Vol 25, No 1 (2022)

The present article deals with experimental and clinical aspects of immuno-hormonal interactions in chemical carcinogenesis i.e., formation of DNA-adducts with chemical carcinogens as a trigger of tumor initiation; synthesis of specific antibodies as markers of human exposure to environmental carcinogens; immunomodulation of chemical carcinogenesis by the specific antibodies in experimental studies; interactions of antibodies against environmental carcinogens with endogenous steroid hormones in human carcinogenesis; immunological interference and inversion of immuno-hormonal interactions by the action of antibodies against environmental carcinogens; immune stimulation of tumor progression in cancer patients. It is shown that antibodies specific to estradiol and progesterone participate in regulation of serum estradiol and progesterone levels in healthy women. Excessive production of antibodies against benzo[a]pyrene is associated with impaired physiological balance between the levels of antibodies to estradiol and progesterone, thus causing disturbed physiological balance between serum estradiol and progesterone. Immuno-hormonal imbalance promotes tumor initiation, its growth and progression. The new approaches to the personalized cancer immunoprediction and immune prevention are discussed. Coordinated synthesis of antibodies against benzo[a]pyrene and estradiol seems to reflect production of DNA-adducts with genotoxic metabolic effects of these compounds manifesting as synergistic carcinogenic effects upon the target cells. Hence, simultaneously increased levels of serum antibodies against benzo[a]pyrene and estradiol in healthy people may be considered an immunological marker of high oncological risk and an reason to use of new immunoprotective tools against polycyclic aromatic hydrocarbons and phytoestrogens. However, ability of these antibodies to raise the blood serum levels of environmental carcinogens and endogenous estradiol, as shown in vitro and in vivo, excludes the opportunity for active cancer immune prevention. Usage of anticarcinogen vaccines aimed for induction of protective secretory antibodies is likely to further increase high levels of procarcinogenic serum antibodies against benzo[a]pyrene and estradiol, followed by additional enhancement of immuno-hormonal imbalance and promotion of carcinogenesis. Development of probiotics transduced with genes encoding human antibodies against environmental carcinogens may present an alternative approach to cancer immune prevention. The antibodies produced by such probiotics would bind appropriate carcinogens and prevent their invasion into the organism, thus inhibiting emergence of DNA-adducts and suppressing synthesis of specific autoantibodies that may promote carcinogenesis. The aim is to substantiate the concept of immuno-hormonal imbalance for the carcinogen-induced hormone-dependent tumors.

Cytokines belong to the class of signaling molecules, being involved into regulation of proliferation, differentiation and effector functions of immunocompetent cells. The ratio of pro- and anti-inflammatory cytokines is important for development of any inflammatory process. IL-1ra, IL-4, IL-10 are among the most important anti-inflammatory cytokines. Their function is to limit and suppress immune response in inflammatory processes of any etiology. In this respect, the aim of this study was to evaluate production of the anti-inflammatory cytokines IL-1ra, IL-4 and IL-10 by peripheral blood cells in children with autoimmune and infectious diseases. Patients and methods: Pediatric parients (2-17 years old) participated in the study including those with juvenile idiopathic arthritis (n = 101); unspecified reactive arthropathy (n = 24); systemic lupus erythematosus (SLE, n = 14); chronic viral hepatitis C (n = 24). 33 healthy children (n = 33) comprised the control group. Heparinized blood samples were diluted with glutamine-containing medium RPMI-1640. Control samples were not treated by any stimulants, and the stimulated samples were supplied with phytohemagglutinin (20 mkg/ml). The samples of diluted blood were incubated for 24 hours (37 °C, 5% CO₂). The supernates were frozen once. The concentrations of IL-1ra, IL-4 and IL-10 in these cell supernatants were determined by ELISA technique (Vector-Best, Russia). Results: It was found that the spontaneous production of anti-inflammatory cytokines (IL-1ra, IL-4 and IL-10) didn’t differ, or was lower in the groups of patients with SLE, juvenile idiopathic arthritis and hepatitis C if compared with control group. SLE is an autoimmune disease, whereas juvenile arthritis is of mixed autoimmune-autoinflammatory etiology. Chronic hepatitis C is a viral disease, but autoimmune responses may manifest at the chronic stage of the disorder. Decreased production of anti-inflammatory cytokines could be an evidence for autoimmune mechanisms of these diseases, despite their different etiology. More intensive spontaneous production of IL-1ra and IL-10 in children with unspecified reactive arthropathy may suggest some compensatory reactions which inhibit development of inflammation in this disorder. IL-1ra, IL-4 and IL-10 production in stimulated cultures didn’t differ between all groups of the patients, or it was lower in comparison with healthy children. Decrease cytokine production in groups of children with different diseases suggests exhausted functional reserve of immunocompetent cells caused by their chronic activation.

