[Use of the cultural variants of Coxsackie A viruses in virological practice].

Coxsackie A viruses belong to the enteroviruses, the isolation of which from infectious materials and further cultivation are possible only when laboratory animals are infected. The authors could adapt the strains of 17 of 23 serotypes of these viruses to RD cell culture. The strains of 8 serotypes were additionally adapted to Vero cell culture. The cultural variants of Coxsackle A viruses were used to prepare immune sera. The Bacterial and Viral Agents Enterprise, M. P. Chumakov Institute of Poliomyelitis and Virus Encephalitides, Russian Academy of Medical Sciences, has set up the production of bacterial and viral drugs based on the cultural variants of 5 Coxsackie A virus serotypes. The cultural variants of 14 Coxsackie A virus serotypes were used to carry out a virus neutralization test. Examination of more than 600 children from Moscow and the Moscow Region showed the wide circulation of individual Coxsackie A virus serotypes. It also demonstrated a drastic reduction in Coxsackie A-7 virus circulation in the past 50 years.

[Toll-like receptor-mediated functional activity of mononuclear cells in children with neutropenia].

AIM: To study the Toll-like receptor (TLR)-mediated functional activity of peripheral blood mononuclear cells (PBMC) from children with different forms of neutropenia. MATERIALS AND METHODS: TLR-mediated functional activity of PBMC was evaluated by production of proinflammatory cytokines--TNF alpha and IFN alpha. Ligands for TLR1/2, TLR 2/6, TLR4, TLR5, and TLR9 were used to stimulate TNF alpha production by PBMC from healthy children and children with neutropenia. Ligands for TLR3, TLR4, TLR7, TLR7/8, TLR8, and TLR9 were used to induce production of IFN alpha. Levels of TNF alpha and IFN alpha were measured in PBMC supernatants by ELISA. The group of patients with neutropenia included 9 children with immune neutropenia and 3 children with congenital neutropenia. Control group consisted of 12 healthy children of the same age range. RESULTS: It was revealed that TLR2, TLR4, and TLR5 ligands have enhanced stimulating effect on TNF alpha production by PBMC of children with congenital neutropenia and had no effect on PBMC of children with immune neutropenia. Children with immune neutropenia are characterized by significantly increased IFN alpha production induced by ligands of TLR3, TLR8, and TLR9. CONCLUSION: Revealed changes of TLR-mediated functional activity of PBMC from children with various forms of neutropenia may play significant role in development and course of infections in these patients.

[Corrective effect of cyclooxygenase inhibitor on functional status of mononuclear cells expressing Toll-like receptors].

AIM: To study the influence of the COX inhibitor--lornoxicam (LX)--on Toll-like receptor (TLR)-mediated production of proinflammatory and anti-inflammatory cytokines by peripheral blood mononuclear cells (PBMC) from healthy subjects and patients with acute pancreatitis (AP) in vitro. MATERIALS AND METHODS: Cytokine production by PBMC of healthy donors was stimulated by TLR1/2 ligand peptidoglycan (PG) and TLR4 ligand lypopolysaccharide (LPS) in presence of LX. Levels of cyotokines (IL-1beta, IL-6, IL-8, IL-10, IL-12, and TNFalpha) were measured by ELISA. Group of patients with acute pancreatitis of toxic etiology included 11 subjects: patients from main group received combined therapy supplemented with NSAID from the oxicam class--LX; patients who received only standard basic treatment formed comparison group. RESULTS: It was found that in vitro LX inhibits production of both proinflammatory and anti-inflammatory cytokines by PBMC of healthy subjects mediated by ligands of TLR1/2 and TLR4. Maximal inhibitory effect of LX was observed when cytokine production was induced through TLR1/2. Patients with AP demonstrated increased production of TNFalpha induced by TLR1/2 and TLR4 ligands. CONCLUSION: LX inhibits TLR-mediated production of both proinflammatory (IL-1, IL-6, IL-8, IL-12, TNFalpha) and anti-inflammatory (IL-10) cytokinesby PBMC of healthy subjects in vitro. Treatment with LX in patients with AP results in diminished effector function of TLR1/2 and TLR4 already during 1st day of the illness and normalization of these indices by 6th day.

[Comparative evaluation of cellular immunity changes in occupational bronchial asthma patients after phytotherapy complexes vs. conventional corticosteroid therapy].

The studies covered changes of phenotypic markers for peripheral immunocytes in occupational bronchial asthma patients under influence of phytotherapy complexes vs. conventional corticosteroid therapy. Phytocomplexes appeared to cause pathogenetically positive changes in phenotypic markers content of peripheral blood.

[Current methods for detection of anti-erythrocytic antibodies in blood donors].

Erythrocytic antibody screening in donors aids in reducing the risk of posttransfusion reactions and complications; donors with serum immune antibodies are taken from donation of plasma and transferred to that of erythrocyte-containing media. The application of highly sensitive tests for antibody screening permits detection of antibodies of wide range and various specificity and immunogenicity. The antibody identification technique makes it possible to establish a priority scale of transfusionally dangerous erythrocytic antigens.

