Monoclonal antibodies against leukocytes in the beluga whale, Delphinapterus leucas
To better understand cetacean immunology, it is important to develop markers that identify specific leukocyte populations. We created a monoclonal antibody (mAb) library against leukocytes of the beluga whale (Delphinapterus leucas), and established five hybridoma clones that produce mAbs. Three of...
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| Published in | JAMSTEC REPORT OF RESEARCH AND DEVELOPMENT Vol. 27; pp. 119 - 126 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English Japanese |
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Yokohama
Japan Agency for Marine-Earth Science and Technology
01.09.2018
Japan Science and Technology Agency |
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| ISSN | 1880-1153 2186-358X |
| DOI | 10.5918/jamstecr.27.119 |
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| Abstract | To better understand cetacean immunology, it is important to develop markers that identify specific leukocyte populations. We created a monoclonal antibody (mAb) library against leukocytes of the beluga whale (Delphinapterus leucas), and established five hybridoma clones that produce mAbs. Three of these mAbs (ID: BW-3C3, BW-2G4, and BW-2G6) react with mononuclear leukocytes including lymphocytes and monocytes. mAb BW-3C3 react to a fraction of the lymphocytes. The mAb-positive cells were identical to cells that also stained with polyclonal anti-whale IgM antibodies, indicating that the mAb BW-3C3 specifically reacts to B lymphocytes. mAb BW-2G4 specifically binds to monocytes that possess a reniformed nucleus. mAb BW-2G6 was found to bind to heterogeneous lymphocytes, namely, anti-whale IgM antibody-positive and -negative lymphocyte populations. This indicates that this mAb reacts with B and non-B lymphocyte fractions. The other two mAbs (ID: BW-4B10 and BW-4G12) react with polymorphonuclear granulocytes. Double staining with Giemsa-eosin showed that mAb BW-4B10 and mAb BW-4G12 specifically identify neutrophils and eosinophils, respectively. This panel of mAbs will be a useful tool for classifying leukocytes and for determining their localization in different tissues, which in turn would contribute to our understanding of cetacean immunology and allow evaluation of leukocyte function in infectious diseases. |
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| AbstractList | To better understand cetacean immunology, it is important to develop markers that identify specific leukocyte populations. We created a monoclonal antibody (mAb) library against leukocytes of the beluga whale (Delphinapterus leucas), and established five hybridoma clones that produce mAbs. Three of these mAbs (ID: BW-3C3, BW-2G4, and BW-2G6) react with mononuclear leukocytes including lymphocytes and monocytes. mAb BW-3C3 react to a fraction of the lymphocytes. The mAb-positive cells were identical to cells that also stained with polyclonal anti-whale IgM antibodies, indicating that the mAb BW-3C3 specifically reacts to B lymphocytes. mAb BW-2G4 specifically binds to monocytes that possess a reniformed nucleus. mAb BW-2G6 was found to bind to heterogeneous lymphocytes, namely, anti-whale IgM antibody-positive and -negative lymphocyte populations. This indicates that this mAb reacts with B and non-B lymphocyte fractions. The other two mAbs (ID: BW-4B10 and BW-4G12) react with polymorphonuclear granulocytes. Double staining with Giemsa-eosin showed that mAb BW-4B10 and mAb BW-4G12 specifically identify neutrophils and eosinophils, respectively. This panel of mAbs will be a useful tool for classifying leukocytes and for determining their localization in different tissues, which in turn would contribute to our understanding of cetacean immunology and allow evaluation of leukocyte function in infectious diseases. |