Measles is a highly contagious viral infection transmitted by airborne droplets, characterized by fever, intoxication and specific rashes on the skin and mucous membranes. Despite the availability of highly effective vaccines and many years of efforts by the world medical community with active immunization of the world’s population against this infection under the auspices of WHO, measles still remains a serious problem. The aim of this work was to investigate the effect of measles infection in adults upon the wide range of lymphocyte subsets and blood cytokine profile in comparison with healthy controls.

The venous blood samples from 50 adult measles patients aged 20 to 55 years, were taken 6±1 days after the onset of skin rash, being compared with blood samples from 50 healthy adults of similar age group. The 200 μL plasma aliquotes resulting from spontaneous sedimentation of the formed elements in an Eppendorf tube were taken, frozen at -30 °C and used within 3 months for the cytokine profile assays. 15 cytokines were tested by multiplex technique (MagPix, BioRad, USA). Mononuclear cells were isolated by gradient centrifugation and immunophenotyped using four-color staining by means of equipment and reagents from BD Biosciences (USA).

In the group of measles patients, activation of innate immunity was revealed, i.e., the IL-1, IL-6, IL-23, IL-31 cytokines and TNF, which belong to early pro-inflammatory cytokines, were significantly increased. In measles patients, a significant increase in cytokines was found, suggesting active participation of epithelial cells in immune response to the measles virus. They produce danger signals (IL-25 and IL-33), inducing the development of adaptive immunity, activate their protective abilities via IL-17F production, and are involved in repair under the influence of IL-22. Some cells of adaptive immunity are infected with the measles virus and die, others actively respond to the viral infection and proliferate, thus leading to changing ratio of their subsets. Hence, the patients showed a significant decrease in T lymphocytes due to a decrease in CD4⁺ cells, an increased percentage of cells in “senescent” and “exhaustion” state, a significant decrease in T_EMRO subpopulations, both among CD4⁺ and CD8⁺ lymphocytes, and an increase in CD8⁺T_CM. The levels of B cell subpopulations (Bm, B1, Breg) in measles patients did not differ from healthy ones, and the level of plasmablasts was significantly increased. The level of CD4⁺ lymphocyte subpopulations and production of their cytokine markers varied greatly. In the patient group, a shift in the type of immune response towards Th2 and Th17 was found, activation of Tfh and Treg was detected, and increased expression of HLA-DR and CD38 activation markers was found.

In response to measles infection, there are several independent, multidirectional processes observed in the patients. On the one hand, the measles virus attacks epithelial cells of mucous membranes and skin and immunocompetent cells, exerting a cytopathic effect and leading to lymphopenia and selective decrease in various lymphocyte subsets. On the other hand, the measles virus initiates activation of both innate and adaptive immunity, thus causing production of the corresponding cytokines, expression of activation markers, and an increase in effector cell subsets. Better understanding of the immunopathogenesis of measles infection and associated immunosuppression will help us to improve vaccination outcomes against this infection and prevent measles-related mortality.

