Figure 1
Figure 1. FLLCs are nonproliferative cells. (A) Distribution of the t(14;18)pos cells within proliferative CD20posCD10pos CXCR4posHoechstpos centroblasts vs CD20posCD10posCXCR4negHoechstneg nonproliferative centrocytes cell sorted from a t(14;18)hi RLN7 sample. Genomic DNA from each GC subsets was tested using a 2-step fluctuation nested PCR assay consisting of 16 replicates. BCL2/IGH translocations were sequenced. Multiple size bands revealed oligoclonality. (B) CFSE-labeled CD20pos purified B cells from RLN4 were stimulated for 4 days. Highly proliferative viable CFSEloDAPIneg and nonproliferative viable CFSEhiDAPIneg B cells were then sorted before quantification of t(14;18) by qPCR. Standard curves were generated from cloned BCL2/IGH (red) and GAPDH (green) PCR products, and it was demonstrated that detection of the BCL2/IGH transcript from the CFSEhi population was above the CFSElo cell population, where it remained below the quantification threshold of 1/25 000 cells. NQ, not quantifiable.

FLLCs are nonproliferative cells. (A) Distribution of the t(14;18)pos cells within proliferative CD20posCD10pos CXCR4posHoechstpos centroblasts vs CD20posCD10posCXCR4negHoechstneg nonproliferative centrocytes cell sorted from a t(14;18)hi RLN7 sample. Genomic DNA from each GC subsets was tested using a 2-step fluctuation nested PCR assay consisting of 16 replicates. BCL2/IGH translocations were sequenced. Multiple size bands revealed oligoclonality. (B) CFSE-labeled CD20pos purified B cells from RLN4 were stimulated for 4 days. Highly proliferative viable CFSEloDAPIneg and nonproliferative viable CFSEhiDAPIneg B cells were then sorted before quantification of t(14;18) by qPCR. Standard curves were generated from cloned BCL2/IGH (red) and GAPDH (green) PCR products, and it was demonstrated that detection of the BCL2/IGH transcript from the CFSEhi population was above the CFSElo cell population, where it remained below the quantification threshold of 1/25 000 cells. NQ, not quantifiable.

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