Protein studies of human non-Hodgkin's B-lymphoma: appraisal by two-dimensional gel electrophoresis

We have utilized two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) coupled with silver stain to identify cellular proteins in human non-Hodgkin's B-lymphoma (NHL). Five cell lines (SKDHL2B, WSU-DLCL2, WSU-NHL, WSU-FSCCL and SKLN1), representing four different NHL maturational stages and a normal Epstein-Barr virus (EBV)-transformed line of B-cell origin (SKLN1) were studied. The NHL lines were immunophenotyped using flow cytometry with lineage associated monoclonal antibodies. Whole cell lysates of the cell lines were subjected to 2-D PAGE analyses. The gels were analyzed with an image scanning computer and the qualitative differences of protein patterns were studied. Results revealed great similarities in patterns of the NHL lines. A master map containing common NHL-protein spots was constructed. When the map of each tumor line was compared to the master map, several protein spots were associated with each NHL-grade. Search for these proteins in the normal EBV-transformed B-cell line showed that only one of the proteins (S3; M(r)/pI 19/5.9) was present. Proteins that were detected in malignant NHL, but not in the normal EBV-line, could provide important information regarding the human NHL B-lymphocyte data-bases. Whether or not these proteins are definite malignant markers to distinguish between different NHL maturational stages needs further exploration through electroblotting and microsequencing.