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    DSMZ cells syrian hamster cells hap t1
    Sequential administration of Ad-OSM and NDV-OSM improves survival of tumor-bearing hamsters and stimulates immune response against cancer cells. a Schematic representation of the experimental setting. Tumor-bearing hamsters were divided into 4 groups ( n = 6) that received the following treatments: Four local administrations of Ad-OSM (2.5 × 10 8 iu/hamster) or NDV-OSM (1 × 10 9 iu/hamster) at days 0, 3, 10 and 13 (Ad/Ad-OSM and NDV/NDV-OSM groups, respectively); or Ad-OSM (2.5 × 10 8 iu/hamster) at days 0 and 3, followed by NDV-OSM (1 × 10 9 iu/hamster) at days 10 and 13 (Ad/NDV-OSM group). b NAbs against Ad (top panel) or NDV (lower panel) were quantified in serum of hamsters at days 11 and 24. Ad/Ad-OSM, NDV/NDV-OSM and Ad/NDV-OSM groups are represented as black, grey and striped bars, respectively. c Progression of tumors in the different groups. Each line represents an individual hamster. d Percentage of surviving animals in each group. e Long-term survivors were sacrificed at day 70 and spleens were collected. Splenocytes were expanded for 5 days in the presence of irradiated <t>HaP-T1</t> cells and IL-2, and then exposed for 8 h with fresh HaP-T1 cells. Expression of IFNγ and FasL was determined by qRT-PCR and compared with splenocytes from a group of untreated tumor-bearing hamsters sacrificed 40 days after cell implantation (saline), or healthy, naïve hamsters. * p
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    Sequential administration of Ad-OSM and NDV-OSM improves survival of tumor-bearing hamsters and stimulates immune response against cancer cells. a Schematic representation of the experimental setting. Tumor-bearing hamsters were divided into 4 groups ( n = 6) that received the following treatments: Four local administrations of Ad-OSM (2.5 × 10 8 iu/hamster) or NDV-OSM (1 × 10 9 iu/hamster) at days 0, 3, 10 and 13 (Ad/Ad-OSM and NDV/NDV-OSM groups, respectively); or Ad-OSM (2.5 × 10 8 iu/hamster) at days 0 and 3, followed by NDV-OSM (1 × 10 9 iu/hamster) at days 10 and 13 (Ad/NDV-OSM group). b NAbs against Ad (top panel) or NDV (lower panel) were quantified in serum of hamsters at days 11 and 24. Ad/Ad-OSM, NDV/NDV-OSM and Ad/NDV-OSM groups are represented as black, grey and striped bars, respectively. c Progression of tumors in the different groups. Each line represents an individual hamster. d Percentage of surviving animals in each group. e Long-term survivors were sacrificed at day 70 and spleens were collected. Splenocytes were expanded for 5 days in the presence of irradiated HaP-T1 cells and IL-2, and then exposed for 8 h with fresh HaP-T1 cells. Expression of IFNγ and FasL was determined by qRT-PCR and compared with splenocytes from a group of untreated tumor-bearing hamsters sacrificed 40 days after cell implantation (saline), or healthy, naïve hamsters. * p

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: Sequential administration of Ad-OSM and NDV-OSM improves survival of tumor-bearing hamsters and stimulates immune response against cancer cells. a Schematic representation of the experimental setting. Tumor-bearing hamsters were divided into 4 groups ( n = 6) that received the following treatments: Four local administrations of Ad-OSM (2.5 × 10 8 iu/hamster) or NDV-OSM (1 × 10 9 iu/hamster) at days 0, 3, 10 and 13 (Ad/Ad-OSM and NDV/NDV-OSM groups, respectively); or Ad-OSM (2.5 × 10 8 iu/hamster) at days 0 and 3, followed by NDV-OSM (1 × 10 9 iu/hamster) at days 10 and 13 (Ad/NDV-OSM group). b NAbs against Ad (top panel) or NDV (lower panel) were quantified in serum of hamsters at days 11 and 24. Ad/Ad-OSM, NDV/NDV-OSM and Ad/NDV-OSM groups are represented as black, grey and striped bars, respectively. c Progression of tumors in the different groups. Each line represents an individual hamster. d Percentage of surviving animals in each group. e Long-term survivors were sacrificed at day 70 and spleens were collected. Splenocytes were expanded for 5 days in the presence of irradiated HaP-T1 cells and IL-2, and then exposed for 8 h with fresh HaP-T1 cells. Expression of IFNγ and FasL was determined by qRT-PCR and compared with splenocytes from a group of untreated tumor-bearing hamsters sacrificed 40 days after cell implantation (saline), or healthy, naïve hamsters. * p