| Author | Nakamura, Yoshimitsu Saito, Chiaki Ohishi, Kazue Yoshida, Takao Maruyama, Tadashi Saito, Masafumi Mishima, Hideki Hongo, Yuki Kakizoe, Yuka Nakazawa, Masatoshi |
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| Cites_doi | 10.1186/s12917-017-0998-3 10.1016/S0165-2427(01)00409-3 10.1016/S0145-305X(97)00021-9 10.1016/j.dci.2008.08.003 10.4236/ojms.2013.34019 10.1016/0076-6879(81)73054-4 10.1016/S0165-2427(03)00158-2 10.1016/0167-5699(91)90109-7 10.1016/S0147-9571(02)00036-X 10.3354/dao02410 10.1292/jvms.16-0076 10.1146/annurev.iy.05.040187.001431 10.1046/j.1469-7580.1999.19440505.x 10.1016/S0269-7491(02)00155-0 10.1007/s00244-011-9651-5 10.1016/0167-5699(94)90193-7 10.1111/j.1748-7692.1994.tb00260.x 10.1201/9781420041637.sec4 10.3390/v6125145 10.1080/15287390490486761 |
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| References_xml | – reference: Bossart, G.D., T.H. Reidarson, L.A. Dierauf, and D.A. Duffield (2001), Clinical Pathology, In: L.A. Dierauf and F.M.D. Gulland (Eds.), CRC handbook of marine mammal medicine, 383-436, CRC Press, Boca Raton. – reference: Kumar, D. and D.F. Cowan (1994), Cross-reactivity of antibodies to human antigens with tissues of the bottlenose dolphin, Tursiops truncatus, using immunoperoxidase techniques, Mar. Mamm. Sci., 10, 188-194. – reference: Testi, R., D. D'Ambrosio, R. De Maria, and A. Santoni (1994), The CD69 receptor: a multipurpose cell-surface trigger for hematopoietic cells, Immunol. Today, 15, 479-483. – reference: Van Bressem, M.F., P.J. Duignan, A. Banyard, M. Barbieri, K.M. Colegrove, S. De Guise, G. Di Guardo, A. Dobson, M. Domingo, D. Fauquier, et al. (2014), Cetaceans morbillivirus: current knowledge and future directions, Viruses, 6, 5145-5181. – reference: Ohishi, K., R. Zenitani, T. Bando, Y. Goto, K. Uchida, T. Maruyama, S. Yamamoto, N. Miyazaki, and Y. Fujise (2003), Pathological and serological evidence of Brucella-infection in baleen whales (Mysticeti) in the western North Pacific, Comp. Immunol. Microbiol. Infect. Dis., 26, 125-136. – reference: Galfre, G. and C. Milstein (1981), Preparation of monoclonal antibodies: Strategies and procedures, Methods in Enzymology, 73, 3-46. – reference: McChesney, M.B. and M.B.A. Oldstone (1987), Viruses perturb lymphocyte functions: selected principles characterizing virus-induced immunosuppression, Annu. Rev. Immunol., 5, 279-304. – reference: Nakamura, Y., M. Konishi, K. Ohishi, C. Kusaka, A. Tame, Y. Hatada, K. Fujikura, M. Nakazawa, M. Fujishima, T. Yoshida, and T. Maruyama (2013), Mucus glycoproteins selectively secreted from bacteriocytes in gill filaments of the deep-sea clam Calyptogena okutanii, Open J. Mar. Sci., 3, 167-174. – reference: Nouri-Shirazi, M., B.F. Bible, M. Zeng, S. Tamjidi, and G.D. Bossart (2017), Phenotyping and comparing the immune cell populations of free-ranging Atlantic bottlenose dolphins (Tursiops truncatus) and dolphins under human care, BMC Vet. Res., 13, 78. – reference: Schaefer, A.M., H.C.W. Stavros, G.D. Bossart, P.A. Fair, J.D. Goldstein, and J.S. Reif (2011), Associations between mercury and hepatic, renal, endocrine, and hematological parameters in Atlantic bottlenose dolphins (Tursiops truncatus) along the eastern coast of Florida and South Carolina, Arch. Environ. Contam. Toxicol., 61, 688-695. – reference: Levin, M., B. Morsey, C. Mori, and S. Guise (2004), Specific non-coplanar PCB-mediated modulation of bottlenose dolphin and beluga whale