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RNA-modulating factors not only regulate multiple steps of cellular RNA metabolism, but also emerge as key effectors of the immune response against invading viral pathogens including human immunodeficiency virus type-1 (HIV-1). However, the cellular RNA-binding proteins involved in the establishment and maintenance of latent HIV-1 reservoirs have not been extensively studied. Here, we screened a panel of 62 cellular RNA-binding proteins and identified NEDD4-binding protein 1 (N4BP1) as a potent interferon-inducible inhibitor of HIV-1 in primary T cells and macrophages. N4BP1 harbours a prototypical PilT N terminus-like RNase domain and inhibits HIV-1 replication by interacting with and degrading viral mRNA species. Following activation of CD4+ T cells, however, N4BP1 undergoes rapid cleavage at Arg 509 by the paracaspase named mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1). Mutational analyses and knockout studies revealed that MALT1-mediated inactivation of N4BP1 facilitates the reactivation of latent HIV-1 proviruses. Taken together, our findings demonstrate that the RNase N4BP1 is an efficient restriction factor of HIV-1 and suggest that inactivation of N4BP1 by induction of MALT1 activation might facilitate elimination of latent HIV-1 reservoirs. The interferon-inducible RNA-binding protein N4BP1 binds to and degrades HIV-1 mRNAs, thus inhibiting viral degradation and promoting viral latency. However, metacaspase-mediated degradation of N4BP1 following activation of CD4+ T cells facilitates HIV-1 latency reversal.

TNFR-associated factor family member–associated NF-κB activator (TANK)–binding kinase 1 (TBK1) is critical for the activation of IFN regulatory factor 3 and type I IFN production upon virus infection. A set of TBK1-binding proteins, 5-azacytidine–induced gene 2 (AZI2; also known as NAP1), TANK, and TBK1-binding protein 1 (TBKBP1), have also been implicated in the production of type I IFNs. Among them, TANK was found to be dispensable for the responses against virus infection. However, physiological roles of AZI2 and TBKBP1 have yet to be clarified. In this study, we found that none of these TBK1-binding proteins is critical for type I IFN production in mice. In contrast, AZI2, but not TBKBP1, is critical for the differentiation of conventional dendritic cells (cDCs) from bone marrow cells in response to GM-CSF. AZI2 controls GM-CSF–induced cell cycling of bone marrow cells via TBK1. GM-CSF–derived DCs from AZI2-deficient mice show severe defects in cytokine production and T cell activation both in vitro and in vivo. Reciprocally, overexpression of AZI2 results in efficient generation of cDCs, and the cells show enhanced T cell activation in response to Ag stimulation. Taken together, AZI2 expression is critical for the generation of cDCs by GM-CSF and can potentially be used to increase the efficiency of immunization by cDCs.

Akirin2, an evolutionarily conserved nuclear protein, is an important factor regulating inflammatory gene transcription in mammalian innate immune cells by bridging the NF-κB and SWI/SNF complexes. Although Akirin is critical for Drosophila immune responses, which totally rely on innate immunity, the mammalian NF-κB system is critical not only for the innate but also for the acquired immune system. Therefore, we investigated the role of mouse Akirin2 in acquired immune cells by ablating Akirin2 function in B lymphocytes. B cell–specific Akirin2-deficient (Cd19Cre/+Akirin2fl/fl) mice showed profound decrease in the splenic follicular (FO) and peritoneal B-1, but not splenic marginal zone (MZ), B cell numbers. However, both Akirin2-deficient FO and MZ B cells showed severe proliferation defect and are prone to undergo apoptosis in response to TLR ligands, CD40, and BCR stimulation. Furthermore, B cell cycling was defective in the absence of Akirin2 owing to impaired expression of genes encoding cyclin D and c-Myc. Additionally, Brg1 recruitment to the Myc and Ccnd2 promoter was severely impaired in Akirin2-deficient B cells. Cd19Cre/+Akirin2fl/fl mice showed impaired in vivo immune responses to T-dependent and -independent Ags. Collectively, these results demonstrate that Akirin2 is critical for the mitogen-induced B cell cycle progression and humoral immune responses by controlling the SWI/SNF complex, further emphasizing the significant function of Akirin2 not only in the innate, but also in adaptive immune cells.

