ゼッショク ラット ニ オケル ケッセイチュウ アミノサン ノウド オヨビ アミラーゼ トリプシン カッセイ ノ キョドウ ニ タイスル ミカク シゲキ ノ エイキョウ

京都大学0048新制・論文博士博士(医学)乙第10802号論医博第1767号新制||医||784(附属図書館)UT51-2001-R781(主査)教授 北 徹, 教授 中尾 一和, 教授 清野 裕学位規則第4条第2項該当Doctor of Medical ScienceKyoto UniversityDA

Circadian and Light-Induced Transcription of Clock Gene Per1 Depends on Histone Acetylation and Deacetylation

Circadian clock genes are regulated through a transcriptional-translational feedback loop. Alterations of the chromatin structure by histone acetyltransferases and histone deacetylases (HDACs) are commonly implicated in the regulation of gene transcription. However, little is known about the transcriptional regulation of mammalian clock genes by chromatin modification. Here, we show that the state of acetylated histones fluctuated in parallel with the rhythm of mouse Per1 (mPer1) or mPer2 expression in fibroblast cells and liver. Mouse CRY1 (mCRY1) repressed transcription with HDACs and mSin3B, which was relieved by the HDAC inhibitor trichostatin A (TSA). In turn, TSA induced endogenous mPer1 expression as well as the acetylation of histones H3 and H4, which interacted with the mPer1 promoter region in fibroblast cells. Moreover, a light pulse stimulated rapid histone acetylation associated with the promoters of mPer1 or mPer2 in the suprachiasmatic nucleus (SCN) and the binding of phospho-CREB in the CRE of mPer1. We also showed that TSA administration into the lateral ventricle induced mPer1 and mPer2 expression in the SCN. Taken together, these data indicate that the rhythmic transcription and light induction of clock genes are regulated by histone acetylation and deacetylation

Binding properties of IgGs to SIV antigens.

<p>The non-NAb cocktail, ten non-NAb IgG lots derived from ten macaques, and CAb were subjected to immunoblotting (ZeptoMetrix). A representative result from two experiments is shown.</p

Predominant nonsynonymous env mutations.

<p>Viral cDNAs encoding Env were amplified from plasma RNAs obtained at 7-9 months (R10-001, R10-004, and R06-029) or 12 months (other animals) and subjected to sequencing analysis. Amino acid substitutions are shown. The asterisk (*) represents a deletion and the double asterisk (**) represents coexistence of multiple deletion patterns. </p

ADCVI activity of the non-NAb cocktail and non-NAb IgG lots.

<p>The reduction in SIV p27 concentration in the supernatant from SIV-infected cell culture with non-NAbs compared to that without antibodies is shown. A representative result, means of duplicate samples, from two experiments is shown.</p

SIV antigen-specific CD8⁺ T-cell responses.

<p>SIV Gag-, Pol-, Vif-, Vpx/Vpr-, Tat-, Rev-, Nef-, and Env-specific CD8⁺ T-cell responses were measured by detection of antigen-specific IFN-γ induction using PBMCs at weeks 26-30 post-challenge.</p

Binding properties of IgGs to SIV virions.

<p>Polyclonal IgGs purified from macaque plasma were subjected to whole virus ELISA using purified SIV_mac239 virions as the antigen. Results on ten IgG lots derived from ten macaques without detectable neutralizing activity (non-NAbs; black lines), five with neutralizing activity (NAbs; red), and a control IgG (CAb; green) are shown in the left panel. Results on the non-NAb cocktail and three non-NAb lots composing the cocktail are in the right. The dotted line represents background absorbance. Time points of plasma sampling are shown in parentheses following the macaque IDs. A representative result, means and SDs of duplicate samples, from two experiments is shown.</p

Association of lymph-node antigens with lower Gag-specific central-memory and higher Env-specific effector-memory CD8+ T-cell frequencies in a macaque AIDS model

Virus-specific CD8+ T cells exert strong suppressive pressure on human/simian immunodeficiency virus (HIV/SIV) replication. These responses have been intensively examined in peripheral blood mononuclear cells (PBMCs) but not fully analyzed in lymph nodes (LNs), where interaction between CD8[+] T cells and HIV/SIV-infected cells occurs. Here, we investigated target antigen specificity of CD8[+] T cells in LNs in a macaque AIDS model. Analysis of virus antigen-specific CD8[+] T-cell responses in the inguinal LNs obtained from twenty rhesus macaques in the chronic phase of SIV infection showed an inverse correlation between viral loads and frequencies of CD8[+] T cells with CD28[+]CD95[+] central memory phenotype targeting the N-terminal half of SIV core antigen (Gag-N). In contrast, analysis of LNs but not PBMCs revealed a positive correlation between viral loads and frequencies of CD8[+] T cells with CD28[−]CD95[+] effector memory phenotype targeting the N-terminal half of SIV envelope (Env-N), soluble antigen. Indeed, LNs with detectable SIV capsid p27 antigen in the germinal center exhibited significantly lower Gag-N-specific CD28[+] CD95[+] CD8[+] T-cell and higher Env-N-specific CD28[−]CD95[+] CD8[+] T-cell responses than those without detectable p27. These results imply that core and envelope antigen-specific CD8[+] T cells show different patterns of interactions with HIV/SIV-infected cells