JP2023175748A - 抗菌療法 - Google Patents
抗菌療法 Download PDFInfo
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- JP2023175748A JP2023175748A JP2023144981A JP2023144981A JP2023175748A JP 2023175748 A JP2023175748 A JP 2023175748A JP 2023144981 A JP2023144981 A JP 2023144981A JP 2023144981 A JP2023144981 A JP 2023144981A JP 2023175748 A JP2023175748 A JP 2023175748A
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Abstract
Description
本出願は、それらの開示が参照により本明細書に組み込まれる、2015年5月5日に出願された米国仮出願第62/157,248号、及び2016年2月26日に出願された米国仮出願第62/300,274号からの、35 U.S.C.§119下での優先権を主張する。
本発明は、助成金第R01AI083358号、助成金第AR067547号及び助成金第HHSN-272201000020C号による政府支援によりなされたものであり、それらの助成金は全て国立衛生研究所によって授与された。政府は、本発明に一定の権利を有する。
本出願は、電子フォーマットの配列表とともに提出されている。配列表は、2016年5月4日に作成されたSequence_ST25.txtという名前のファイルとして提供され、サイズは45Kbである。配列表の電子フォーマットの情報は、その全体において参照により本明細書に組み込まれる。
本開示は、感染を治療し、並びに皮膚及び粘膜の微生物叢を調節して、ディスバイオシス(dysbiosis)に関連し又はディスバイオシスによって悪化する疾患又は障害を治療するための方法及び組成物に関する。
配列番号2のaa32からaa61まで-KCSWWNASCHLGNNGKICTVSHECAAGCNL(配列番号56)
配列番号4のaa29からaa66まで-ATPTITTSSATCGGIIVAASAAQCPTLACSSRCGKRKK(配列番号57)
を含むアミノ酸鎖を含むホゴシジンの成熟形態を意味する。
MKTQRNGHSLGRWSLVLLLLGLVMPLAIIAQVLSYKEAVLRAIDGINQRSSDANLYRLLDLDPRPTMDGDPDTPKPVSFTVKETVCPRTTQQSPEDCDFKKDGLVKRCMGTVTLNQARGSFDISCDKDNKRFALLGDFFRKSKEKIGKEFKRIVQRIDDFLRNLVPRTES (配列番号5)
CoNS株及び抗菌ペプチドの単離。
アトピー性皮膚炎を有する大人の患者及び年齢が一致した非アトピー対象を動員した。人口統計学的データを表1に示す。ヒト対象を伴う全ての実験は、施設によって承認されたIRBプロトコールに従って実施された。
細菌株のFAME分析。細菌細胞抽出物の鹸化及びメチル化された試料中に存在する脂肪酸メチルエステルの相対存在量によって表すことができる全細菌細胞(主に細胞膜)の脂質組成は、各菌株にきわめてほぼ固有であるため、同定された菌株を脂肪酸メチルエステル(FAME)分析に供した。細菌株を標準的な技術に従って培養し、回収した。細胞を鹸化及びメチル化に供し、その後、ガスクロマトグラフィー用の移動相溶媒中に抽出した。試料をロードし、機器製造者の指示書に従って実施した。得られたクロマトグラムを図11~19に示す。
共生細菌は黄色ブドウ球菌によるコロニー形成から皮膚を保護する。AMPを産生する共生CoNSと、黄色ブドウ球菌によるコロニー形成との間の関係を明らかにし、これらの菌株によって産生される活性ペプチドを同定した後、これらの細菌の存在が黄色ブドウ球菌によるコロニー形成を減少させるかどうかについて試験する実験を行った。臨床的CoNS単離物は、所定量の黄色ブドウ球菌が最初に適用された、消毒したブタ皮膚の表面に適用された。黄色ブドウ球菌の生存率の有意な低下が、正常ヒト皮膚上の細菌密度の推定値(1×105CFU/cm2)と一致する密度でスタフィロコッカス・ホミニスA9の単回適用後に見られた(図20A)。適用前に死滅させ、洗われたスタフィロコッカス・ホミニスA9の適用、又は培養において抗菌活性を示さなかった他のスタフィロコッカス・ホミニス菌株の使用は、黄色ブドウ球菌の生存に影響を及ぼさなかった。同様に、所定量の黄色ブドウ球菌が適用されたマウスの背中への活性スタフィロコッカス・ホミニスの単回適用は、皮膚上の黄色ブドウ球菌の生存を減少させた(図20B)。対照的に、同様の密度での不活性株の適用は黄色ブドウ球菌を阻害しなかった。
ex vivoでのブタ皮膚及びマウスへの抗菌性CoNSの移植。凍結ブタ皮膚シートをLoretta Tomlin Animal Technologies(Livermore、CA)から入手し、3%クロロキシレノールを用いて外科用ブラシで消毒した。皮膚シートを2.5cm×2.5cmに切断し、滅菌PBSで20回超すすぎ、クロロキシレノール残留物を除去した。細菌適用の少なくとも24時間前に、ランダムに選択されたC57BL6雌性6週齢マウスの背中の皮膚を剃毛し、脱毛クリームで処理し、水ですすいだ。剃毛した皮膚をアルコールスワブで2回洗浄して、もともとコロニー形成していた細菌を除去した。生存動物の実験を含むすべての実験は、実験動物のケア及び使用ガイドラインの承認に従った。
自家微生物叢移植。AD患者への自家微生物叢移植(AMT)のアプローチは、米国食品医薬品局(FDA)によって正式に承認されており、このプロトコールは治験新薬申請(IND)として申請されている(UCSD承認番号15786)。スクリーニングの来院時に、両肘前窩の病変部位に対して黄色ブドウ球菌保菌者であるAD患者をスクリーニングした。一方、皮膚の細菌を、AD患者の上腕の非病変皮膚から拭き取ることによって取得し、黄色ブドウ球菌に対して抗菌活性を生じさせるCoNS株をスクリーニングした。抗菌性CoNS分離株の種は、全長16S rRNA遺伝子のサンガー配列決定によって同定された。患者が移植療法を受けた2回目の来院まで、CoNS分離株のグリセロールストックを-80℃で保存した。各CoNS株をTSB中で一晩個別に増やした。各CoNS株は、細菌の生存率に影響を与えないことが確認された皮膚保湿剤中に1×107 CFU/gで製剤化された。抗菌活性を有する表皮ブドウ球菌又はスタフィロコッカス・ホミニスの単一の菌株のみが3人の患者から単離された。これらの場合において、CoNSの単一株が製剤化された。3つ及び2つの抗菌性スタフィロコッカス・ホミニス又は表皮ブドウ球菌の菌株が2人のAD患者から単離された(図20D)。これらの場合において、各CoNSの等しいCFUが107 CFU/gの総濃度で製剤化された。2回目の来院で、関連する領域を測定し、両前腕の病変部位上に生存している黄色ブドウ球菌の基準CFUを上記のように定量した。一方の腕を10mg/cm2でAMT製剤を用いて処置し、1×105 CFU/cm2のCoNSを得た。他方の腕は、同量の保湿剤で処置しただけであった。全ての処置を二重盲検法で行い、全ての結果を分析した後には盲検化しなかった。対象は、入浴、シャワー、運動、又は腕へのいかなる外用製品の適用も避け、次回の来院まで他方の腕に適用されたCoNSの交差汚染を避けるために、清潔な長袖のシャツを身に着けた。3回目の来院時に、関与領域における黄色ブドウ球菌CFUを測定した。ビヒクル腕とAMT腕の間の黄色ブドウ球菌の生存率の差異は、[AMT(χ時間)-ビヒクル(χ時間)]/AMT(基準)として計算され、黄色ブドウ球菌CFU Δ%(χ=0時間又は24時間)を得た(図20E)。
