Inactivation of herpes simplex type 1 gene vector on immobilized metal affinity chromatography: Oxidative damage by hydroxyl free radicals and its prevention

Jiang, Canping; Ataai, Mohammad M.; Ozuer, Ali; Krisky, David; Wechuck, James B.; Pornsuwan, Soraya; Pourarian, Fazard

doi:10.1002/bit.20943

Cited by 7 publications

(4 citation statements)

References 49 publications

(66 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…27 Physical titers in genome copies/ml were determined by real-time quantitative PCR for the ICP47 gene, as described. 51 Plasmids. pCEA-HveA for expression of scCEA-HveA was constructed by ligation of two PCR products.…”

Section: Methodsmentioning

confidence: 99%

Bispecific Adapter-Mediated Retargeting of a Receptor-Restricted HSV-1 Vector to CEA-Bearing Tumor Cells

et al. 2011

View full text Add to dashboard Cite

The safety and efficacy of viral therapies for solid tumors can be enhanced by redirecting the virus infection to tumor-specific cell-surface markers. Successful retargeting of herpes simplex virus type 1 (HSV-1) has been achieved using vectors that carry a modified envelope glycoprotein D (gD) engineered to interact directly with novel receptors. In addition, soluble bridging molecules (adapters) have been used to link gD indirectly to cell-specific receptors. Here, we describe the development of an adapter connecting gD to the common tumor antigen carcinoembryonic antigen (CEA). The adapter consisted of a CEA-specific single-chain antibody fused to the gD-binding region of the gD receptor, herpes virus entry mediator (HVEM). We used this adapter in combination with a vector that is detargeted for recognition of the widely expressed gD receptor nectin-1, but retains an intact binding region for the less common HVEM. We show that the adapter enabled infection of HSV-resistant Chinese hamster ovary (CHO) cells expressing ectopic CEA and nectin-1/CEA-bearing human gastric carcinoma cells that are resistant to the vector alone. We observed cell-to-cell spread following adapter-mediated infection in vitro and reduced tumor growth in vivo, indicating that this method of vector retargeting may provide a novel strategy for tumor-specific delivery of tumoricidal HSV.

show abstract

Section: Methodsmentioning

confidence: 99%

Bispecific Adapter-Mediated Retargeting of a Receptor-Restricted HSV-1 Vector to CEA-Bearing Tumor Cells

et al. 2011

View full text Add to dashboard Cite

show abstract

“…We determined the titers of the different viral preparations on J1.1-2 and B78H1 cells and their receptor-bearing derivatives by using a standard plaque formation assay and normalized the results, expressed as PFU/ml, to genome titers (gc per ml) as determined by real-time qPCR for the viral ICP47 gene (22). As shown in Fig.…”

Section: Establishment Of Cell Lines Expressing Hvem or Nectin-1mentioning

confidence: 99%

Generation of Herpesvirus Entry Mediator (HVEM)-Restricted Herpes Simplex Virus Type 1 Mutant Viruses: Resistance of HVEM-Expressing Cells and Identification of Mutations That Rescue Nectin-1 Recognition

