[VACCINE-ASSOCIATED PARALYTIC POLIOMYELITIS IN RUSSIAN FEDERATION DURING THE PERIOD OF CHANGES IN VACCINATION SCHEDULE (2006-2013 yy.)].

The results of virologic testing of clinical materials and epidemiological analysis of vaccine-associated paralytic poliomyelitis (VAPP) cases obtained in 2006-2013 during AFP surveillance are presented. Among the 2976 cases of AFP 30 cases were VAPP. 15 cases were observed in OPV recipients, whereas 15 cases were observed in non-vaccinated contacts. The age of the patients varied from 4 months to 5.5 years (13.6 ± 12.4 months old). Children younger than 1 year constituted 63.3% of the group; boys were dominant (73.3%); 53.3% of children were vaccinated with OPV; the time period between receipt of OPV and onset of palsy was from 2 to 32 days (18.7 ± 8.2). Lower paraparesis was documented in 48.3% of patients; lower monoparesis in 37.9%; upper monoparesis, in 6.9%; tetraparesis with bulbar syndrome, in 6%. The majority of the patients (85.7%) had an unfavorable premorbid status. The violations of the humoral immunity were found in 73.9% cases: CVID (52.9%), hypogammaglobulinemia (41.2%); selective lgA deflciency (5.9%). In 70.6% cases damage to humoral immunity was combined with poor premorbid status. The most frequently observed (76%, p < 0.05) represented the single type of poliovirus--type 2 (44%) and type 3 (32%). All strains were of the vaccine origin, the divergence from the homotypic Sabin strains fell within the region of the gene encoding VPI protein, which did not exceed 0.5% of nucleotide substitutions except vaccine derived poliovirus type 2--multiple recombinant (type 2/type 3/ type 2/type 1) with the degree of the divergence of 1.44% isolated from 6-month old unvaccinated child (RUS08063034001). The frequency of the VAPP cases was a total of 1 case per 3.4 million doses of distributed OPV in 2006-2013; 2.2 cases per 1 million of newborns were observed. This frequency decreased after the introduction of the sequential scheme of vaccination (IPV, OPV) in 2008-2013 as compared with the period of exclusive use of OPV in 2006-2007: 1 case per 4.9 million doses, 1.4 cases per million newborns and 1 case per 1.9 million doses, 4.9 cases per 1 million newborns, respectively. The study has been financed from Russian Federation budget within the framework of the Program for eradication of poliomyelitis in the Russian Federation, WHO Polio eradication initiative, WHO's European Regional Bureau, Russian Foundation for Basic Research (project No. 15-15-00147).

[Level of sIgA, IgG, IgA in vaginal secretion in women with chronic inflammatory disease of small pelvis organs].

AIM: Determine levels of sIgA, IgG, IgA in vaginal secretion and saliva of women of reproductive age with chronic inflammatory diseases of small pelvis organs (IDSPO) at exacerbation stage and remission period. MATERIALS AND METHODS: Clinical-laboratory and gynecological examination of 105 women was carried out. Based on the results obtained 3 groups were formed: patients with IDSPO at exacerbation stage; patients at remission stage; clinically healthy women. sIgA, IgG, IgA parameters were studied in vaginal secretion and saliva in women with IDSPO at exacerbation stage and remission period by radial immune diffusion in gel by Manchini method. RESULTS: An increase of immunoglobulin level in vaginal secretion of women with IDSPO at remission period and a sharper increase of these parameters during exacerbation of the disease compared with women of the control group were detected. During analysis of sIgA, IgG, IgA levels in saliva in the same groups of women the results were obtained that give evidence that the presence of IDSPO and local immune reaction do not lead to the changes of these parameters. CONCLUSION: The obtained parameters on the dependence of an increase of immunoglobulin levels in vaginal secretions and the degree of intensity of the inflammatory process give basis to use them with the aim of additional diagnostics.

[Expression of TLR2 and TLR9 genes by epithelial cells of cervical canal mucous membrane in women with inflammatory diseases of small pelvis organs].