Glaucoma is widely known to have a progressive course and occupy a leading place among the causes of vision loss and blindness. Increased intraocular pressure is the key harmful factor among the causes of glaucoma occurrence. In some cases, however, the progressive disease is also observed at normal values of ophthalmic tonus. Early diagnosis of glaucoma will allow for timely therapy, which in turn will reduce the risk of complications and prevent neuroopticopathy progression. According to the literature data, the pathogenesis of primary open-angle glaucoma is associated with nitric oxide (NO), due to imbalance between endothelium-produced vasoconstrictors and vasodilators, especially, endotelin-1 and nitric oxide. Decreased NO level combined with endotelin-1 hyperproduction is associated with development and progression of a number of ocular disorders including glaucomatous atrophy of the optic nerve. Since nitric oxide is produced by endothelial NO-synthase (eNOS), one may assume that eNOS is involved in pathogenesis of neurodegenerative changes in primary open-angle glaucoma. However, despite numerous studies on the pathogenesis of glaucoma, the distinct factors of innate immune response remain poorly studied. The purpose of the present study was a search for association between polymorphic markers (C774T, T786C, Glu298Asp) of the eNOS gene and the risk of primary open-angle glaucoma among the Perm Region residents.

Peripheral blood of patients with primary open-angle glaucoma (the main group) and cataract without glaucoma (a comparison group) was used as initial biomaterial. In comparison group, arterial hypertension was most often encountered as concomitant pathology. Genomic DNA was first isolated from the blood samples, followed by rt-PCR using reagent kits for determining C774T, T786C, Glu298Asp polymorphic markers in the eNOS gene.

The prevalence of polymorphic variants of the innate immunity genes T786C, C774T and Glu298Asp of the eNOS gene was analyzed in patients with primary open-angle glaucoma. There were no significant differences in the distribution of genotypes and alleles of eNOS gene for the C774T and Glu298Asp polymorphic markers. An increased frequency of homozygous TT genotype was found, along with decreased occurrence of C allele at the polymorphic T786C locus of the eNOS gene, as well as a trend for decreased frequency of the TC and CC genotypes. Arterial hypertension potentiated the negative effect of increased intraocular pressure upon the glaucoma-associated optic neuropathy. Conclusions. The studied changes in genotypes and allelic frequencies of eNOS gene may be regarded as risk factors that increase probability of the primary open-angle glaucoma and predict severity of the disease.

Arterial hypertension is an urgent health problem worldwide causing an increase in temporary and permanent disability, invalidity and mortality due to cardiovascular diseases. Researchers recognize the multifactorial nature of arterial hypertension, but environmental factors are of particular potential importance. The working conditions at oil production enterprises are characterized by a more pronounced influence of these factors which may predispose for early development of disadaptation disorders, functional changes in immune and humoral regulation, and, finally, for increased risk of cardiovascular diseases in people engaged in oil production. The aim of the present work was to study the features of immunity and humoral risk factors of arterial hypertension in hypertensive employees at an oil-producing enterprise. To this purpose, a comparative analysis of lymphocyte subpopulations (CD3⁺CD4⁺, CD3⁺CD25⁺), markers of apoptosis (CD3⁺CD95⁺, TNFR, p53, Bax, Annexin V-FITC+7AAD), phagocytic activity of leukocytes (absolute phagocytosis index), and the levels of vascular humoral factors (nitric oxide and homocysteine) was performed in employees of an oil production enterprise exposed to adverse production factors. The observational group consisted of employees with established episodes of increased blood pressure. A comparison group consisted of individuals without clinical manifestations of cardiovascular disease. As a result of the clinical and laboratory examination of employees at the oil-producing enterprise with arterial hypertension, some functional changes in immune regulation were revealed. This group was characterized by a significantly (p < 0.05) decreased CD3⁺CD4⁺, and CD3⁺CD25⁺ lymphocyte contents, along with increased levels of regulatory CD127 lymphocytes against the comparison group (p < 0.05). The workers at an oil production enterprise with arterial hypertension are characterized by decreased (p < 0.05) phagocytic activity of peripheral blood leukocytes using the criteria of absolute phagocytosis. We found some signs of inhibited lymphocyte apoptosis (p < 0.05), i.e., a decrease in CD95⁺, TNFR, and p53 over the background values, as well as increased Bax levels over the comparison group (p < 0.05). However, the content of TNFR and p53 significantly (p < 0.05) exceeded the reference level, regardless of previous arterial hypertension episodes. Development of the high blood pressure episodes among the employees at oi-producing plant showed a significant association (p < 0.05) with elevated levels of homocysteine and nitric oxide concentrations which are known to induce endothelial dysfunction, atherogenesis and, hence, a persistent increase in blood pressure.