同種異系移植。SH-A9、SH-C2、SE-A11、AMT1、AMT2、AMT3、AMT4-C2、AMT4-G1、AMT4-D12、AMT5-C5、AMT5-G6及び/又はSE-MO34菌株の105 CFU/gは、細菌生存率に影響を与えないことが確認されている皮膚保湿剤中に製剤化される。治療対象は、アトピー性皮膚炎及び/又は活性な黄色ブドウ球菌感染の存在に基づいて同定される。対象は、3日間、入浴、シャワー、運動、又は患部へのいかなる外用製品の適用も避けるように指示される。適合した患者は、7日間後、処置領域における黄色ブドウ球菌レベルの有意な減少を示す(回収可能な黄色ブドウ球菌コロニーカウントにおいて3log又はそれ以上の減少)。適合した患者は、初期治療後、少なくとも14日間継続している黄色ブドウ球菌感染及び/又はアトピー性皮膚炎の症状の臨床的に観察可能な減少を示す。
表皮ブドウ球菌によって産生されたAMPの精製。代表的な抗菌性表皮ブドウ球菌A11株由来の滅菌馴化培地を用いて、正常な皮膚上の抗菌活性を有する分子をさらに同定し、これはアトピーにおいて存在量が低かった。活性物を硫酸アンモニウム(70%飽和)により沈殿させ、H2Oに溶解し、Sep-Pakカートリッジ(Waters Co)に適用した。活性画分をH2O中の40%アセトニトリルで溶出し、HiTrap(登録商標)SP(GE Healthcare Life Sciences)分離に供し、活性物を500mM NaClで溶出した。第三段階のHPLC精製は、0.1%(v/v)TFA中の5%~50%のアセトニトリルの線形勾配で0.8mL/分にてCapCel Pak C8(5μm、300Å、4.6×250mm)(Shiseido Co.)を用いて行われた。
SEQUENCE LISTING
<110> The Regents of the University of California
<120> ANTIMICROBIAL THERAPY
<130> PA23-456
<140>
<141> 2016-05-05
<150> US 62/157,248
<151> 2015-05-05
<150> US 62/300,274
<151> 2016-02-26
<160> 57
<170> PatentIn version 3.5
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<211> 186
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(186)
<400> 1
atg agt aaa tta gaa cta ctt aat gaa tct aaa gca aat tat ctt gaa 48
Met Ser Lys Leu Glu Leu Leu Asn Glu Ser Lys Ala Asn Tyr Leu Glu
1 5 10 15
aaa ctt act gat gaa aaa att gaa gaa acg gaa gca tac ggc ggt aaa 96
Lys Leu Thr Asp Glu Lys Ile Glu Glu Thr Glu Ala Tyr Gly Gly Lys
20 25 30
tgt tct tgg tgg aat gca tca tgt cat tta gga aat aat ggg aaa att 144
Cys Ser Trp Trp Asn Ala Ser Cys His Leu Gly Asn Asn Gly Lys Ile
35 40 45
tgt aca gtt tct cat gag tgt gca gca gga tgt aat tta taa 186
Cys Thr Val Ser His Glu Cys Ala Ala Gly Cys Asn Leu
50 55 60
<210> 2
<211> 61
<212> PRT
<213> Staphylococcus hominis
<400> 2
Met Ser Lys Leu Glu Leu Leu Asn Glu Ser Lys Ala Asn Tyr Leu Glu
1 5 10 15
Lys Leu Thr Asp Glu Lys Ile Glu Glu Thr Glu Ala Tyr Gly Gly Lys
20 25 30
Cys Ser Trp Trp Asn Ala Ser Cys His Leu Gly Asn Asn Gly Lys Ile
35 40 45
Cys Thr Val Ser His Glu Cys Ala Ala Gly Cys Asn Leu
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<210> 3
<211> 201
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(201)
<400> 3
atg ttt agt aaa aat ttc caa aga aat gaa aag atg gaa aat act ttg 48
Met Phe Ser Lys Asn Phe Gln Arg Asn Glu Lys Met Glu Asn Thr Leu
1 5 10 15
aaa aag gta agt tca gct aat gat gtg aat gga gga gct aca ccg act 96
Lys Lys Val Ser Ser Ala Asn Asp Val Asn Gly Gly Ala Thr Pro Thr
20 25 30
att act aca tct tca gca act tgt ggt ggt att att gtt gcg gca agt 144
Ile Thr Thr Ser Ser Ala Thr Cys Gly Gly Ile Ile Val Ala Ala Ser
35 40 45
gct gct cag tgt ccg aca tta gct tgc tct tct aga tgt gga aaa aga 192
Ala Ala Gln Cys Pro Thr Leu Ala Cys Ser Ser Arg Cys Gly Lys Arg
50 55 60
