scholarly journals In vivo bioluminescence imaging of Burkholderia mallei respiratory infection and treatment in the mouse model

2011 ◽  
Vol 2 ◽  
Author(s):  
Shane Massey
PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0120925 ◽  
Author(s):  
Binje Vick ◽  
Maja Rothenberg ◽  
Nadine Sandhöfer ◽  
Michela Carlet ◽  
Cornelia Finkenzeller ◽  
...  

2011 ◽  
Vol 30 (3) ◽  
pp. 335-340 ◽  
Author(s):  
Jonathan R. Pribaz ◽  
Nicholas M. Bernthal ◽  
Fabrizio Billi ◽  
John S. Cho ◽  
Romela Irene Ramos ◽  
...  

2013 ◽  
Vol 12 (3) ◽  
pp. 7290.2012.00029 ◽  
Author(s):  
Monika A. Jarzabek ◽  
Peter C. Huszthy ◽  
Kai O. Skaftnesmo ◽  
Emmet McCormack ◽  
Patrick Dicker ◽  
...  

PLoS ONE ◽  
2017 ◽  
Vol 12 (8) ◽  
pp. e0183469 ◽  
Author(s):  
Marisa R. Buchakjian ◽  
Nicole M. Merritt ◽  
Devon L. Moose ◽  
Adam J. Dupuy ◽  
Munir R. Tanas ◽  
...  

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1110-1110
Author(s):  
Martin Chopra ◽  
Simone S Riedel ◽  
Viktoria von Krosigk ◽  
Carina A Bäuerlein ◽  
Christian Brede ◽  
...  

Abstract Abstract 1110 The cytokine tumor necrosis factor-α (TNF) has pleiotropic functions both in normal physiology and disease. TNF and its relative lymphotoxin-α (LT) signal by activating two cell surface receptors TNFR1 and TNFR2. TNFR1 is expressed on most cells whereas TNFR2 is mainly expressed in cells of the hematopoietic system. TNF-TNFR interactions were shown to play a major role in graft-versus-leukemia effect and in the immunosurveillance of solid tumors. To study the contribution of TNF-TNFR interactions on tumor cell progression we employed a syngenic B16 melanoma mouse model combined with in vivo bioluminescence imaging. Firefly luciferase-transgenic B16 melanoma cells were injected intravenously into syngenic albino C57BL/6 hosts. The host mice were either of wildtype, TNF, LT, TNFR1, TNFR2 knockout or TNFR1R2 double knockout genotype. The localization and expansion of the B16 cells was monitored by in vivo bioluminescence imaging for up to 14 days. On days 15, mice were sacrificed and internal organs were imaged ex vivo to further elucidate the organ-specific tumor burden. B16 tumors were primarily found in the lungs of all genotypes. All female knockout genotypes displayed a higher lung tumor burden than wildtype mice. In male mice, only TNF knockout presented enhanced tumor cell signals. Following ex vivo imaging we evaluated the pulmonary infiltration of NK1.1 or NKp46, CD8, CD4 and CD4/CD25/Foxp3 regulatory T cells by flow cytometry and immunofluorescence microscopy. Compared to wildtype mice, more regulatory T cells infiltrated the lungs of female TNFR1 knockout mice (200%). In LT knockout mice, very few NK cells (<20%) but more CD4+ cells (160%) infiltrated the lungs. Only subtle changes occurred in the other deficient mouse strains. However, these changes were independent of the presence of tumor cells and could also be found in normal knockout mice without B16 tumors. Within sections of tumor-bearing lungs, we found that TNF and all three TNFR knockouts exhibited less CD8+ cells within tumors than did wildtype or LT knockout mice. The number of CD8+ cells in normal lung tissue was not altered across the different genotypes. The deficit in NK cells of LT knockout mice was confirmed by histology. The enhanced tumor progression in all knockout mice could be a secondary effect due to their altered immune phenotype rather than to the loss of TNF-TNFR interactions. To circumvent this potential experimental bias and to further assess the influence of the loss of expression of parts of the TNF/TNFR-system in immune cells only, we generated bone marrow chimeras by reconstituting lethally irradiated female wildtype mice with bone marrow derived from TNF, LT, TNFR1 or TNFR2 knockout mice. Tumor cell signals in these chimeric mice progressed more than in normal wildtype mice. In contrast to the first set of experiments with knockout mice, we found that mice reconstituted with either TNF or TNFR2 knockout bone marrow presented less tumor cell signal than did mice reconstituted with wildtype bone marrow. TNF-TNFR interactions between immune cells appear to exhibit pro-tumorigenic functions in our mouse model. These results show that TNF-TNFR interactions are an important step in tumor cell progression and that the outcome of these interactions differs, depending on whether immune or parenchymal cells are deficient in TNF-TNFR signalling components. Disclosures: No relevant conflicts of interest to declare.


