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El tratamiento de las celulas E5 con interferón aumenta el total de MHC I, pero no el de superficie.
Ambos interferón-b y interferón-g (IFN-b o g-IFN) aumentan la actividad de transcripción del promotor del gen de la cadena pesada de MHC I (Agrawal y Kishore, 2000) conduciendo a mayores niveles de expresión de la cadena pesada. Hemos demostrado anteriormente que en las celulas E5 hay menos ARNm y proteina de la cadena pesada de MHC I (Ashra ® et al., 2002). Para ver si algun paso en la biosíntesis de la cadena pesada está permanentemente inhibido por E5, tratamos celulas PalF, no-E5 y E5, ya sea con b-o g-IFN o el control de sobrenadante 2DG . El tratamiento con IFN, pero no con 2DG, incremento de forma notable la producción de la cadena pesada de MHC I por aproximadamente la misma medida en todas las célulares evaluadas por immunoblotting (Figura 5). Este muestra que las células E5 responden a IFN y que la ruta biosintética de la cadena pesada de MHC I no esta inhibida irreversiblemente por E5. Las células tratadas con IFN También se analizaron por FACS para determinar si aumento la producción de la cadena pesada llevando a un aumento correspondiente de MHC I en la superficie. La expresión de MHC I en la superficie se incrementó sólo en células PalF y no-E5 pero no en las celulas E5, o en células que expresan el hypertransforming 4-E5 N17A mutante (Figura 5b), a pesar del aumento en el total de MHC I. Estos resultados muestran inequívocamente que E5 inhibe el transporte de
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Treatment of E5 cells with interferon increases total but
not surface MHC I
Both b-interferon and g-interferon (b-IFN or g-IFN)
increase the transcription activity of the MHC I
heavy chain gene promoter (Agrawal and Kishore,
2000) leading to higher expression levels of heavy
chain. We have previously shown that in E5 cells
there is less MHC I heavy chain mRNA and protein
(Ashra® et al., 2002). To see if the biosynthetic
pathway of heavy chain is permanently inhibited by
E5, we treated PalF, no-E5 and E5 cells with either
b- or g-IFN or the control supernatant 2DG.
Treatment with either IFN, but not with 2DG,
noticeably increased production of the MHC I heavy
chain to approximately the same extent in all cell
lines as assessed by immunoblotting (Figure 5a). This
shows that E5 cells are responsive to IFN and the
biosynthetic pathway of MHC I heavy chain is not
irreversibly inhibited by E5. The IFN-treated cells
were also analysed by FACS to ascertain whether the
increased production of heavy chain led to a
corresponding increase in surface MHC I. MHC I
surface expression increased only in PalF and no-E5
cells but not in E5 cells, or in cells expressing the
hypertransforming 4-E5 N17A mutant (Figure 5b),
despite the increase in total MHC I. These results
unequivocally show that E5 inhibits the transport of
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Last Update: 2011-08-29 |
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