CYTOKINE EXPRESSION PROFILES OF PERITONEAL FLUID IN THE PATIENTS WITH UTERINE LEIOMYOMA

Abstract. Uterine leiomyoma (UL) is a hormone-dependent benign tumor of uterus. Social significance of UL is stipulated by its high rate among fertile females. Scarce data exist about the impact of cytokines in UL progression. Th1/Th2 paradigm is one of crucial points in modern immunology. Evaluation of cytokines involved into either type of immune response is of special significance for studying the diseases accompanied by the changes of extracellular matrix, e.g., leiomyomas. In present study, we analyzed peritoneal fluids from UL patients, with multiplex detection of IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, GM-CSF, IFN-γ and TNF-α (Th1/Th2 panel), by means of a Bio-Plex® instrument (Bio-Rad, USA). Twenty-seven patients were observed in our study (20 patients with UL, and 7 myoma-free women (a group of comparison). The mean age of the patients was 43.5±0.6 years. The duration of UL ranged from 0 to 18 years. As a result, the levels of IL-10, GM-CSF, IFN-γ and TNF-α in patients with long-existing UL (over 5 years) were significantly higher (p<0,05) than in group with a disease story of <5 years. IFN-γ values in peritoneal fluid patients with UL did inversely correlate with uterine size. Moreover, the levels of IFN-γ in patients with smaller uterine volume (<8 weeks of pregnancy) were increased in relation to the group with larger tumor size. IL-10 contents were increased in the patients with adenomyosis, rapid and slow growth of UL, and in both types of tumor (simple and proliferative). Increased IL-5 levels were observed in the patients with single tumor nodules (as related to the patients bearing multiple nodes, and comparison group). Furthermore, intramural and subserosal location of nodes was characterized by increased levels of IL-5. In the patients free of adenomyosis, IL-5 value was increased against the comparison group. The changes in IL-2, IL-4, IL-12 and IL-13 levels in patients with UL were not statistically significant

LOCAL APPLICATION OF RECOMBINANT INTERFERON-ALFA2 FOR TREATMENT OF RECURRENT RESPIRATORY PAPILLOMATOSIS

Abstract. Recurrent respiratory papillomatosis (RRP) is the most frequently occurring tumour of the upper airways associated with a human papilloma virus (HPV). The aim of this study was to reveal some features of systemic and local immunity in RRP, to investigate clinical and immunological efficiency of local treatment with recombinant interferon-α (rIFNα), and to determine clinical and laboratory indications to it’s administration. The study included forty-one patients with confirmed RRP. Their examination included histological examination of papillomas, detection of HPV DNA in papilloma tissues using PCR technique, phenotyping of circulating lymphocytes (CD3+, CD4+, CD8+, CD25+, HLA-DR+) by means of flow cytometry. The levels of IFNγ, TNFα, GM-CSF, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13 in laryngeal secretions were quantified by a multiplex immunoassay. In all cases, we revealed an initially decreased functional activity of Т-lymphocytes, as well as low contents of Т-killer and NK-cells. In laryngeal secretions, increased values of Th1-type-specific cytokines (IFNγ and TNFα) were found. Besides that, high levels of local IL-4 were detected thus being typical to alternative Th2-type response. Single inhalations 1000 000 ME of «Interal» or «Roferon» preparations were administered daily (a total of 10-15 millions ME per therapeutic course). Thirteen patients received the treatment after surgery, as an adjuvant therapy, and eleven patients underwent monotherapy. Complete tumor regression of tumors following this monotherapy was observed in 45,5% of the patients, whereas partial regression was registered in 45%. The effect was mostly expressed in frequently recurring juvenile papillomatosis with aggressive course and histological pattern of actively proliferating papilloma. In the patients with complete tumor regression, high initial levels of TNFα and IL-4/IFNг ratios were revealed initially in laryngeal secretions. When rIFNб was administered, an increase in Th1-type cytokines (IFNγ, IL-12, IL-2) and GM-CSF were revealed, as well as decrease of initially high levels of IL-10, like as lowered levels of IL-4 and IL-13. In cases of partial tumor regression, the levels of IFNα, IL-12, IL-2 and GM-CSF were decreased against initial values, and in the course of treatment they fluctuated within normal reference values. Cancellation of the therapy was followed by increase in IL-4 and IL-13 levels. Hence, the local rIFNα therapy is effective in controlling aggressive recurrent respiratory papillomatosis. Initial increase in TNFα and IL-4/IFNγ ratios in laryngeal secretions may be considered as favorable predictor of anticipated therapy, whereas increased levels of IFNα, IL-2 and IL-12 together with decrease (normalization) of IL-4/IFNγ ratio may reflect the efficiency of the treatment performed

