after treatment, despite its significant growth from 189.77±23.77 to 261.61±70.82 pg/ml, (р<0.001), while in the comparison group it increased to almost the norm values, from 330.50±39.25 to 465.66±74.51 pg/ml, (р>0.1). Increase in the IL-4 level before the treatment was observed during the research in children of the main group and the comparison group compared with that of healthy children. The indices constituted 10.17±0.29 pg/ml and 7.66±0.32 pg/ml versus 3.11±0.29 pg/ml, р<0.001. However, IL-4 level was 1.3 times higher in the main group of children than in the comparison group, p <0.001. After treatment IL-4 content index significantly decreased in children of the main group but remained above the norm level (from 10.17±0.29 to 7.21±0.69 versus 3.11±0.29 pg/ml, р<0.001), while it normalized in the comparison group from 7.66±0.32 to 5.19±0.51 versus 3.11±0.29 pg/ml, (р>0.1). IL-2, IL-4, IFN-γ and β2-MG concentration in serum of children from the studied groups who suffered from pneumonia is shown in the table 1. Table 1 IL-2, IL-4, IFN-γ and β2-MG concentration in serum of children from the studied groups who suffered from pneumonia, (М±m) Indices Control group Main group (n=40) Comparison group(n=40) (n=20) before the after the before the after the treatment treatment treatment treatment IL-2, 512.15± 189.77± 261.61± 330.50± 465.66± pg/ml 67.26 23.77* 70.82 *, º 39.25*,º 74.51º, ⌂ IL -4, 3.11±0.29 10.17±0.29* 7.21± 7.66±0.32*,º 5.19±0.51º, ⌂ *, º , ∆ pg/ml 0.69 β2-MG, 7.59±0.73 3.01±0.30* 5.33± 4.92±0.43*,º 7.21±0.78º, ⌂ µg/ml 0.76*, º , ∆ * IFN-γ, 6.96±0.28 2.90±0.23 3.98± 9.09±0.28*,º 7.26±0.81*, ⌂ *, º , ∆ pg/ml 0.41 Note. Differences are reliable concerning the index: * – in the children of the control group (p <0.05-0.001); º – in the children of the main group and the comparison group; ∆ – in the children of the main group before and after the treatment; ⌂ – in the children of the comparison group before and after the treatment. During the study of β2-MG concentration, it was found to be reduced before treatment in the children of the main group compared to healthy children. The indices constituted 3.01±0.30 µg/ml versus 7.59±0.73 µg/ml (р<0.00). β2MG concentration was also reduced in children of the comparison group compared to the norm comprising 4.92±0.43 µg/ml versus 7.59±0.73 µg/ml (р<0.001). However, its indices were 1.6 times higher than in children of the main group (р<0.001). Its normalization was not observed in children of the main group after treatment despite its significant increase (5.33±0.76 versus 7.59±0.73 µg/ml, р<0.01). However, β2-MG practically normalized in children of the comparison group comprising 7.21±0.78 versus 7.59±0.73 µg/ml, (р>0.1). One of the major mechanisms of body antiviral defense is interferon system which is directly involved in the regulation of immune response and immunity maturation. The course and outcome of disease depends on the speed and efficiency of its inclusion into the process of immune defense. Thus, IFN-γ level was significantly reduced in children of the main group and reasonably differed from the index of healthy children constituting 2.90±0.23pg/ml versus 6.96±0.28 pg/ml, (р<0.001). This indicated interferonogenesis significant inhibition in this category of children in response to frequent ARD. A notable increase in serum concentrations of IFN-γ was observed in children of the comparison group compared to healthy children constituting 9.09±0.28 pg/ml versus 6.96±0.28 pg/ml (р<0.001). Indices in the group of healthy children were 3.1 times higher than in children of the main group, (р<0.001). Normalization of IFN-γ was not

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observed in children of the main group after the treatment, despite its significant increase (from 2.90±0.23 to 3.98±0.41 versus 6.96±0.28 pg/ml, р<0.01). Decrease in its level to almost normal values was noted in the comparison group (from 9.09±0.28 to 7.26±0.81 versus 6.96±0.28 pg/ml, (р>0.01)). Pneumonia in children who often suffer from ARD is accompanied by cytokines imbalance indicating the inhibition of Th1 differentiation and Th2 type activation, insufficient induced production of proinflammatory cytokines for the development of adequate inflammatory process. Violation of any of these parts may be one of pathogenic or beneficial factors of frequent ARD development determining the course, complications and prognosis of the disease. Reduced level of β2-MH in blood serum is also a marker of suppression of T-cell immunity. Maintenance of cytokines imbalance and decreased β2-MG content in these children after discharge from hospital is the basis for finding efficient ways to optimize their treatment.

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Conclusions Cytokines imbalance was detected in children with who often suffered from ARD due to the sharp reduction of IL-2 and IFN-γ. This indicated depletion of the immune system reserves, its switching from T-cell to B-cell part, which cannot provide adequate antiviral and antibacterial body response. Sharp decrease in β2-MG level in blood serum of these children is a rather informative serological marker of Tcell immunity suppression.

Prospects for further research Cytokines imbalance remained in these children after completion of inpatient treatment in comparison with children who were sick not very frequently. This is the basis for finding efficient ways to optimize their treatment. References 1. Aleshyna RM. Syndrome of secondary immune deficiency: clinical and laboratory characteristics. Klinichna imunolohiia. Alerholohiia. Infektolohiia. 2007; 2 (07): 17-20. 2. Antypkin YuH, Turetska AO. Community-acquired pneumonia in children. Zdorovie zhenshchiny. 2008; 4: 159161. 3. Banadyha NV, Tomashivska TV. Cellular and humoral immunity in infants with community-acquired pneumonia. Sovremennaya pediatriya. 2008; 2(19): 36-38. 4. Berezhnyi VV. Immunocorrection in pediatrics. Nova medytsyna. 2005; 5: 54-59. 5. Koroid NV, Zaplatnikov AL, Minhalimova HA, Hlukhareva NS. Community-acquired pneumonia in children: diagnosis and treatment. Russkiy meditsinskiy zhurnal. 2011; 22: 22–27. 6. Ivardava MI. Immunomodulators in the treatment of acute respiratory infections in sickly children. Voprosy sovremennoi pediatriyi. 2011; 10(3): 103-107. 7. Kovalchuk LV, Suslikov VL. Immune reactivity under the conditions of natural zinc deficiency. Imunologiya. 2008; 6: 336-339. 8. Mavrutenkov VV. Diagnostic value of β2-microglobulin determination in serum of patients with acute upper respiratory tract diseases. Medychni perspectyvy. 2005; Х(3): 66-68. 9. Netrebenko OK, Shcheplyagina LA. Immunonutrients in children’s diets. Pediatriya. 2006; 6: 6-14. 10. Pochyvalov AV, Pogorelova EI. Sickly children and new opportunities of immunomodulating therapy. Detskie infektsii. 2010; 1: 50-53.



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