2019 ◽  
Vol 11 ◽  
Author(s):  
Ambrose A. Dunn-Meynell ◽  
Peter Dowling ◽  
Michelle Marchese ◽  
Esther Rodriguez ◽  
Benjamin Blumberg ◽  
...  

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1809-1809
Author(s):  
Sabrina Kraus ◽  
Martin Chopra ◽  
Christian Brede ◽  
Simone S Riedel ◽  
Mike Friedrich ◽  
...  

Abstract Abstract 1809 Various imaging platforms are well established in hematology research. Nevertheless, the three-dimensional architecture of the bone marrow and tumor growth within this microenvironment remain largely uncharacterized. To date the major hindrance to microscopically image tumor engraftment and the immune response in the bone marrow on a single cell level is the compact structure of the bone that is almost impossible to image through. Therefore, we developed a novel bioluminescent mouse model that recapitulates the clinical characteristics of MM using the new human UMM3 cell line (CD38+, CD56+, CD138+, CD19−, CD20−), from the pleural effusion of a patient with an IgG lambda myeloma (ISS stage I) as well as the well-characterized RPMI8226 cell line transduced to express eGFP and firefly luciferase (UMM3eGFPluc and RPMIeGFPluc). 1×106 MM cells were injected intravenously into NOD.Cg-Prkdcscid IL2rg (NSG) mice and disease progression and bone marrow (BM) engraftment were monitored twice weekly by in vivo bioluminescence imaging. Both cell lines homed to the BM compartment, reflecting MM pathophysiology. Histological analysis confirmed BM engraftment and showed multiple osteolytic lesions for both UMM3 and RPMI cells. Since we were interested in imaging the interactions between human MM cells and the bone marrow microenvironment on a single cell level, we employed the multi-color LSFM after decalcification, specific deep-tissue antibody staining and clearing of the bone structures. With this innovative microscopy technique, we were able to establish a novel tool to display tumor cell engraftment in the bone marrow compartment in three dimensions through the intact bone. We recorded 1500 optical sections for three individual channels each (488, 532, and 647 nm) with an increment of 5μm which allows scanning the whole bone marrow compartment of the sternum within minutes in single-cell resolution. Using higher magnification enabled us to even visualize subcellular components within the bone marrow. Moreover, tissue autofluorescene, recorded mainly in the 488 nm channel, displayed detailed microanatomical structures which allowed for the localization of individual cells within their anatomical context. We could establish protocols for various fluorophore-coupled antibodies and successfully stained CD138+ cells in relation to CD3+ cells and to the microenvironment in the bone marrow. The CD138-positive cells infiltrated the bone marrow in a number of small clusters and comprising about 15% of cellular elements in total. Ex vivo bioluminescence imaging of the sternum from UMM3 tumor-bearing mice revealed massive infiltration of luciferase-expressing cells into the bone marrow compartment. This could also be confirmed by flow cytometrical analysis of bone marrow cells which showed eGFP+hCD138+ cells. We have successfully introduced a novel technique to study MM cell engraftment and progression in a humanized mouse model. We were able to track the tumor cells both in the living animal by in vivo bioluminescence imaging and on single-cell resolution by multi-color LSFM within the intact bone. Our model may lead to better insights into the pathogenesis of MM and could serve as a model for preclinical testing of new therapeutic approaches for the treatment of MM patients. Disclosures: No relevant conflicts of interest to declare.


2012 ◽  
Vol 12 (1) ◽  
pp. 112 ◽  
Author(s):  
Alfredo G Torres ◽  
Roberto J Cieza ◽  
Maricarmen Rojas-Lopez ◽  
Carla A Blumentritt ◽  
Cristiane S Souza ◽  
...  

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