scholarly journals Strain-specific metastatic phenotypes in pheochromocytoma allograft mice

2018 ◽  
Vol 25 (12) ◽  
pp. 993-1004 ◽  
Author(s):  
Martin Ullrich ◽  
Josephine Liers ◽  
Mirko Peitzsch ◽  
Anja Feldmann ◽  
Ralf Bergmann ◽  
...  
Keyword(s):  
Nude Mice ◽  
Nk Cell ◽  
Somatostatin Receptor ◽  
Metastatic Spread ◽  
Mouse Strains ◽  
Radiotracer Uptake ◽  
Positron Emission ◽  
Liquid Chromatography Tandem Mass ◽  
Urinary Dopamine

Somatostatin receptor-targeting endoradiotherapy offers potential for treating metastatic pheochromocytomas and paragangliomas, an approach likely to benefit from combination radiosensitization therapy. To provide reliable preclinical in vivo models of metastatic disease, this study characterized the metastatic spread of luciferase-expressing mouse pheochromocytoma (MPC) cells in mouse strains with different immunologic conditions. Bioluminescence imaging showed that, in contrast to subcutaneous non-metastatic engraftment of luciferase-expressing MPC cells in NMRI-nude mice, intravenous cell injection provided only suboptimal metastatic spread in both NMRI-nude mice and hairless SCID (SHO) mice. Treatment of NMRI-nude mice with anti-Asialo GM1 serum enhanced metastatic spread due to substantial depletion of natural killer (NK) cells. However, reproducible metastatic spread was only observed in NK cell-defective SCID/beige mice and in hairless immunocompetent SKH1 mice bearing disseminated or liver metastases, respectively. Liquid chromatography tandem mass spectrometry of urine samples showed that subcutaneous and metastasized tumor models exhibit comparable renal monoamine excretion profiles characterized by increasing urinary dopamine, 3-methoxytyramine, norepinephrine and normetanephrine. Metastases-related epinephrine and metanephrine were only detectable in SCID/beige mice. Positron emission tomography and immunohistochemistry revealed that all metastases maintained somatostatin receptor-specific radiotracer uptake and immunoreactivity, respectively. In conclusion, we demonstrate that intravenous injection of luciferase-expressing MPC cells into SCID/beige and SKH1 mice provides reproducible and clinically relevant spread of catecholamine-producing and somatostatin receptor-positive metastases. These standardized preclinical models allow for precise monitoring of disease progression and should facilitate further investigations on theranostic approaches against metastatic pheochromocytomas and paragangliomas.

scholarly journals Soluble and Exosome-Bound α-Galactosylceramide Mediate Preferential Proliferation of Educated NK Cells with Increased Anti-Tumor Capacity

Cancers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 298
Author(s):  
Arnika K. Wagner ◽  
Ulf Gehrmann ◽  
Stefanie Hiltbrunner ◽  
Valentina Carannante ◽  
Thuy T. Luu ◽  
...  
Keyword(s):  
Nk Cells ◽  
Cancer Immunotherapy ◽  
Nk Cell ◽  
Mouse Strains ◽  
Target Cells ◽  
Class I Mhc ◽  
Mhc I ◽  
Cell Responses ◽  
Missing Self

Natural killer (NK) cells can kill target cells via the recognition of stress molecules and down-regulation of major histocompatibility complex class I (MHC-I). Some NK cells are educated to recognize and kill cells that have lost their MHC-I expression, e.g., tumor or virus-infected cells. A desired property of cancer immunotherapy is, therefore, to activate educated NK cells during anti-tumor responses in vivo. We here analyze NK cell responses to α-galactosylceramide (αGC), a potent activator of invariant NKT (iNKT) cells, or to exosomes loaded with αGC. In mouse strains which express different MHC-I alleles using an extended NK cell flow cytometry panel, we show that αGC induces a biased NK cell proliferation of educated NK cells. Importantly, iNKT cell-induced activation of NK cells selectively increased in vivo missing self-responses, leading to more effective rejection of tumor cells. Exosomes from antigen-presenting cells are attractive anti-cancer therapy tools as they may induce both innate and adaptive immune responses, thereby addressing the hurdle of tumor heterogeneity. Adding αGC to antigen-loaded dendritic-cell-derived exosomes also led to an increase in missing self-responses in addition to boosted T and B cell responses. This study manifests αGC as an attractive adjuvant in cancer immunotherapy, as it increases the functional capacity of educated NK cells and enhances the innate, missing self-based antitumor response.