aaa aaa taa 201
Lys Lys
65
<210> 4
<211> 66
<212> PRT
<213> Staphylococcus hominis
<400> 4
Met Phe Ser Lys Asn Phe Gln Arg Asn Glu Lys Met Glu Asn Thr Leu
1 5 10 15
Lys Lys Val Ser Ser Ala Asn Asp Val Asn Gly Gly Ala Thr Pro Thr
20 25 30
Ile Thr Thr Ser Ser Ala Thr Cys Gly Gly Ile Ile Val Ala Ala Ser
35 40 45
Ala Ala Gln Cys Pro Thr Leu Ala Cys Ser Ser Arg Cys Gly Lys Arg
50 55 60
Lys Lys
65
<210> 5
<211> 170
<212> PRT
<213> Homo sapiens
<400> 5
Met Lys Thr Gln Arg Asn Gly His Ser Leu Gly Arg Trp Ser Leu Val
1 5 10 15
Leu Leu Leu Leu Gly Leu Val Met Pro Leu Ala Ile Ile Ala Gln Val
20 25 30
Leu Ser Tyr Lys Glu Ala Val Leu Arg Ala Ile Asp Gly Ile Asn Gln
35 40 45
Arg Ser Ser Asp Ala Asn Leu Tyr Arg Leu Leu Asp Leu Asp Pro Arg
50 55 60
Pro Thr Met Asp Gly Asp Pro Asp Thr Pro Lys Pro Val Ser Phe Thr
65 70 75 80
Val Lys Glu Thr Val Cys Pro Arg Thr Thr Gln Gln Ser Pro Glu Asp
85 90 95
Cys Asp Phe Lys Lys Asp Gly Leu Val Lys Arg Cys Met Gly Thr Val
100 105 110
Thr Leu Asn Gln Ala Arg Gly Ser Phe Asp Ile Ser Cys Asp Lys Asp
115 120 125
Asn Lys Arg Phe Ala Leu Leu Gly Asp Phe Phe Arg Lys Ser Lys Glu
130 135 140
Lys Ile Gly Lys Glu Phe Lys Arg Ile Val Gln Arg Ile Asp Asp Phe
145 150 155 160
Leu Arg Asn Leu Val Pro Arg Thr Glu Ser
165 170
<210> 6
<211> 5
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<223> Synthetic linker
<400> 6
Gly Gly Gly Gly Ser
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<210> 7
<211> 22
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<213> Artificial Sequence
<220>
<223> Synthetic linker
<400> 7
Gly Gly Gly Gly Gly Gly Ser Met Phe Gly Gly Ala Lys Lys Arg Ser
1 5 10 15
Gly Gly Gly Gly Gly Gly
20
<210> 8
<211> 20
<212> DNA
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<210> 9
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<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus aureus-specific PCR primer
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<210> 10
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis-specific PCR primer
<400> 10
tcagcagttg aaggacagat 20
<210> 11
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis-specific PCR primer
<400> 11
ccagaacaat gaatggttaa gg 22
<210> 12
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
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<400> 12
tttgggctac acacgtgcta caatggacaa 30
<210> 13
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus-genus specific 16S sequence PCR primer
<400> 13
aacaacttta tgggatttgc wtga 24
<210> 14
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Universal sequence of bacterial 16S rRNA PCR primer
<400> 14
agagtttgga tcmtggctca g 21
<210> 15
<211> 17
<212> DNA
<213> Artificial Sequence
<220>
<223> Universal sequence of bacterial 16S rRNA PCR primer
<400> 15
aaggaggtgw tccarcc 17
<210> 16
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epidermn PCR primer
<400> 16
gattcaggag ctgaaccaag a 21
<210> 17
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epidermn PCR primer
<400> 17
ttgaagccct gccaatctaa 20
<210> 18
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis pep5 PCR primer
