Construction of conditionally replicating adenovirus expressing staphylococcal enterotoxin A gene: potential usefulness for anti-tumor therapies.

Abstract

OBJECTIVE:

The aim of this study was to construct a conditionally replicating adenovirus pPE3-SEA expressing staphylococcal enterotoxin A (SEA) gene.

MATERIALS AND METHODS:

A full-length SEA gene fragment was cloned into pENTR12 plasmid to obtain a recombinant viral plasmid pENTR12-SEA. The pENTR12-SEA plasmid was co-transfected into HEK293 cells along with pPE3-ccdB, which encoded for the virus backbone, to generate recombinant adenovirus pPE3-SEA vector. Amplified pPE3-SEA vectors were purified, and viral titer was determined using the 50% tissue culture infective dose method.

RESULTS:

The PCR, restriction enzyme digestion, and sequence analyses proved successful construction of replicating oncolytic adenovirus pENTR12-SEA and recombinant SEA expressing oncolytic adenovirus pPE3-SEA. The viral titer was 2.5 × 1010 pfu/ml.

CONCLUSIONS:

We successfully constructed conditionally replicating adenovirus pPE3-SEA which can be utilized for experimental studies of tumor-targeted therapies.

• Authors: P-Y Zhang, L Hao, Z-G Zhang, B-Z Dong, D Yang, X-L Wang, X-J Xuan, Z Yan, L Qing, Z-D Shi, D Liu, C-H Han
• Journal: European review for medical and pharmacological sciences (Eur Rev Med Pharmacol Sci) Vol. 18 Issue 16 Pg. 2258-63 (Aug 2014) ISSN: 2284-0729 [Electronic] Italy
• PMID: 25219823 (Publication Type: Journal Article)

Chemical References

• Enterotoxins
• enterotoxin A, Staphylococcal

Topics

• Adenoviridae (genetics, physiology)
• Enterotoxins (genetics)
• Genetic Therapy
• Genetic Vectors
• HEK293 Cells
• Humans
• Neoplasms (therapy)
• Transfection
• Virus Replication