RESUMO
The clinical use of lymphocytes engineered to express high affinity T-cell receptors (TCRs) specific for two broadly expressed tumor-associated antigens is strongly limited by MHC-restricted fratricide of lymphocytes and TCR-mediated killing of hematopoietic stem cells. Specific clinical applications must therefore be conceived to bypass these limitations.
RESUMO
Objective To define which hyaluronan synthase (HAS), of three hyaluronan synthesizing enzymes HAS-1, HAS-2, and HAS-3, is primarily responsible for hyaluronan synthesis and extracellular matrix/extracellular coat formation in human peritoneal mesothelial cells (HPMCs) . Methods As a prerequisite study, the expression of each HAS mRNA in cultured HPMCs was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Only the expression of HAS-2 and HAS-3 mRNA could be detected. The level of HAS-2 mRNA expression was about 10 fold higher than that of HAS-3. HAS-2 specific antisense oligonucleotide was then transfected into cultured HPMCs by lipofectamine. After 0, 8, 24, and 48 hours, the expression of HAS-2 mRNA was detected by RT-PCR and the extracellular coat was measured by particle exclusion test. Results 8 hours and 24 hours after transfection, the expression of HAS-2 mRNA in HPMCs decreased by 58% and 89% respectively; 48 hours after transfection, the expression of HAS-2 mRNA in HPMCs partially restored to 25% of the normal level. Correspondingly, 24 hours after transfection, the extracellular matrix/extracellular coat in HPMCs almost completely disappeared. However, as control, sense and reverse oligonucleotides showed no effect. Conclusion HAS-2 plays a leading role in HPMCs hyaluronan synthesis and the formation of extracellualr matrix/extracellular coat.