Skip to main content

Cytogenetic Methods in Chronic Lymphocytic Leukemia

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 730))

Abstract

In chronic lymphocytic leukemia of the B-lineage (B-CLL), cytogenetic alterations are highly relevant for prognosis and therapeutic decisions. With conventional techniques, chromosome banding analysis in CLL has been hampered by the low quality of metaphases and low rates of cytogenetic alterations due to a low in vitro proliferation rate of CLL cells. Thus, interphase fluorescence in situ hybridization (FISH) has become the standard technique for cytogenetic analysis in CLL. However, interphase FISH is not able to provide an overview on the whole karyotype. In order to improve chromosome banding analysis in CLL, specific stimulation techniques have been developed. These either use CD40 ligand or oligonucleotides (e.g., CpG) and IL-2 in combination. With the respective techniques, metaphase cultivation is successful in >90% of CLL cases and aberrant karyotypes can be detected in nearly 90% of CLL cases. This has allowed the detection of new clinically relevant subgroups (e.g., complex aberrant karyotype cases) and a more differentiated picture of distinct cytogenetic subtypes, e.g., cases with a 13q deletion. Efforts should continue to define the value of chromosomal banding in CLL focusing as well on the interaction with already established techniques such as interphase FISH or immunophenotyping.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Hallek M, Stahel RA, Greil R. ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of chronic lymphocytic leukemia. Ann Oncol 2005; 16 Suppl 1: i50–1.

    Article  PubMed  Google Scholar 

  2. Hallek M, Cheson BD, Catovsky D, Caligaris-Cappio F, Dighiero G, Döhner H et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia (IWCLL) updating the National Cancer Institute-Working Group (NCI-WG) 1996 guidelines. Blood 2008; 111(12): 5446–56.

    Article  PubMed  CAS  Google Scholar 

  3. Shanafelt TD, Call TG. Current approach to diagnosis and management of chronic lymphocytic leukemia. Mayo Clin Proc 2004; 79(3): 388–98.

    Article  PubMed  Google Scholar 

  4. Oscier DG, Gardiner AC, Mould SJ, Glide S, Davis ZA, Ibbotson RE et al. Multivariate analysis of prognostic factors in CLL: clinical stage, IGVH gene mutational status, and loss or mutation of the p53 gene are independent prognostic factors. Blood 2002; 100(4): 1177–84.

    PubMed  CAS  Google Scholar 

  5. Juliusson G, Oscier DG, Fitchett M, Ross FM, Stockdill G, Mackie MJ et al. Prognostic subgroups in B-cell chronic lymphocytic leukemia defined by specific chromosomal abnormalities. N Engl J Med 1990; 323(11): 720–4.

    Article  PubMed  CAS  Google Scholar 

  6. Döhner H, Stilgenbauer S, Benner A, Leupolt E, Kröber A, Bullinger L et al. Genomic aber­rations and survival in chronic lymphocytic leukemia. N Engl J Med 2000; 343(26): 1910–6.

    Article  PubMed  CAS  Google Scholar 

  7. Mayr C, Speicher MR, Kofler DM, Buhmann R, Strehl J, Busch R et al. Chromosomal translocations are associated with poor prognosis in chronic lymphocytic leukemia. Blood 2006; 107(2): 742–51.

    Article  PubMed  CAS  Google Scholar 

  8. Buhmann R, Kurzeder C, Rehklau J, Westhaus D, Bursch S, Hiddemann W et al. CD40L stimulation enhances the ability of conventional metaphase cytogenetics to detect chromosome aberrations in B-cell chronic lymphocytic leukaemia cells. Br J Haematol 2002; 118(4): 968–75.

    Article  PubMed  CAS  Google Scholar 

  9. Hallaert DY, Jaspers A, van Noesel CJ, van Oers MH, Kater AP, Eldering E. c-Abl kinase inhibitors overcome CD40-mediated drug resistance in CLL: implications for therapeutic targeting of chemoresistant niches. Blood 2008; 112(13): 5141–9.

