Cytokine treatment of human bone marrow activates anti- XXXX effector cells: monitoring of purging by polymerase chain reaction and DNA analysis.

@entity137 @entity2940 @entity66 @entity5 @entity310 @entity8680 @entity1778 @entity7420 @entity379 @entity1199 @entity405
The aims of this study were to investigate the role of cytokines ( @entity5 necrosis factor alpha ( @entity310 ), @entity379 ( @entity379 ) and @entity1199 ( @entity1199 ) in augmenting graft-versus- @entity1778 (GVL). We have investigated the effector cells involved in GVL, by studying the role of these cells in purging of the cell line K562 in short-term bone marrow cultures. The effect of the addition in vitro of @entity7420 was also studied. Monitoring of purging was achieved by @entity137 assays, DNA analysis and the use of the polymerase chain reaction for the detection of bcr/abl transcripts in the Philadelphia positive (Ph+) K562 cell line. Supernatants from @entity1199 -treated and non-treated bone marrow were tested for cytokine production ( @entity310 and @entity379 ). The results have shown that the main cytotoxic effector cells in the bone marrow generated by @entity1199 have the @entity2940 + @entity405 + phenotype. Overnight incubation of bone marrow was sufficient to generate cytotoxic cells as measured by @entity8680 (Cr51) release assays. Measurable levels of @entity310 but not @entity379 were also detected in supernatants. Addition of @entity310 and @entity379 to the @entity1199 in the bone marrow cultures augmented the @entity137 but tended to inhibit progenitor cell growth as measured by granulocyte-macrophage colony-forming unit (GM-CFU) and erythroid blast-forming unit (BFU-e) assays. An estimate of the purging of the marrow could also be achieved by DNA analysis of K562 DNA in bone marrow. The bcr/abl transcript could still be detected by PCR analysis in marrow containing 1% K562 and treated with @entity1199 for 24 h, but by 6 days of incubation the bcr/abl transcript was weak or undectable. The results suggest that although reduction in the proportion of @entity1778 in contaminated marrow can be detected after incubation with @entity1199 for 24 h, complete elimination of minimal @entity66 requires longer incubation times.