Abstract 460: 64 Cu-DOTATATE for in vivo Positron Emission Tomography Imaging of Somatostatin Receptor 2 Expressing Macrophages in a Göttingen Minipig Model of Atherosclerosis

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Sune Folke Pedersen ◽  
Trine Pagh Ludvigsen ◽  
Andreas Vegge ◽  
Camilla Schumacher-Petersen ◽  
Rasmus Sejersten Ripa ◽  
...  
Keyword(s):  
Positron Emission Tomography ◽  
Abdominal Aorta ◽  
Right Atrium ◽  
Plasma Lipids ◽  
Somatostatin Receptor ◽  
Emission Tomography ◽  
Activated Macrophages ◽  
Lean Control ◽  
Positron Emission

Background: Somatostatin receptor subtype 2 (SSTR 2 ) is expressed by activated macrophages which are important effector cells in atherogenesis and a major constituent of atherosclerotic plaques. SSTR 2 can be targeted non-invasively in vivo using the tracer 64 Cu-DOTATATE and positron emission tomography (PET). Methods: Male castrated Göttingen minipigs were treated according to two diet regimes for 43 weeks: chow (lean control; n=1) or high-fat/high-cholesterol diet (HFHC, obese group; n=2). Plasma lipids: total cholesterol (TC) and triglycerides (TG) were measured and SSTR 2 expression was assessed using hybrid positron emission tomography/magnetic resonance (PET/MR) imaging of the abdominal aorta, one hour (range: 60-63 minutes) post injection with 200 MBq (range 202-214 MBq) of 64 Cu-DOTATATE. Consectutive regions of interest (ROIs) were drawn guided by MR to include vessel wall and lumen of the abdominal aorta to calculate standardized uptake values (SUVs). Target-to-background (TBR) ratios were calculated using right atrium SUVs for blood pool correction (SUV vessel /SUV right atrium = TBR) and reported as mean and maximal values (TBR mean ; TBR max ). Results: Three minipigs (age 17 months) were included and PET/MR was completed in all animals. Mean uptake of 64 Cu-DOTATATE was lowest for the abdominal aorta in the lean control: TBR mean = 0.44 and TBR max = 0.73 (range: TBR mean = 0.29 - 0.62; TBR max = 0.49 - 1.10) and highest in the HFHC group: TBR mean = 0.66 and TBR max = 1.28 (range: TBR mean = 0.26 - 1.38; TBR max = 0.39 - 3.19). Plasma lipids: lean control: TC = 1.34 mmol/L; TG = 0.38 mmol/L; HFHC group (mean values): TC = 18.7mmol/L (range: 11.6 - 25.8mmol/L); TG = 0.6 mmol/L (range: 0.37 - 0.83mmol/L). Conclusion: Non-invasive 64 Cu-DOTATATE PET/MR is feasible for assessment of SSTR 2 expression in a Göttingen minipig model of diet-induced atherosclerosis. Additional studies are needed including assessment of histology findings and gene expression to confirm the presence of activated macrophages in order to validate the use of 64 Cu-DOTATATE PET/MR in this model.

scholarly journals Mice Deficient in Interleukin-4 (IL-4) or IL-4 Receptor α Have Higher Resistance to Sporozoite Infection with Plasmodium berghei (ANKA) than Do Naive Wild-Type Mice