AIM: Determine subpopulation composition of blood lymphocytes and the level of expression of TLR2 and TLR9 by epithelial cells of cervical canal mucous membrane in women of reproductive age with inflammatory disease of small pelvis organs (IDSPO) at exacerbation stage and remission period. MATERIALS AND METHODS: Clinical-laboratory and gynecological examination of 105 women was carried out and 3 groups were formed based on the results: patients at IDSPO exacerbation stage; patients at remission stage; clinically healthy women. By using real time PCR, TLR2, TLR9 gene expression levels were determined in epithelial cells of cervical canal mucous membrane in women of all the 3 groups. Subpopulation composition of blood lymphocytes was determined by flow cytofluorimetry by using monoclonal antibodies with CD45+ CD3+ -T-cell, CD45+ CD3+ CD4+ -T-helper, CD45+, CD3+, CD8+ -T-suppressors-cytotoxic killers, CD45+, CD3-, CD16+, CD56+ natural killers, CD45+, CD3-, CD19+ -B-lymphocytes. Immune fluorescence reaction evaluation was carried out in flow cytofluorimeter Cytomics FC 500 (Becton Coulter, USA). RESULTS: The level of expression of TLR2 gene in the studied groups of patients was established not to differ significantly from parameters in the comparison groups, however it should be noted that this parameter in women with IDSPO at exacerbation stage (causative agents of the infectious process--ureaplasma, staphylococcus, candida) was somewhat higher than in the comparison group. Significantly high level of TLR9 gene expression in cervical canal epithelial cells was detected to correlate with the presence of infectious causative agents. In the group of women with exacerbation of the infectious process the expression of TLR9 was 14.5 times higher compared with the group of women without IDSPO. Among groups of women with IDSPO significant differences in relation to control group in relative and absolute levels of CD3+ T-lymphocytes; CD4+ T-helpers; CD8+ cytotoxic killer T-suppressors, B-lymphocytes compared with the same parameters in clinically healthy women were not detected. CONCLUSION: The increase of TLR9 gene expression level in cervical canal cells of women with IDSPO may serve as an additional diagnostic feature of the presence and degree of severity of the disease.

[The role of immunodeficiencies in development of complications during vaccination of children with BCG vaccine].

AIM: Study of the interrelation between the presence of immune deficiency and development of complications during vaccination of newborns with BCG vaccine. MATERIALS AND METHODS: In 24 children with complications of vaccine process in the form of cold abscess and lymphadenitis indicators of lymphocyte subpopulation levels were studied by flow cytofluorimetry on Beckman Coulter cytofluoriemter by using monoclonal antibodies with markers CD45+CD3+ - T-cell, CD45+CD3+CD4+ - T-helpers, CD45+CD3+CD8+ - T-supressors-cytotoxic killers, CD45+CD3 CD16+CD56+ - natural killers, CD45+CD3-CD19+ - B-lymphocytes. The level of IgG, IgA, IgM in sera was determined by immune diffusion method in agar by Mancini. RESULTS: In 4 children selective deficiency of IgA, in 5 - hyper-IgM syndrome was detected, which is an innate immunodeficiency and is characterized by the lack of sera IgA, reduction of IgG level and increase of IgM. In 9 children a reduction of CD16+ natural killer lymphocytes was detected, in some cases combined with a reduction of CD8+ T-supressors-cytotoxic killers. CONCLUSION: The reason of development of complications during BCG administration is the presence of immunodeficiency in children. In these children severe course of the vaccine process, presence of axillary lymphadenitis was observed, therapy of these children continued from 4 to 6 months.

[Serological activity of vaccine and circulating Bordetella pertussis strains].