<400> 18
ctgatgaact tgaacctcaa actg 24
<210> 19
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis pep5 PCR primer
<400> 19
gacactgtaa ataaacgcgt agc 23
<210> 20
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epicidin280 PCR primer
<400> 20
gcaactagac aggtatgtcc taaa 24
<210> 21
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epicidin280 PCR primer
<400> 21
catctaagat taaatgaggg tggtt 25
<210> 22
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epilancin K7 PCR primer
<400> 22
taagtccgca atctgctagt g 21
<210> 23
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus epidermidis epilancin K7 PCR primer
<400> 23
cagtaatatt gcaaccgcat gt 22
<210> 24
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus hominis Hogocidin-alpha PCR primer
<400> 24
atgagtaaat tagaactact taatg 25
<210> 25
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> Staphylococcus hominis Hogocidin-alpha PCR primer
<400> 25
ttataaatta catcctgctg cacac 25
<210> 26
<211> 8
<212> PRT
<213> Staphylococcus hominis
<400> 26
Lys Cys Ser Asp Asp Asn Ala Ala
1 5
<210> 27
<211> 138
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(138)
<400> 27
atg gat att ata aaa gta aat aaa aca gaa aga atg aat gat aat aga 48
Met Asp Ile Ile Lys Val Asn Lys Thr Glu Arg Met Asn Asp Asn Arg
1 5 10 15
aaa att gta atg att ttt tct tta tac gat aca ttt ttt aac gct act 96
Lys Ile Val Met Ile Phe Ser Leu Tyr Asp Thr Phe Phe Asn Ala Thr
20 25 30
aat aca cat aag cta aag agt atg aag ctt aat gcg aaa taa 138
Asn Thr His Lys Leu Lys Ser Met Lys Leu Asn Ala Lys
35 40 45
<210> 28
<211> 45
<212> PRT
<213> Staphylococcus hominis
<400> 28
Met Asp Ile Ile Lys Val Asn Lys Thr Glu Arg Met Asn Asp Asn Arg
1 5 10 15
Lys Ile Val Met Ile Phe Ser Leu Tyr Asp Thr Phe Phe Asn Ala Thr
20 25 30
Asn Thr His Lys Leu Lys Ser Met Lys Leu Asn Ala Lys
35 40 45
<210> 29
<211> 168
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(168)
<400> 29
atg gta tgg atc cat ggt ggt ggt aac tta ggt ggt gct ggc tta gaa 48
Met Val Trp Ile His Gly Gly Gly Asn Leu Gly Gly Ala Gly Leu Glu
1 5 10 15
gat gct ttt gat ggt aat act tta gct aaa cat aca tca aaa att aaa 96
Asp Ala Phe Asp Gly Asn Thr Leu Ala Lys His Thr Ser Lys Ile Lys
20 25 30
tat gtt ttt gga aat ttg caa cct gaa aac cat tat gat gat att gat 144
Tyr Val Phe Gly Asn Leu Gln Pro Glu Asn His Tyr Asp Asp Ile Asp
35 40 45
ata ata atc tca aaa caa tta taa 168
Ile Ile Ile Ser Lys Gln Leu
50 55
<210> 30
<211> 55
<212> PRT
<213> Staphylococcus epidermidis
<400> 30
Met Val Trp Ile His Gly Gly Gly Asn Leu Gly Gly Ala Gly Leu Glu
1 5 10 15
Asp Ala Phe Asp Gly Asn Thr Leu Ala Lys His Thr Ser Lys Ile Lys
20 25 30
Tyr Val Phe Gly Asn Leu Gln Pro Glu Asn His Tyr Asp Asp Ile Asp
35 40 45
Ile Ile Ile Ser Lys Gln Leu
50 55
<210> 31
<211> 207
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(207)
<400> 31
atg att tca gtg ata ctg ccg atg gaa gaa ata att att gca ata ata 48
Met Ile Ser Val Ile Leu Pro Met Glu Glu Ile Ile Ile Ala Ile Ile
1 5 10 15
aat aac gac tta ggc cat tta att ttt gag aat aaa aaa aat agt ggg 96
Asn Asn Asp Leu Gly His Leu Ile Phe Glu Asn Lys Lys Asn Ser Gly