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: Irradiation, Expressing, Quantitative RT-PCR

    Immunosuppression abrogates the antitumor effect of OVs expressing OSM. The experimental setting is as described in Fig. 6 . Treatment groups ( n = 6/7) included control (saline solution injection); cyclophosphamide (CP); sequential administration of Ad-OSM and NDV-OSM (Ad/NDV-OSM) and the same treatment in combination with cyclophosphamide (Ad/NDV-OSM + CP). a Progression of tumors in the different groups. Each line represents an individual hamster. The numbers indicate the number of animals with complete response/total. In the Ad/NDV-OSM group the individuals with progressive disease, partial response and complete response are indicated by red, blue and green lines, respectively. b Long-term survivors from the Ad/NDV-OSM were subjected to a subcutaneous re-challenge with HaP-T1 cells. The graphs indicate the progression of tumors in each animal. A group of untreated hamsters bearing pancreatic tumors were used as a control

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: Immunosuppression abrogates the antitumor effect of OVs expressing OSM. The experimental setting is as described in Fig. 6 . Treatment groups ( n = 6/7) included control (saline solution injection); cyclophosphamide (CP); sequential administration of Ad-OSM and NDV-OSM (Ad/NDV-OSM) and the same treatment in combination with cyclophosphamide (Ad/NDV-OSM + CP). a Progression of tumors in the different groups. Each line represents an individual hamster. The numbers indicate the number of animals with complete response/total. In the Ad/NDV-OSM group the individuals with progressive disease, partial response and complete response are indicated by red, blue and green lines, respectively. b Long-term survivors from the Ad/NDV-OSM were subjected to a subcutaneous re-challenge with HaP-T1 cells. The graphs indicate the progression of tumors in each animal. A group of untreated hamsters bearing pancreatic tumors were used as a control

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: Expressing, Injection

    Ad-OSM and NDV-OSM replicate in human and hamster PDAC cells. a Schematic representation of the main viruses used in this study. ITR, adenovirus inverted terminal repeat; ΔE1A 922–947 indicates the nucleotides deleted from the E1A gene; ΔE3 6.7/gp19K indicates the deletion of these genes in the E3 region; GFP, enhanced green fluorescence protein; TK, thymidine kinase from HSV-1; NP, P, M, F, HN and L are NDV genes. b - e Quantification of infectious particles produced at the indicated times in PANC-1 and HaP-T1 cells infected with the different viruses (representative results of at least 2 experiments performed in triplicate). Black columns correspond to cells treated with the hypoxia-mimetic drug CoCl 2. * p

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: Ad-OSM and NDV-OSM replicate in human and hamster PDAC cells. a Schematic representation of the main viruses used in this study. ITR, adenovirus inverted terminal repeat; ΔE1A 922–947 indicates the nucleotides deleted from the E1A gene; ΔE3 6.7/gp19K indicates the deletion of these genes in the E3 region; GFP, enhanced green fluorescence protein; TK, thymidine kinase from HSV-1; NP, P, M, F, HN and L are NDV genes. b - e Quantification of infectious particles produced at the indicated times in PANC-1 and HaP-T1 cells infected with the different viruses (representative results of at least 2 experiments performed in triplicate). Black columns correspond to cells treated with the hypoxia-mimetic drug CoCl 2. * p

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: Fluorescence, Produced, Infection