2004 ◽  
Vol 72 (1) ◽  
pp. 322-331 ◽  
Author(s):  
Michael Saeftel ◽  
Andreas Krueger ◽  
Sandra Arriens ◽  
Volker Heussler ◽  
Paul Racz ◽  
...  
Keyword(s):  
Nk Cells ◽  
Plasmodium Berghei ◽  
Nk Cell ◽  
Interleukin 4 ◽  
Mouse Strains ◽  
No Production ◽  
Liver Stage ◽  
Innate Immune Mechanism ◽  
Ifn Γ

ABSTRACT BALB/c interleukin-4 (IL-4−/−) or IL-4 receptor-α (IL-4rα−/−) knockout (KO) mice were used to assess the roles of the IL-4 and IL-13 pathways during infections with the blood or liver stages of plasmodium in murine malaria. Intraperitoneal infection with the blood-stage erythrocytes of Plasmodium berghei (ANKA) resulted in 100% mortality within 24 days in BALB/c mice, as well as in the mutant mouse strains. However, when infected intravenously with the sporozoite liver stage, 60 to 80% of IL-4−/− and IL-4rα−/− mice survived, whereas all BALB/c mice succumbed with high parasitemia. Compared to infected BALB/c controls, the surviving KO mice showed increased NK cell numbers and expression of inducible nitric oxide synthase (iNOS) in the liver and were able to eliminate parasites early during infection. In vivo blockade of NO resulted in 100% mortality of sporozoite-infected KO mice. In vivo depletion of NK cells also resulted in 80 to 100% mortality, with a significant reduction in gamma interferon (IFN-γ) production in the liver. These results suggest that IFN-γ-producing NK cells are critical in host resistance against the sporozoite liver stage by inducing NO production, an effective killing effector molecule against Plasmodium. The absence of IL-4-mediated functions increases the protective innate immune mechanism identified above, which results in immunity against P. berghei infection in these mice, with no major role for IL-13.

scholarly journals Selective Down-Regulation of the NKG2D Ligand H60 by Mouse Cytomegalovirus m155 Glycoprotein

2005 ◽  
Vol 79 (5) ◽  
pp. 2920-2930 ◽  
Author(s):  
Milena Hasan ◽  
Astrid Krmpotic ◽  
Zsolt Ruzsics ◽  
Ivan Bubic ◽  
Tihana Lenac ◽  
...  
Keyword(s):  
Gene Family ◽  
Nk Cell ◽  
Cell Receptor ◽  
Mouse Strains ◽  
Mcmv Infection ◽  
Partial Restoration ◽  
Golgi Compartment ◽  
Infected Cells ◽  
Nkg2d Receptor

ABSTRACT Both human and mouse cytomegaloviruses (CMVs) encode proteins that inhibit the activation of NK cells by down-regulating cellular ligands for the activating NK cell receptor NKG2D. Up to now, three ligands for the NKG2D receptor, named RAE-1, H60, and MULT-1, have been identified in mice. The resistance of mouse strains to murine CMV (MCMV) infection is determined by their ability to generate an effective NK cell response. The MCMV gene m152, a member of the m145 gene family, down-regulates the expression of RAE-1 in order to avoid NK cell control in vivo. Here we report that the m155 gene, another member of the m145 gene family, encodes a protein that interferes with the expression of H60 on the surfaces of infected cells. Deletion of the m155 gene leads to an only partial restoration of H60 expression on the cell surface, suggesting the involvement of another, so far unknown, viral inhibitor. In spite of this, an m155 deletion mutant virus shows NK cell-dependent attenuation in vivo. The acquisition of endo-β-N-acetylglucosaminidase H resistance and the preserved half-life of H60 in MCMV-infected cells indicate that the m155-mediated effect must take place in a compartment after H60 exits from the ERGIC-cis-Golgi compartment.

scholarly journals Cooperation between cytotoxic and helper T lymphocytes in protection against lethal Sendai virus infection. Protection by T cells is MHC-restricted and MHC-regulated; a model for MHC-disease associations.