20 25 30
ttt tct ttt ttc ata ata aaa cct ttc ata act aat att tat ttt cta 144
Phe Ser Phe Phe Ile Ile Lys Pro Phe Ile Thr Asn Ile Tyr Phe Leu
35 40 45
tca ggt att aaa aaa att tta caa aaa cag aga aaa tat tat acg atg 192
Ser Gly Ile Lys Lys Ile Leu Gln Lys Gln Arg Lys Tyr Tyr Thr Met
50 55 60
cta ata aaa gta taa 207
Leu Ile Lys Val
65
<210> 32
<211> 68
<212> PRT
<213> Staphylococcus epidermidis
<400> 32
Met Ile Ser Val Ile Leu Pro Met Glu Glu Ile Ile Ile Ala Ile Ile
1 5 10 15
Asn Asn Asp Leu Gly His Leu Ile Phe Glu Asn Lys Lys Asn Ser Gly
20 25 30
Phe Ser Phe Phe Ile Ile Lys Pro Phe Ile Thr Asn Ile Tyr Phe Leu
35 40 45
Ser Gly Ile Lys Lys Ile Leu Gln Lys Gln Arg Lys Tyr Tyr Thr Met
50 55 60
Leu Ile Lys Val
65
<210> 33
<211> 210
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(210)
<400> 33
atg aac ata tac tta aaa gta att tta act tct tta ttt ttt gct tta 48
Met Asn Ile Tyr Leu Lys Val Ile Leu Thr Ser Leu Phe Phe Ala Leu
1 5 10 15
ata att ttt att gta act tat ata acg act aag caa tgg gga aca tcg 96
Ile Ile Phe Ile Val Thr Tyr Ile Thr Thr Lys Gln Trp Gly Thr Ser
20 25 30
tta ggt ttt tca tct tta tca ttt atc ggt aac ttt att tac gat tat 144
Leu Gly Phe Ser Ser Leu Ser Phe Ile Gly Asn Phe Ile Tyr Asp Tyr
35 40 45
tca acg aaa tta agt gat aaa aaa tat gaa aaa aga ata aat agc aac 192
Ser Thr Lys Leu Ser Asp Lys Lys Tyr Glu Lys Arg Ile Asn Ser Asn
50 55 60
aaa aaa gat aaa ctt tag 210
Lys Lys Asp Lys Leu
65
<210> 34
<211> 69
<212> PRT
<213> Staphylococcus epidermidis
<400> 34
Met Asn Ile Tyr Leu Lys Val Ile Leu Thr Ser Leu Phe Phe Ala Leu
1 5 10 15
Ile Ile Phe Ile Val Thr Tyr Ile Thr Thr Lys Gln Trp Gly Thr Ser
20 25 30
Leu Gly Phe Ser Ser Leu Ser Phe Ile Gly Asn Phe Ile Tyr Asp Tyr
35 40 45
Ser Thr Lys Leu Ser Asp Lys Lys Tyr Glu Lys Arg Ile Asn Ser Asn
50 55 60
Lys Lys Asp Lys Leu
65
<210> 35
<211> 183
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(183)
<400> 35
atg aaa aat aac aaa aat tta ttt gat tta gaa att aaa aaa gaa aca 48
Met Lys Asn Asn Lys Asn Leu Phe Asp Leu Glu Ile Lys Lys Glu Thr
1 5 10 15
agt caa aac act gat gaa ctt gaa cct caa act gct gga cca gcg att 96
Ser Gln Asn Thr Asp Glu Leu Glu Pro Gln Thr Ala Gly Pro Ala Ile
20 25 30
aga gct tct gtg aaa caa tgt cag aaa act ttg aaa gct acg cgt tta 144
Arg Ala Ser Val Lys Gln Cys Gln Lys Thr Leu Lys Ala Thr Arg Leu
35 40 45
ttt aca gtg tct tgc aaa gga aaa aac gga tgt aaa tag 183
Phe Thr Val Ser Cys Lys Gly Lys Asn Gly Cys Lys
50 55 60
<210> 36
<211> 60
<212> PRT
<213> Staphylococcus epidermidis
<400> 36
Met Lys Asn Asn Lys Asn Leu Phe Asp Leu Glu Ile Lys Lys Glu Thr
1 5 10 15
Ser Gln Asn Thr Asp Glu Leu Glu Pro Gln Thr Ala Gly Pro Ala Ile
20 25 30
Arg Ala Ser Val Lys Gln Cys Gln Lys Thr Leu Lys Ala Thr Arg Leu
35 40 45
Phe Thr Val Ser Cys Lys Gly Lys Asn Gly Cys Lys
50 55 60
<210> 37
<211> 171
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(171)
<400> 37
atg gaa aac aaa aaa gat tta ttt gat tta gaa atc aaa aaa gat aat 48
Met Glu Asn Lys Lys Asp Leu Phe Asp Leu Glu Ile Lys Lys Asp Asn
1 5 10 15
atg gaa aat aat aat gaa tta gaa gct caa tct ctt ggt cct gca att 96
