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Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells.

Publication ,  Journal Article
Hwang, K-K; Chen, X; Kozink, DM; Gustilo, M; Marshall, DJ; Whitesides, JF; Liao, H-X; Catera, R; Chu, CC; Yan, X-J; Luftig, MA; Haynes, BF ...
Published in: Blood
February 16, 2012

B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the clonal expansion of CD5-expressing B lymphocytes that produce mAbs often reactive with microbial or autoantigens. Long-term culture of B-CLL clones would permit the collection and characterization of B-CLL mAbs to study antigen specificity and of B-CLL DNA to investigate molecular mechanisms promoting the disease. However, the derivation of long-term cell lines (eg, by EBV), has not been efficient. We have improved the efficiency of EBV B-CLL transformation of CpG oligonucleotide-stimulated cells by incubating patient peripheral blood mononuclear cells in the presence of an irradiated mouse macrophage cell line, J774A.1. Using this approach, peripheral blood mononuclear cells isolated from 13 of 21 B-CLL patients were transformed as documented by IGHV-D-J sequencing. Four clones grew and retained CD5 expression in culture for 2 to 4 months. However, despite documentation of EBV infection by expression of EBNA2 and LMP1, B-CLL cells died after removal of macrophage feeder cells. Nevertheless, using electrofusion technology, we generated 6 stable hetero-hybridoma cell lines from EBV-transformed B-CLL cells, and these hetero-hybridomas produced immunoglobulin. Thus, we have established enhanced methods of B-CLL culture that will enable broader interrogation of B-CLL cells at the genetic and protein levels.

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Published In

Blood

DOI

EISSN

1528-0020

Publication Date

February 16, 2012

Volume

119

Issue

7

Start / End Page

e35 / e44

Location

United States

Related Subject Headings

  • Viral Proteins
  • Up-Regulation
  • Mice
  • Macrophages
  • Leukemia, Lymphocytic, Chronic, B-Cell
  • Immunology
  • Humans
  • Herpesvirus 4, Human
  • Feeder Cells
  • Epstein-Barr Virus Nuclear Antigens
 

Citation

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Hwang, K.-K., Chen, X., Kozink, D. M., Gustilo, M., Marshall, D. J., Whitesides, J. F., … Chiorazzi, N. (2012). Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells. Blood, 119(7), e35–e44. https://doi.org/10.1182/blood-2011-08-371203
Hwang, Kwan-Ki, Xi Chen, Daniel M. Kozink, Marietta Gustilo, Dawn J. Marshall, John F. Whitesides, Hua-Xin Liao, et al. “Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells.Blood 119, no. 7 (February 16, 2012): e35–44. https://doi.org/10.1182/blood-2011-08-371203.
Hwang K-K, Chen X, Kozink DM, Gustilo M, Marshall DJ, Whitesides JF, et al. Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells. Blood. 2012 Feb 16;119(7):e35–44.
Hwang, Kwan-Ki, et al. “Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells.Blood, vol. 119, no. 7, Feb. 2012, pp. e35–44. Pubmed, doi:10.1182/blood-2011-08-371203.
Hwang K-K, Chen X, Kozink DM, Gustilo M, Marshall DJ, Whitesides JF, Liao H-X, Catera R, Chu CC, Yan X-J, Luftig MA, Haynes BF, Chiorazzi N. Enhanced outgrowth of EBV-transformed chronic lymphocytic leukemia B cells mediated by coculture with macrophage feeder cells. Blood. 2012 Feb 16;119(7):e35–e44.

Published In

Blood

DOI

EISSN

1528-0020

Publication Date

February 16, 2012

Volume

119

Issue

7

Start / End Page

e35 / e44

Location

United States

Related Subject Headings

  • Viral Proteins
  • Up-Regulation
  • Mice
  • Macrophages
  • Leukemia, Lymphocytic, Chronic, B-Cell
  • Immunology
  • Humans
  • Herpesvirus 4, Human
  • Feeder Cells
  • Epstein-Barr Virus Nuclear Antigens