RESUMO
According to the data of the Federal Service on Customers' Rights Protection and Human Well-being Surveillance, the enzyme-linked immunosorbent assay (ELISA) detects the tick-borne encephalitis virus (TBEV) in ticks more often than the polymerase chain reaction (PCR). The goal of this work was to compare TBEV detection efficiency in the ixodid ticks of different species with the commercial kits based on ELISA and real-time PCR. Ticks of five species were parenterally infected with 2-6 IgPFU of the European or Siberian TBEV subtypes. We formed randomized and encoded series of infected and intact ticks of different species, and in "blind" experiment analyzed the ticks on the TBEV presence with the kits based on ELISA and real-time PCR. ELISA and real-time PCR effectiveness of the TBEV detection in ticks was not affected by gender, species of ticks or presence of blood meal. The kits based on ELISA were less sensitive than those based on real-time PCR. ELISA effectiveness depended on the TBEV subtype. The presence of the false positive reactions and sensitivity of ELISA were affected by the protocols of reaction. The problem of the different TBEV prevalence in the field-collected ticks obtained with various methods remains to be studied.
Assuntos
Antígenos Virais/imunologia , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/normas , Ixodidae/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Animais , Antígenos Virais/genética , Vírus da Encefalite Transmitidos por Carrapatos/genética , Vírus da Encefalite Transmitidos por Carrapatos/imunologia , Reações Falso-Positivas , Feminino , Masculino , Camundongos , Coelhos , Sensibilidade e Especificidade , Carga ViralRESUMO
The content of cytokines, the levels of antibodies (Ab) to proinflammatory cytokines and fetoproteins (FP) in serum, as well as the effects of fitohemagglutinin and FP on the level of cell production of cytokines into the conditioned medium were studied in relation to the pattern of a response of mononuclear cells (MNC) to FP. With the positive reaction of MNC to FP, estimated by an increment of CD9* cells and detectable in grades 2-3 dysplasias, the anti-inflammatory effect with lower anti-inflammatory cytokines was shown to be achieved due to elevated Ab levels, which may compensate for the low content of IL-4 and IL-10. With FP, there was an increase in the cell production of IL-10 that is known to stimulate antibody formation in the early phase of tumor growth, as evidenced by the association of the levels of Ab to FP with the grade of dysplasia.
Assuntos
Formação de Anticorpos/imunologia , Autoanticorpos/imunologia , Leucócitos Mononucleares/imunologia , Tuberculose Pulmonar/imunologia , alfa-Fetoproteínas/imunologia , Adulto , Idoso , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Tuberculose Pulmonar/patologiaRESUMO
A representative collection was obtained containing 68 monoclonal antibodies (MAB) to Toxoplasma gondii antigens, which was characterized by the binding with the below fractions of tochizoites in the immune-enzyme assay (IEA) and immunoblotting (IB): membrane (MEM), somatic (water-soluble, SOM) and excretory-secretory (ES). Most of MABs were produced to MEM antigens (43), 6 MABs reacted with the somatic fraction, and 3 MABs reacted with both fractions. Two MABs to ES antigen were detected in the latter group. An analysis of MABs in concurrent IEA and IB revealed the immune-dominant proteins of the MEM and SOM fractions of antibodies to T. gondii tochizoites (p30 and p27, respectively). The presence of 2 non-overlapping antigenic determinants was shown for p30. Further research would detect MABs that could be used in the diagnosis of toxoplasmosis.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos de Protozoários/imunologia , Toxoplasma/imunologia , Animais , Membrana Celular/imunologia , Epitopos , Immunoblotting/métodos , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The hybrid gene of influenza virus hemagglutinin (HA) of the H1-subtype, carrying the sequence coding for the fragment of H3-subtype antigenic site B, was constructed. The product of expression of this gene in E. coli was obtained as a fusion protein with beta-galactosidase. The chimeric protein was shown to retain the antigenic properties of HA of H1-subtype and to interact specifically with antibodies against the synthetic peptide corresponding to the B site fragment of HA of the H3-subtype.
Assuntos
Expressão Gênica , Genes Sintéticos , Hemaglutininas Virais/biossíntese , Vírus da Influenza A/imunologia , Proteínas Recombinantes/biossíntese , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Epitopos/imunologia , Escherichia coli/genética , Genes Virais , Hemaglutininas Virais/genética , Hemaglutininas Virais/imunologia , Vírus da Influenza A/genética , Dados de Sequência Molecular , Conformação Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologiaAssuntos
Antígenos Virais/genética , Vírus da Encefalite Transmitidos por Carrapatos/genética , Regulação Viral da Expressão Gênica/genética , Variação Genética/genética , Recombinação Genética/genética , Vaccinia virus/genética , Antígenos Virais/análise , Vírus da Encefalite Transmitidos por Carrapatos/imunologia , Genes Virais , Vetores Genéticos/genética , Plasmídeos/genética , Regiões Promotoras Genéticas/genética , Vaccinia virus/imunologiaAssuntos
Regulação Viral da Expressão Gênica/genética , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/genética , Imunização/métodos , Precursores de Proteínas/genética , Vacinas Sintéticas/genética , Vaccinia virus/genética , Proteínas do Envelope Viral/genética , Vacinas Virais/genética , Animais , Formação de Anticorpos/genética , Formação de Anticorpos/imunologia , Regulação Viral da Expressão Gênica/imunologia , Genes Virais/genética , Antígenos de Superfície da Hepatite B/imunologia , Imunidade Celular/genética , Imunidade Celular/imunologia , Macaca mulatta , Camundongos , Camundongos Endogâmicos BALB C , Precursores de Proteínas/imunologia , Coelhos , Vacinas Sintéticas/imunologia , Vaccinia virus/imunologia , Proteínas do Envelope Viral/imunologia , Vacinas Virais/imunologiaRESUMO
The full-length copy of the hemagglutinin gene of influenza virus was inserted into M13 phage DNA. The DNA sequence coding for the hydrophobic prepeptide was removed from the gene by oligonucleotide-directed mutagenesis. The possibilities of expression of the full-length and mutant genes in E. coli were investigated. The beta-galactosidase-hemagglutinin fusion proteins were isolated. The fusion proteins exhibited specific binding to antiviral antibodies. This binding could be competitively inhibited by excess of viral hemagglutinin, demonstrating that these fusion proteins contained antigenic determinants of hemagglutinin.