PATTERN OF LOCAL IMMUNE ALTERATIONS IN THE PATIENTS WITH CHRONIC ADENOIDITIS, AND AN OPPORTUNITY OF THEIR CORRECTION WITH IMUNOFAN

Abstract. The issue of chronic adenoiditis, due to its medical and social significance, is in focus of attention for otolaryngologists, pediatricians, immunologists. According to various works, chronic adenoiditis in children comprise 20 to 56% of upper respiratory diseases. Chronic adenoiditis is characterized by relative resistance to conventional therapy, and, in pronounced cases, by low reversibility of pathological events. The studies performed have shown that the patients with chronic adenoiditis show some features of local and general immune deficiency that were exhibited as decreased serum IgA concentration and local sIgA secretion, as well as insufficient income of IgG from vascular system to the area of inflammation in patients with chronic adenoiditis. These local immune deficiencies were considered to be an indication for local immunotherapy. Dynamics of clinical symptoms and changes in immunoglobulin profile has shown that clinical efficiency of Imunofan therapy was dependent not solely on the initial clinical features of disease, but on the local immunity state. Analysis of initial immunoglobulin concentrations in nasopharyngeal lavage from the patients showing differential clinical response have demonstrated that therapeutic effect of Imunofan was most pronounced in the patients with initially moderate activity of inflammatory process, deficient IgG income from vascular system, decreased local IgA synthesis in the inflammatory foci. Certain clinical features of chronic adenoiditis, i.e., clinical course without hypertrophy of pharyngeal tonsils, local immune deficiency (decreased local IgA secretion and deficient IgG income from blood), with a background of moderate inflammatory events, may be used as a rationale for administration of local immunotherapy with Imunofan to these patients. (Med. Immunol., 2008, vol. 10, N 6, pp 551-562)

СYTOKINES IN OROPHARYNGEAL LAVAGES FROM THE PATIENTS WITH CHRONIC ADENOIDITIS IN THE COURSE OF LOCAL MONOTHERAPY WITH IMUNOPHAN

Abstract. Since decades, the aspects of chronic adenoiditis draw attention of specialists in ORL. From different data, prevalence of chronic adenoiditis is 20 to 56 per cent among children with diseases of upper respiratory ways. Chronic adenoiditis is relatively resistant to conventional therapy, and irreversible clinical course is revealed in more severe cases. The studies performed have shown that the patients with chronic adenoiditis exhibit signs of local immune deficiency that manifested into decreased local secretion of IL-4, IL-10 and GM-CSF, thus being considered as an indication for local immunocorrection procedures. To this purpose, we used Imunophan as a spray for intranasal application. Dynamics of clinical symptoms and local immune parameters showed that the efficiency of Imunophan treatment was dependent of both initial clinical features of the disorder, and on cytokine levels in oropharyngeal lavages. Therapeutic effect of Imunophan was more expressed in the patients with initial predomination of Th1 immune response and cell-type immune reactions in the area of inflammation. A set of certain clinical and laboratory data, i.e., absence of enlarged pharyngeal glands, predominance of local Th1 type response and cellular type of immune response may be used for monitoring before administration of local immunotherapy with Imunophan in the patients with chronic adenoiditis. (Med. Immunol., 2008, vol. 10, N 2-3, pp 261-268)