Met Glu Asn Asn Asn Glu Leu Glu Ala Gln Ser Leu Gly Pro Ala Ile
20 25 30
aag gca act aga cag gta tgt cct aaa gca aca cgt ttt gtt aca gtt 144
Lys Ala Thr Arg Gln Val Cys Pro Lys Ala Thr Arg Phe Val Thr Val
35 40 45
tct tgt aaa aaa agt gat tgt caa tag 171
Ser Cys Lys Lys Ser Asp Cys Gln
50 55
<210> 38
<211> 56
<212> PRT
<213> Staphylococcus hominis
<400> 38
Met Glu Asn Lys Lys Asp Leu Phe Asp Leu Glu Ile Lys Lys Asp Asn
1 5 10 15
Met Glu Asn Asn Asn Glu Leu Glu Ala Gln Ser Leu Gly Pro Ala Ile
20 25 30
Lys Ala Thr Arg Gln Val Cys Pro Lys Ala Thr Arg Phe Val Thr Val
35 40 45
Ser Cys Lys Lys Ser Asp Cys Gln
50 55
<210> 39
<211> 135
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(135)
<400> 39
atg aaa gtt gtt aaa gaa aag aaa gaa ctt ttt gat ctt gac gtt aaa 48
Met Lys Val Val Lys Glu Lys Lys Glu Leu Phe Asp Leu Asp Val Lys
1 5 10 15
gta aat gcg aga gac atg aat aat tca gaa tca ggt cca cct aat aca 96
Val Asn Ala Arg Asp Met Asn Asn Ser Glu Ser Gly Pro Pro Asn Thr
20 25 30
agt tta ata tgg tgt acg gat gga tgc gct aaa cgg taa 135
Ser Leu Ile Trp Cys Thr Asp Gly Cys Ala Lys Arg
35 40
<210> 40
<211> 44
<212> PRT
<213> Staphylococcus hominis
<400> 40
Met Lys Val Val Lys Glu Lys Lys Glu Leu Phe Asp Leu Asp Val Lys
1 5 10 15
Val Asn Ala Arg Asp Met Asn Asn Ser Glu Ser Gly Pro Pro Asn Thr
20 25 30
Ser Leu Ile Trp Cys Thr Asp Gly Cys Ala Lys Arg
35 40
<210> 41
<211> 138
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(138)
<400> 41
atg gat att ata aaa gta aat aaa aca gaa aga atg aat gat aat aga 48
Met Asp Ile Ile Lys Val Asn Lys Thr Glu Arg Met Asn Asp Asn Arg
1 5 10 15
aaa att gta atg att ttt tct tta tac gat aca ttt ttt aac gct act 96
Lys Ile Val Met Ile Phe Ser Leu Tyr Asp Thr Phe Phe Asn Ala Thr
20 25 30
aat aca cat aag cta aag agt atg aag ctt aat gcg aaa taa 138
Asn Thr His Lys Leu Lys Ser Met Lys Leu Asn Ala Lys
35 40 45
<210> 42
<211> 45
<212> PRT
<213> Staphylococcus hominis
<400> 42
Met Asp Ile Ile Lys Val Asn Lys Thr Glu Arg Met Asn Asp Asn Arg
1 5 10 15
Lys Ile Val Met Ile Phe Ser Leu Tyr Asp Thr Phe Phe Asn Ala Thr
20 25 30
Asn Thr His Lys Leu Lys Ser Met Lys Leu Asn Ala Lys
35 40 45
<210> 43
<211> 201
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(201)
<400> 43
atg agt aat aaa gat tta gaa tta ttt aat aca gcc ggt gat tta ata 48
Met Ser Asn Lys Asp Leu Glu Leu Phe Asn Thr Ala Gly Asp Leu Ile
1 5 10 15
caa gaa tta aaa gat ggt gac cta aat atc cat tta tat ggt gaa tcg 96
Gln Glu Leu Lys Asp Gly Asp Leu Asn Ile His Leu Tyr Gly Glu Ser
20 25 30
gaa att aga aaa aaa tct ttc tct caa aaa aca ggg aat gat ggg aaa 144
Glu Ile Arg Lys Lys Ser Phe Ser Gln Lys Thr Gly Asn Asp Gly Lys
35 40 45
cat tgt aca att act tgg gaa tgt tct ata tgt cct act aaa act tgt 192
His Cys Thr Ile Thr Trp Glu Cys Ser Ile Cys Pro Thr Lys Thr Cys
50 55 60
tgg tgc taa 201
Trp Cys
65
<210> 44
<211> 66
<212> PRT
<213> Staphylococcus hominis
<400> 44
Met Ser Asn Lys Asp Leu Glu Leu Phe Asn Thr Ala Gly Asp Leu Ile
1 5 10 15
Gln Glu Leu Lys Asp Gly Asp Leu Asn Ile His Leu Tyr Gly Glu Ser
20 25 30
Glu Ile Arg Lys Lys Ser Phe Ser Gln Lys Thr Gly Asn Asp Gly Lys
35 40 45
