Correlation between presence of clonal rearrangements of immunoglobulin heavy chain genes and B-cell antigen in patients with nodular sclerosis and mixed cellularity Hodgkin's disease

Abstract

S 47 null phenotype. In addition to CD21 and CD20 antigens, RS cells are found to be positive for a number of antibodies detecting FDRC-associated antigens such as HJ2, 2B44, CNA.42, CD23, anti-acid cysteine protease inhibitor (ACPI: gift from Professor Rinne) and both cell types possess Fc receptor for IgG. The hypothesis is further backed by the findings of CD15 antigen expression by occasional RS-like dysplastic FDRC in Castleman's disease which is characterized by hyperplasia of FDRC. Epstein-Barr virus (EBV) nucleic acids are found in approximately 50% of classic Hodgkin's disease. Double labelling techniques show the presence of EBV-positive RS cells within FDRC network. The relation of RS cells to FDRC is further supported by the recent observation of a case of pseudo-inflammatory tumour rich in FDRCs which were found to be EBV positive. Interestingly, that tumour showed EBV-positive FDRC showing a RS-like feature. The FDRC origin of RS cells could explain the frequent germline configuration of ig and TCR genes in lymph nodes involved by Hodgkin's disease. Whether FDRC might be the only cells involved in conversion to RS cells by loss or gain of antigens remains to be determined. Correlation between presence of clonal rearrangements of immunoglobulin heavy chain genes and B-cell antigen in patients with nodular sclerosis and mixed cellularity Hodgkin's disease A. Orazi, B. Jiang, C-H. Lee, G. English, G. Cattorettii*, K. John and R. S. Neiman, Division of Pathology and Laboratory Medicine. Indiana University School c?[ Medicine, Indianapolis, Indiana 46202, and *Division ~?/ Surgical Pathology, Columbia University, New York, NK USA Southern blot analysis of Hodgkin's disease (HD), although often comprised by the small number of abnormal cells present in the tissue, have tended to favor a B-cell derivation of the Hodgkin's and Reed Sternberg (HRS) cells. Recent immunohistochemical findings in cases of nodular sclerosis iNS) and mixed cellularity (MC) HD (Schmid et al, Am J Pathol, 139:701, 1991) and the description of the integration of Epstein-Barr virus (EBV) in HRS cells have provided further evidence in favor of a B-cell origin in most of these cases. 18 frozen and 29 paraffin embedded sections of lymph node specimens from 29 patients with pretreatment HD (22 NS-HD and 7 MC-HD) were studies by molecular analysis and immunohistochemistry to determine the phenotype of HRS cells. All selected cases showed classic morphology and CD45-, CD30+, CD15+, BLA.36+ HRS cells, in 11/ 29 (38%) cases HRS cells were reactive with at least one B-cell marker (CD20, HM57, MB2), 7/29 (24%) cases showed reactivity with the T-cell marker CD3, and 11/ 29 (38%) cases displayed a 'null' phenotype. By using a polymerase chain reaction (PCR) and consensus primers for the V and J regions of the immunoglobulin heavy chain (IgH) gene, we were able to detect B-cell clonality in 9/18 (50%) frozen samples of HD analyzed. IgH gene rearrangement was present in 8/15 (53%) NS-HD and in 1/3 (33%) MC-HD. In 5/9 (56%) of these cases HRS cells were reactive with at least one B-cell marker while one case expressed the T-cell marker CD3. The other cases with IgH rearrangement showed a 'null' immunophenotype. IgH gene analysis was negative in all remaining CD3 + cases and in two other case that expressed B-cell markers by immunohistology. Our results show that PCR represents a specific and sensitive technique for the detection of IgH gene rearrangement in cases of HD. The results also confirm a lymphoid B-cell derivation of HRS cells in a high proportion of the cases. Configuration of Ig in DNA from Sternberg-Reed cells H. Stein, M. Hummel, J. Tamaru and K. Ziemann The nature of Hodgkin and Reed-Sternberg (HRS) cells remains in question. Immunophenotypic studies favour a relation to the lymphoid lineage with the existence of Band T-cell types. However, studies on the detection of antigen (AG) receptor gene rearrangement provided inconsistent results. They concur in that rearranged lg and T-cell receptor (TCR) genes are not demonstrable in most Hodgkin's disease (HD) cases. To clarify whether this is because of the insensitivity of the method of detection or due to a real absence of clonal lg heavy chain (IgH) rearrangements, a polymerase chain reaction (PCR) method with high sensitivity was applied to DNA extracted fu tissue sections and from single HRS cells picked from tissue sections. The results obtained show that, in contrast to the data reporled by Pfreudschuh's group, a VDJ rean'angement is demonstrable in the majority of HD cases with B antigen-positive HRS cells. No cases with T-cell antigen-positive HRS cells harboured detectable clonal VDJ rearrangements. Of 10 sequenced rearranged IgH genes, the VH segment of six cases contained considerable somatic mutations. These results suggest that the HRS cells of cases with rearranged IgH genes are B-cell related and correspond in their differentiation stage either to naive pregerminal centre B-cells or (more commonly) to germinal centre/ post-germinal centre-derived memory B-cells. Molecular findings in HD distinctive gene expression in isolated RS cells H. Grisser, T. Ariyasu and E. Mack, The Ontario Cancer Institute, Princess Margaret Hospital, Department ()f Oncologic Pathology, 500 Sherbourne Street, Toronto, Ontario, Canada M4X IK9 and University of Toronto, Department of Pathology, Toronto, Ontario, Canada Characterization of PCR-amplified cDNAs (PCR-cDNA) from lymph node-derived, single cells previously has confirmed their lymphoid origin. Gene expression profiles are similar among RS cells from the same specimen but vary among different HD cases of the same subtype. A subtractive PCR-cDNA library approach is now used