The single nucleotide polymorphism (SNP) of aldehyde dehydrogenase-2 (ALDH2) codon 487, GAA (Glu) or AAA (Lys), was examined using green fluorescent protein (GFP)-display, an electrophoretic detection method for single amino acid changes. Although no shift in migration between the GFP-ALDH (Glu487) and GFP-ALDH (Lys487) fusion proteins was observed on SDS/urea gel, the two migrated to different positions when tagged with Asp. The SNP analysis was performed with GFP-ALDH-Asp3, and GFP-ALDH-Asp3 constructed from donors having the codon GAA/GAA, GAA/AAA or AAA/AAA was detected as different patterns as expected. GFP-display is potentially a unique method in SNP analysis, which does not require any special equipment or chemicals.

Transcription of inflammatory genes in innate immune cells is coordinately regulated by transcription factors, including NF-κB, and chromatin modifiers. However, it remains unclear how microbial sensing initiates chromatin remodeling. Here, we show that Akirin2, an evolutionarily conserved nuclear protein, bridges NF-κB and the chromatin remodeling SWI/SNF complex by interacting with BRG1-Associated Factor 60 (BAF60) proteins as well as IκB-ζ, which forms a complex with the NF-κB p50 subunit. These interactions are essential for Toll-like receptor-, RIG-I-, and Listeria-mediated expression of proinflammatory genes including Il6 and Il12b in macrophages. Consistently, effective clearance of Listeria infection required Akirin2. Furthermore, Akirin2 and IκB-ζ recruitment to the Il6 promoter depend upon the presence of IκB-ζ and Akirin2, respectively, for regulation of chromatin remodeling. BAF60 proteins were also essential for the induction of Il6 in response to LPS stimulation. Collectively, the IκB-ζ-Akirin2-BAF60 complex physically links the NF-κB and SWI/SNF complexes in innate immune cell activation. By recruiting SWI/SNF chromatin remodellers to IκB-ζ, transcriptional coactivator for NF-κB, the conserved nuclear protein Akirin2 stimulates pro-inflammatory gene promoters in mouse macrophages during innate immune responses to viral or bacterial infection.

昭和63年1月より12月までの1年間に, 隣接する宇部市と小野田市の2市で皮膚科医と麻酔科医を受診した帯状疱疹患者は1002例であつた。この1002例の患者につき統計的観察を行つた。その結果, 1)季節的には夏期に多く, 春にも少し多い傾向がみられた。2)女子にやや多く(男:女=1:1.4), 年齢的には50歳∼70歳代に最も多かつたが, 10歳代にも小さな山があり2峰性を示した。3)発生部位は胸·上肢が多かつた。4)基礎疾患を有するものは年齢別で大きな差がみられ10歳未満で7.9%, 10歳代で5.6%, 60歳代で35.4%, 70歳代で53.3%であつた。5)再罹患例は7例あり, うち1例は3度目の罹患であつた。6)発病後1ヵ月以上疼痛が続いた症例は52例(5.2%)であり, 70歳代が最も多く20例であつた。7)汎発性帯状疱疹は30例あり, 60歳代, 70歳代に多かつたが小児にも2例みられた。8)Hunt症候群を併発したものは7例, 9)眼疾患を併発したものは5例, 10)複発性帯状疱疹は3例であつた。11)悪性腫瘍を伴つた帯状疱疹患者は16例(1.6%)であつた。

Specific interaction between green fluorescent protein (GFP)-tagged human alpha- or gamma-enolase(97-242) (alpha or gammaENO(97-242)) and the rhodamine-labeled DNA fragment containing the c-myc P2 promoter was detected by a fluorescence resonance energy transfer (FRET)-based assay, designated as a "real-time FRET assay." The approach of donor (GFP) and acceptor (rhodamine) was caused by the association between ENO(97-242) and the c-myc P2 promoter, and the time-dependent increase in fluorescence intensity of the reaction mixture was observed at ex=400 nm and em=590 nm. The relative affinity (R(as)) of ENO(97-242) mutants to the wild type was investigated with a real-time FRET assay, and it was clarified that the amino acids that participated in the interaction existed comparatively broadly. Although it was difficult to measure the absolute value of the affinity for the binding protein by using this method, it was possible to investigate the relative affinity of mutants for the wild type. A real-time FRET assay using the GFP-tagged protein could be used as not only a qualitative, but also as a quantitative analysis, this being the best for investigating the key amino acids in binding proteins.