His Cys Thr Ile Thr Trp Glu Cys Ser Ile Cys Pro Thr Lys Thr Cys
50 55 60
Trp Cys
65
<210> 45
<211> 117
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(117)
<400> 45
atg gga act tca gag gta aga aaa gga aaa gga ggc ggt ttt agt acc 48
Met Gly Thr Ser Glu Val Arg Lys Gly Lys Gly Gly Gly Phe Ser Thr
1 5 10 15
gta acc gtt gta aca cca att gta ccg aca tcg aag tgt gcc tca att 96
Val Thr Val Val Thr Pro Ile Val Pro Thr Ser Lys Cys Ala Ser Ile
20 25 30
gta aaa cca tgt aac aaa taa 117
Val Lys Pro Cys Asn Lys
35
<210> 46
<211> 38
<212> PRT
<213> Staphylococcus hominis
<400> 46
Met Gly Thr Ser Glu Val Arg Lys Gly Lys Gly Gly Gly Phe Ser Thr
1 5 10 15
Val Thr Val Val Thr Pro Ile Val Pro Thr Ser Lys Cys Ala Ser Ile
20 25 30
Val Lys Pro Cys Asn Lys
35
<210> 47
<211> 165
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(165)
<400> 47
atg act aaa ata act aaa gat gat ttg aaa aag att aca gaa aat cgt 48
Met Thr Lys Ile Thr Lys Asp Asp Leu Lys Lys Ile Thr Glu Asn Arg
1 5 10 15
att gaa gca cgt aca cat cca acc gtt gtt cct gta agt gct gct gta 96
Ile Glu Ala Arg Thr His Pro Thr Val Val Pro Val Ser Ala Ala Val
20 25 30
tgc gga gtt gct act aaa tta gta cca aca tcg aaa tgt gct tca att 144
Cys Gly Val Ala Thr Lys Leu Val Pro Thr Ser Lys Cys Ala Ser Ile
35 40 45
gta aaa cca tgt aat aaa taa 165
Val Lys Pro Cys Asn Lys
50
<210> 48
<211> 54
<212> PRT
<213> Staphylococcus hominis
<400> 48
Met Thr Lys Ile Thr Lys Asp Asp Leu Lys Lys Ile Thr Glu Asn Arg
1 5 10 15
Ile Glu Ala Arg Thr His Pro Thr Val Val Pro Val Ser Ala Ala Val
20 25 30
Cys Gly Val Ala Thr Lys Leu Val Pro Thr Ser Lys Cys Ala Ser Ile
35 40 45
Val Lys Pro Cys Asn Lys
50
<210> 49
<211> 522
<212> DNA
<213> Staphylococcus hominis
<220>
<221> CDS
<222> (1)..(522)
<400> 49
atg act ata gat tta gta atg ata ctt att att ttg atg tat atg ata 48
Met Thr Ile Asp Leu Val Met Ile Leu Ile Ile Leu Met Tyr Met Ile
1 5 10 15
att ggt ttt aga aga ggc ctt tgg ctg aat agt ctt cat ttg tcg tct 96
Ile Gly Phe Arg Arg Gly Leu Trp Leu Asn Ser Leu His Leu Ser Ser
20 25 30
aca ctt gtc tca cta ttc att gcg cat cgt ttt tac caa tat ata tca 144
Thr Leu Val Ser Leu Phe Ile Ala His Arg Phe Tyr Gln Tyr Ile Ser
35 40 45
aaa caa atg att gtt ttt gtt cca ttt cct aaa aca gtt gct ttt gac 192
Lys Gln Met Ile Val Phe Val Pro Phe Pro Lys Thr Val Ala Phe Asp
50 55 60
acg cac ttc gca ttt caa tac cat gat gta caa caa cgt ttt gat act 240
Thr His Phe Ala Phe Gln Tyr His Asp Val Gln Gln Arg Phe Asp Thr
65 70 75 80
att gtg gca ttt tta tgt att gct ttt ata agt aag ttg ctt tta tat 288
Ile Val Ala Phe Leu Cys Ile Ala Phe Ile Ser Lys Leu Leu Leu Tyr
85 90 95
ctt att att gta act ttt gat aat ata gtg tca tat cat aat att cat 336
Leu Ile Ile Val Thr Phe Asp Asn Ile Val Ser Tyr His Asn Ile His
100 105 110
gtt aca agt cga ata ttg gga agc gta tta ggt agt att gca agt gtg 384
Val Thr Ser Arg Ile Leu Gly Ser Val Leu Gly Ser Ile Ala Ser Val
115 120 125
att gta ctg caa ctt gtt tta tat tta gta tct tta tat cct aat gaa 432
Ile Val Leu Gln Leu Val Leu Tyr Leu Val Ser Leu Tyr Pro Asn Glu
130 135 140
tgg att caa gaa agt tta aaa tac ggt tat tta agc cat att att cta 480