    NDV-OSM shows stronger oncolytic effect than Ad-OSM on PDAC cells in vitro. a Percentage of transduced (GFP + ) HaP-T1 cells 24 h after infection with Ad-GFP and NDV-GFP at the indicated MOIs. b-d cells were infected at different MOIs (X-axis, logarithmic scale) and the percentage of viable cells remaining in the monolayer was quantified 5 days later by crystal violet staining (representative results of at least 2 experiments with 4 samples per point). Uninfected cells were used as a reference (100 % survival). e HaP-T1 cells were co-infected with equal MOIs of Ad-OSM and NDV-OSM, or infected with NDV-OSM only. The change in cytotoxic effect (enhancement or inhibition) is represented as a positive or negative percentage, respectively, considering 0 the value obtained by single NDV-OSM infection at each MOI

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: NDV-OSM shows stronger oncolytic effect than Ad-OSM on PDAC cells in vitro. a Percentage of transduced (GFP + ) HaP-T1 cells 24 h after infection with Ad-GFP and NDV-GFP at the indicated MOIs. b-d cells were infected at different MOIs (X-axis, logarithmic scale) and the percentage of viable cells remaining in the monolayer was quantified 5 days later by crystal violet staining (representative results of at least 2 experiments with 4 samples per point). Uninfected cells were used as a reference (100 % survival). e HaP-T1 cells were co-infected with equal MOIs of Ad-OSM and NDV-OSM, or infected with NDV-OSM only. The change in cytotoxic effect (enhancement or inhibition) is represented as a positive or negative percentage, respectively, considering 0 the value obtained by single NDV-OSM infection at each MOI

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: In Vitro, Infection, Staining, Inhibition

    Ad-OSM and NDV-OSM show limited antitumor effect as monotherapy. Tumors were established by intrapancreatic inoculation of HaP-T1 cells. a Left panel shows the histopathological examination of a representative tumor (Hematoxylin/Eosin staining, magnification x400). Note the infiltration of normal pancreas and engulfment of secretory acini (white arrows). Right panel shows a schematic representation of the experimental setting. Treatments started 2 weeks after cell implantation (referred to as day 0) by intratumoral (i.t.) injection of Ad-OSM or NDV-OSM ( n = 6-12). Control animals received the same volume of saline solution ( n = 12-16). A second dose of each virus or saline was administered at day 3. All hamsters were sacrificed one month after the initiation of treatment. b Concentration of hOSM and type I IFN in the serum of hamsters 24 h after treatment with the indicated viruses, determined by ELISA and bioassay, respectively. Viral doses correspond to i.u. (x10 8 ). c Apart from the efficacy study, an independent group of tumor-bearing hamsters ( n = 5) received Ad-OSM at 2.5 × 10 8 i.u. at days 1 and 3 (arrows), and serum hOSM concentration was quantified right before and 24 h after each virus administration. d Individual tumor volumes and group averages determined by necropsy one month after initiation of treatments in the efficacy study. n.d., not determined due to early death of animals. e Additional groups of hamsters ( n = 5) received Ad-OSM (2.5 × 10 8 i.u/hamster), NDV-OSM (1 × 10 9 iu/hamster) or saline (control), and were sacrificed 48 h after treatment for histopathological examination of tumors. Microphotographs show Hematoxylin/Eosin staining, magnification X100). The percentage of necrotic area in each group ( n = 5) is represented in the right panel. * p