1986 ◽  
Vol 164 (3) ◽  
pp. 723-738 ◽  
Author(s):  
W M Kast ◽  
A M Bronkhorst ◽  
L P de Waal ◽  
C J Melief
Keyword(s):  
T Cell ◽  
Nk Cell ◽  
Sendai Virus ◽  
Lethal Dose ◽  
Cell Activity ◽  
Mouse Strains ◽  
Class I Mhc ◽  
Virus Dose ◽  
Disease Associations

The in vivo importance of class I MHC regulation of the Tc response to a natural pathogenic agent of high virulence was studied on the basis of our previous demonstration of a major difference in the capacity to generate a Sendai virus-specific Tc response between C57BL/6 (B6, H-2b) mice and H-2Kb mutant B6.C-H-2bm1 (bm 1) mice. These two mouse strains differ from each other only in three amino acids in the crucial H-2Kb restriction element for this response. bm 1 mice, in contrast to B6 mice, are Tc nonresponders against this virus, but show Sendai-specific T cell proliferation, antibody production, and DTH reactions, as well as NK cell activity, equal to those of B6 mice. B6, Sendai Tc-deficient bm 1 and T cell-deficient B6 nu/nu mice differ from each other in susceptibility to lethal pneumonia induced by i.n. inoculation of virulent Sendai virus. The lethal dose (LD50) in B6 mice averaged 152 TCID50, in bm 1 mice, 14 TCID50 and in B6 nu/nu mice 0.5 TCID50. The importance of Tc was also shown by the complete protection of B6 nu/nu mice against infection with a lethal virus dose by i.v. injection of a Sendai virus-specific, IL-2-dependent and H-2Kb-restricted B6 Tc clone. In vivo protection by this Tc clone was H-2Kb-restricted. Apart from Tc, an important role for virus-specific Th cells is evident from the difference in susceptibility between bm 1 and B6 nu/nu mice. This conclusion was supported by the demonstration that the mean survival time of B6 nu/nu and bm 1 nu/nu mice could be significantly prolonged, in an I-Ab-restricted manner, by the injection of in vitro-propagated, Sendai-specific B6 or bm 1 Th clones after a lethal dose of Sendai virus, and by the demonstration that inoculation of these Th clones provided help to virus-specific Tc by means of IL-2 production. Strikingly, Th and Tc cooperate in anti-Sendai virus immunity, since permanent survival of lethally infected nu/nu mice was only achieved by inoculation of a mixture of Tc and Th clones or a mixture of a Tc clone and rIL-2. This study provides a unique model for the study of MHC-disease associations.

OS06.5A Fluorescence image-guided resection of intracranial meningioma: an experimental in vivo study on nude mice

2021 ◽  
Vol 23 (Supplement_2) ◽  
pp. ii9-ii10
Author(s):  
S Linsler ◽  
S Senger ◽  
S Müller ◽  
A Müller ◽  
J Oertel
Keyword(s):  
Tumor Growth ◽  
Nude Mice ◽  
Aminolevulinic Acid ◽  
Somatostatin Receptor ◽  
Brain Slices ◽  
Fluorescence Microscope ◽  
Intracranial Meningioma ◽  
Tumor Mass ◽  
Image Guided

