The present invention relates to the field of (vector) vaccines, and especially to novel EHV's having an inactivation of UL18 and/or UL8. The present invention further concerns related expression cassettes and vectors, which are suitable to express genes of interest, especially antigen encoding sequences. The viral vectors of the present invention are useful for producing an immunogenic composition or vaccine.

The present invention relates to virus growth media that improve the yield of alpha-herpesviruses (e.g., HSV-2) grown in cell cultures. The growth media of the invention include two additives, a disaccharide and a lipid mixture, that can be added to serum-free or serum-enriched growth media to improve the efficiency of virus production. The invention further provides methods of producing alpha-herpesviruses (e.g., HSV-2) in such growth media.

The present invention relates to the field of (vector) vaccines, and especially to novel EHV's having an inactivation of UL18 and/or UL8. The present invention further concerns related expression cassettes and vectors, which are suitable to express genes of interest, especially antigen encoding sequences. The viral vectors of the present invention are useful for producing an immunogenic composition or vaccine.

Kaposi's sarcoma-associated herpesvirus (KSHV) is an opportunistic pathogen causing Kaposi's sarcoma. It is capable of establishing latent infection, which can be reactivated to engage lytic infection for progeny production. KSHV contains a ?165 kilobase DNA genome predicted to encode at least 90 open reading frames (ORFs). In this report, we generated 91 KSHV mutants, each characterized by the disruption of a single viral ORF. The growth of these mutants in cultured cells was examined to systematically investigate the necessity of each ORF for viral latency, reactivation, and lytic replication. Salient aspects are (a) 44 ORFs are essential for viral lytic replication in cultured cells and 47 are nonessential; (b) KSHV reactivation can be positively or negatively regulated by specific viral ORFs; and (c) ORFs identified to regulate viral reactivation encode functions modulating both innate and adaptive immune responses. The intersection of viral immunomodulatory genes controlling reactivation suggests that KSHV engages in a concerted effort to communicate and respond to the host immune system for reactivation and replication using a viral sensory network. Our results imply a novel mechanism in which reactivation of KSHV is actively controlled by the virus in response to its surrounding environment, leading to the opportunistic nature of viral diseases that are strongly correlated to the host's immune status and conditions.

The present invention discloses a cyprinid herpesvirus II-sensitive brain tissue cell line of Carassius auratus gibelio and establishing method and use thereof. The cell line is deposited in China Center for Type Culture Collection under an accession number of CCTCC No: C2013179. The brain tissue cell line of Carassius auratus gibelio is in good growth state and sensitive to CyHV-2 that is presently hardly cultured with ordinary fish cell lines. After six passages of CyHV-2 in GiCB cells, viral nucleic acid can still be detected and a cytopathic effect is stable. When ultrathin microscopic sections are prepared from cells having the cytopathic effect, considerable mature CyHV-2 virions and their replication process can be observed in GiCB cells. The construction method of the brain tissue cell line of Carassius auratus gibelio of the present invention has high repeatability, scientific and reasonable conditions.

This application discloses a method for purifying pDNA, particularly pDNA that that can be used to produce RNA, the RNA preferably encoding a therapeutic or immunogenic peptide or polypeptide. The pDNA can be grown in a bacteria such as E. coli by culturing or fermenting bacteria containing the plasmid and obtaining and purifying the pDNA. The present method allows the pDNA to be obtained in high yield and with high purity. In one embodiment of the invention, the level of all non-pDNA materials can be significantly reduced by the process. In some embodiments, the ratio of supercoiled plasmid DNA (scDNA) to non-supercoiled pDNA (non-scDNA, such as open circular plasmid DNA (ocDNA)) can be increased by one or more process steps that separate or allow for separation of scDNA and ocDNA or process steps that increase the amount of scDNA to ocDNA.

This application describes a modified herpes simplex virus type 1 (HSV-1), capable of being efficiently produced in suspension cell culture, and a method of producing HSV-1 vectors in suspension cell culture.