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: Ad-OSM and NDV-OSM show limited antitumor effect as monotherapy. Tumors were established by intrapancreatic inoculation of HaP-T1 cells. a Left panel shows the histopathological examination of a representative tumor (Hematoxylin/Eosin staining, magnification x400). Note the infiltration of normal pancreas and engulfment of secretory acini (white arrows). Right panel shows a schematic representation of the experimental setting. Treatments started 2 weeks after cell implantation (referred to as day 0) by intratumoral (i.t.) injection of Ad-OSM or NDV-OSM ( n = 6-12). Control animals received the same volume of saline solution ( n = 12-16). A second dose of each virus or saline was administered at day 3. All hamsters were sacrificed one month after the initiation of treatment. b Concentration of hOSM and type I IFN in the serum of hamsters 24 h after treatment with the indicated viruses, determined by ELISA and bioassay, respectively. Viral doses correspond to i.u. (x10 8 ). c Apart from the efficacy study, an independent group of tumor-bearing hamsters ( n = 5) received Ad-OSM at 2.5 × 10 8 i.u. at days 1 and 3 (arrows), and serum hOSM concentration was quantified right before and 24 h after each virus administration. d Individual tumor volumes and group averages determined by necropsy one month after initiation of treatments in the efficacy study. n.d., not determined due to early death of animals. e Additional groups of hamsters ( n = 5) received Ad-OSM (2.5 × 10 8 i.u/hamster), NDV-OSM (1 × 10 9 iu/hamster) or saline (control), and were sacrificed 48 h after treatment for histopathological examination of tumors. Microphotographs show Hematoxylin/Eosin staining, magnification X100). The percentage of necrotic area in each group ( n = 5) is represented in the right panel. * p

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: Staining, Injection, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Human OSM is biologically active on PANC-1 and HaP-T1 cell lines. a Western blot analysis of STAT3 activation in HaP-T1 cells treated with 20 ng/ml of recombinant hOSM at the indicated times. b mRNA levels of IL-15Rα and ICAM1, determined by qRT-PCR in HaP-T1 and PANC-1 cell lines treated with 20 ng/ml of recombinant hOSM at the indicated times. c Concentration of hOSM in the conditioned medium of HaP-T1 cells infected for 24 h with the indicated MOIs of Ad-OSM and NDV-OSM, determined by ELISA. d Western blot analysis of STAT1 activation in HuH-7 cells treated with conditioned medium from HaP-T1 cells infected or not with Ad-OSM, NDV-OSM or their respective GFP-expressing controls. e mRNA levels of IL-15Rα and ICAM1, determined by qRT-PCR, in HaP-T1 and PANC-1 cell lines treated for 24 h with the same conditioned media (representative results of at least 2 experiments performed in triplicate). ** p

    Journal: Molecular Cancer

    Article Title: Enhanced therapeutic effect using sequential administration of antigenically distinct oncolytic viruses expressing oncostatin M in a Syrian hamster orthotopic pancreatic cancer model

    doi: 10.1186/s12943-015-0479-x

    Figure Lengend Snippet: Human OSM is biologically active on PANC-1 and HaP-T1 cell lines. a Western blot analysis of STAT3 activation in HaP-T1 cells treated with 20 ng/ml of recombinant hOSM at the indicated times. b mRNA levels of IL-15Rα and ICAM1, determined by qRT-PCR in HaP-T1 and PANC-1 cell lines treated with 20 ng/ml of recombinant hOSM at the indicated times. c Concentration of hOSM in the conditioned medium of HaP-T1 cells infected for 24 h with the indicated MOIs of Ad-OSM and NDV-OSM, determined by ELISA. d Western blot analysis of STAT1 activation in HuH-7 cells treated with conditioned medium from HaP-T1 cells infected or not with Ad-OSM, NDV-OSM or their respective GFP-expressing controls. e mRNA levels of IL-15Rα and ICAM1, determined by qRT-PCR, in HaP-T1 and PANC-1 cell lines treated for 24 h with the same conditioned media (representative results of at least 2 experiments performed in triplicate). ** p

    Article Snippet: Cells Syrian hamster cells HaP-T1 (DSMZ ACC 222) and H2T (courtesy of Dr. CM Townsend, University of Texas Medical Branch, TX, USA), and human cell lines HuH-7 (JCRB Genebank, Japan), Hep2 (ATCC CCL-23), A549 (ATCC CCL-185), BxPC-3 (ATCC CRL-1687) and PANC-1 (ATCC CRL-1469), were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10 % fetal calf serum, 100 μg/ml streptomycin and 100 units/ml penicillin.

    Techniques: Western Blot, Activation Assay, Recombinant, Quantitative RT-PCR, Concentration Assay, Infection, Enzyme-linked Immunosorbent Assay, Expressing