RESUMO
Graft-versus-host disease (GvHD) is a common life-threatening complication that can occur following allogeneic hematopoietic stem cell transplantation. This occurs if donor T cells recognize the host as foreign. During acute GvHD (aGVHD), activated T cells utilize glycolysis as the main source of energy generation. Therefore, inhibition of T cell glycolysis is a potential treatment strategy for aGVHD. In the present study, the effects of the combination of the glycolysis inhibitor 3-bromopyruvate (3-BrPA) and the mTOR inhibitor rapamycin (RAPA) on a mode of aGVHD were explored. In vitro mixed lymphocyte culture model was established by using splenocytes from C57BL/6 (H-2b) mice as responder and inactivated splenocytes from BALB/c (H-2d) mice as stimulator. In this model, 3-BrPA treatment (0-100 µmol/l) was found to suppress cell viability, increase cell apoptosis and reduce IFN-γ secretion, in a concentration-dependent manner. 3-BrPA treatment (0-100 µmol/l) was found to suppress cell viability, increase cell apoptosis and reduce IFN-γ secretion, in a concentration-dependent manner. In addition, combined treatment with 3-BrPA (0-100 µmol/l) alongside RAPA (20 µmol/l) exhibited synergistic effects on inhibiting cell viability and IFN-γ production, compared with those following either treatment alone. An aGVHD model was established by injection of bone marrow cells and spleen cells from the donor-C57BL/6(H-2b) mice to the receptor-BALB/c(H-2d) mice which were underwent total body irradiation first. In the aGVHD model, 3-BrPA (10 mg/kg/day), RAPA (2.5 and 5 mg/kg/day) and both in combination (5 and 2.5 mg/kg/day for 3-BrPA and RAPA, respectively) were all found to alleviate the damage caused by aGVHD, in addition to prolonging the survival time of mice with acute GvHD. In particular, the combined 3-BrPA and RAPA treatment resulted in the highest median survival time among all groups tested. In addition, the effects induced by combined 3-BrPA and RAPA treatment were found to be comparable to those in the 5 mg/kg/day RAPA group but superior to the 3-BrPA group with regards to the cumulative survival profile, GvHD score and lung histological score. The 3-BrPA and RAPA combination group also exhibited the lowest IFN-γ levels among all groups. Therefore, the combination of inhibiting both glycolysis and mTOR activity is a promising strategy for acute GvHD prevention.
RESUMO
This study aimed to confirm the presence of Blocking Factors (BFs) in Mixed Lymphocyte Culture (MLC) from female normal reproducer and sub-fertile rabbit inoculated with two injection of the allogenic lymphotherapy (LIT) to analyze its effect on rate fertility and pregnancy success. The BFs measuring was done intervening MLC with MTT-Formazan non-radioactive technique. It was demonstrated BFs presence in MLC in female rabbit groups. In sub-fertile female reproducers treated with allogenic lymphotherapy a significant increase in the level of FBs after every LIT was observed, as well as a rate fertility increase. Furthermore, it was established that BFs act on cell proliferation inhibiting the MLC of other species, clearly indicating that the inhibit effect of the BFs is inter-specific and no intraspecific as had sustain until now.
Esta investigación se basó en un modelo experimental de origen animal, dirigido a comprobar la existencia de factores bloqueadores (FBs) del cultivo mixto de linfocitos (CML) en grupos de conejas reproductoras normales y subfértiles. A los animales de experimentación se les aplicó dos dosis de Linfoterapia (LIT) alogénica, con el fin de analizar sus efectos en el aumento de la tasa de fertilidad y del éxito gestacional. La medición de los FBs se realizó mediante CML con la técnica no radioactiva MTT-Formazan. Se comprobó la existencia de FBs del CML en todos los grupos de conejas estudiados. En conejas reproductoras subfértiles tratadas con LIT alogénica se observó un incremento significativo de los niveles de los FBs después de cada LIT, así como el aumento en la tasa de fertilidad de las mismas. Además, se estableció que los FBs de proliferación celular actúan inhibiendo el CML de otras especies, lo que indica claramente que el efecto inhibitorio de FBs es interespecífico y no intraespecífico como se ha sostenido hasta ahora.