[Effect of toll-like receptors (TLR) ligands on in vitro synthesis of proinflammatory cytokines by peripheral blood mononuclear cells from healthy persons and from patients with common variable immunodeficiency].

Production of citokines of tumor-necrosis factor (TNF)-alpha and interferon (IFN)-alpha by peripheral blood mononuclear cells (PBMC) from healthy men and from patients with common variable immunodeficiency (CVID) after stimulation with zymosan, lypopolysaccharides, flagellin and CpG, which are ligands of TLR 2/6, TLR 4, TLR 5, TLR 9 respectively, was studied in vitro. In healthy men production of TNF-alpha varied between individuals, whereas synthesis of IFN-alpha was similar. Spontaneous production of TNF-alpha by PBMC in patients with CVID was increased and accompanied by decrease in TNF-alpha production stimulated by each analyzed ligands except CpG. Observed changes in TLR-dependent TNF-alpha production can play important role in pathogenesis of CVID.

Analysis of toll-like receptor-dependent production of proinflammatory cytokines in vitro by human peripheral blood mononuclears of donors and patients with primary immunodeficiency.

The production of TNF-alpha and IFN-alpha cytokines by peripheral blood mononuclears in response to stimulation by TLR2/6, TLR4, TLR5, TLR9 ligands (zymosan, LPS, flagellin, and CpG-oligodeoxynucleotide, respectively) was studied in donors and patients with common variable immunodeficiency. Individual characteristics of TNF-alpha production by mononuclears were revealed in donors. Reduced stimulated production of TNF-alpha in response to stimulation with TLR4 and TLR5 ligands in vitro was detected in patients with common variable immunodeficiency.

[Complement 3 in hypertension and arteriosclerosis]. / Tretii komponent komplementa pri gipertonicheskoi bolezni i ateroskleroze.

The serum levels, residual functional activity (RFA), phenotype of complement component 3 (C3) were determined in 20 patients with hypertensive disease (HD), 11 with documented coronary atherosclerosis (CA), and 11 with HD + CA. The sera from 21 apparently healthy subjects were used as a control. There was a higher frequency of the allotype C3F in the HD, CA, and HD + CA groups (0.400, 0.417, and 0.364, respectively) than in the controls. The HD + CA group showed a significant (p less than 0.05) decrease in RFA of C3 as compared to the HD and CA groups. There were significant differences in the serum C3 concentrations and RFA in allotype C3F carriers between the patients with CA and HD. The findings suggest that allotype C3F carriage may be a factor predisposing to accelerated progression of CA in HD.

[Effects of atrial natriuretic factor on proliferative response and natural cytotoxicity of human lymphocytes]. / Vliianie atrial'nogo natriiureticheskogo faktora na proliferatvnyi otvet i estestvennuiu tsitotoksichnost' limfotsitov cheloveka.

Effect of synthetic analogues of atrial natriuretic factor (ANF) on proliferative response and natural cytotoxic lymphocytes of human subjects was investigated in vitro. ANF-III and ANF-IV increased blast transformation lymphocytes induced by a Con-A suboptimal dose. The increase of cells activity was comparable with the effect of interleukin-2, added at a dose of 50-100 unit/ml.

Anticardiolipin antibodies in Sneddon's syndrome.

We studied 24 patients (18 women, 6 men), aged 29 to 54, with Sneddon's syndrome. The clinical picture of Sneddon's syndrome was characterized by cerebrovascular disorders, livedo reticularis, disturbance of peripheral circulation, arterial hypertension, cardiac pathology (ischemic heart disease, heart murmurs), complicated obstetric history in women, and disturbed sexual function in men. In 6 of 17 examined patients with Sneddon's syndrome there was a high concentration of anticardiolipin antibodies (ACA) but no antibodies to native DNA and LE cells. The course of the disease in the patients with a high ACA level, when compared with normal ACA level patients, was characterized by a more rapid progression and more severe clinical manifestations. The study demonstrates the similarity of clinical symptoms and immunologic disturbances in Sneddon's syndrome and the antiphospholipid syndrome and suggests the importance of ACA in the pathogenesis of some cases of Sneddon's syndrome.

[Study of lymphocyte subpopulations and the expression of activation markers in diabetes mellitus types 1 and 2]. / Izuchenie subpopuliatsii limfotsitov i ékspressii markerov aktivatsii pri sakharnom diabete 1-go i 2-go tipov.

Diabetic patients with the disease duration less than 1 year (group 1 eight patients with diabetes mellitus type I and group 2 seven patients with diabetes mellitus type II) and 8 healthy donors were examined for subpopulations of lymphocytes (CD 3-, CD 4-, CD 8-, CD 20-positive cells, CD4/CD8), expression of activation markers on peripheral blood mononuclears investigated with monoclonal antibodies of BMA and OKT series, and serum neopterin concentration. Group I patients had low CD4/CD8 and increased number of CD 8 cells, 1a/DR-positive lymphocytes (28.6 +/- 7%), OKT9-positive lymphocytes (8.0 +/- 4.7%), activated neopterin synthesis registered neither in group 2 patients nor donors. The number of CD 3 and CD 4 cells was similar in the diabetics and donors. B-lymphocyte level in group 1 patients was on the decrease. Unbalance in lymphocyte subpopulations, increased expression of activation markers and of serum neopterin can be noted in viral infection reflecting impairment of immunoregulating mechanisms in diabetes mellitus type I.

