THE VALUE OF IMMUNOHISTOCHEMISTRY
IN THE ASSESSMENT OF BONE MARROW DISORDERS

Course Director: Attilio Orazi, M.D., FRCPath. (Engl.) Indiana University School of Medicine James Whitcomb Riley Hospital for Children Indianapolis, Indiana Dennis P. O'Malley, M.D. Indiana University School of Medicine James Whitcomb Riley Hospital for Children Indianapolis, Indiana Click here for the complete PowerPoint presentation. You may go directly to any major heading of the text and reference section via the table of contents below. Once you display any section, you may also view the text sequentially by using the 'Next' and 'Previous' buttons. Introduction The assessment of bone marrow morphology has historically been left in the hands of the hematologist. The hematologist has a greater interest in bone marrow diseases than do many pathologists. Moreover, by the development of special Romanovsky type stains the hematologist has had an excellent method of examining bone marrow cells, whereas conventional histopathology and H&E stains have been inadequate to assess the subtleties of bone marrow morphology. There is no question that the ideal study of bone marrow involves examination of both marrow smears and tissue sections. Each is complementary to the other. However, pathologists in general have not taken advantage of the techniques that are now available which we believe can provide greater insight into and knowledge regarding bone marrow disorders. There are two reasons for this. The first has to do with technical phenomena. Although issues regarding adequate fixation and sectioning are not directly the theme of this short course, it is important to note that high quality tissue sections can reveal a significant amount of information that is not usually obtained unless one is careful about the adequacy of fixation and the quality of sectioning. We do not believe that it is necessary to employ such techniques as plastic embedding to obtain high quality sections of bone marrow if careful attention is paid to these technical issues. The second reason that bone marrow aspirates have been more reliable than sections historically in assessing bone marrow proliferations, is that live cells are necessary to perform needed ancillary cytochemical and immunologic tests. However, with the rapid recent advances in immunohistochemistry, the advantage of having living cells has become less significant. The growing battery of paraffin-reactive immunohistological reagents, coupled with newer techniques mainly based on the use of heat-induced antigen retrieval (e.g. microwave oven boiling) have opened up a new vista to the study of bone marrow by interested pathologists. This course is meant as an introduction to the use of immunohistochemistry in bone marrow sections. It is based largely on personal experience at the National Tumor Institute of Milan, Italy, Indiana University School of Medicine, Indianapolis, IN, and College of Physicians and Surgeons of Columbia University, New York, NY. SECTION I Immunohistology Reagents Commonly Used to Study Bone Marrow Disorders Hemoglobin von Willebrand Factor (Factor VIII-Related Antigen) Myeloperoxidase CD68 Lysozyme CD34 TdT CD99 CD45RB (Leucocyte Common Antigen) CD20 (L-26) CD79a (HM57) Other Markers Used to Study B-cell Proliferations: CD5, CD23, CD10 Cyclin D1 BCL-2 CD138 (Syndecan-1) Anti-lambda and Anti-kappa Light Chain Anti-immunoglobin Heavy Chain (IgG, IgA, IgM) DBA.44 Tartrate Resistant Acid Phosphatase CD3 Other T-cell Markers: CD2, CD7, TIA-1 CD43 CD30 (BerH2) CD15 CD1a and S-100 CD56 (N-CAM) CD117 (c-KIT) p53 Protein Proliferation-associated Markers: PC10 And MIB-1 Apoptosis by In-situ End Labeling Normal Bone Marrow: Immunohistochemical Identification of Different Cell Components SECTION II Specific Diagnostic Problems Malignant Lymphomas (See Tables 9-20) B-cell Neoplasms Immunosecretory Disorders T-cell Neoplasms Hodgkin's Disease (See Tables 21,22) Lymphoid Follicles and Lymphoid Hyperplasia vs. Lymphoma (See Tables 23,24) Histiocytic/dendritic-cell Neoplasms and Related Disorders (See Table 25) Systemic Mastocytosis Acute Leukemias (See Tables 26-35) Minimal Residual Acute Leukemic Disease (See Tables 36-39) Myelodysplastic Syndromes and Myeloproliferative Disorders (See Tables 40-58) Myelodysplastic/Myeloproliferative Diseases (See Tables 59-62) The Use of Bone Marrow Biopsy to Document the Proliferative Effects of Hematopoietic Growth Factors on Bone Marrow Cells (See Tables 63-68) Metastatic Malignancy from Unknown Primary Source (See Tables 69-71) SECTION III Investigational Approaches Applied to Bone Marrow Biopsies p53 overexpression by immunohistochemistry Apoptosis by in-situ end labeling Angiogenesis by immunohistochemistry Opinions stated and/or conclusions reached in this syllabus are the responsibility of the authors and are not necessarily endorsed by the Academy.