FEATURES OF LOCAL mRNA SYNTHESIS FOR SOME CC- AND CXC-CHEMOKINES AND THEIR RECEPTORS IN ENDOMETRIAL HYPERPLASIA

Аbstract.  Endometrial  hyperplasia  (EH)  represents  an  excessive  increase  in  thickness  and  volume  of proliferating endometrium accompanied by altered glandular structure. This disorder is higly prevalent among women in their premenopausal period. There exist only scarce data concerning possible role of chemokines and their receptors in EH pathogenesis and clinical course. Hence, the aim of our study was to analyze mRNA expression  of  several  key  chemokines  and  their  receptors  in  endometrial  tissue  samples  from  EH  patients. This work included sixty-three women with disturbed menstrual  cycle  and/or  pathological  changes  of endometrium, as assessed by sonographic studies. The patients were 32 to 61 years old (a mean of 48.4±0.6 years). The levels of mRNA expression were determined by  gene-specific  PCR  in  a  semiquantitative  manner,  whereas promoter genotypes of matrix metalloproteinases (ММР1 -16071G/2G and ММР3 -11715А/6A) were identified by means of allele-specific PCR. Results of the study included a significant increase of mRNA for MIP-1α, eotaxin 2, along with decreased amounts of mRNA for CCR-3 (a specific receptor for eotaxins), in polyps developing from hyperplastic endometrium. MIP-1α synthesis fades away with increasing age. An increased level of MIP-1β was shown in prolonged and recurrent disturbances of menstrual cycle, whereas elevation of MIP-1α and CXCR-1 was registered in cases of multiple pregnancies. In threatening abortions, an increase of MIP-1β gene expression was revealed. Hence, the local chemokine system reacts to inflammatory and hemorrhagic complications with increased mRNA expression of certain chemokine genes. Determination of the chemokine mRNA levels, as well as their receptors in patients with endometrial hyperplasia may reflect a general background of this disorder. (Med. Immunol., 2011, vol. 13, N 2-3, pp 189-196

PLASMA CYTOKINES LEVELS IN PATIENTS UNDERGOING LONG-TERM HAEMODIALYSIS

Аbstract.  Patients  with  end-stage  renal  disease  need  their  kidney  functions  to  be  replaced.  Chronic haemodialysis represents a most common method of such substitution treatment. This procedure results in successful survival of such patients for years. Chronic haemodialysis is accompanied by a complication which is known as β2-microglobulin amyloidosis. In this case, amyloid substance consisting of β2-microglobulin (β2-MG) accumulates in bones, ligaments and joints. Biological causes of β2-MG amyloidosis are still not established. To elucidate the role of inflammation in the pathogenesis of β2-MG amyloidosis, the levels of  IL-2,  IL-4,  IL-6,  IL-8,  IL-10,  GM-CSF,  IFNγ, TNFα were quantified in plasma of patients undergoing  long-term haemodialysis. Mean amounts of all the mentioned cytokines in haemodialysis patients proved to be significantly higher than in control group consisting of healthy subjects. When comparing a group receiving standard  dialysis  procedure  versus  a  subgroup  receiving  haemodiafiltration,  a  single  reliable  difference  was revealed for GM-CSF levels (p < 0.04), without any differences shown for other cytokines. With increasing terms of chronic haemodialysis, the levels of IL-2, IL-4, IL-6, IL-8, GM-CSF, IFNγ, TNFα were increased, or, at least, they did not decrease. After three years of dialysis, IL-10 concentrations were statistically indistinguishable from normal levels. In patients undergoing haemodiafiltration, plasma levels of IL-2, IL-4, IL-8, GM-CSF, IFNγ, TNFα did not drop with increasing terms of dialysis. The levels of IL-6 and IL-10 decreased after three years of dialysis, to near-normal levels.In general, these results suggest that IL-10 and IL-6 may be regarded as candidates for further studies as potential markers of β2-microglobulin amyloidosis. (Med. Immunol., 2011, vol. 13, N 2-3, pp 211-218

