Enzymatic characterization of blood lymphocytes in various clinical and pathogenetic variants of respiratory allergy

A. Lazareva, O. Kolenchukova, S. Smirnova
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Abstract

Allergy is a sufficient social and economic issue of modern times. Altered immunity in allergic disorders is based, mainly, on the lymphocyte disturbances.Immune characteristics depend both on populations and subpopulational profile of immune cells, and on intrinsic intensity and specific features of theirintracellular metabolism. Interest to studies of intracellular metabolism of lymphocytes id determined by high-scale energetic and plastic processes aimed for support of immune homeostasis. The aim of this study was to evaluate the state of intracellular metabolism in peripheral blood lymphocytes from the patients with respiratory allergies of different genesis and respiratory affection.The study included patients with various clinical variants of respiratory allergy (n = 152) at the age of 21 to 63 years old, and virtually healthy blood donors (n = 209), comparable for age and sex. Within these cohorts, we have separately analyzed, e.g., respiratory atopy (atopic rhinosinusitis and atopic bronchial asthma), as well as respiratory pseudoatopy (polypous rhinosinusitis and asthmatic triad). Allergic disorders of upper respiratory ways were diagnosed in a complex clinical examination by allergologist/immunologist and otorhinolaryngologists. Bronchial asthma verification was based on current GINA criteria (2014). We used m standard common clinical methods and specific allergological diagnostics, e.g., allergological anamnesis, skin prick tests, with different non-infectious allergens, measurement of total and specific IgE’s with ELISA method. The parameters of intracellular metabolism of peripheral blood lymphocytes were determined with bioluminescent technique with bacterial luciferase. Actifity of NAD(P) and NAD(P)H enzymes was measured.Dehydrogenase activities in lymphocytes were expressed as enzyme un its (EU, 1 unit = 1 mcmol/min) per 104 cells.Certain changes of intracellular activities in peripheral lymphocytes are revealed, dependent on genesis and origin of allergic inflammation, and affection level of respiratory ways. In respiratory atopy (atopic rhinosinusitis and atopic bronchial asthma), irrelevant on the level of respiratory affection, the activities of intracellular enzymes suggest increased plastic processes that are maximally pronounced in atopic bronchial asthma. In respiratory pseudoatopy (polypous rhinosinusitis and asthmatic triad) the metabolic changes of lymphocytes presume activation of both plastic and energetic processes, with decreased intensity in asthmatic triad condition. Independent on genesis of respiratory allergic inflammation, we have determined low activity of NAD(P)-dependent glutamate dehydrogenase, and NAD(H)-dependent lactate dehydrogenase in allergic inflammation of upper respiratory ways (atopic rhinosinusitis and polypous). Its activity is statistically higher in bronchial asthma (atopic bronchial asthma and asthmatic triad.
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血液淋巴细胞在呼吸道过敏的各种临床和致病变异中的酶学特征
过敏是一个充分的社会和经济问题的现代。过敏性疾病的免疫改变主要是基于淋巴细胞紊乱。免疫特性既取决于免疫细胞的群体和亚群特征,也取决于其细胞内代谢的内在强度和特定特征。对淋巴细胞细胞内代谢的研究兴趣是由旨在支持免疫稳态的大规模能量和可塑性过程决定的。本研究的目的是评估不同病因和呼吸道影响的呼吸道过敏患者外周血淋巴细胞的细胞内代谢状态。该研究包括21至63岁的各种呼吸道过敏临床变异患者(n=152),以及在年龄和性别上具有可比性的几乎健康的献血者(n=209)。在这些队列中,我们分别分析了呼吸性特应性(特应性鼻窦炎和特应性支气管哮喘),以及呼吸性假性特应性疾病(息肉性鼻窦炎和哮喘三联征)。过敏学家/免疫学家和耳鼻喉科医生在复杂的临床检查中诊断出上呼吸道过敏性疾病。支气管哮喘验证基于当前的GINA标准(2014)。我们使用了标准的常见临床方法和特定的过敏性诊断方法,如过敏性记忆、皮肤点刺试验、不同的非传染性过敏原、ELISA法测量总IgE和特异性IgE。用细菌萤光素酶生物发光技术测定外周血淋巴细胞的细胞内代谢参数。测定NAD(P)和NAD(P)H酶的活性。淋巴细胞中的脱氢酶活性表示为每104个细胞中的酶un-its(EU,1单位=1mcmol/min)。揭示了外周淋巴细胞细胞内活性的某些变化,这取决于过敏性炎症的发生和起源,以及呼吸方式的影响程度。在呼吸性特应性疾病(特应性鼻窦炎和特应性支气管哮喘)中,与呼吸影响水平无关,细胞内酶的活性表明可塑性过程增加,在特应性哮喘中最为明显。在呼吸性假性特应性(息肉性鼻窦炎和哮喘三联征)中,淋巴细胞的代谢变化假定可塑性和能量过程的激活,哮喘三联征的强度降低。与呼吸道过敏性炎症的发生无关,我们已经确定了NAD(P)依赖性谷氨酸脱氢酶和NAD(H)依赖性乳酸脱氢酶在上呼吸道过敏性炎(特应性鼻窦炎和息肉)中的低活性。其活性在支气管哮喘(特应性支气管哮喘和哮喘三联征)中具有统计学意义。
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