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Amplifying Igh translocations

Nature Immunology volume 6, page 117 (2005)Cite this article

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PCR has been used to amplify chromosome translocation breakpoints between Myc and Igh switch regions in B cell tumors and mouse tissues1,2,3,4. However, the Igh switch region is both long and redundant and this is a particularly challenging part of the genome to amplify accurately. To assay for Igh-Myc translocations Unniraman et al.5 used primers specific for Myc combined with consensus primers that hybridize in multiple locations in Igh Sμ and Sα switch region repeats and did 80 cycles of nested amplification followed by Southern blotting for detection5. When we did this assay using DNA extracted from lymph nodes of interleukin 6 (Il6)-transgenic mice, which have Igh-Myc translocations, we found that multiple species were present in ethidium bromide–stained gels and that some of the minor bands hybridized with internal Myc-specific probes (data shown). In contrast, nested PCR with a combination of Myc and Igh constant region–specific primers produced discrete bands in ethidium bromide–stained gels that also hybridized with the same internal Myc-specific probe1,2,3,4 (data shown). Using the 80-cycle consensus primer PCR assay, Unniraman et al.5 were able to amplify Igh-Myc hybrid DNA from 30% of the gut samples from activation-induced deaminase (AID)–deficient mice but not from kidney or brain samples. The tissue samples that were positive were reproducibly positive only 40% of the time5. Nevertheless, cloning showed that some of the amplicons were hybrid molecules of Myc and Igh5. Unniraman et al.5 suggest that these amplicons come from B cells that experienced AID-independent Igh-Myc translocations and that these cell populations were expanded by pristane injection in an unknown but AID-dependent way. However, these amplicons are also found in mouse strains that fail to develop pristane-induced plasmacytoma and are therefore not predictive of plasmacytoma development1.

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Figure 1: Igh-Myc PCR and Southern blotting.

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Authors and Affiliations

  1. The Rockefeller University, Howard Hughes Medical Institute, Laboratory of Molecular Immunology, New York, 10021, New York, USA

    Almudena R Ramiro, Mila Jankovic & Michel C Nussenzweig

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  1. Almudena R Ramiro
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  2. Mila Jankovic
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  3. Michel C Nussenzweig
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Correspondence to Michel C Nussenzweig.

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Ramiro, A., Jankovic, M. & Nussenzweig, M. Amplifying Igh translocations. Nat Immunol 6, 117 (2005). https://doi.org/10.1038/ni0205-117

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