DYNAMICS OF CYTOKINE PROFILE IN PATIENTS WITH RHEUMATOID ARTHRITIS AS INFLUENCED BY INFLIXIMAB (REMIKEID) ADMINISTRATION

Abstract. Rheumatoid arthritis (RA) is a chronic autoimmune systemic disease with predominantly destructive lesions of peripheral joints, with prevalence of 0.6 to 1.6% in general population. An important pathogenetic role in this disease is now attributed to imbalance between pro- and antiinflammatory cytokines. Clinical introduction of biological preparations, such as Infliximab (monoclonal antibodies to TNFα) within last years have changed therapeutic approach to treatment of rheumatic diseases. The aim of our research was to evaluate dynamics of pro- and antiinflammatory cytokine profile in the patients with rheumatoid arthritis (RА) during combined therapy with Infliximab and Methotrexate (MT). The study included 30 patients (27 females, 3 males, mean age of 52.5±2.0 years) who received combined therapy with МТ and Infliximab (Inx). Inx was initially injected at a single dose of 3 mg/kg intravenously, followed by administration 2 and 6 weeks later, and then repeated every 8 weeks. Regular examination of the patients included clinical and laboratory studies (ESR, levels of IL-6, IL-8, TNFα, IL-4, IL-10, GSM-CSF, IFNγ). Levels of antibodies against Infliximab in the groups of RА patients were determined before treatment and 22 weeks later. Efficiency of the therapy was estimated according to DAS28 3V Index and to HAQ Questionnaire.Upon decreased activity of disease, as assessed by DAS28, and improvement of HAQ parameters, a marked decrease in proinflammatory cytokine levels (IL-6, IL-8, TNFα) was detected, that confirming a pathogenetic significance of cytokine in RА patients. In patients with marked clinical effect (group I), an initially normal contents of TNFα was found in blood serum, and this group showed better response to Infliximab therapy, than groups II and III (resp., moderate and absent response) with initially high contents of TNFα and other cytokines, that was proven by correlations with ACR criteria and HAQ functional index. These events were accompanied by more significant improvement of RА course and increased functional abilities of joints. In patients from group III (absence of clinical effects), the level of antibodies to Infliximab before therapy was high, and it was increased by 22 week of treatment. It was shown that, in cases of initially high levels of endogenous anti-TNFα antibodies, clinical response to Infliximab therapy is likely to be reduced. Thus, it is possible to suggest that determination of initial TNFα and IL-10 levels, as well as starting levels of antibodies to Infliximab, and their changes in the course of therapy can be used as immunological parameters, thus allowing to predict the responses to Infliximab therapy. (Med. Immunol., 2008, vol. 10, N 2-3, pp 251-260)

IMPACT OF SOME CYTOKINES TO EFFICIENCY OF INFERTILITY TREATMENT BY MEANS OF IN VITRO FERTILIZATION

Abstract. Present study defines clinical, hormonal and immunological factors allowing to predict outcomes of IVF cycles in patients with combined forms of infertility at their reproductive age. Indices of gonadotrophic ovarian stimulation in IVF cycles in patients of reproductive age with combined forms of infertility depend on their age and cytokine contents in follicular fluid. Effectiveness of gonadotropin-induced ovarian stimulation is inversely dependent on the patients’ age, its increase is related to increased contents of pro-inflammatory cytokines in follicular fluid and increased expression of CCR1, along with decreased expression of CCR3 chemokine receptors in endometrium. Effectiveness of gonadotropin-induced ovarian stimulation depends on concentration of cytokines in the follicular fluid, which is argued by presence of a significant direct correlation between the number of growing follicles, oocytes and local contents of IL-10, IL-15, eotaxin, MIP-1β, RANTES, TNFα, IFNγ, and GMCSF. Free testosterone levels show a positive correlation with numbers of oocytes obtained from the ovarian punctate, and CXCR2 and CCR3 expression in endometrium, as well as a negative correlation was found with IL-4 contents in follicular fluid. In patients with negative IVF cycle outcomes, levels of IL-13 in follicular fluid upon ovarian stimulation proved to be twofold higher than those in patients with successful pregnancy after IVF. Prognosis for pregnancy in a cycle of treatment in patients with combined forms of infertility of reproductive age is determined by a patients’ story, and a variety of clinical, hormonal and embryological factors which can increase predictive accuracy of the IVF outcomes. To determine a probability of successful pregnancy after IVF cycle, an appropriate prognostic index is recommended, with a sensitivity levels of 93% and specificity of 89%

