Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies |
| |
| Affiliation: | 1. Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), P.O Box 4059, Kelvin Grove, Queensland, Australia;2. Faculty of Science, Health, Education and Engineering, University of Sunshine Coast (USC), P.O Box, 4556, Sippy Downs, Queensland, Australia;3. The Walter Eliza Hall Institute of Medical Research, 1G Royal Parade, 3052, The University of Melbourne, Parkville, Victoria, Australia;1. Center for Research on Intracellular Bacteria, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland;2. Children''s Hospital of Geneva, University Hospitals of Geneva and Medical School of the University Hospitals of Geneva and Medical School of the University of Geneva, Switzerland;3. St Mary''s Hospital, London, United Kingdom;1. Section of Pulmonary, Critical Care Medicine, and Allergy/Immunology, LSU Health Sciences Center, New Orleans, United States;2. The Research Institute for Children, Children’s Hospital, New Orleans and Department of Chemistry, University of New Orleans, United States;3. Department of Microbiology, Immunology, and Parasitology, LSU Health Sciences Center, New Orleans, United States;4. Department of Immunology, University of Pittsburgh, United States;1. Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan;2. Division of Molecular Oncology, National Cancer Center Research Institute, Tokyo, Japan;3. Department of Frontier Life Science, Nagasaki University Graduate School of Biomedical Science, Nagasaki, Japan;4. Laboratory of Virus Control, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan;5. Department of Laboratory Medicine, Kawasaki Medical School, Kurashiki, Japan;6. Department of Gastroenterology and Hematology, Faculty of Medicine, University of Miyazaki, Miyazaki, Japan;7. Department of Hematology and Immunology, Kagoshima University Hospital, Kagoshima, Japan;8. Laboratory of DNA Information Analysis, Human Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo, Japan;9. Department of Advanced Diagnosis, Clinical Research Center, Nagoya Medical Center, Nagoya, Japan;10. Department of Hematology, Kumamoto University School of Medicine, Kumamoto, Japan;11. Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto, Japan;12. Department of Hematology, Sasebo City General Hospital, Sasebo, Japan;13. Department of Hematology, Atomic Bomb Disease and Hibakusya Medicine Unit, Atomic Bomb Disease Institute, Nagasaki University, Nagasaki, Japan;14. Department of Hematology, National Cancer Center Hospital, Tokyo, Japan;15. Division of Hematology, Department of Medicine, Showa University School of Medicine, Tokyo, Japan;16. Department of Hematology and Oncology, Kanazawa University Hospital, Kanazawa, Japan;17. Division of Hematology, Department of Internal Medicine, Tokyo Metropolitan Ohtsuka Hospital, Tokyo, Japan;18. Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan;19. Laboratory of Molecular Medicine, Human Genome Center, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan;20. Division of Cancer Genomics, National Cancer Center Research Institute, Tokyo, Japan;21. Department of Hematology, Imamura General Hospital, Kagoshima, Japan;22. Department of Advanced Medical Innovation, St. Marianna University Graduate School of Medicine, Kawasaki, Japan;1. Key Lab of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China;2. Tongren Polytechnic College, Tongren 554300, Guizhou, China;3. National and Local Engineering Research Centre for Separation and Purification Ethnic Chinese Veterinary Herbs, Tongren 554300, Guizhou, China;1. School of Electronic and Information Engineering, Xi’an Jiaotong University, Xi’an, China;2. School of Electrical Engineering, Xi’an Jiaotong University, Xi’an, China;1. School of Medical and Applied Sciences, Central Queensland University, Rockhampton, Queensland 4702, Australia;2. School of Science and Health, Western Sydney University, Hawkesbury, Penrith, NSW 2751, Australia |
| |
| Abstract: | The koala (Phascolarctos cinereus) is an arboreal herbivorous marsupial that is an Australian icon. Koalas in many parts of Australia are under multiple threats including habitat destruction, dog attacks, vehicular accidents, and infectious diseases such as Chlamydia spp. and the koala retrovirus (KoRV), which may contribute to the incidence of lymphoma and leukaemia in this species. Due to a lack of koala-specific immune reagents and assays there is currently no way to adequately analyse the immune response in healthy, diseased or vaccinated animals. This paper reports the production and characterisation of the first anti-koala CD4 monoclonal antibody (mAb). The koala CD4 gene was identified and used to develop recombinant proteins for mAb production. Fluorochrome-conjugated anti-CD4 mAb was used to measure the levels of CD4+ lymphocytes collected from koala spleens (41.1%, range 20–45.1%) lymph nodes (36.3%, range 19–55.9%) and peripheral blood (23.8%, range 17.3–35%) by flow cytometry. Biotin-conjugated anti-CD4 mAb was used for western blot to determine an approximate size of 52 kDa for the koala CD4 molecule and used in immunohistochemistry to identify CD4+ cells in the paracortical region and germinal centres of spleen and lymph nodes. Using the anti-CD4 mab we showed that CD4 cells from vaccinated, but not control, koalas proliferated following in vitro stimulation with UV-inactivated Chlamydia pecorum and recombinant chlamydial antigens. Since CD4+ T cells have been shown to play a pivotal role in clearing chlamydial infection in both human and mouse infections, using this novel antibody will help determine the role CD4+ T cells play in protection against chlamydial infection in koalas and also enhance our knowledge of how KoRV affects the koala immune system. |
| |
| Keywords: | Koala Chlamydia CD4 T cell Monoclonal antibody |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
