Issn Print: 0192-8562
Publication Date: 1997/07/01
#672 Identification of genes involved in peripheral B-cell maturation
Excerpt
The functional histology of peripheral lymphoid tissues has recently been refined and five subsets of B-cells have been defined using FACS analysis with a combination of cell surface markers. These subsets (Bm1,2,3,4,5) correspond to progressive stages of lymphoid development (follicular mantle - Bm1 and Bm2, germinal center -Bm3 [centroblast] and Bm4 [centrocyte], and memory cells- Bm5). In order to isolate genes specific to the germinal center, representational difference analysis (RDA-subtractive hybridization with PCR amplification) was applied to subtract follicular mantle (Bm1 and Bm2) from germinal center (Bm3) cDNA. Since the sorted B-cell subsets are in limiting supply, RNA from the cell lines B12, HeLa and Jurkat as well as total tonsil was screened with RNase protection assays using probes made from the isolated gene segments of interest to confirm differential expression. B1-2 is a Burkitt's lymphoma line similar to Bm3 cells. HeLa is a cervical carcinoma line and Jurkat is derived from T-cell lymphoma. Of 25 cDNA fragments screened so far 5 appear to be differentially expressed (on in tonsil and BL2 but not in HeLa). These gene segments have significant homology to sequenced clones in the EST data base. We are currently screening a panel of lymphoma and leukemia cell lines to better define the distribution of these genes in normal as well as malignant B-cell populations.
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