Expression of recombinant adeno-associated viral containing human thyrotropin receptor A subunit in vivo and in vitro / 西安交通大学学报(医学版)
Journal of Xi'an Jiaotong University(Medical Sciences)
;
(6): 678-682, 2020.
Artigo
em Chinês
| WPRIM
| ID: wpr-843844
ABSTRACT
Objective:
To investigate the expression and expression efficiency of recombinant adeno-associated viral containing human thyrotropin receptor A subunit (rAAV2/9-hTSHR289-IRES-ZS Green) in mouse embryonic fibroblasts (NIH3T3) or in mice.Methods:
NIH3T3 was infected by rAAV2/9-hTSHR289-IRES-ZsGreen and its TSHR289 protein expression was detected by Western blotting. Female BALB/c mice were intramuscularly injected with different doses of rAAV2/9-hTSHR289-IRES-ZsGreen; the expressions of the target protein in different organs were determined by immunofluorescence while the serum TSHR antibody (TRAb) titer was determined by radioimmunoassay.Results:
NIH3T3 cells transfected with rAAV2/9-hTSHR289-IRES-ZsGreen expressed hTSHR289 protein both at 48 h and 72 h. The expression of target protein at 48 h or 72 h was significantly higher than that at 24 h (P<0.05). There were different levels of TSHR289 expression in leg skeletal muscle, heart, liver and spleen under fluorescence microscope. The results of radioimmunoassay showed that the higher dose injection produced a higher titer of TSHR antibody, but only the TRAb level in high dose and control injection exhibited a statistical difference (P<0.05) at week 4, week 8 and week 12. Furthermore, strong antibody response was observed in the mice injected with high dose and medium dose of rAAV2/9-hTSHR289-IRES-ZsGreen at week 4 and gradually weakened between 4 and 8 weeks. In addition, the antibody lasted for 12 weeks.Results:
rAAV2/9-hTSHR289-IRES-ZsGreen can highly express hTSHR289 protein in vivo and in vitro, and the hTSHR289 protein displays strong immunogenicity. It indicates that rAAV2/9-hTSHR289-IRES-ZsGreen might become a more effective means of preparing Graves' disease model.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Journal of Xi'an Jiaotong University(Medical Sciences)
Ano de publicação:
2020
Tipo de documento:
Artigo
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