AIM: To study activity of vaccine and circulating strains of Bordetella pertussis in serological reactions with serum samples from healthy vaccinated children and children with pertussis infection. MATERIALS AND METHODS: One hundred forty-six serum samples from children with pertussis infection as well as 158 samples from healthy vaccinated children aged 3 - 5 years old were studied. Serologic activity of 3 vaccine strains and 7 strains of B. pertussis isolated from patients with pertussis in 2001 - 2005 against sera of children with pertussis infection or vaccinated children was assessed with hemagglutination assay (HA), radial gel immunodiffusion (RGI), and immunoelectrophoresis (IEP). RESULTS: In HA both serum samples of infected and vaccinated children were equally active in agglutination of microbial preparations prepared from vaccine or recently isolated strains of B. pertussis. RGI assay showed that 81 - 84% of serum samples from infected children and 17 -19% of samples from healthy vaccinated children reacted with vaccine strains, and 81 - 85% of samples from infected children and 16 - 20% of samples from healthy vaccinated children reacted with circulating strains: Sera from patients with pertussis formed identical lines of precipitation with vaccine and circulating strains in RGI assay and three types of precipitation arches profile in IEP. Sera from healthy vaccinated children formed identical precipitation arches with vaccine and circulating strains in RGI assay and one type of precipitation arches profile in IEP. CONCLUSION: Antibodies of patients with pertussis were equally active against vaccine and circulating strains of B. pertussis. Antibodies of vaccinated children were also equally active against vaccine and circulating strains although revealed more narrow spectrum of antigens compared to children with pertussis infection.

[Isotypes of anti-pertussis antibodies in pertussis patients at different stages of the disease].

AIM: Evaluation of anti-pertussis antibodies in pertussis patients at different stages after the onset of the disease. MATERIALS AND METHODS: Levels of IgG, IgG1, IgG2, IgG3, IgG4, IgA and IgM antibodies against the antigen complex of pertussis were evaluated by enzyme immunoassay (EIA). Sera samples were analyzed from 208 pertussis patients examined from week 1 to 10 after the onset of the disease. RESULTS: 51%, 82% and 86% pertussis patients, and 67%, 72% and 78% patients examined from week 1 to 3 after the onset of the disease had increased levels of IgM, IgA and IgG antibodies respectively. 85%, 70%, 74% and 68% pertussis patients, and 76%, 57%, 87%, 57% patients examined from week 1 to 3 after the onset of the disease had increased IgG1, IgG2, IgG3 and IgG4 levels respectively. 92% of all examined pertussis patients and 83% of patients examined from week 1 to 3 after the onset of the disease had an overall increase of anti-pertussis antibody levels. Increased level of IgM antibodies was detected predominately from week 1 to 5 after the onset of the disease. Most of the patients examined from week 3 to 10 after the onset of the disease had increased levels of IgA, IgG, IgG1, IgG2 and IgG4 antibodies, and IgG3 antibody level was increased predominately in patients examined from week 2 to 6 after the onset of the disease. CONCLUSION: Serological indicators of pertussis measured by EIA were observed in 83% of the patients examined at the early stages after the onset of the disease. Simultaneous measurement of IgA, IgG and IgM antibody levels is the most effective approach for serological diagnostics of pertussis due anti-pertussis antibodies isotype composition heterogeneity at different stages after the onset of the disease. Increased levels of IgM and IgG3 antibodies are serologic indicators of the acute phase of pertussis infection.

[Delayed results of vaccination of children with recurrent respiratory infections against measles and mumps].

AIM: Assessment of results of immunization against measles and mumps in children with recurrent respiratory infections. MATERIALS AND METHODS: Levels of IgG against measles and mumps viruses were measured using enzyme immunoassay. Two hundred and twelve serum samples obtained from 6 groups of children with 20 - 45 persons (boys and girls) in each were tested. Children of various ages were presented in each group. Also, immunologic parameters were measured in all children. RESULTS: It was established that mean antibody titers to measles and mumps viruses did not change during 10 years. In more than 50% of children correlation between high and low titers of antibodies of different specificity was found. CONCLUSION: Recurrent respiratory illnesses determine complex regulation of transition of memory cells into cells secreting antibodies to measles and mumps viruses.

[Monitoring of antibodies against diphtheria, tetanus and pertussis in pregnant women].

