[Immunosuppressive factors produced by a T cell line derived from XXXX ].

@entity981 @entity3208 @entity1356 @entity310 @entity4909 @entity35063 @entity2975 @entity1199 @entity405 @entity214
The supernatant of @entity405 + cells, isolated from a permanent lymphoblastoid cell clone established from a long term culture of a T cell @entity981 , contained two distinct molecules with suppressive activity on @entity35063 -induced PBMC proliferation. This clone does not produce @entity310 , @entity4909 , alpha-IFN, gamma-IFN, @entity1356 , @entity1199 and has not natural killer activity. In the attempt to purify and biochemically characterize the lymphoblastic cell line-derived T-cell SFs, a multi-step chromatographic separation has been used. Two different peaks of biologic activity have been separated by HPLC gel permeation in the range of 100-120 Kd and 75-85 Kd referred to as high molecular weight suppressor factor HMWSF, and low molecular weight suppressor factor LMWSF, respectively. These fractions were then concentrated, dialyzed and further purified by anion exchange HPLC. This chromatographic step allowed us to considerably purify the two SFs. The biologically active fractions derived from the previous chromatographic step were eventually subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by @entity2975 gel electrophoresis in @entity3208 ( @entity214 -PAGE): a single band corresponding to 115 Kd was observed for HMWSF, while LMWSF yielded a single band at 80 Kd. The isoelectric points (pI) of the different SFs was determined by flat-bed isoelectric-focusing: the HMWSF yielded a single band at pI 7.4, while a much lower pI was observed for LMWSF, 3.5-3.6. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4 degrees C and 7-10 days at -80 degrees C.