    Article  PubMed  CAS  Google Scholar 

  10. Decker T, Schneller F, Hipp S, Miething C, Jahn T, Duyster J, Peschel C. Cell cycle progression of chronic lymphocytic leukemia cells is controlled by cyclin D2, cyclin D3, cyclin-dependent kinase (cdk) 4 and the cdk inhibitor p27. Leukemia 2002; 16(3): 327–34.

    Article  PubMed  CAS  Google Scholar 

  11. Dicker F, Schnittger S, Haferlach T, Kern W, Schoch C. Immunostimulatory oligonucleotide-induced metaphase cytogenetics detect chromosomal aberrations in 80% of CLL patients: a study of 132 CLL cases with correlation to FISH, IgVH status, and CD38 expression. Blood 2006; 108(9): 3152–60.

    Article  PubMed  CAS  Google Scholar 

  12. Gahrton G, Robert KH, Friberg K, Zech L, Bird AG. Nonrandom chromosomal aberrations in chronic lymphocytic leukemia revealed by polyclonal B-cell-mitogen stimulation. Blood 1980; 56(4): 640–7.

    PubMed  CAS  Google Scholar 

  13. Stilgenbauer S, Bullinger L, Lichter P, Döhner H. Genetics of chronic lymphocytic leukemia: genomic aberrations and V(H) gene mutation status in pathogenesis and clinical course. Leukemia 2002; 16(6): 993–1007.

    Article  PubMed  CAS  Google Scholar 

  14. Dicker F, Herholz H, Schnittger S, Nakao A, Patten N, Wu L et al. The detection of TP53 mutations in chronic lymphocytic leukemia independently predicts rapid disease progression and is highly correlated with a complex aberrant karyotype. Leukemia 2009; 23(1): 117–24.

    Article  PubMed  CAS  Google Scholar 

  15. Haferlach C, Dicker F, Schnittger S, Kern W, Haferlach T. Comprehensive genetic characterization of CLL: a study on 506 cases analysed with chromosome banding analysis, interphase FISH, IgV(H) status and immunophenotyping. Leukemia 2007; 21(12): 2442–51.

    Article  PubMed  CAS  Google Scholar 

  16. Wu X, Nowakowski G, Smoley S, Arendt B, Peterson M, Van Dyke D et al. Cytogenetic analysis of normal human B cells following CpG stimulation: implications for interpretation of CpG induced CLL metaphase analysis. ASH annual meeting abstracts; Blood 2008; 112: #3124.

    Google Scholar 

  17. Put N, Konings P, Rack K, Jamar M, Van Roy N, Libouton JM et al. Improved detection of chromosomal abnormalities in chronic lymphocytic leukemia by conventional ­cytogenetics using CpG oligonucleotide and interleukin-2 stimulation: a Belgian multicentric study. Genes Chromosomes Cancer 2009; 48(10): 843–53.

    Article  PubMed  CAS  Google Scholar 

  18. Schoch C, Kern W, Kohlmann A, Hiddemann W, Schnittger S, Haferlach T. Acute myeloid leukemia with a complex aberrant karyotype is a distinct biological entity characterized by genomic imbalances and a specific gene expression profile. Genes Chromosomes Cancer 2005; 43(3): 227–38.

    Article  PubMed  CAS  Google Scholar 

  19. Struski S, Gervais C, Helias C, Herbrecht R, Audhuy B, Mauvieux L. Stimulation of B-cell lymphoproliferations with CpG-oligonucleotide DSP30 plus IL-2 is more effective than with TPA to detect clonal abnormalities. Leukemia 2009; 23(3): 617–9.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Claudia Haferlach .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Haferlach, C., Bacher, U. (2011). Cytogenetic Methods in Chronic Lymphocytic Leukemia. In: Campbell, L. (eds) Cancer Cytogenetics. Methods in Molecular Biology, vol 730. Humana Press. https://doi.org/10.1007/978-1-61779-074-4_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-074-4_9

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-073-7

  • Online ISBN: 978-1-61779-074-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics