Use of a negative selectable marker for rapid selection of recombinant vaccinia virus

White, Stacy D.; Conwell, Kip; Langland, Jeffrey; Jacobs, Bertram L.

doi:10.2144/000113667

Cited by 16 publications

(18 citation statements)

References 22 publications

(18 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A negative selection cassette, PNR, was inserted into the TK locus of the virus by linear in vivo recombination. The cassette expresses the coumermycin dimerization domain of Escherichia coli gyrase B fused to the catalytic domain of mammalian dsRNA-dependent protein kinase, making the virus sensitive to coumermycin (described in [ 32 ]) following insertion; also in the cassette is a neomycin resistance gene. Plaques from the IVR were screened for neomycin resistance to identify viruses containing the cassette: NYVAC-KC-PNR.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Targeting HIV-1 Env gp140 to LOX-1 Elicits Immune Responses in Rhesus Macaques

Zurawski

Flamar

et al. 2016

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Improved antigenicity against HIV-1 envelope (Env) protein is needed to elicit vaccine-induced protective immunity in humans. Here we describe the first tests in non-human primates (NHPs) of Env gp140 protein fused to a humanized anti-LOX-1 recombinant antibody for delivering Env directly to LOX-1-bearing antigen presenting cells, especially dendritic cells (DC). LOX-1, or 1ectin-like oxidized low-density lipoprotein (LDL) receptor-1, is expressed on various antigen presenting cells and endothelial cells, and is involved in promoting humoral immune responses. The anti-LOX-1 Env gp140 fusion protein was tested for priming immune responses and boosting responses in animals primed with replication competent NYVAC-KC Env gp140 vaccinia virus. Anti-LOX-1 Env gp140 vaccination elicited robust cellular and humoral responses when used for either priming or boosting immunity. Co-administration with Poly ICLC, a TLR3 agonist, was superior to GLA, a TLR4 agonist. Both CD4+ and CD8+ Env-specific T cell responses were elicited by anti-LOX-1 Env gp140, but in particular the CD4+ T cells were multifunctional and directed to multiple epitopes. Serum IgG and IgA antibody responses induced by anti-LOX-1 Env gp140 against various gp140 domains were cross-reactive across HIV-1 clades; however, the sera neutralized only HIV-1 bearing sequences most similar to the clade C 96ZM651 Env gp140 carried by the anti-LOX-1 vehicle. These data, as well as the safety of this protein vaccine, justify further exploration of this DC-targeting vaccine approach for protective immunity against HIV-1.

show abstract

Section: Methodsmentioning

confidence: 99%

“…Neutralization was measured as a function of reductions in luciferase (Luc) reporter gene expression after a single round of infection in TZM-bl cells as described [ 32 ]. TZM-bl cells were obtained from the NIH AIDS Research and Reference Reagent Program, contributed by John Kappes and Xiaoyun Wu.…”

Section: Methodsmentioning

confidence: 99%

Targeting HIV-1 Env gp140 to LOX-1 Elicits Immune Responses in Rhesus Macaques

Zurawski

Flamar

et al. 2016

PLoS ONE

Self Cite

View full text Add to dashboard Cite

show abstract

“…These were selected to include proteins with different functions and timing of expression. To achieve this, we constructed 3 different ASFV libraries, a gene library for DNA vaccination ( 15 – 17 ), a recombinant vaccinia virus (rVACV) library ( 18 – 20 ), and a library for expression and capture of individual recombinant proteins using an Escherichia coli in vitro transcription and translation system. We immunized pigs with pools of these ASFV antigens delivered by a DNA prime and recombinant vaccinia virus boost and ranked the immune responses to the individual captured proteins.…”

Section: Introductionmentioning

confidence: 99%

Immunization of Pigs by DNA Prime and Recombinant Vaccinia Virus Boost To Identify and Rank African Swine Fever Virus Immunogenic and Protective Proteins

Jancovich

Chapman

Hansen

et al. 2018

J Virol

View full text Add to dashboard Cite

African swine fever virus (ASFV) causes an acute hemorrhagic fever in domestic pigs, with high socioeconomic impact. No vaccine is available, limiting options for control. Although live attenuated ASFV can induce up to 100% protection against lethal challenge, little is known of the antigens which induce this protective response. To identify additional ASFV immunogenic and potentially protective antigens, we cloned 47 viral genes in individual plasmids for gene vaccination and in recombinant vaccinia viruses. These antigens were selected to include proteins with different functions and timing of expression. Pools of up to 22 antigens were delivered by DNA prime and recombinant vaccinia virus boost to groups of pigs. Responses of immune lymphocytes from pigs to individual recombinant proteins and to ASFV were measured by interferon gamma enzyme-linked immunosorbent spot (ELISpot) assays to identify a subset of the antigens that consistently induced the highest responses. All 47 antigens were then delivered to pigs by DNA prime and recombinant vaccinia virus boost, and pigs were challenged with a lethal dose of ASFV isolate Georgia 2007/1. Although pigs developed clinical and pathological signs consistent with acute ASFV, viral genome levels were significantly reduced in blood and several lymph tissues in those pigs immunized with vectors expressing ASFV antigens compared with the levels in control pigs.IMPORTANCE The lack of a vaccine limits the options to control African swine fever. Advances have been made in the development of genetically modified live attenuated ASFV that can induce protection against challenge. However, there may be safety issues relating to the use of these in the field. There is little information about ASFV antigens that can induce a protective immune response against challenge. We carried out a large screen of 30% of ASFV antigens by delivering individual genes in different pools to pigs by DNA immunization prime and recombinant vaccinia virus boost. The responses in immunized pigs to these individual antigens were compared to identify the most immunogenic. Lethal challenge of pigs immunized with a pool of antigens resulted in reduced levels of virus in blood and lymph tissues compared to those in pigs immunized with control vectors. Novel immunogenic ASFV proteins have been identified for further testing as vaccine candidates.

show abstract

“…The technology was developed in the 1980s [34,35], and has since been in widespread use. Several selection procedures have been developed, which include sorting on the basis of thymidine kinase positive or negative status [34,35] and antibiotic resistance [36,37]. To add to the versatility of the methodology, linear DNA such as a polymerase chain reaction product, may be used instead of plasmid DNA.…”

Section: Recombinant Viral Vectors For Vaccinationmentioning

confidence: 99%

Gene Transfer for Prophylaxis and Therapy of Viral Infections

Arbuthnot

2015

Gene Therapy for Viral Infections

View full text Add to dashboard Cite

Use of a negative selectable marker for rapid selection of recombinant vaccinia virus

Cited by 16 publications

References 22 publications

Targeting HIV-1 Env gp140 to LOX-1 Elicits Immune Responses in Rhesus Macaques

Targeting HIV-1 Env gp140 to LOX-1 Elicits Immune Responses in Rhesus Macaques

Immunization of Pigs by DNA Prime and Recombinant Vaccinia Virus Boost To Identify and Rank African Swine Fever Virus Immunogenic and Protective Proteins

Gene Transfer for Prophylaxis and Therapy of Viral Infections

Contact Info

Product

Resources

About