Characterization of a Novel Monoclonal Antibody (27H2) Recognizing Human CD34 Class III Epitope
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.
Abstract
Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.
참고문헌 (27)
- Civin CI, "Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells", J Immunol, 133, pp.157~165 (1984)
- Tindle RW, "A novel monoclonal antibody BI-3C5 recognises myeloblasts and non-B non-T lymphoblasts in acute leukaemias and CGL blast crises, and reacts with immature cells in normal bone marrow", Leuk Res, 9, pp.1~9 (1985)
- Watt SM, "Distribution and epitope analysis of the cell membrane glycoprotein (HPCA-1) associated with human hemopoietic progenitor cells", Leukemia, 1, pp.417~426 (1987)
- Beschorner WE, "Localization of hematopoietic progenitor cells in tissue with the anti-My-10 monoclonal antibody", Am J Pathol, 119, pp.1~4 (1985)
- Fina L, "Expression of the CD34 gene in vascular endothelial cells", Blood, 75, pp.2417~2426 (1990)
- Civin CI, "Purification and expansion of human hematopoietic stem", progenitor cells, 1995, pp.770, 91~98
- Greaves MF, "Molecular features of CD34: a hemopoietic progenitor cell-associated molecule", Leukemia, 6(Suppl 1), pp.31~36 (1992)
- Sutherland DR, "The CD34 antigen: structure, biology, and potential clinical applications", J Hematother, 1, pp.115~129 (1992)
- Sutherland DR, "The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering", J Hematother, 5, pp.213~226 (1996)
- Siena S, "Flow cytometry to estimate circulating hematopoietic progenitors for autologous transplantation: comparative analysis of different CD34 monoclonal antibodies", Haematologica, 76, pp.330~333 (1991)
- Katz FE, "Identification of a membrane glycoprotein associated with haemopoietic progenitor cells", Leuk Res, 9, pp.191~198 (1985)
- Shpall EJ, "Transplantation of autologous CD34+ hematopoietic progenitor cells into breast cancer patients following high-dose chemotherapy", J Clin Oncol, 12, pp.28~32 (1994)
- Vescio R, "Multicenter phase III trial to evaluate CD34(＋) selected versus unselected autologous peripheral blood progenitor cell transplantation in multiple myeloma", Blood, 93, pp.1858~1868 (1999)
- Civin CI, "M7.1. Report on the CD34 cluster workshop, Leukocyte typing IV. White cell differentiation antigens", Proceedings of the 4th International Workshop and Conference, pp.818~825 (1989), Oxford University Press
- Greaves MF, "Report on the CD34 cluster Workshop, In Leukocyte Typing V", pp.840~846 (1995), Oxford University Press
- Nishio H, "MC7. CD34 workshop panel report, In Leucocyte typing VI. White cell differentiation antigens", Proceedings of the 6th International Workshop and Conference, pp.974~984 (1997), Garland Publishing Inc.
- Titley I, "Extent of variability inherent in measurements of CD34-positive cells in different human haemopoietic tissues", Bone Marrow Transplant, 16, pp.611~616 (1995)
- Titley I, "Extent of variability inherent in measurements of CD34-positive cells in different human haemopoietic tissues", Bone Marrow Transplant, 16, pp.611~616 (1995)
- Cannovo N, "Significant diversity of the reactivity of monoclonal CD34 antibodies requires an extended classification of the CD34 epitopes", Lab Hematol, 3, pp.253~260 (1997)
- Serke S, "Expression of class I, II and III epitopes of the CD34 antigen by normal and leukemic hemopoietic cells", Cytometry, 26, pp.154~160 (1996)
- Steen R, "Differences in the distribution of CD34 epitopes on normal haemopoietic progenitor cells and leukaemic blast cells", Br J Haematol, 94, pp.597~605 (1996)
- Lanza F, "CD34＋ leukemic cells assessed by different CD34 monoclonal antibodies", Leuk Lymphoma, 18(Suppl 1), pp.25~30 (1995)
- Weinberg DS, "QBEnd10 (CD34) antibody is unsuitable for routine use in the ISHAGE CD34＋ cell determination assay", J Hematother, 5, pp.599~603 (1996)
- Macey MG, "How should CD34＋ cells be analysed? A study of three classes of antibody and five leucocyte preparation procedures", J Immunol Methods, 204, pp.175~188 (1997)
- Sovalat H, "Comparative analysis of class I, II and III epitope-detecting CD34 monoclonal antibodies by quantitative flow cytometry", Hematol Cell Ther, 40, pp.259~268 (1998)
- Sutherland DR, "The ISHAGE Guidelines for CD34＋ cell determination: Applications in autologous and allogenic blood and marrow transplantation. In: Bone Marrow Transplantation: A Practical Approach", pp.239~246 (2000), Churchill Livingstone
- Lanza F, "Structural and functional features of the CD34 antigen: an update", J Biol Regul Homeost Agents, 15, pp.1~13 (2001)