RESUMO
The limitation of traditional Ad vectors result in wide application of capsid-incorporation of antigens into adenovirus capsid proteins, but usually it can't rescue virus successfully when we engineered the hypervariable regions (HVRs) of hexon in adenovirus serotype 3(Ad3) vector. So we deleted or retained some amino acids in HVR1, HVR2, HVR5, HVR7 predicted by bioinformatics, constructed recombinant Ad3 vector pBRAddeltaE3GFP-mHexon, and transfected it into AD293 cell to confirm the influence on the virus rescue. These data of amino acids that can be deleted or retained in the HVRs of Ad3 vector should provide operating foundation for antigen capsid-incorporation strategy in human adenovirus serotype 3, and also lay the groundwork for application of expressing foreign antigens in the hexon of human adenovirus serotype 3 as a platform of multivalent vaccine vectors.
Assuntos
Adenovírus Humanos/genética , Biologia Computacional , Engenharia Genética/métodos , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/genética , Adenovírus Humanos/imunologia , Sequência de Aminoácidos , Linhagem Celular , DNA Recombinante/genética , Vetores Genéticos/genética , Humanos , Modelos Moleculares , Dados de Sequência Molecular , Plasmídeos/genética , Conformação Proteica , Especificidade da Espécie , Proteínas do Envelope Viral/imunologiaRESUMO
AIMS: Gender-related phenotypes in the cardiovascular system have been observed in various genetically modified mice. Here, we report that cardiac functions are significantly improved only in male CD38-null mice and we explore the potential mechanisms of the sexual dimorphism mediated by CD38 deficiency. MAIN METHODS: Cardiac functions of mice were measured by pressure-volume conductance catheter technique and echocardiography. Serum sex steroids were determined by radioimmunoassay. Relative mRNA levels of myocardial contractile-associated proteins in cardiomyocytes were analyzed by real-time PCR analysis. To clarify the effects of testosterone on the sexual dimorphism, flutamide, an androgen receptor antagonist, was subcutaneously infused into the male null mice for 6 weeks with an osmotic mini-pump. KEY FINDINGS: The myocardial contractility, contraction and relaxation velocities were significantly enhanced only in male CD38-null mice, in which the levels of serum testosterone were markedly elevated. The elevated testosterone levels in the null mice were correlated to an obvious decrease in expression of androgen receptor and dramatic increases in expressions of major genes involved in myocardial contraction, including ryanodine receptor type 2 (RyR2), sarcoplasmic reticular Ca(2+) ATPase (SERCA2) and Na(+)/Ca(2+)-exchanger protein 1 (NCX1), and α myosin heavy chain (α-MHC). More importantly, all of the alternations that were observed in the male null mice were almost completely restored by flutamide administration. SIGNIFICANCE: Elevated serum level of testosterone in the male CD38(-/-) mice enhances cardiac functions through upregulation of major calcium regulatory proteins, which improve our understanding on sex disparities and molecular mechanisms in the incidence and manifestation of heart diseases.