A method for rapid and simultaneous determination of 19 kinds of organophosphorus pesticides was developed by FPD-GC with two kinds of capillary columns (DB-5MS and DB-210). Sample preparation of the 19 pesticides in agricultural products was systematically established as follows. 1) Pesticides in vegetables and fruits were extracted with acetone, re-extracted with dichloromethane, and determined by GC. 2) Pesticides in onion were extracted with acetone by homogenation of a frozen sample in dilute phosphoric acid, and re-extracted with dichloromethane. 3) Pesticides in soybean, redbean and hulled rice were extracted with 30% aqueous acetone, then cleaned up with zinc acetate in order to remove some of the components, including fatty acids, as a precipitate, and re-extracted with dichloromethane. 4) Pesticides in green tea were extracted and re-extracted by the same method as in 3). Then the dichloromethane extract was concentrated to dryness, and the residue obtained was extracted with n-hexane in order to remove insoluble components.Recoveries and coefficients of variation of the pesticides in all agricultural products were in the ranges of 46.7-120% and 0.1-14.9%, respectively.

1.羽色などの外部形態に基づく繁殖期のカルガモの雌雄判別を試みるため,栃木県内で得られた80個体を材料に,雌雄による外部形態の違いを観察した。2.上尾筒,下尾羽及び腹に雌雄の差異が認められた。中央尾翼,翼鏡,三列風切及びみずかきの色にも変異が認められたが,雌雄の違いによるものであるとは判断できなかった。3.カルガモの雌雄判別を行う場合,上尾筒,下尾筒及び腹が有効な部位であるといえる。4.野外においてカルガモを観察する場合,至近距離であれば雌雄を判別することは可能であると思われる。

ヒヨドリの雌雄を判別する方法を確立するために,外部形態部位の計測値を基に,判別分析を行った。分析には,神奈川県三浦半島および東京周辺で捕獲されたもので,捕獲直後に計測したもの(標本A,C),関東～関西の各地で採集された後,冷凍保存されていたもの(標本B),北海道～九州の各地で採集され,山階鳥研標本室に保存されていたもの(標本D)を用いた。1.標本A～Cでは,測定した10部位の大きさと体重の平均値は,ほとんどの場合で,雄の方が雌よりも有意に大きく,体のサイズにおける明瞭な性的二型を示した。2.各計測値の標本ごとの平均値と分散を標本間で比較した結果,捕獲後冷凍保存されていた標本Bの一部の部位を除くと,標本A～Cは同じ母集団からの抽出標本と考えられた。それに対して,乾燥標本(標本D)はこれらとは異なった母集団からのものと結論された。3.同一シーズンに同一場所から捕獲された標本Aについて,判別分析を行った結果,翼長のみを用いた場合には,誤判別率は0.08であったが,変数の数を増やしていくと,誤判別率は低下した。しかし,〓蹠長,鼻孔前端長,全長,翼開長,全頭長,嘴幅の6変数を用いたときは,測定した全変数(11変数)を用いた場合とほぼ同程度の判別効率(誤判別率0.021)を示した。4.より一般的な判別関数を得るために,各地で得られた標本も加えて判別分析を行った結果,尾長,〓蹠長,鼻孔前端長,翼開長,全頭長の5変数を用いることで,誤判別率0.043の判別関数が得られた。また,この代替として,自然翼長,尾長,〓蹠長,嘴峰長,全頭長の5変数を用いた場合には,誤判別率は0.055であった。したがって,これらの関数を用いて,野外で捕獲したヒヨドリの雌雄を判別できる。5.標本A～Cで得られた判別関数を乾燥標本(標本D)に適用したところ,誤判別率は0.26であった。このため,乾燥標本の雌雄判別は,別途検討する必要がある。