Journal: Molecular Therapy

Article Title: Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF

doi: 10.1038/mt.2010.161

Figure Lengend Snippet: Ad5/3-D24-GMCSF induces cell killing and expression of functionally active GMCSF in vitro. Viability of (a) MDA-MB-436 and (b) A549 cells after infection with Ad5luc1, Ad5wt, and Ad5/3-D24-GMCSF. (c) GMCSF secretion by A549 cells after infection with 100 VP/cell Ad5/3-D24-GMCSF. (d) TF1 cells, whose viability is dependent on functional human GMCSF (hGMCSF) were cultured in the presence of 2 ng/ml hGMCSF or indicated amount of filtered supernatant from Ad5/3-D24-GMCSF-infected cells. Normal growth media was used as negative control. Viability of cells was determined after 5 days; viability of cells with commercial hGMSCF was set as 100%. hGMCSF, human granulocyte–macrophage colony–stimulating factor; VP, virus particles.

Article Snippet: 20 , 30 Ad5wt is wild-type Ad5 strain Ad300 from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, In Vitro, Infection, Functional Assay, Cell Culture, Negative Control, Virus

Journal: Molecular Therapy

Article Title: Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF

doi: 10.1038/mt.2010.161

Figure Lengend Snippet: Antitumor efficacy and tumor selectivity of Ad5/3-D24-GMCSF in immune-competent Syrian hamsters. Syrian hamsters were inoculated subcutaneously with HapT1 cells. (a,b) Tumors were injected with 1 × 108 VP/tumor on days indicated by arrows, 1/5 of dose was given intravenously on day 1. NaCl was used as mock treatment. *P ≤ 0.05 against mock. In b the effect of low-dose cyclophosphamide (CP) in conjunction with Ad5/3-D24-GMCSF was tested and found to increase efficacy. Figure indicates tumor volumes recorded 15 days after injection of virus. (c,d) HapT1 tumors were grown and injected once with 1 × 108 VP/tumor Ad5/3-D24-GMCSF. (c) Virus replication was studied with quantitative PCR. To evaluate tumor selectivity of the virus, livers of non-tumor-bearing hamsters were injected and no replication was seen. Viral E4 copy number was normalized to genomic DNA with GAPDH primers. (d) Human GMCSF concentration was measured in virus injected tumors, serum and livers of tumor-bearing hamsters and livers of non-tumor-bearing hamsters injected into the liver. Data are expressed as mean ± SEM, *P ≤ 0.05 and ***P ≤ 0.005 versus the 0.5-hour time point. GMCSF, granulocyte–macrophage colony–stimulating factor; VP, virus particles.

Article Snippet: 20 , 30 Ad5wt is wild-type Ad5 strain Ad300 from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Injection, Virus, Real-time Polymerase Chain Reaction, Concentration Assay

Journal: Molecular Therapy

Article Title: Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF

doi: 10.1038/mt.2010.161

Figure Lengend Snippet: Radiological responses to treatment with Ad5/3-D24-GMCSF and survival after treatment. (a) Pre- and (b) post-treatment computed tomography scans of K75 with complete resolution of pleural effusion, O129 with reduction of peritoneal tumors, and I98 illustrating reduction of a liver metastases (note: post-treatments scan for this patient is a magnetic resonance image). (c) Kaplan–Meier analysis of the survival of patients. Censored refers to patients who were still alive at the time of submission of the manuscript.

Article Snippet: 20 , 30 Ad5wt is wild-type Ad5 strain Ad300 from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Computed Tomography

Journal: Molecular Therapy

Article Title: Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF

doi: 10.1038/mt.2010.161

Figure Lengend Snippet: Ad5/3-D24-GMCSF treatment influences adenovirus- and tumor-specific cytotoxic T-lymphocytes. Total peripheral blood mononuclear cells were isolated from treated patients before and a month after treatment and pulsed with (a) an adenovirus type 5 penton-derived peptide pool and (b) a survivin-derived peptide pool. Interferon-γ ELISPOT was performed. The value before treatment was assigned 100 and the value after treatment is expressed relative to that. Spot forming colonies (SFC) are expressed as a mean of triplicate experiments, and background (SFC without peptide) was subtracted. Patients were grouped depending on the change in SFC after treatment; increased (>50%), no change, and decreased (>50%).

