title: Roles of Bcl-2 and caspase-9 and -3 in CD30-induced human eosinophil apoptosis.
abstract: OBJECTIVE
  Activation of cell surface CD30 by immobilized anti-CD30 monoclonal antibodies (mAb) induces strong apoptosis in human eosinophils. This anti-CD30 mAb-induced eosinophil apoptosis is inhibited by the addition of inhibitors of p38, ERK1/2 mitogen-activated protein kinases, and phosphatidylinositol 3-kinase. However, there is little data investigating the role of Bcl-2 and caspases in eosinophil apoptosis induced by anti-CD30 mAb. We sought to determine whether anti-CD30 mAb induces human eosinophil apoptosis via Bcl-2 and caspase pathways.
  
  
  METHODS
  Peripheral blood was drawn from 37 healthy volunteers. The CD30 expression on eosinophils was measured at various time points. Eosinophils were then cultured in plates precoated with anti-CD30 mAb (clone Ber-H8), isotype control immunoglobulin G1, interleukin (IL)-5, or dexamethasone. Western blot analysis was performed to determine the expression of Bcl-2, procaspase-8, -9, and -3, and caspase-8, -9, and -3 after cross-linking of CD30. Human eosinophils were also cultured in plates precoated with anti-CD30 mAb (clone Ber-H8) in the presence or absence of caspase-9 or -3 inhibitors. Eosinophil apoptosis was assessed using flow cytometry.
  
  
  RESULTS
  The addition of anti-CD30 mAb significantly increased eosinophil apoptosis compared with controls. In western blot analysis, the addition of anti-CD30 mAb significantly decreased the expression of Bcl-2 and procaspase-9 and -3 and increased the expression of caspase-9 and -3. The addition of caspase-9 or -3 inhibitors decreased anti-CD30 mAb-induced human eosinophil apoptosis. Procaspase-8 or caspase-8 expression was not changed in response to various stimuli.
  
  
  CONCLUSIONS
  Anti-CD30 mAb-induced human eosinophil apoptosis is likely to be mediated through Bcl-2 and caspase-9 and -3.
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Hye Jin
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Eun-Kyoung
authors.initials: EK
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Young Eun
authors.initials: YE
authors.lastname: Seo
authors.affiliation: Department of Medicine, The Graduate School, Yonsei University, Seoul, Republic of Korea; Department of Pediatrics, CHA University School of Medicine, Seoul, Republic of Korea.
authors.forename: Youn Ho
authors.initials: YH
authors.lastname: Shin
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Hwan Soo
authors.initials: HS
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Yoon Hong
authors.initials: YH
authors.lastname: Chun
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Jong-Seo
authors.initials: JS
authors.lastname: Yoon
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
authors.forename: Hyun Hee
authors.initials: HH
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, CHA University School of Medicine, Seoul, Republic of Korea.
authors.forename: Man Yong
authors.initials: MY
authors.lastname: Han
authors.affiliation: Asthma and Allergy Center, Department of Pediatrics, Inje University Sanggye Paik Hospital, Seoul, Republic of Korea.
authors.forename: Chang-Keun
authors.initials: CK
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Kyu-Earn
authors.initials: KE
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Seoul National University Hospital, Seoul, Republic of Korea.
authors.forename: Young Yull
authors.initials: YY
authors.lastname: Koh
authors.affiliation: Department of Pediatrics, School of Medicine, The Catholic University of Korea, Seoul, Republic of Korea. Electronic address: jintackk@catholic.ac.kr.
authors.forename: Jin Tack
authors.initials: JT
authors.lastname: Kim
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology / Q000502:physiology*; D017209:Apoptosis / Q000276:immunology*; D015153:Blotting, Western; D053148:Caspase 3 / Q000502:physiology*; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000502:physiology*; D020169:Caspases; D053471:Caspases, Initiator; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D004804:Eosinophils / Q000166:cytology / Q000276:immunology*; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D015848:Interleukin-5 / Q000378:metabolism; D017730:Ki-1 Antigen / Q000096:biosynthesis / Q000276:immunology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; C000595707:BCL2 protein, human; C508599:IL5 protein, human; D015848:Interleukin-5; D017730:Ki-1 Antigen; D019253:Proto-Oncogene Proteins c-bcl-2; C504717:SGN-30 monoclonal antibody; D019869:Phosphatidylinositol 3-Kinases; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases; D053471:Caspases, Initiator
keywords: Bcl-2; apoptosis; caspases; eosinophils