AIM: To assess antibody levels against diphtheria, pertussis, and tetanus in pregnant women. MATERIALS AND METHODS: One hundred and two virtually healthy pregnant women aged 18-35 years were studied. Antibodies to diphtheria and tetanus were measured in passive hemagglutination reaction with diphtheria and tetanus diagnostic kits. Antibodies to Bordetella pertussis antigens were determined in hemagglutination assay (HA) and in enzyme immunoassay (EIA). RESULTS: Protective titers of anti-diphtheria and anti-tetanus antibodies were detected in 91.2% and 94.1% of participants respectively, whereas high titers--in 24.5% and 27.4% respectively. Low levels of IgG to B. pertussis antigens measured by EIA were observed in 70.6% of participants whereas moderate and high levels--in 22.5% and 6.9% respectively. Conditionally protective levels of anti-pertussis antibodies measured by HA were detected in 10.8% of participants. CONCLUSION: Obtained results demonstrate high level of protection of pregnant women against diphtheria and tetanus and low level of anti-pertussis immunity.

[Distribution of specific IgG subclasses in patients with pertussis and in healthy persons].

AIM: To assess level of IgG1, IgG2, IgG3, and IgG4 to complex of antigens of Bordetella pertussis in patients with whooping cough and healthy children and adults. MATERIALS AND METHODS: Levels of anti-pertussis IgG subclasses in sera of patients with pertussis and healthy children and adults were measured with solid phase immunoenzyme assay using peroxidase-conjugated monoclonal antibodies to human IgG1, IgG2, IgG3, and IgG4. RESULTS: In patients with pertussis, IgG1-IgG3-IgG2-IgG4 type of distribution of subclasses with predominance of IgG1 and IgG3 was revealed. In healthy children and adults the character type of subclasses distribution was IgG1-IgG2-IgG4 with absent or low level of IgG3. CONCLUSION: Detection of specific IgG3 mainly in patients with pertussis allows to consider them as a reliable serological sign of pertussis infection.

[Humoral immunity against pertussis and its prevalence in population].

Levels of IgG and IgA to complex of Bordetella pertussis antigens were assessed in 503 healthy children aged 1 - 14 years, 75 adolescents aged 15 - 17 years, and in 504 adults aged 18 - 54 years. The highest level of IgG was observed in children aged < 5 years. In older age groups progressive decrease of IgG level was noted, which more most prominent in 9 - 11 year-olds with subsequent stabilization of the level in adolescents and adults. Significant heterogeneity of IgG level was noted in all age groups. Rate of detection of increased IgA level correlated with age-related decrease of IgG level and increased from 2 - 5% in children aged 1 - 5 years to 12 - 16% in older children and adults. Obtained data point to low levels of immunity against pertussis in older children, adolescents and adults and high undetected incidence of pertussis in studied population.

[Improvement of pertussis diagnostics in adults with prolonged cough].

AIM: To study clinical and laboratory data and levels of IgG, IgG1, IgG2, IgG3, IgG4, IgA and IgM to Bordetella pertussis complex of antigens in adults with prolonged cough. MATERIALS AND METHODS: Antibody levels to Bordetella pertussis complex of antigens were measured by solid phase immunoenzyme assay. Clinical and laboratory methods included CBC, chest X-ray, measurement of respiratory function, allergologic tests. RESULTS: In 16 out of 75 studied patients (21%) serological signs that are characteristic for current pertussis infection (increased levels of specific IgG and IgA as well as IgG1 - IgG3 - IgG2 - IgG4 type of distribution of specific IgG subclasses) were observed. Clinical and laboratory parameters--course of disease, characteristics of cough, results of CBC--corresponded to diagnosis of pertussis. In other studied patients levels of specific antibodies did not differ from levels observed in healthy persons and observed clinical signs corresponded to other respiratory diseases. CONCLUSION: Obtained results prove the high incidence of pertussis in adults, its essential importance as etiologic factor of prolonged cough and high informative value of serologic tests.

[Diagnostic value of IgG, IgA, and IgM to Bordetella pertussis antigens in patients with pertussis].

