INSITU DETECTION OF HUMAN IG LIGHT-CHAIN MESSENGER-RNA ON FORMALIN-FIXED AND PARAFFIN-EMBEDDED TISSUE-SECTIONS USING DIGOXIGENIN-LABELED RNA PROBES

Digoxigenin-labelled RNA probes complementary to human immunoglobulin (Ig) kappa and lambda light-chain mRNAs were produced by in vitro transcription. Using these probes, several existing in situ hybridization protocols were studied. By modifying and optimizing pretreatment procedures, which include hybridization, stringency washings and probe detection, a simplified non-radioactive in situ hybridization method for Ig light-chain mRNAs was developed. The light-chain signals were consistently identified in plasma cells, germinal centrocytes, centroblasts and immunoblasts in formalin-fixed and paraffin-embedded sections of lymphoid tissues. Monotypic light-chain mRNA was demonstrated in archival cases of kappa or lambda light-chain-restricted B-cell lymphoma. Background staining was found to be negligible in all the tissues tested. These results indicate that the in situ hybridization methodology described in this study is specific and sensitive for the detection of Ig light-chain mRNAs and has practical value in routine histology.