Comparative immunological study on hematopoietic organ in dolphin

• Project/Area Number: 20780145
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: General fisheries
• Research Institution: Nihon University
• Principal Investigator: ITOU Takuya Nihon University, 生物資源科学部, 准教授 (20307820)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2010: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000); Fiscal Year 2009: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2008: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: イルカ / 造血 / モノクローナル抗体 / 好中球 / 好酸球 / 比較免疫学 / 造血細胞 / 造血幹細胞

Research Abstract

The monoclonal antibody(DN1), which specifically reacts with the bottlenose dolphin neutrophils, was produced. The DN1 was useful for high-purity isolation of the dolphin granulocytes. The CD34, which specifically develops in the hematopoietic cells, were molecularly cloned in the bottlenose dolphin. The CD34 mRNA was strongly expressed in the bone marrow of the dolphin. Morphological observation has shown that the humerus is a source of hematopoietic tissue in dolphin.

Research Products

• [Journal Article] Molecular cloning and expression of bottlenose dolphin CD34. Vet (2011)
  • Author(s): Segawa T, Itou T, Echigoya Y, Suzuki M, Koie H, Sakai T.
  • Journal Title: Immunol. Immunopathol Volume: 139 Pages: 303-307
  • Peer Reviewed
• [Journal Article] Molecular cloning and expression of bottlenose dolphin CD34. (2011)
  • Author(s): Segawa T, Itou T, Echigoya Y, Suzuki M, Koie H, Sakai T.
  • Journal Title: Vet.Immunol.Immunopathol. 139 Pages: 303-307
  • Peer Reviewed
• [Journal Article] Molecular cloning and expression of bottlenose dolphin CD34. (2011)
  • Author(s): Segawa T, et al.
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 139 Pages: 303-307
  • Peer Reviewed
• [Journal Article] Bone marrow biopsy from the flipper of a dolphin (2010)
  • Author(s): Itou T, Koie H, Segawa T, Kato M, Yanagisawa M, Ueda K, Kuwano R, Suzuki M, Moritomo T, Sakai T.
  • Journal Title: Vet. J Volume: 185 Pages: 216-217
  • Peer Reviewed
• [Journal Article] Bone marrow biopsy from the flipper of a dolphin. (2010)
  • Author(s): Itou T, Koie H, Segawa T, Kato M, Yanagisawa M, Ueda K, Kuwano R, Suzuki M, Moritomo T, Sakai T.
  • Journal Title: Vet.J. 185 Pages: 216-217
  • Peer Reviewed
• [Journal Article] Bone marrow biopsy from the flipper of a dolphin. (2010)
  • Author(s): Itou T, et al.
  • Journal Title: The Veterinary Journal Volume: 185 Pages: 216-217
  • Peer Reviewed
• [Journal Article] Production of monoclonal antibody specific for bottlenose dolphin neutrophils and its application to cell separation (2009)
  • Author(s): Kato M, Itou T, Nagatsuka N, Sakai T.
  • Journal Title: Dev. Comp. Immunol Volume: 33 Pages: 14-17
  • Peer Reviewed
• [Journal Article] Production of monoclonal antibody specific for bottlenose dolphin neutrophils and its application to cell separation. (2009)
  • Author(s): Kato M, Itou T, Nagatsuka N, Sakai T.
  • Journal Title: Dev.Comp.Immunol. 33 Pages: 14-17
  • Peer Reviewed
• [Journal Article] Production of monoclonal antibody specific for bottlenose dolphin neutrophils and its application to cell senaration (2009)
  • Author(s): Kato M, et.al.
  • Journal Title: Dev. Comp. Immunol 33 Pages: 14-17
  • Peer Reviewed
• [Journal Article] Bone marrow biopsy from the flipper of a dolphin.
  • Author(s): Itou T, et al.
  • Journal Title: The Veterinary Journal (In press)
  • Peer Reviewed
• [Presentation] イルカ骨髄細胞の分化能および増殖能の解析 (2011)
  • Author(s): 伊藤琢也
  • Organizer: 日本大学幹細胞フォーラム
  • Place of Presentation: 日本大学会館
  • Year and Date: 2011-01-22
• [Presentation] ハンドウイルカCD34の分子クローニングと発現解析 (2010)
  • Author(s): 瀬川太雄
  • Organizer: 第150回日本獣医学会学術集会
  • Place of Presentation: 帯広畜産大学
  • Year and Date: 2010-09-18
• [Presentation] ハンドウイルカCD34の分子クローニングと発現解析 (2010)
  • Author(s): 瀬川太雄, 他
  • Organizer: 第150回日本獣医学会学術集会
  • Place of Presentation: 帯広畜産大学キャンパス
  • Year and Date: 2010-09-18
• [Presentation] イルカの造血は骨髄で行われている (2009)
  • Author(s): 伊藤琢也
  • Organizer: 日本比較免疫学会第21回学術集会
  • Place of Presentation: 日本大学生物資源科学部
  • Year and Date: 2009-08-04
• [Presentation] イルカの造血は骨髄で行われている (2009)
  • Author(s): 伊藤琢也, 他
  • Organizer: 日本比較免疫学会第21回学術集会
  • Place of Presentation: 日本大学生物資源学部
  • Year and Date: 2009-08-04
• [Presentation] ハンドウイルカ好中球に対する特異的モノクローナル抗体の作出とそれを用いた血球分離法の確立 (2008)
  • Author(s): 伊藤琢也
  • Organizer: 第146回日本獣医学会学術集会
  • Place of Presentation: 宮崎大学(シーガイア)
  • Year and Date: 2008-09-25
• [Presentation] コロニー形成能および分化能の解析によるイルカ骨髄における造血細胞の証明
  • Author(s): 瀬川太雄
  • Organizer: 第152回日本獣医学会学術集会(東日本大震災の為中止)