EXPRESSION OF mRNAS FOR CHEMOKINES AND CHEMOKINE RECEPTORS IN THE SKIN FROM PATIENTS WITH PSORIASIS

Abstract. Some issues in etiology and pathogenesis of psoriasis are poorly studied. Therefore, a search for new potential markers is actual for diagnostics of psoriasis in less clear cases. In this study, an attempt was undertaken to evaluate contribution of some chemokines and appropriate receptors into pathogenesis of psoriasis. The main group consisted of the patients with psoriatic arthritis (n = 20) and psoriasis vulgaris (n = 9). A group of comparison consisted of patients with sclerodermia (n = 4), and a control group was represented by healthy persons (n = 9). The specimens were taken from visually normal and affected skin areas from psoriatic patients obtained by punch biopsy. Expression of the following chemokines was performed: CCL3/MIP-1α, CCL4/MIP-1β, CCL5/RANTES, CCL11/eotaxin, CCL24/eotaxin-2, CXCL8/IL-8 and their receptors (CCR1, CCR3, CCR5, CXCR1, CXCR2). In cases with PASI values < 10, an increased expression of the following genes was revealed for CCL11/eotaxin (p = 0.03), CXCR1 (р = 0.008), CXCR2 (р = 0.0006) in virtually intact skin and affected skin areas, as well as increased gene expression of CCL24/eotaxin 2 (p = 0.009), CCL5/RANTES (p = 0.05) in visually normal skin.With PASI values of 10 to 20, an increased gene expression was found for CCL11/eotaxin (p = 0.005), CCL24/eotaxin 2 (p = 0.02), CCL5/RANTES (p = 0.01), CXCR1 (р = 0.0009), CXCR2 (р = 0.002) in skin biopsies from visually healthy and affected skin, as well as increased expression CXCL8 (IL-8) (p = 0.005) in visually normal skin. In cases with PASI > 20, an increased expression of CCL11/eotaxin (p = 0.001), CCL24/eotaxin 2 (p = 0.001), CCL3/MIP-1α (р = 0.02), CXCR1 (p = 0.0001), CXCR2 (p = 0.001) was detected in visually healthy skin samples and affected skin of the patients, as well as higher expression of CCL4/MIP-1β (р = 0.03) in affected skin areas. A reverse correlation was revealed between expression of chemokines, i.e., CCL24/eotaxin 2 (r = –0,94, p = 0.005), CCL3/MIP-1α (r = –0,94, p = 0.005), CCL4/MIP-1β (r = –0,85, p = 0.03) and their receptors: ССR5 (r = –0,79, p = 0.005) and CXCR2 (r = –0,94, p = 0.005) in visually normal skin of the patients with psoriasis and PASI values < 10.A direct correlation was found between expression of mRNAs for CCL11/eotaxin (r = 0.69, p = 0.04) in visually healthy skin from psoriatic patients, and CCR5 (r = 0.82, p = 0.006) in affected skin of the patients with psoriasis and PASI values of 10 to 20. In the group of patients with PASI values over 20, no correlations were detectable. These data allow us of concluding aboutan important contribution of chemokine system to pathogenesis of psoriasis. (Med. Immunol., vol. 10, N 4-5, pp 337-346)

EVALUATION AND CLINICAL SIGNIFICANCE OF CHEMOKINES AND THEIR SPECIFIC RECEPTORS IN PATIENTS WITH BRONCHIAL ASTHMA

Abstract. A group of patients with bronchial asthma (BA) was under investigation, being classified according to their disease activity, and exacerbation risk factors. We studied expression of specific mRNAs for chemokines and their receptors in nasopharyngeal mucosa (n = 70) and amounts of those chemokines in blood serum (n = 59). In the patients with BA exacerbations after acute respiratory infections, an increased expression of eotaxin, MIP-1α, MIP-1β, CXCR1 mRNAs was observed, along with decreased CCR3 mRNA, as compared to a control group. In blood serum of these patients, a decreased levels of eotaxin and MIP-1β mRNA was accompanied by increase of MIP-1β and eotaxin-2 amounts. For the patients with allergen-provoked BA attacks, an increased expression of eotaxin, eotaxin-2, MIP-1β, CXCR1, CXCR1 was revealed, as well as decreased CCR1 mRNA expression, whereas serum samples showed significantly decreased contents of eotaxin and MIP-1β, along with increase in MIP-1α, RANTES and eotaxin-2 mRNAs. In BA remission state, a decreased mRNA expression was found for eotaxin, CCR1 and CXCR2 was observed, with increased serum contents of MIP-1α, RANTES and eotaxin-2. Hence, the changes of chemokine system detected in BA may be sought as markers of the disease exacerbation, depending on its primary origin, as well as potential biomarkers of the disease itself