Abstract BACKGROUND The use of photodynamic agents in malignant cranial tumor surgery is quite common. For example five-aminolevulinic acid (5-ALA)-induced porphyrins in malignant gliomas are potent photosensitizers. Until today there is no comparable selective fluorescent substance available for meningiomas. Nevertheless, there is a demand for intraoperative fluorescent identification of e.g. invasive skull base meningiomas to increase radicality. This study was established to investigate fluorescent image-guided resection with somatostatin receptor labelled fluorescence dye for intracranial meningioma in the nude mice. MATERIAL AND METHODS Primary meningioma cell culture samples were stereotactically implanted subdural into 20 nude mice. 90 days after inoculation of the cells, a cranial MRI with contrast agent revealed tumor growth. After detection of tumor mass in MRI, FAM-TOC5,6-Carboxyfluoresceine-Tyr3-Octreotide was injected intravenously and tumor mass was hereafter resected under visualization via fluorescence microscope and endoscope. After attempted total resection, animal were sacrificed brain slices were obtained and histologically analysed to verify the resection extent. RESULTS In 18 mice tumor growth was detected in MRI after 90 days of inoculation. The tumor mass could be clearly identified with fluorescence microscope and endoscope after injecting FAM-TOC5,6-Carboxyfluoresceine-Tyr3-Octreotide. The tumor margins could be better visualized. After fluorescence-guided resection no remaining tumor could be identified in histological analysis. CONCLUSION This study describes for the first time the use of FAM-TOC5,6-Carboxyfluoresceine-Tyr3-Octreotide and demonstrates its value of fluorescent identification of meningioma cells in vivo. Furthermore, the authors established a new experimental animal model for fluorescence meningioma surgery.

Intravenous, Intraperitoneal, and Subcutaneous Routes of Administering 64Cu-DOTA-HB22.7 Display Equivalent Tumor Targeting Ability.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2359-2359
Author(s):  
Shiloh M. Martin ◽  
Robert T. O’Donnell ◽  
Hayes McKnight ◽  
Julie L. Sutcliff ◽  
Joseph M. Tuscano
Keyword(s):  
Nude Mice ◽  
Tumor Targeting ◽  
Emission Tomography ◽  
New Agents ◽  
Routes Of Administration ◽  
Positron Emission ◽  
Specific Tumor ◽  
Human Lymphoma ◽  
Targeting Ability

Abstract BACKGROUND: Chemotherapy for Non-Hodgkin’s Lymphoma (NHL) is initially effective, but often limited by toxicity and resistance. New agents for the treatment of NHL are needed. HB22.7 is a monoclonal antibody that binds CD22 and has previously been shown to reduce human lymphoma xenograft volume in nude mice. OBJECTIVES: (1) Develop a 64Cu-DOTA-HB22.7 antibody, for in vivo imaging and potential therapy of NHL. (2) Determine if tumor targeting of 64Cu-DOTA-HB22.7 is equivalent between intravenous (IV), intraperitoneal (IP), and subcutaneous (SQ) routes of administration. METHODS: DOTA-HB22.7’s ability to bind CD22+ cells was assessed by flow cytometry. 64Cu-DOTA-HB22.7 was injected IV, IP, or SQ into xenograft-bearing nude mice and tumor targeting assessed by positron emission tomography (PET) and biodistribution assay. Pharmacokinetics were determined by measuring radioactivity of blood samples. Serum was analyzed by radio-TLC. RESULTS: DOTA conjugation does not affect HB22.7’s ability to bind CD22. 64Cu-DOTA-HB22.7 demonstrates specific tumor targeting at 24 and 48 hrs. Targeting is equivalent regardless of route of administration. Pharmacokinetics demonstrate that 64Cu-DOTA-HB22.7 can access the bloodstream (and thus, target tumor) in IP or SQ injections. By 48 hrs, blood 64Cu levels are (a) equivalent, regardless of injection route and (b) below peak levels, indicating clearance from the circulation. Serum analysis shows that right-shifted TLC peaks, possibly 64Cu-DOTA-HB22.7 metabolites, are present only in the IV injected group at 48 hrs. CONCLUSION: These findings establish 64Cu-DOTA-HB22.7 as an NHL-specific imaging agent and indicate its potential for use in radioimmunotherapy. These findings provide evidence that more accessible routes of administration can achieve equivalent targeting results, and may lead to more efficient and accurate administration of antibody-based therapeutics in mice.