Comparative study of IgG, IgA, and IgM levels to complex of antigens (CA) of vaccine strain No. 475 and separate antigens of Bordetella pertussis: pertussis toxin (PT), filamentous hemagglutinin (FHA), lypopolysaccharide (LPS), agglutinogens 1 (Aggl.1) and 2 (Aggl.2) was performed by ELISA in 80 patients with pertussis and 80 healthy vaccinated children. Antibodies to mentioned antigens were detected both in ill and healthy children but their levels were remarkably higher in patients. The most reliable serologic marker of pertussis was IgA, which were detected in the majority of patients. Detection rates of this class of antibodies to CA, PT, FHA, LPS, Aggl.1, and Aggl.2 were 91%, 77.5%, 69%, 59%, 80%, and 12%, respectively. Elevated levels of specific IgA were registered in 5% of healthy children. Obtained results showed high information value of detection of the IgA and IgG antibodies to CA, PT, FHA, and Aggl.1 using ELISA for pertussis diagnostics. Simplicity and economy of the CA obtainment allow to recommend CA-based ELISA for serologic diagnostics of pertussis.

[Paralytic poliomyelitis in Russian Federation in 1998-2005].

From 1998 through 2005 3,294 cases of acute flaccid paralysis (AFP) including 93 cases with clinical picture of poliomyelitis were registered in Russian Federation. From the latter cases 91 were classified as vaccine-associated paralytic poliomyelitis (VAPP): 66 were VAPP cases in oral poliomyelitis vaccine (OPV) recipients and 25--VAPP cases in contacts. VAPP rate was 1 case per 1.6 million of distributed OPV doses, 1 case per 2.2 million doses for OPV recipients, and 1 case per 186,000 doses for recipients of 1st OPV dose in children aged < 1 year. Majority of VAPP cases in recipients occurred after 1st dose (89.4%) and in contacts--in non-vaccinated children (76%). Mean interval between OPV administration and onset of VAPP in recipients was 21 days. Children aged < 1 year were predominant among VAPP cases (92.4% among recipient VAPP cases, and 80% among contact VAPP cases). Majority of the patients had unfavorable health status including defects of immunity. Most of poliovirus strains isolated from VAPP cases belonged to type 3 (52.9%) whereas to type 2 and 1--29.8% and 17.4% of strains respectively. All VAPP cases were associated with vaccine-derived polioviruses. A highly diverged poliovirus type 1 (2.65% of nucleotide substitutions in VP1 region) was isolated from patient with contact VAPP. Formation of poliovirus-neutralizing serum antibodies in children with VAPP including persons with immunodeficiency reflects the ability of the organism to produce specific antiviral immune response.

[Cellular and humoral immunodeficiency in children with vaccine-associated paralytic poliomyelitis].

Markers of humoral and cellular immunity in 16 patients with vaccine-associated paralytic poliomyelitis (VAPP) were evaluated. Signs of immunodeficiency (decrease of T- and B-lymphocytes counts, impaired synthesis of immunoglobulins, defects of phagocytosis, decrease of NK number) were revealed in all of the patients. Majority of them (81.3%) had defects in humoral immunity. Decrease of CD31, CD4+ and CD8+ was detected in 86.7, 35.7 and 91.7% of the patients respectively. Study of serum immunoglobulins performed in 15 patients showed decrease of IgG, IgM and IgA levels in 6 (40%), 1 (6.7%) and 6 (40%) of the patients respectively. Agammaglobulinemia was diagnosed in one patient in which only trace quantities of IgA and IgG were detected and IgM level was well below the normal. Congenital deficiency of IgA was diagnosed in 3 children. Majority of the children (11 from 12) had comorbidities (frequent respiratory infections, dermatitis, changes of intestinal microflora). Thus, immunocompromised condition of a child is a risk factor for VAPP after administration of alive oral poliovaccine.

[Effect of the sialic acids of mucin on its ability to increase the virulence of serotype A meningococci]. / Vliianie sialovykh kislot mutsina na ego sposobnost' povyshat' virulentnost' meningokokka serogruppy A.

The influence of the content of bound sialic acids in mucin on its capacity to decrease the infective dose of meningococcal culture has been studied; as a result, the direct relationship with a high degree of correlation between these two characteristics has been revealed. The direct relationship between the content of bound sialic acids and the viscosity of aqueous mucin solutions has also been revealed in this study. The above-mentioned biological activity of mucin is not observed in the presence of free sialic acids.