Article Snippet: 20 , 30 Ad5wt is wild-type Ad5 strain Ad300 from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Isolation, Derivative Assay, Enzyme-linked Immunospot

Journal: Molecular Therapy

Article Title: Treatment of Cancer Patients With a Serotype 5/3 Chimeric Oncolytic Adenovirus Expressing GMCSF

doi: 10.1038/mt.2010.161

Figure Lengend Snippet: 5/3 chimeric adenoviruses are able to transfect and kill cells in ex vivo ascites and pleural effusion samples. Before treatment cells from (a) pleural effusion of V136 and (b) ascites of K75 were infected with Ad5luc1 (Ad5 capsid) and Ad5/3luc1 (Ad5/3 capsid) at 5,000 VP/cell and analyzed for luciferase expression (expressed as relative light units, RLU). ***P < 0.005 against Ad5luc1. Cells from pretreatment samples of pleural effusion of (c) V136 and (d) M137 were infected with 100 VP/cell Ad5/3-D24-GMCSF or the nonreplicative control. Viability of cells was analyzed 6 days later. ***P < 0.005 against uninfected cells. (e) Copy numbers of Ad5/3-D24-GMCSF were analyzed with quantitative PCR from serum samples and a post-treatment ascites sample of O82. NA, not analyzed; VP, virus particles.

Article Snippet: 20 , 30 Ad5wt is wild-type Ad5 strain Ad300 from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Ex Vivo, Infection, Luciferase, Expressing, Control, Real-time Polymerase Chain Reaction, Virus

Journal: Biologicals : journal of the International Association of Biological Standardization

Article Title: Evaluation of next-generation sequencing performance for in vitro detection of viruses in biological products.

doi: 10.1016/j.biologicals.2023.101739

Figure Lengend Snippet: Fig. 2. Overview of NGS evaluation for in vitro detection of viruses. Step 1: Sample preparation. Total RNA was extracted from Ad5-infected cells and non-infected cells. Total RNAs from Ad5-infected and non-infected cells were mixed at a ratio of 1:10 to 1:108. Step 2: Sequencing. The library was prepared for HiSeq X Ten and GridION. Step 3: Analysis. The flow of the data analysis and references used in this study are shown.

Article Snippet: Human Ad5 (Strain: Adenoid 75, VR-1516) was obtained from ATCC.

Techniques: In Vitro, Sample Prep, Infection, Sequencing

Journal: Biologicals : journal of the International Association of Biological Standardization

Article Title: Evaluation of next-generation sequencing performance for in vitro detection of viruses in biological products.

doi: 10.1016/j.biologicals.2023.101739

Figure Lengend Snippet: Fig. 1. The infectivity of Ad5 in Vero cells. Vero cells were infected with Ad5 at an MOI 0.01 and 0.1 for 1h. At 1, 4, and 7 days after infection, the cells were observed using microscopy. Total RNA was extracted from Ad5-infected cells at each time point, and the number of copies of Ad5 mRNA was determined using RT-qPCR.

Article Snippet: Human Ad5 (Strain: Adenoid 75, VR-1516) was obtained from ATCC.

Techniques: Infection, Microscopy, Quantitative RT-PCR

Journal: Microbiology Spectrum

Article Title: A novel simian adenovirus-vectored COVID-19 vaccine elicits effective mucosal and systemic immunity in mice by intranasal and intramuscular vaccination regimens

doi: 10.1128/spectrum.01794-23

Figure Lengend Snippet: Production of Sad23L-nCoV-S and Ad5-nCoV-S vaccines. (a) Images of Sad23L-nCoV-S and (b) Ad5-nCoV-S vaccine stains by negatively stained with 2% phosphotungstic acid under transmission electron microscope (TEM). (c) Immunofluorescence staining for SARS-CoV-2 S protein in adenovirus-vectored vaccine infected HEK293A cells. (d) Western blot analysis for Sad23L-nCoV-S or Ad5-nCoV-S vaccine infected HEK-293A cells by rabbit polyclonal antibody specific to RBD of SARS-CoV-2.

Article Snippet: HEK293 cells infected with Sad23L-nCoV-S and Ad5-nCoV-S vaccine strains or vector controls were fixed in 4% cold paraformaldehyde, blocked, and incubated with mouse monoclonal antibody to SARS-CoV-2 S protein (Sino Biological, China) in 1% BSA/PBST at 4°C overnight.

Techniques: Staining, Transmission Assay, Microscopy, Immunofluorescence, Infection, Western Blot