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Effect and Mechanism of HDAC11 on Inducing Tolerance

Author: LianZuoZuo
Tutor: GongJianPing；WangXiaoBo
School: Chongqing Medical University
Course: Surgery
Keywords: Histone deacetylase 11 Kupffer cells Interleukin- 10 Antigen-presenting Immune tolerance
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract

Purpose of the histone deacetylase enzyme 11 (histone deacetylase, HDAC11) and cell division cycle, nervous system development, tumorigenesis and immune function regulation, and is closely linked. New research shows that inhibition of monocyte-macrophage cell RAW264.7 HDAC11 activity can promote the expression of the cells endogenous interleukin-10 (interleukin 10, IL-10), thereby inducing immune tolerance generated. This study was designed by observing the inhibition of lipopolysaccharide Kupffer cells (KCs) the HDAC11 active presenting its endogenous IL-10 expression and cell surface antigen expression of related molecules, to explore inhibition of HDAC11 induction KCS immune tolerance generated the mechanism. BALB / c mice the KCS by the density gradient centrifugation separated the adherent cells RPMI1640 culture medium containing 20% ??fetal calf serum (FCS) for 24 h were randomly divided into three groups: (1) the control group (blank group , continue to RPMI1640 culture 20h), (2) negative control group (group of negative control vector transfected with blank plasmid), (3) plasmid group (transfected of HDAC11 short hairpin RNA plasmid (HDAC11-shRNA)), three groups were given lipopolysaccharide. 3h, Real time-PCR and Western-blot method to detect the of KCS in HDAC11 and IL-10 gene and protein expression; flow cytometry KCS surface molecules MHC-II, B7-1, B7-2 and CD40 expression ; identifiable ovalbumin (ovalbumin, OVA) transgenic mice spleen T cells and the three sets of co-cultured KCs 3d after 3H-TdR notation T cell proliferation; detected by enzyme-linked immunosorbent assay (ELISA) the culture supernatant of IL-2 content. (1) Real-time PCR results: HDAC11 and IL-10 2-delta Ct value as the expression level of this value and GAPDH ratio of X ± S on behalf of its relative gene expression levels. HDAC11 plasmid group was the lowest, 0.076 ± 0.007, compared with two control groups (blank control group 0.145 ± 0.012, the negative group 0.141 ± 0.013), the difference was statistically significant (P lt; 0.05); IL-10 plasmid The highest group was 0.084 ± 0.006, compared with the two control groups (blank control group 0.051 ± 0.004, the negative group 0.052 ± 0.004), a statistically significant difference (P lt; 0.05). (2) Western-blot results: HDAC11 and IL-10 bands and GAPDH gray value of the ratio of X ± S on behalf of its protein relative expression levels. Real-time PCR results similar HDAC11 plasmid group was the lowest, 0.132 ± 0.012, compared with the two control groups (blank control group 0.373 ± 0.023, the negative group 0.365 ± 0.019), the difference was statistically significant (P lt; 0.05 ); whereas IL-10 expression plasmid, 0.218 ± 0.018, compared with the two control groups (blank control group 0.096 ± 0.009, the negative group 0.101 ± 0.011), a statistically significant difference (P lt; 0.05) (3) Flow cytometry results: plasmid group KC surface of antigen-presenting related molecules was lower than the control group and negative control group. (4) co-cultured T cell proliferation results: plasmid group the lowest CPM value is 2163 ± 146, compared with the two control groups (blank group, 3654 ± 165, negative group 3509 ± 142), the difference was statistically significant (P lt; 0.05). (5) ELISA results: plasmid group co-culture supernatant IL-2 content was the lowest, 463.7 ± 38.9 pg / mL, compared with the two control groups (blank control group 879.2 ± 65.5 pg / mL, negative group 854.4 ± 67.3 pg / mL), the difference was statistically significant (P lt; 0.05) The data was no significant difference between the two control groups (P gt; 0.05) Conclusion The experiments show that inhibition of HDAC11 KCs can promote expression of endogenous IL-10 overexpression, thereby further inhibiting the KCs antigen-presenting ability of cultured T cells activation and proliferation of immune tolerance induced by generating. HDAC11 may play an important role in the liver this immune privileged organ in the treatment of advanced liver disease endotoxemia induced liver transplantation immune tolerance in animal experiments and clinical application provides a good pre-cell experimental basis for the later application HDAC11.

Effect and Mechanism of HDAC11 on Inducing Tolerance

Abstract

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