phagocytosis upon in vitro exposure, J. Toxicol. Environ. Health A, 67, 1517-1535. – reference: Sidorenko, S.P. and E.A. Clark (1993), Characterization of a cell surface glycoprotein IPO-3, expressed on activated human B and T lymphocytes, J. Immunol., 151, 4614-4624. – reference: Nakata, H., A. Sakakibara, M. Kanoh, S. Kudo, H. Watanabe, N. Nagai, N. Miyazaki, Y. Asano, and S. Tanabe (2002), Evaluation of mitogen-induced responses in marine mammal and human lymphocytes by in-vitro exposure of butyltins and non-ortho coplanar PCBs, Environ. Pollut., 120, 245-253. – reference: Hein, W.R. and C.R. Mackay (1991), Prominence of gamma delta T cells in the ruminant immune system, Immunol. Today, 12, 30-34. – reference: De Guise, S., K. Erickson, M. Blanchard, L. DiMolfetto, H.D. Lepper, J. Wang, J.L. Stott, and D.A. Ferrick (2002), Monoclonal antibodies to lymphocyte surface antigens for cetacean homologues to CD2, CD19 and CD21, Vet. Immunol. Immunopathol., 84, 209-221. – reference: Jaber, J.R., A. Fernandez, P. Herraez, A. Espinosa de los Monteros, G.A. Ramirez, P.M. Garcia, T. Fernandez, M. Arbelo, and J. Perez (2003), Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax-embedded tissues, Vet. Immunol. Immunopathol., 96, 65-72. – reference: Kato, M., T. Itou, N. Nagatsuka, and T. Sakai (2009), Production of monoclonal antibody specific for bottlenose dolphin neutrophils and its application to cell separation, Dev. Comp. Immunol., 33, 14-17. – reference: Cowan, D.F. and T.L. Smith (1999), Morphology of the lymphoid organs of the bottlenose dolphin, Tursiops truncatus, J. Anat., 194, 505-517. – reference: Ohishi, K., T. Bando, E. Abe, Y. Kawai, Y. Fujise, and T. Maruyama (2016), Long-term and large-scale epidemiology of Brucella infection in baleen whales and sperm whales in the western North Pacific and Antarctic oceans, J. Vet. Med. Sci., 78, 1457-1464. – reference: De Guise, S., J. Bernier, D. Martineau, P. Beland, and M. Fournier (1997), Phenotyping of beluga whale blood lymphocytes using monoclonal antibodies, Dev. Comp. Immunol., 21, 425-433. – reference: Bossart, G.D., T.A. Romano, M.M. Peden-Adams, A. Schaefer, S. McCulloch, J.D. Goldstein, C.D. Rice, J.T. Saliki, P.A. Fair, and J.S. Reif (2011), Clinicoimmunopathologic findings in Atlantic bottlenose dolphins Tursiops truncatus with positive cetacean morbillivirus antibody titers, Dis. Aquat. Organ., 97, 103-112. – ident: 15 doi: 10.1186/s12917-017-0998-3 – ident: 5 doi: 10.1016/S0165-2427(01)00409-3 – ident: 4 doi: 10.1016/S0145-305X(97)00021-9 – ident: 9 doi: 10.1016/j.dci.2008.08.003 – ident: 13 doi: 10.4236/ojms.2013.34019 – ident: 6 doi: 10.1016/0076-6879(81)73054-4 – ident: 8 doi: 10.1016/S0165-2427(03)00158-2 – ident: 7 doi: 10.1016/0167-5699(91)90109-7 – ident: 19 – ident: 16 doi: 10.1016/S0147-9571(02)00036-X – ident: 2 doi: 10.3354/dao02410 – ident: 17 doi: 10.1292/jvms.16-0076 – ident: 12 doi: 10.1146/annurev.iy.05.040187.001431 – ident: 3 doi: 10.1046/j.1469-7580.1999.19440505.x – ident: 14 doi: 10.1016/S0269-7491(02)00155-0 – ident: 18 doi: 10.1007/s00244-011-9651-5 – ident: 20 doi: 10.1016/0167-5699(94)90193-7 – ident: 10 doi: 10.1111/j.1748-7692.1994.tb00260.x – ident: 1 doi: 10.1201/9781420041637.sec4 – ident: 21 doi: 10.3390/v6125145 – ident: 11 doi: 10.1080/15287390490486761 |
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| SubjectTerms | Aquatic mammals beluga whale cetacean Immunology Infectious diseases leukocyte Lymphocytes Monoclonal antibodies monoclonal antibody top predator |
| Title | Monoclonal antibodies against leukocytes in the beluga whale, Delphinapterus leucas |
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