RESUMO
AIM: To investigate the immunogenicity of a novel DNA vaccine, pSW3891/HBc, based on HBV core gene in Balb/c mice. METHODS: A novel DNA vaccine, pSW3891/HBc, encoding HBV core gene was constructed using a vector plasmid pSW3891. Balb/c mice were immunized with either pSW3891/HBc or empty vector DNA via gene gun. IgG anti-HBc responses in mouse sera were demonstrated by ELISA. Specific cytotoxicity of cytotoxic T lymphocytes (CTLs) of mice was quantitatively measured by lactate dehydrogenase release assay. RESULTS: HBcAg was expressed effectively in 293T cell line transiently transfected with pSW3891/HBc. Strong IgG anti-HBc responses were elicited in mice immunized with pSW3891/HBc. The end-point titers of anti-HBc reached the highest 1:97,200, 4 wk after the third immunization. The specific CTL killing with the highest specific lysis reached 73.25% at effector:target ratio of 20:1 in mice that received pSW3891/HBc DNA vaccine. CONCLUSION: pSW3891/HBc vaccination elicits specific anti-HBc response and induces HBc-specific CTL response in immunized Balb/c mice.
Assuntos
Antígenos do Núcleo do Vírus da Hepatite B/genética , Vacinas contra Hepatite B/genética , Hepatite B Crônica/terapia , Vacinas de DNA/genética , Animais , Feminino , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Vacinas contra Hepatite B/imunologia , Vacinas contra Hepatite B/farmacologia , Hepatite B Crônica/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos , Linfócitos T Citotóxicos/imunologia , Vacinas de DNA/imunologia , Vacinas de DNA/farmacologiaRESUMO
OBJECTIVES: To observe immunogenicity of new DNA vaccine encoding for hepatitis B virus core antigen (HBcAg). METHODS: A new DNA vaccine (pSW3891/HBc) encoding for hepatitis B virus core antigen was constructed using plasmid pSW3891 which can be used in human. Control and experiment groups of Balb/c mice were immunized with pSW3891 or pSW3891/HBc by gene gun. Anti-HBc in sera of mice was tested by ELISA (enzyme linked immune sorbent assay). Specific cytotoxicity of cytotoxic T lymphocyte (CTL) of mice was detected by LDH release assay. RESULTS: pSW3891/HBc can express in 293T cell line effectively. Mice immunized with pSW3891/HBc showed strong anti-HBc response and specific high cytotoxicity of CTL. CONCLUSION: pSW3891/HBc induced significantly humoral and cellular immune responses in Balb/c mice.
Assuntos
Anticorpos Anti-Hepatite B/sangue , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Vacinas contra Hepatite B/imunologia , Vacinas de DNA/imunologia , Animais , Antígenos do Núcleo do Vírus da Hepatite B/genética , Imunidade Celular , Camundongos , Camundongos Endogâmicos BALB C , Linfócitos T Citotóxicos/imunologiaRESUMO
OBJECTIVE: Although many antigens have been investigated, the method for the bile canaliculus staining using optical microscopy needs to be improved. The aim of the present study was to assess the expression pattern of a candidate marker, CD25, in normal and diseased liver tissue. METHODS: Immunohistochemistry, immunofluorescence, and immune electron microscopy assays were performed with 41 liver sections and 2 different anti-CD25 monoclonal antibodies. A polyclonal antibody against carcinoembryonic antigen (CEA) was also used to stain bile canaliculus as a control. CD25 expression levels in normal and diseased liver tissue were also determined. RESULTS: CD25 was predominantly localized at the bile canaliculus of adult and infantile liver, evidenced by both immunohistochemistry and immunofluorescence assays. The electron microscopy assay showed that there were obvious amorphous electron-dense deposits at the bile canaliculus. In contrast, the CEA-positive area included bile canaliculus as well as basolateral aspects of hepatocytes. CD25 expression levels did not differ significantly among different disease states. CONCLUSION: This study provides the first evidence that CD25 is a novel marker of bile canaliculus. Characteristics of CD25 expression may shed light on immunohistochemistry and immunofluorescence analysis of bile canaliculus in both basic and clinical hepatic investigations.
Assuntos
Canalículos Biliares/metabolismo , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Canalículos Biliares/patologia , Biomarcadores/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Imunofluorescência , Rejeição de Enxerto/metabolismo , Rejeição de Enxerto/patologia , Hepatite B/metabolismo , Hepatite B/patologia , Humanos , Imuno-Histoquímica/métodos , Cirrose Hepática Biliar/metabolismo , Cirrose Hepática Biliar/patologia , Hepatopatias/metabolismo , Hepatopatias/patologia , Transplante de Fígado , Microscopia Imunoeletrônica/métodosRESUMO
OBJECTIVE: To construct an apparatus for the oxygen uptake measurement of rats exposed to hypobaric hypoxia at different simulated altitude. METHODS: The capacity of this apparatus was about 0.01 m3. It included animal experimental cabin, reference cabin, altimeter, altitude vertical velocity indicator, pressure difference inductor and oxygen compensator, low scale manometer, soda lime and calcium chloride, small fan, thermometer, circulating water system and vacuum pump. The oxygen uptake of the rats at 6 000 m, 4 000 m and 1 000 m simulated altitude was measured using this apparatus. RESULTS: The oxygen uptake of the rats at 50 m, 4 000 m and 6 000 m simulated altitude was (24.4 +/- 2.1), (10.8 +/- 2.0) and (8.8 +/- 1.6) ml O2/(kg x min) respectively (average +/- s, n = 10). The oxygen uptake decreased as altitude increased. CONCLUSION: This apparatus can be used to measure the oxygen uptake of the rats at different simulated altitude.