• [Presentation] コロニー形成能および分化能の解析によるイルカ骨髄における造血細胞の証明
  • Author(s): 瀬川太雄
  • Organizer: 第152回日本獣医学会学術集会
  • Place of Presentation: (学会中止、抄録のみ)
• [Remarks] 国際雑誌における研究内容(雑誌論文(2))のゲスト論説Vet. J. 185, 101-102, 2010. Guest Editorial. By Daniel Martineau
  • URL: http://dx.doi.org/10.1016/j.tvjl.2009.12.026
• [Remarks] A new approach to bone biopsy in cetaceans
• [Remarks] In this issue of The Veterinary Journal, Itou et al.(2010) report a convenient method to sample bone marrow in cetaceans. This is a welcome addition to the veterinary armamentarium, because up to now veterinarians have been reluctant to carry out this procedure in cetaceans. Because the iliac crest and femur, the sampling sites used in terrestrial mammals, are absent in cetaceans, bone marrow is usually obtained from vertebrae, a site prone to iatrogenic damage. With the new proposed approach, practitioners have an additional diagnostic tool when confronted with the most common indications for bone marrow biopsy in cetaceans, such as anemia with no evidence of regeneration, regenerative anemia unresponsive to treatment, or, more rarely, an abnormal leukogram raising suspicions of leukemia/lymphoma.
• [Remarks] If biopsies of bone marrow are carried out more often, a better knowledge of the changes associated with physiological states such as aging will be gained. Bone marrow biopsy will also help to elucidate the etiology of new intriguing clinical conditions. For instance, anemia with various degrees of splenic and hepatic hemosiderosis is occasionally seen in cetaceans kept in aquaria. Some of these cases are probably due to chronic bacterial infections(D. Martineau, personal observation ; G. Bossart, personal communication). Other cases of anemia with hemosiderosis might be caused by chronic inflammatory states induced by bacteria newly described in cetaceans, such as Bartonella henselae and Brucella ceti, whose pathogenicity remains obscure ([Gonzalez-Barrientos et al., 2009] and [Maggi et al., 2008]). These intracellular bacteria are phylogenetically close and are both potential zoonotic agents(Wattam et al., 2009). They are known to infect bone marrow in terrestrial mammals and humans(Franco et al., 2007). In people, B. henselae is thought to infect and persist in erythrocytic progenitors within the bone marrow (Greub and Raoult, 2002 ; Florin et al., 2008). Whether Brucella spp. or Bartonella spp. infect the bone marrow has not been determined in cetaceans.
• [Remarks] Biopsy of bone marrow could also be useful in marine ecotoxicology to define the biological significance(e. g. toxicity) of environmental contaminants. Worldwide, free-ranging cetaceans are contaminated and variously intoxicated by polychlorinated biphenyls and polycyclic aromatic hydrocarbons, which are known immunosuppressive and/or carcinogenic compounds ([Martineau et al., 1987],[Martineau et al., 1988], [Marsili et al., 2001] and [Wells et al., 2005]). Bone marrow is one of the target organs for these contaminants ([Thurmond et al., 1999] and [Galvan et al., 2006]). New contaminants such as perfluorinated compounds, polybrominated diphenylether and their derivatives may also have an effect on the bone marrow (Yamamoto et al., 2006).
• [Remarks] Bone marrow stromal cells(BMSC) have been recently employed in the exciting new fields of cell and gene therapy. In human and veterinary medicine, BMSCs have been used as work horses for the expression of therapeutic proteins because they can be conveniently sampled in large numbers from the bone marrow of patients. For instance, autologous BMSCs genetically engineered to produce human, murine or canine erythropoietin have been sampled, engineered in vitro with the EPO gene from the corresponding species, and expanded in vitro. Subsequently, the EPO-engineered BMSCs have been implanted subcutaneously in a commercially available matrix in mice and dogs. The production and release of EPO has elevated hematocrit in these animals for weeks and even months. In addition, BMSC can differentiate in cardiomyocytes, myoblasts, osteoblasts, endothelial cells and other cell types. BMSCs are also able to induce neovascularisation. For these reasons, they have been used to repair necrotic cardiac muscle, and large bone defects in experimental 'regenerative' medicine(Fontaine et al., 2004 ; reviewed in Eliopoulos et al., 2008). Consequently, easy access to cetacean bone marrow(and consequently to BMSCs) could open the way to cell therapy in cetaceans.
• [Remarks] References Itou et al., 2010, Bone marrow biopsy from the flipper of a dolphin. The Veterinary Journal, 185(2010), pp. 216-217.他省略