19,651 research outputs found

    Immune cell depletion during IAV infection in tumour-bearing mice.

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    Tumours were subcutaneously (s.c.) transplanted and mice were infected with 150 PFU/mL influenza A/PR/8/34 (IAV) as described. 200μg of depleting antibodies against NK1.1 (a), CSF1R (b), CD4 (c) or CD8 (d) were applied i.p. on days 4, 7 and 10 after tumour cell transplantation, respectively. Depletion was confirmed by flow cytometry. Representative dot plots of tumours 12 days after tumour cell transplantation are shown. (TIF)</p

    Reproductive Biology of Anadromous Rainbow Smelt, \u3cem\u3eOsmerus mordax\u3c/em\u3e, in the Ipswich Bay Area, Massachusetts

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    The reproductive biology of anadromous rainbow smelt, Osmerus mordax, was investigated in the Parker River and Essex Bay systems from 1977 through 1979. Variations in growth and seasonal energy content of body tissue of young-of-the-year (YOY) anadromous smelt were examined to investigate their relationships to spawning of precocious (age I) fish. By late fall of their first year, YOY that will spawn precociously in the spring have developing gonads, are longer in total length and have a higher energy content (kcals/g) of body tissue than non-precocious fish. The number of fish spawning at age I increased during the spawning season, with male precocious spawners consistently outnumbering female precocious fish. Several aspects of the reproductive ecology of rainbow smelt were examined during the 1979 spawning season. Results from two field experiments indicated that egg survival was positively correlated with water velocity (up to 60 - 80 cm/s). Smelt spawned many more eggs (by a factor of 12 - 15) on aquatic vegetation (Podostemum ceratophyllum abrotanoides) than on two smooth-surfaced substrates (ceramic tile and gravel/ rubble). Survival to hatching on the vegetation was approximately 10% compared to a 1% rate on the other surfaces. In addition, diameter of water-hardened, unfertilized eggs was positively correlated with female total length and prolarval size at hatching. These reproductive characteristics are discussed relative to the life history strategy of anadromous rainbow smelt in the Parker River-Plum Island Sound syste

    CD4<sup>+</sup> T cells are not altered in the tumour upon infection.

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    B16 tumours were transplanted and mice were infected as described in Fig 1. Tumours were analysed by flow cytometry 12 days post transplantation. (a) Frequencies of viable CD4+ T cells. (b) Frequencies of CD43 or Granzyme B (GzmB) of CD43 expressing cells of CD4+ T cells. Data from 2 experiments are shown. Error bars represent SEM. (TIF)</p

    IAV infection suppresses tumour growth dose dependently.

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    1 x 105 B16F1 (B16) tumour cells were transplanted subcutaneously (s.c.) into the right flank per mouse. 4 days after tumour transplantation, mice were infected intranasally with Influenza A/PR/8/34 (IAV). The tumour volume was measured everyday once palpable upon infection. Data from 2 experiments with 3–4 mice per group per experiment. Error bars represent SEM. Statistical tests on tumour growth development were performed as Two-way-ANOVA followed by Tukey’s multiple comparisons test. **** = p (TIF)</p

    IAV infection impedes differentiation of T-cell exhaustion.

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    B16F1 (B16) tumour-bearing mice were infected with Influenza A/PR/8/34 (IAV) (150 PFU/mL) as described. 8 days post infection mice were sacrificed for flow cytometry analysis. (a) Frequencies of PD-1 expressing gp100-specific CD8+ T-cells in the tumour. (b) Representative dot plots of CD8+ T-cells expressing PD-1 and TIM-3. (c) Frequencies of PD-1high TIM-3+ or PD-1int TIM-3- of gp100-specific CD8+ T-cells in the tumour. (d) Frequencies of Granzyme B (GzmB) expressing cells of the populations described in c. (e) Frequencies of Thymocyte selection-associated high mobility group box protein (TOX) expressing gp100-specific CD8+ T-cells in the tumour. Data from 2 experiments are shown. Error bars represent SEM. Statistical tests between two groups were performed as Student’s t-tests. * = p<0.05, ** = p<0.01, *** = p<0.001.</p

    IAV infection suppresses tumour growth.

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    (a-d) 1 x 105 B16F1 (B16) tumour cells were transplanted subcutaneously (s.c.) into the right flank per mouse. 4 days after tumour transplantation, mice were infected intranasally with 150 PFU/mL Influenza A/PR/8/34 (IAV). (a) Viral load was measured by M1 expression relative to RPS9 expression in the lungs of infected mice with or without B16 tumour 8 days post infection (dpi). n = 9 for B16+IAV, n = 6 for IAV (b) Viral load as measured from homogenized lung supernatants by counting plaque forming units (PFU) from plaque assay 8 dpi. (c) Body weight change after IAV infection. Data from 2 experiments with 4–6 mice per group per experiment. (d-g) 1 x 105 B16 (d and g), CT26 (e) or 5 x 105 Lewis Lung Carcinoma (LLC) (f) tumour cells were transplanted s.c. and mice were infected as described in a) with (d-f) IAV or (g) inactivated IAV. The tumour volume was measured everyday once palpable upon infection. d&e: Data from 4 experiments with 3–4 mice per group per experiment, f: n = 4, g: Data from 2 experiments with 4 mice per group per experiment. (h) Viral load as measured by M1 expression relative to RPS9 expression in the tumour or lung of infected mice 12 days post tumour transplantation. (i) B16 tumours were transplanted s.c. with 1 x 105 tumour cells and mice were infected intravenously with Friend Virus (FV) 4 days post tumour transplantation. The tumour volume was scored everyday once palpable. n = 4. Error bars represent SEM. Statistical tests on tumour growth development were performed as Two-way-ANOVA, in Sidak’s multiple comparisons test when two groups were compared or Tukey’s multiple comparisons test when more than two groups were compared. Viral titres were compared with non-parametric t tests. * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001, ns = not significant.</p