[Disorders of humoral immunity in hypertension]. / Narushenie gumoral'nogo immuniteta pri gipertonicheskoi bolezni.

Examination included 51 patients with essential hypertension (EH) of an uncomplicated course, labile (IB-IIA stage, according to A. L. Myasnikov's classification) and stable (IIB stage) hypertension. Clinical characteristics were given to the stages and duration of EH, body weight of the patients, arterial hypertension (AH) heredity and the AP level. Immunological examination included determination of the concentration of the basic classes of immunoglobulins IgG, IgA, IgM, circulating immune complexes (CIC), concentration of IgE and beta 2-microglobulins. It was revealed that EH development is attended by an increased concentration of immunoglobulins, primarily of IgA (23 per cent), IgE (31 per cent) and CIC (21 per cent), which is associated, to a certain degree, with a factor of AH hereditary aggravation.

[Immunoregulatory disorders and activation of the immune system in dilated cardiomyopathy]. / Narushenie immunoreguliatsii i aktivatsiia sistemy immuniteta pri dilatatsionnoi kardiomiopatii.

The level of immunoregulatory lymphocytic subpopulations (CD3-, CD4-, CD8-cells, CD4/SD8 ratio, IaDr-positive cells, SD16-cells), lymphocyte natural cytotoxicity (NCT), the concentration of serum interferon (IF), beta 2-microglobulin (beta 2-M) and neopterin were investigated in 30 patients with dilatative cardiomyopathy (DCMP). Immunoregulatory lymphocytic subpopulations were defined by indirect immunofluorescence using monoclonal antibodies to differentiating markers of BMA and OKT series. A decrease in SD4/SD8 ratio less than 1 was observed in 10 patients, an increase in SD4/SD8 ratio over 2.3 was observed in 5 patients. A significant rise of IaDr-positive cells, a decrease in NCT, an increase in the concentration of IF, beta 2-M and neopterin were observed. There was correlation of an increase in IF concentration, the number of IaDr-positive cells and an increase in the level of neopterin and beta 2-M. Signs of activation of the immune system were more frequently noted in patients with a decreased (less than 1) or increased CD4/CD8 ratio. The role of disorders of immunoregulation and activation of the immune system in DCMP pathogenesis was discussed.

[Use of monoclonal antibodies for the intratypic differentiation of strains of poliomyelitis type I virus]. / Ispol'zovanie monoklonal'nykh antitel dlia vnutritipovoi differentsiatsii shtammov virusa poliomielita tipa I.

Six clones of mouse hybridomas producing highly specific antibodies to the vaccine LSc 2ab strain of poliomyelitis virus type I have been derived. The clone 10C5 antibodies were used for intratypic serodifferentiation of 75 poliomyelitis virus type I strains from the collection of the Institute of Poliomyelitis and Viral Encephalitides of the USSR Acad. Med. Sci. which had been isolated in the USSR from children with paralytic poliomyelitis in 1950-1983. General large-scale vaccination campaigns for children using live poliomyelitis vaccine (SLV) had started in 1959. Among 12 strains isolated in 1950-1958, only 2 strains could be neutralized with clone 10C5 antibody. These 2 strains were evidently of the vaccine origin since they had been isolated in 1957 in Leningrad where at that time first large-scale trials of SLV had already been initiated (Smorodintsev, A.A. in: "Live Vaccine against Poliomyelitis", Leningrad, 1960, 42-60). In the period after the beginning of large-scale vaccination campaigns (1959-1983), 11 out of 26 strains and 25 out of 36 strains isolated in areas of the moderate and hot climate, respectively, showed antigenic relationship with the vaccine LSc 2ab strain. The portion of vaccine-like strains isolated in different years beginning since 1959 was sufficiently stable comprising about half of the total number of poliomyelitis virus type I strains examined.

[Enterovirus type 71 pathogenicity for laboratory animals]. / Patogennost énterovirusa tipa 71 dlia laboratornykh zhivotnykh.

Enterovirus type 71 (E71) strains isolated from specimens of the brain and feces of children during an outbreak of poliomyelitis-like infection in Bulgaria in 1975 produce paralytic disease in newborn and adult cotton rats, newborn white mice, and monkeys. By the type of the myo- and neurotropic properties manifested in animal experiments the Bulgarian E71 strains are very close to neuropathogenic strains of Coxsackie A7, A14, and A16 viruses. The infection induced by the Bulgarian E71 strains in green monkeys was by clinical symptoms (tremor, convulsions, death), the speed of the development of the process and the type of morphological lesions particularly similar to the clinical and morphological manifestations of infection observed in most severe cases in children during the outbreak in Bulgaria. Inoculation of newborn and adult cotton rats, newborn white mice and Macaca rhesus monkeys with large doses of the prototype BrCr strain of enterovirus 71 (10(5.0)--10(6) TCD50) caused no clinically manifest disease of the animals.