Uchida

Shah²,

Ozuer³

et al. 2009

J Virol

Self Cite

View full text Add to dashboard Cite

Both initial infection and cell-to-cell spread by herpes simplex virus type 1 (HSV-1) require the interaction of the viral glycoprotein D (gD) with an entry receptor on the cell surface. The two major HSV entry receptors, herpesvirus entry mediator (HVEM) and nectin-1, mediate infection independently but are coexpressed on a variety of cells. To determine if both receptors are active in these instances, we have established mutant viruses that are selectively impaired for recognition of one or the other receptor. In plaque assays, these viruses showed approximately 1,000-fold selectivity for the matched receptor over the mismatched receptor. Separate assays showed that each virus is impaired for both infection and spread through the mismatched receptor. We tested several human tumor cell lines for susceptibility to these viruses and observed that HT29 colon carcinoma cells are susceptible to infection by nectin-1-restricted virus but are highly resistant to HVEM-restricted virus infection, despite readily detectable HVEM expression on the cell surface. HVEM cDNA isolated from HT29 cells rendered HSV-resistant cells permissive for infection by the HVEM-restricted virus, suggesting that HT29 cells lack a cofactor for HVEM-mediated infection or express an HVEM-specific inhibitory factor. Passaging of HVEM-restricted virus on nectin-1-expressing cells yielded a set of gD missense mutations that each restored functional recognition of nectin-1. These mutations identify residues that likely play a role in shaping the nectin-1 binding site of gD. Our findings illustrate the utility of these receptor-restricted viruses in studying the early events in HSV infection.Entry of herpes simplex virus type 1 (HSV-1) into cells requires the envelope glycoprotein D (gD), gB, gH, and gL (5, 13, 21, 27). After virion adsorption to cell surface proteoglycans, gD binds to one of its receptors, herpesvirus entry mediator (HVEM, or HveA), nectin-1 (HveC), or 3-O-sulfated heparan sulfate (3-O-S HS) (19,32,43). Receptor binding results in a conformational change in gD, which is believed to activate either or both gB and gH as mediators of fusion between the viral envelope and the plasma membrane or an endosomal membrane (7,18,20,26,31,34,35). Fusion leads to nucleocapsid release into the cytoplasm for subsequent viral genome delivery to the nucleus. Since productive HSV-1 infection is set in motion by the gD-receptor interaction, cellular susceptibility to HSV-1 is initially determined by the presence of one or more functional gD receptors on the cell surface.In addition to supporting HSV-1 entry, gD receptors also support HSV-1 cell-to-cell spread (8,41). However, it is not clear whether they act by the same mechanism in entry and spread. For example, gD mutants that allow gD-receptor-independent virus spread but not receptor-independent entry have been isolated (39). Likewise, gD is required for entry of the related alphaherpesvirus pseudorabies virus (PRV) but not for PRV lateral spread (2,36,37,40). Two additional glycoproteins, gE an...

show abstract

“…However, these studies were carried out with vaccine purified using centrifugation-based methods which are not readily scaled for commercial production. Indeed, a number of groups have defined laboratory-scale procedures for purification of herpes viruses based upon centrifugation [28], [29], gradients [30], [31], [32], [33], filtration [34] and affinity chromatography [35], [36].…”

Section: Introductionmentioning

confidence: 99%

High-Purity Preparation of HSV-2 Vaccine Candidate ACAM529 Is Immunogenic and Efficacious In Vivo

et al. 2013

View full text Add to dashboard Cite

Genital herpes is a sexually transmitted infection (STI) caused by herpes simplex virus 2 (HSV-2) and to a lesser extent herpes simplex virus 1 (HSV-1). Infection by HSV-2 is life-long and is associated with significant cost to healthcare systems and social stigma despite the highly prevalent nature of the disease. For instance, the proportion of HSV-2 seropositive to seronegative adults is approximately 1 in 5 in the US and greater than 4 in 5 in some areas of sub-Saharan Africa. The replication-defective vaccine strain virus dl5-29 was re-derived using cells appropriate for GMP manufacturing and renamed ACAM529. Immunization with dl5-29 was previously reported to be protective both in mice and in guinea pigs, however these studies were performed with vaccine that was purified using methods that cannot be scaled for manufacturing of clinical material. Here we describe methods which serve as a major step towards preparation of ACAM529 which may be suitable for testing in humans. ACAM529 can be harvested from infected cell culture of the trans-complementing cell line AV529 clone 19 (AV529-19) without mechanical cell disruption. ACAM529 may then be purified with respect to host cell DNA and proteins by a novel purification scheme, which includes a combination of endonuclease treatment, depth filtration, anion-exchange chromatography and ultrafiltration/diafiltration (UF/DF). The resultant virus retains infectivity and is ∼ 200-fold more pure with respect to host cell DNA and proteins than is ACAM529 purified by ultracentrifugation. Additionally, we describe a side-by-side comparison of chromatography-purified ACAM529 with sucrose cushion-purified ACAM529, which shows that both preparations are equally immunogenic and protective when tested in vivo.

show abstract

Inactivation of herpes simplex type 1 gene vector on immobilized metal affinity chromatography: Oxidative damage by hydroxyl free radicals and its prevention

Cited by 7 publications

References 49 publications

Bispecific Adapter-Mediated Retargeting of a Receptor-Restricted HSV-1 Vector to CEA-Bearing Tumor Cells

Bispecific Adapter-Mediated Retargeting of a Receptor-Restricted HSV-1 Vector to CEA-Bearing Tumor Cells

Generation of Herpesvirus Entry Mediator (HVEM)-Restricted Herpes Simplex Virus Type 1 Mutant Viruses: Resistance of HVEM-Expressing Cells and Identification of Mutations That Rescue Nectin-1 Recognition

High-Purity Preparation of HSV-2 Vaccine Candidate ACAM529 Is Immunogenic and Efficacious In Vivo

Contact Info

Product

Resources

About