title: In situ determination of B-cell heavy chain and kappa/lambda light chain expression patterns: methodology and clinical utility.
abstract: The authors describe a reverse transcriptase in situ polymerase chain reaction (RT in situ PCR) method that allows the determination of B-cell clonality as defined by expression of kappa and lambda mRNA as well as the different VH families that comprise the entire heavy chain (IgH) sequence using paraffin-embedded, formalin-fixed tissue. Polyclonal expression of B-cell light and heavy chains was documented in two reactive lymph nodes and a spleen in the same histologic distribution as the B-cell marker CD20. Monoclonal expression of kappa versus lambda mRNA was documented in 11 cases of B-cell lymphoma and was corroborated in 5 cases by flow cytometry; each case showed monoclonal expression of IgH. The authors analyzed seven additional tissues where a definitive diagnosis of B-cell lymphoma could not be rendered based on histologic, immunohistologic. and clinical analysis. RT in situ PCR for IgH and kappa versus lambda expression differentiated the B-cell lymphomas (n = 2) from reactive B-cell processes (n = 3) and from two cases of B-cell-rich T-cell lymphoma. The described RT in situ PCR methodology allows routine determination of the monoclonal versus multiclonal expression patterns of B cells, which will facilitate the diagnosis of B-cell lymphomas and aid in understanding the pathogenesis of B-cell malignancies using archival, paraffin-embedded tissues.
authors.affiliation: Department of Pathology, Ohio State University School of Medicine and Public Health, Columbus, USA.
authors.forename: C
authors.initials: C
authors.lastname: Morrison
authors.forename: P
authors.initials: P
authors.lastname: Porcu
authors.forename: M A
authors.initials: MA
authors.lastname: Caliguiri
authors.forename: G J
authors.initials: GJ
authors.lastname: Nuovo
mesh_terms: D018951:Antigens, CD20 / Q000032:analysis; D001402:B-Lymphocytes / Q000473:pathology; D002999:Clone Cells; D017931:DNA Primers / Q000737:chemistry; D003937:Diagnosis, Differential; D005260:Female; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000235:genetics*; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000378:metabolism / Q000473:pathology; D016393:Lymphoma, B-Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018442:Lymphoma, B-Cell, Marginal Zone / Q000235:genetics / Q000473:pathology; D016399:Lymphoma, T-Cell / Q000235:genetics / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000378:metabolism / Q000473:pathology
chemical_list: D018951:Antigens, CD20; D017931:DNA Primers; D012333:RNA, Messenger; D012334:RNA, Neoplasm