    Association between a pyroelectric infrared sensor monitoring system and a 3-dimensional accelerometer to assess movement in preweaning dairy calves

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    The objective of this study was to correlate movement assessed by a pyroelectric infrared sensor system in preweaning dairy calves with lying and standing time assessed by a 3D accelerometer considering the temperature-humidity index (THI). A total of 35 dairy calves (1–7 d of age) were enrolled in the study and 20 calves were included in the final analyses. The lying and standing time of the calves was monitored with a 3D accelerometer (Hobo Pendant G Data Logger, Onset Computer Corporation, USA), which was used as the gold standard reference. The infrared sensor monitoring system (IMS; Calf Monitoring System, Futuro Farming GmbH, Germany) was fixed to the fence of the calf hutch within the calf's reach. Temperature-humidity was monitored with 2 validated THI sensors inside and on outside of each calf hutch. Additionally, one THI sensor was located near the calf hutches. The observation period lasted 14 consecutive days. The average standing time assessed by the 3D accelerometer was 13.4 ± 12.7 (mean ± standard deviation) min/h and the average lying time was 46.6 (±12.7) min/h. The median (25th percentile; 75th percentile) number of movements measured by the IMS was 360 (60; 919) movements per hour. Number of movements per hour measured by the IMS was compared with data obtained with a validated 3D accelerometer. The Pearson correlation coefficient between both standing and lying time and the number of movements was r = 0.85 and r = −0.85, respectively. The Pearson correlation coefficients were only slightly influenced by THI (low THI [72]: r = 0.81). Our data show that the number of movements of dairy calves measured by IMS were highly correlated with the chosen gold standard reference method. High THI slightly affects the measurement accuracy of IMS

    Blockade of CXCR3 reverses tumour growth restriction by IAV infection.

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    B16 tumour-bearing mice were infected with Influenza A/PR/8/34 (IAV) (150 PFU/mL) as described. 8 days post infection mice were sacrificed for flow cytometry or Luminex analysis. (a) Chemokines were analysed via flow cytometry. Mean fluorescence intensity (MFI) of the respective chemokine receptors was measured on CD8+ T cells in the tumour. Data from two individual experiments are shown. (b) Absolute CXCL10 levels in tumour and lung were measured by Luminex analysis 12 days post tumour cell injection. (c) B16 tumour-bearing mice were infected with IAV and treated with 200 μg anti-CXCR3 antibodies on days 4, 7 and 10 post tumour cell injection. Data from 3 experiments with 3–4 mice per group per experiment. (d) Absolute cell numbers of gp100-specific CD8+ T-cells in the lung from mice treated as described in c). (e) Frequencies of CD43 and Granzyme B (GzmB) expressing CD8+ T-cells in the tumour. Error bars represent SEM. Statistical tests between two groups were performed as non-parametric Mann-Whitney t-tests. Statistical tests with more than two groups were performed as One-way-ANOVA, followed by Kruskal-Wallis multiple comparison test. Statistical tests on tumour growth development were performed as Two-way-ANOVA, in Tukey’s multiple comparisons test. * = p<0.05, ** = p<0.01.</p

    IAV infection strengthens cancer-specific CD8<sup>+</sup> T-cell immunity.

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    B16 tumour-bearing mice were infected with IAV (150 PFU/mL) as described. 8 days post infection mice were sacrificed for flow cytometry analysis. (a) Representative histograms of analysis of CD43, Granzyme B (GzmB), and Ki67 expression in the tumour. (b) Frequencies of CD43, GzmB of CD43+ or Ki67 expressing CD8+ T-cells in the tumour. (c) Representative dot plots of gp100-tetramer+ CD8+ T-cells in the tumour. (d) Frequencies of gp100-specific CD8+ T-cells in the tumour. (e) Frequencies of CD43, GzmB of CD43+ or Ki67 expressing gp100-specific CD8+ T-cells in the tumour. (f) Representative dot plots of gp100-tetramer+ CD8+ T-cells in the lung. (g) Frequencies of gp100-specific CD8+ T-cells in the lung. (h) Frequencies of CD43, GzmB of CD43+ or Ki67 expressing gp100-specific CD8+ T-cells in the lung. (i) CD8+ T cells were isolated from tumours 12 days after tumour transplantation and co-cultured with eFluor670 stained B16 cells. After 18 h of co-culture, the frequency of remaining viable tumour cells was quantified using flow cytometry, and cytotoxic capacity was calculated in comparison to tumour cell-only wells (n  =  4 co-cultures per group). (j) Mean fluorescence intensity (MFI) of Interferon (IFN)γ expressing gp100-specific CD8+ T-cells. (k) B16 tumour-bearing mice were treated with 250 μg anti-IFNγ antibodies on days 4, 6, 8 and 10 post tumour cell injection. Data from 2 experiments with 4 mice per group per experiment. Error bars represent SEM. Statistical tests between two groups were performed as Student’s t-tests. Statistical test on tumour growth development was performed as Two-way-ANOVA, followed by Tukey’s multiple comparisons test. * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001.</p
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