RESUMO
Cationic PLG nanoparticles and liposome were prepared and used as package molecules to pack up pUC18-CpG. The effects of the packed pUC18-CpG on the cellular and humoral immune responses were detected in the mice that were inoculated with pig paratyphoid vaccine. The results showed that compared with the control, the amount of IgG and the titre of specific antibody were significantly increased in the sera of mice immunized with the CpG plasmid entrapped by cationic PLG nanoparticles; the proliferation and induced IL-2 bioactivity of lymphocytes were significantly enhanced in the spleen of the immunized mice; the stimulatory effect of cationic PLG nanoparticles was similar to or stronger than that of cationic liposome. These indicated that cationic PLG nanoparticle could be employed as an effective package molecule to promote the immunostimulatory effect of pUC18-CpG.
Assuntos
Animais , Masculino , Camundongos , Adjuvantes Imunológicos , Farmacologia , Imunoglobulina G , Sangue , Interleucina-2 , Sangue , Lipossomos , Camundongos Endogâmicos BALB C , Nanoestruturas , Oligodesoxirribonucleotídeos , Farmacologia , Suínos , Vacinas Tíficas-Paratíficas , Alergia e ImunologiaRESUMO
The experiment was conducted to prepare chitosan nanoparticles (CNP), to entrap VRMIL-2 with CNP, the eukaryotic VR1020 expression plasmid containing murine IL-2 gene (mlL-2), and to investigate the expression in vivo and the regulatory effect of mIL-2 on immune-response and immuno-protection in mice inoculated muscularly with CNP entrapped VMIL-2 at 21 days old. The results showed that IgG, IgM and IgA contents increased to different degrees in the sera from the inoculated mice, which were remarkably higher than those of the controls inoculated VR1020 packed with CNP (P<0.05); so were the IL-2, IL-4 and IL-6 contents in the sera of the immunized mice. The number of white blood cells and lymphocytes significantly increased respectively in the vaccinated mice, compared with those of controls. These mice were orally challenged with virulent E. coli 35 days post-inoculation, and all the immune responses were significantly higher than those of the control except the number of neutrophils. The mice inoculated with VRMIL-2 survived healthily, while the mice of control group were ill with the evident lesions. Although there are no remarkable differences between the cellular and humoral immune indexes of mice inoculated with CNP-VRMIL-2 and nude VRMIL-2 (P>0.05), the dosage of CNP-VRMIL-2 is only one fifth of the VRMIL-2. These indicated that entrapment of mIL-2 gene with chitosan nanoparticles could remarkably enhance the expression of mIL-2 in vivo, and significantly raise the levels of cellular and humoral immune, and increase the resistance of mice against E. coli infection. The results suggested that chitosan nanoparticles and IL-2 gene could be used as an effective immunoenhancer to increase the immunity of animals against infection.
Assuntos
Animais , Feminino , Camundongos , Adjuvantes Imunológicos , Farmacologia , Quitosana , Farmacologia , Infecções por Escherichia coli , Alergia e Imunologia , Imunoglobulina G , Sangue , Interleucina-2 , Genética , Farmacologia , NanoestruturasRESUMO
The main component of this pill is 2-Octadecanoic acid-4-Palmitic acid-2, 4-Pentanediyl ester separated from chloroform extract of neem oil. The microcapsules coated by the re-curdle method were fabricated with an average particle size of 100-180 microm. The morphological characteristics, incorporation efficiency, carrier reclamation efficiency of the microcapsule were investigated. Kunming mice were used in the experiment, and the anti-fertility effect of the microcapsule on the histology and apoptosis was studied by light and electron microscopy and the flow cytometry. The data obtained clearly indicated that the microcapsule could lead to the payload of medicine, the incorporation efficiency being 90%. After the microcapsules were given to the male mice orally, its anti-fertility effect came into being and could keep the mice in a state of reversible infertility for a long time. The results of histological study and flow cytometry indicate that the mechanism of its anti-fertility effect involves mainly the inhibition of sperm motility and the arrest of spermatogenic process.
Assuntos
Animais , Masculino , Camundongos , Cápsulas , Anticoncepcionais Masculinos , Farmacologia , Composição de Medicamentos , Glicerídeos , Química , Ácidos Palmíticos , Farmacologia , Tamanho da Partícula , Motilidade dos Espermatozoides , Espermatogênese , Ácidos Esteáricos , Farmacologia , Terpenos , QuímicaRESUMO
To construct the ss/HBsAg protein gene-engineering vaccine for developing the diagnosis and cure tumors in clinical medicine and promoting the growth in animal husbandry production. A pair of primers were designed according separately to the sequence of Somatostatin gene(S14) and HBsAg gene. Their gene fragments were separately amplified by using PCR and cloned, following sequencing, the DNA fragments were inserted into pBluescript vector. Then the ss/HBsAg chimera was constructed and was cloned into pPICZaA plasmid, and transformed into electroporated Pichia pastoris. High yield protein expression was obtained. Expressed protein was proved with high specificity and it's molecular weigh was about 28 KD identified by SDS-PAGE and Western blot.
Assuntos
Fusão Gênica Artificial , Clonagem Molecular , Expressão Gênica , Vetores Genéticos , Antígenos de Superfície da Hepatite B , Genética , Pichia , Metabolismo , Proteínas Recombinantes de Fusão , Somatostatina , GenéticaRESUMO
pFH23. These studies provide the possibility of further research on the expression of recombinant antigenic genes, the immunity of their expressed products and the protection of animals.