scholarly journals Genetic Control Of Natural Killing and In Vivo Tumor Elimination by the Chok Locus

1998 ◽  
Vol 188 (12) ◽  
pp. 2243-2256 ◽  
Author(s):  
Azza H. Idris ◽  
Koho Iizuka ◽  
Hamish R.C. Smith ◽  
Anthony A. Scalzo ◽  
Wayne M. Yokoyama
Keyword(s):  
Nk Cells ◽  
Cho Cells ◽  
Molecular Mechanisms ◽  
Nk Cell ◽  
Interleukin 2 ◽  
Chinese Hamster ◽  
Mouse Strains ◽  
Xenograft Rejection ◽  
Natural Killing

The molecular mechanisms underlying target recognition during natural killing are not well understood. One approach to dissect the complexities of natural killer (NK) cell recognition is through exploitation of genetic differences among inbred mouse strains. In this study, we determined that interleukin 2–activated BALB/c-derived NK cells could not lyse Chinese hamster ovary (CHO) cells as efficiently as C57BL/6-derived NK cells, despite equivalent capacity to kill other targets. This strain-determined difference was also exhibited by freshly isolated NK cells, and was determined to be independent of host major histocompatibility haplotype. Furthermore, CHO killing did not correlate with expression of NK1.1 or 2B4 activation molecules. Genetic mapping studies revealed linkage between the locus influencing CHO killing, termed Chok, and loci encoded within the NK gene complex (NKC), suggesting that Chok encodes an NK cell receptor specific for CHO cells. In vivo assays recapitulated the in vitro data, and both studies determined that Chok regulates an NK perforin–dependent cytotoxic process. These results may have implications for the role of NK cells in xenograft rejection. Our genetic analysis suggests Chok is a single locus that affects NK cell–mediated cytotoxicity similar to other NKC loci that also regulate the complex activity of NK cells.

scholarly journals MicroRNA-20a mediates the cytotoxicity of natural killer cells in endometriosis via ERG/HLX/STAT4/perforin axis

2019 ◽  
Author(s):  
Li-Juan Chen ◽  
Bin Hu ◽  
Zhi-Qiang Han ◽  
Jian Ni ◽  
Yong-Ming Zhou ◽  
...  
Keyword(s):  
Nk Cells ◽  
Natural Killer ◽  
Nude Mice ◽  
Nk Cell ◽  
Protective Role ◽  
Cell Cytotoxicity ◽  
Nk Cell Cytotoxicity

Abstract Background: Intriguingly, microRNA-20a (miR-20a) has been recently witnessed to be involved in the pathogenesis of endometriosis (EMS) but the molecular mechanism controlled by miR-20a is to be undefined. The present study is designed to probe into how miR-20a acts to regulate the cytotoxicity of natural killer (NK) cells. Methods: Most of all, consistent with the clinical determination in EMS patients, miR-20a was determined to be down-regulated in NK cells isolated from nude mice. miR-20a could specifically bind to ERG and negatively regulates its expression in NK cells. Additionally, shRNA-mediated silencing of ERG decreased the expression of HLX. HLX up-regulated STAT4 by inducing proteasome degradation and inhibited NK cell cytotoxicity. Results: Of great importance, forced expression of miR-20a consequently induced NK cell cytotoxicity in vitro by increasing perforin expression via enhancement of STAT4 that was caused by impairing the binding of ERG to HLX enhancer. The in vivo experiments further confirmed the promoting role of miR-20a in the cytotoxicity of NK cells isolated from EMS nude mice and subsequent protective role of miR-20a against EMS-induced endometrial injury. Conclusion: The aforementioned data suggest that miR-20a potentiates the cytotoxicity of NK via up-regulating perforin mediated by ERG/HLX/STAT4, highlighting potential novel mechanisms associated with EMS progression.

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