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      Authentication of Primordial Characteristics of the CLBL-1 Cell Line Prove the Integrity of a Canine B-Cell Lymphoma in a Murine In Vivo Model

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          Abstract

          Cell lines are key tools in cancer research allowing the generation of neoplasias in animal models resembling the initial tumours able to mimic the original neoplasias closely in vivo. Canine lymphoma is the major hematopoietic malignancy in dogs and considered as a valuable spontaneous large animal model for human Non-Hodgkin's Lymphoma (NHL). Herein we describe the establishment and characterisation of an in vivo model using the canine B-cell lymphoma cell line CLBL-1 analysing the stability of the induced tumours and the ability to resemble the original material. CLBL-1 was injected into Rag2 −/−γ c −/− mice. The generated tumor material was analysed by immunophenotyping and histopathology and used to establish the cell line CLBL-1M. Both cell lines were karyotyped for detection of chromosomal aberrations. Additionally, CLBL-1 was stimulated with IL-2 and DSP30 as described for primary canine B-cell lymphomas and NHL to examine the stimulatory effect on cell proliferation. CLBL-1 in vivo application resulted in lymphoma-like disease and tumor formation. Immunophenotypic analysis of tumorous material showed expression of CD45 +, MHCII +, CD11a + and CD79αcy +. PARR analysis showed positivity for IgH indicating a monoclonal character. These cytogenetic, molecular, immunophenotypical and histological characterisations of the in vivo model reveal that the induced tumours and thereof generated cell line resemble closely the original material. After DSP30 and IL-2 stimulation, CLBL-1 showed to respond in the same way as primary material. The herein described CLBL-1 in vivo model provides a highly stable tool for B-cell lymphoma research in veterinary and human medicine allowing various further in vivo studies.

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          Most cited references 35

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          RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement.

          We have generated mice that carry a germline mutation in which a large portion of the RAG-2 coding region is deleted. Homozygous mutants are viable but fail to produce mature B or T lymphocytes. Very immature lymphoid cells were present in primary lymphoid organs of mutant animals as defined by surface marker analyses and Abelson murine leukemia virus (A-MuLV) transformation assays. However, these cells did not rearrange their immunoglobulin or T cell receptor loci. Lack of V(D)J recombination activity in mutant pre-B cell lines could be restored by introduction of a functional RAG-2 expression vector. Therefore, loss of RAG-2 function in vivo results in total inability to initiate V(D)J rearrangement, leading to a novel severe combined immune deficient (SCID) phenotype. Because the SCID phenotype was the only obvious abnormality detected in RAG-2 mutant mice, RAG-2 function and V(D)J recombinase activity, per se, are not required for development of cells other than lymphocytes.
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            Defective lymphoid development in mice lacking expression of the common cytokine receptor gamma chain.

            The common gamma chain (gamma c) of the IL-2, IL-4, IL-7, IL-9, and IL-15 receptors is defective in humans with XSCID. Mice lacking gamma c expression had hypoplastic thymuses; the thymocytes responded to gamma c-independent mitogens, but not gamma c-dependent stimuli. Splenic T cells were diminished at 3 weeks of age, but CD4+ T cells markedly increased by 4 weeks. B cells were greatly diminished in contrast with the situation in XSCID. NK cells, gamma delta intestinal intraepithelial lymphocytes, dendritic epidermal T cells, peripheral lymph nodes, and gut-associated lymphoid tissue were absent. These findings underscore the importance of gamma c in lymphoid development. Moreover, differences in humans and mice lacking gamma c expression indicate species-specific differences in the roles of gamma c-dependent cytokines or in the existence of redundant pathways. These mice provide an important model for studying the pathophysiology provide an important model for studying the pathophysiology of and gene therapy for human XSCID.
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              Spontaneously occurring tumors of companion animals as models for human cancer.

               D Vail,  E Macewen (1999)
              Spontaneous tumors in companion animals (dog and cat) offer a unique opportunity as models for human cancer biology and translational cancer therapeutics. The relatively high incidence of some cancers, similar biologic behavior, large body size, comparable responses to cytotoxic agents, and shorter overall lifespan are the factors that contribute to the advantages of the companion animal model. The tumor types that offer the best comparative interest include lymphoma/leukemia, osteosarcoma, STS, melanoma, and mammary tumors. With the increase in new therapeutic agents (traditional chemotherapy, gene therapy, biologic agents, etc.), the companion animal model can provide useful populations to test new agents where efficacy and toxicity can be examined.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                28 June 2012
                : 7
                : 6
                Affiliations
                [1 ]Central Laboratory, Department of Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria
                [2 ]Small Animal Clinic and Research Cluster of Excellence ‘REBIRTH’, University of Veterinary Medicine Hannover, Hannover, Lower Saxony, Germany
                [3 ]VetBioBank, VetCore Facility for Research, University of Veterinary Medicine Vienna, Vienna, Austria
                [4 ]Institute of Pathology, Department of Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria
                [5 ]Department of Biomedical Sciences, Translational Oncology, Institute for Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Vienna, Austria
                [6 ]Department of Pathobiology, Institute of Immunology, University of Veterinary Medicine Vienna, Vienna, Austria
                National Cancer Institute, United States of America
                Author notes

                Conceived and designed the experiments: IS IN AS HME. Performed the experiments: BCR S. Willenbrock NRB IW AFB S. Wagner BK SE. Analyzed the data: BCR S. Willenbrock NRB IW AFB S. Wagner BK SE. Contributed reagents/materials/analysis tools: IS IN AS HME. Wrote the paper: BCR S. Willenbrock SE HME.

                Article
                PONE-D-12-04827
                10.1371/journal.pone.0040078
                3386195
                22761949
                Rütgen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                Counts
                Pages: 13
                Categories
                Research Article
                Biology
                Genetics
                Cytogenetics
                Cytogenetic Analysis
                Cancer Genetics
                Immunology
                Immune Cells
                B Cells
                Model Organisms
                Animal Models
                Mouse
                Medicine
                Oncology
                Basic Cancer Research
                Tumor Physiology
                Veterinary Science
                Animal Types
                Laboratory Animals
                Small Animals
                Veterinary Medicine
                Veterinary Oncology

                Uncategorized

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