Original Paper
MYC expression and distribution in normal mature lymphoid cells
Giorgio Cattoretti,
Corresponding Author
Giorgio Cattoretti
Department of Pathology, AO San Gerardo, Monza, Italy
Department of Surgical Sciences, Universitá degli Studi di Milano Bicocca, Milano, Italy
Correspondence to: Professor Giorgio Cattoretti, Department of Pathology, University of Milano-Bicocca and San Gerardo Hospital, Via Pergolesi 33, 20900 Monza (MB), Italy. e-mail: [email protected]Search for more papers by this authorGiorgio Cattoretti,
Corresponding Author
Giorgio Cattoretti
Department of Pathology, AO San Gerardo, Monza, Italy
Department of Surgical Sciences, Universitá degli Studi di Milano Bicocca, Milano, Italy
Correspondence to: Professor Giorgio Cattoretti, Department of Pathology, University of Milano-Bicocca and San Gerardo Hospital, Via Pergolesi 33, 20900 Monza (MB), Italy. e-mail: [email protected]Search for more papers by this authorNo conflicts of interest were declared.
Abstract
The distribution of the product of the proto-oncogene MYC in lymphoid tissue has not been established in three decades, due to a combination of factors including low abundance, short half-life, and antibody sensitivity and specificity. We sought to validate antibodies in order to define the expression and distribution of MYC in mature normal lymphoid cells by multiparametric immunophenotyping. Having validated two antibodies for flow cytometry and for immunohistochemistry, we analysed normal tonsil tissue. MYC is expressed predominantly in B cells, some of which are interfollicular large, activated, and cycling CD30+, IRF4+, AID± blasts. Follicular mantle, isotype-switched memory B cells and FcRH4/IRTA1+ B cells express MYC in a wide range of levels and are small non-proliferating CDKN1B/p27-positive or -negative resting B lymphocytes. Germinal centre founder cells, CD30+ BCL6± AID± germinal centre blasts, and a population of GC cells in the apical light zone express MYC. MYC is expressed in all phases of the cell cycle in activated and mature B cells, but rarely in other lymphoid types and only partially fulfils the predictions derived from extractive and ex vivo experiments of the past 30 years. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Supporting Information
| Filename | Description |
|---|---|
| path4141-sup-0001-Figure S1.tifTIFF image, 434.2 KB | Figure S1. Flow cytometric detection of MYC staining. A representative experiment on tonsil cells is shown. CD20-gated B-lymphocytes (inset, top left) are stained for IgD and CD38, separating the B cells in germinal center (1), GC founder cells (2), memory B cells (3) and follicular mantle B cells (4). Each of these subsets was gated and independently stained for negative control (grey shadow), MYC (continuous line) or BCL6 as a positive control (dashed line). |
| path4141-sup-0002-Figure S2.tifTIFF image, 5.7 MB | Figure S2. Control staining for specificity and subcellular localization. A) and B) show a low magnification (10x, A) and high magnification (40x, B) image of a fixed and embedded tonsil section, stained with an identical protocol as the ones shown in Figure 3, where the MYC specific antibody was substituted with an identically diluted rabbit Ig. An asterisk in B marks a Germinal Center. Note the absence of specific staining. C) a serial section of the specimen shown in Figure 4A, was double stained in immunofluorescence with an identical protocol, except that both the primary antibodies were substituted by identically diluted negative mouse and rabbit Igs. Note the absence of specific staining. D) A fixed and embedded tonsil section was stained in double immunofluorescence with the protocol described for one nuclear antigen, PU.1 (red) highly expressed in the Germinal Center on the left, and for IRF4 (green) nuclear and cytoplasmic. Note the exclusive mutual distribution on the left and on the right, except the small follicular mantle B cells (asterisk, center) which co-express both at low levels both (yellow). In the inset is shown the mutually exclusive distribution of a nuclear antigen (Oct2, green) and a cytoplasmic antigen (CD68, red). None of the images show nuclear counterstain except C (DAPI). |
| path4141-sup-0003-Table S1.pdfPDF document, 53.9 KB | Table S1. Supplemental antibodies used. |
| path4141-sup-0004-Appendix S1.docWord document, 31.5 KB | Appendix S1. Immunohistochemistry |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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