[Isolation of modified allergens from bee venom]. / Poluchenie modifitsirovannykh allergenov pchelinogo iada.

To obtain modified bee venom (BV) allergens, covalent binding of BV with previously carboxylated polyethylene glycol (PEG) has been used. The conjugation of BV and PEG has been achieved by means of carbodiimide. Thus 4 kinds of the conjugate with BV/PEG ratio ranging from 30:1 to 63:1 have been obtained. The study has shown that chemical treatment in the process of this reaction, dialysis and chromatography does not lead to a decrease in the specific activity of BV, while lyophilization produces such an effect. The above method for the modification of BV allergen, used with a view to obtaining high molecular compositions, is reproducible, ensures sufficient yield (about 30%), and permits obtaining conjugates with specified BV/PEG ratios.

[Protective activity of antigenic preparations isolated by different methods from Streptococcus pneumoniae serotype 3]. / Izuchenie zashchitnoi aktivnosti antigennykh preparatov, vydelennykh razlichnymi metodami iz Streptococcus pneumoniae serotipa 3.

The specific protective activity of antigenic preparations isolated from Str. pneumoniae culture by sedimentation with 96% ethanol or acetone was demonstrated in experiments with the active immunization of mice and their subsequent challenge with a virulent culture. The protective activity of antigenic preparations, expressed in micrograms, was shown to be directly related to the protein content of these preparations and inversely related to their carbohydrate content. The deproteinization of antigenic complexes isolated from Str. pneumoniae grown both in solid and in liquid culture media was accompanied by an increase in their immunogenicity.

[A radioimmunological method for determining IgA-class antimeningococcal secretory antibodies in human saliva]. / Radioimmunologicheskii metod opredeleniia antimeningokokkovykh sekretornykh antitel IgA-klassa v sliune cheloveka.

The data on the content of secretory IgA antibodies to group A Neisseria meningitidis protein antigen in the saliva of persons, both having had contact with N. meningitidis culture and having had no such contact, are presented. The results were obtained by the method of radioimmunoassay, developed specially for the determination of N. meningitidis protein antigen.

[Local immunity in acute dysentery. II. The content of antibody-forming cells in the large intestine mucosa in acute Flexner dysentery in the dynamics of the disease]. / O mestnom immunitete pri ostroi dizenterii. Soobshchenie II. Soderzhanie antiteloobrazuiushchikh kletok v slizistoi tolstoi kishki u bol'nykh ostroi dizenteriei Fleksnera v dinamike zabolevaniia.

Antibody-forming cells were detected in the large intestine of patients with acute Flexner's dysentery by means of the modified Jerne - Nordin method of hemolysis in agar. This method allowed one to determine the classes of immunoglobulins produced by the cells contained in tissue microspecimens obtained by the biopsy of the intestinal mucosa. The maximum amount of antibody-containing cells could be detected on days 7-12 of the disease. The content of antibody-forming cells was shown to depend on the severity of dysentery, the duration of the disease and the therapeutic methods used in the process of treatment. IgA was found to be the most frequent antigen type.

[Evaluation of the immunogenicity of meningococcal vaccines in experiments on mice]. / Otsenka immunogennosti meningokokkovykh vaktsin v opytakh na myshakh.

The immunogenicity of 2 meningococcal vaccines, multicomponent vaccine produced at the Mechnikov Research Institute for Vaccines and Sera in Moscow and polysaccharide vaccine obtained from Merck Sharp & Dohme (USA), was evaluated on experimental meningococcal sepsis in mice, produced by the injection of meningococcal culture in mucin suspension. The protective effect of these 2 vaccines, expressed in terms of ED50, was 0.28 +/- 0.12 for the multicomponent vaccine and 0.25 +/- 0.24 for the polysaccharide vaccine; the challenge dose used in the test was 10 LD50 of the culture. The multicomponent vaccine gave the maximum immunological effect in a dose of 8 micrograms, while higher or lower doses induced a lesser increase in antibody titer and thus gave lower protection to mice against infection.