Trp Ile Gln Glu Ser Leu Lys Tyr Gly Tyr Leu Ser His Ile Ile Leu
145 150 155 160
ttt aag atg ccg ttt tta tca tct tat ata cta aat tta taa 522
Phe Lys Met Pro Phe Leu Ser Ser Tyr Ile Leu Asn Leu
165 170
<210> 50
<211> 173
<212> PRT
<213> Staphylococcus hominis
<400> 50
Met Thr Ile Asp Leu Val Met Ile Leu Ile Ile Leu Met Tyr Met Ile
1 5 10 15
Ile Gly Phe Arg Arg Gly Leu Trp Leu Asn Ser Leu His Leu Ser Ser
20 25 30
Thr Leu Val Ser Leu Phe Ile Ala His Arg Phe Tyr Gln Tyr Ile Ser
35 40 45
Lys Gln Met Ile Val Phe Val Pro Phe Pro Lys Thr Val Ala Phe Asp
50 55 60
Thr His Phe Ala Phe Gln Tyr His Asp Val Gln Gln Arg Phe Asp Thr
65 70 75 80
Ile Val Ala Phe Leu Cys Ile Ala Phe Ile Ser Lys Leu Leu Leu Tyr
85 90 95
Leu Ile Ile Val Thr Phe Asp Asn Ile Val Ser Tyr His Asn Ile His
100 105 110
Val Thr Ser Arg Ile Leu Gly Ser Val Leu Gly Ser Ile Ala Ser Val
115 120 125
Ile Val Leu Gln Leu Val Leu Tyr Leu Val Ser Leu Tyr Pro Asn Glu
130 135 140
Trp Ile Gln Glu Ser Leu Lys Tyr Gly Tyr Leu Ser His Ile Ile Leu
145 150 155 160
Phe Lys Met Pro Phe Leu Ser Ser Tyr Ile Leu Asn Leu
165 170
<210> 51
<211> 522
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(522)
<400> 51
atg ctc att gat ata gtt gtt ctt ctt att att tgt tac ttt ata gtg 48
Met Leu Ile Asp Ile Val Val Leu Leu Ile Ile Cys Tyr Phe Ile Val
1 5 10 15
ata ggg ttt cgt aga ggt att tgg tta tcg ata ttg cac ttt gct tct 96
Ile Gly Phe Arg Arg Gly Ile Trp Leu Ser Ile Leu His Phe Ala Ser
20 25 30
tca att gta tct tta tat att gcg tca caa cat tat caa tcg att gcg 144
Ser Ile Val Ser Leu Tyr Ile Ala Ser Gln His Tyr Gln Ser Ile Ala
35 40 45
caa cgt tta gtt gta ttt gtg cca ttt ccg aaa acg gtg gcg ttt gat 192
Gln Arg Leu Val Val Phe Val Pro Phe Pro Lys Thr Val Ala Phe Asp
50 55 60
atg gtc tat act ata cct tat gat cat ttg caa tac aga ttt gaa aaa 240
Met Val Tyr Thr Ile Pro Tyr Asp His Leu Gln Tyr Arg Phe Glu Lys
65 70 75 80
gtg ata gca ttt att ata ata ttt ggt atg tgt aag ctt att ttg tat 288
Val Ile Ala Phe Ile Ile Ile Phe Gly Met Cys Lys Leu Ile Leu Tyr
85 90 95
cta gtt gtt gtt aca ttt gat aat ata ata acg tat aaa aag ata cat 336
Leu Val Val Val Thr Phe Asp Asn Ile Ile Thr Tyr Lys Lys Ile His
100 105 110
tta gta agt cgg ata tcg agt gtc gtt ttg agt atc ata tcg gtt ttt 384
Leu Val Ser Arg Ile Ser Ser Val Val Leu Ser Ile Ile Ser Val Phe
115 120 125
ata tat tta caa att gga ctt tat tta tta tcg cta tat ccg cat tca 432
Ile Tyr Leu Gln Ile Gly Leu Tyr Leu Leu Ser Leu Tyr Pro His Ser
130 135 140
ttt ata cag tac caa tta tct caa tcg cta gta agt cga gtt gtg att 480
Phe Ile Gln Tyr Gln Leu Ser Gln Ser Leu Val Ser Arg Val Val Ile
145 150 155 160
gaa caa att cct tat tta tca caa ttt att tta aat tta taa 522
Glu Gln Ile Pro Tyr Leu Ser Gln Phe Ile Leu Asn Leu
165 170
<210> 52
<211> 173
<212> PRT
<213> Staphylococcus epidermidis
<400> 52
Met Leu Ile Asp Ile Val Val Leu Leu Ile Ile Cys Tyr Phe Ile Val
1 5 10 15
Ile Gly Phe Arg Arg Gly Ile Trp Leu Ser Ile Leu His Phe Ala Ser
20 25 30
Ser Ile Val Ser Leu Tyr Ile Ala Ser Gln His Tyr Gln Ser Ile Ala
35 40 45
Gln Arg Leu Val Val Phe Val Pro Phe Pro Lys Thr Val Ala Phe Asp
50 55 60
Met Val Tyr Thr Ile Pro Tyr Asp His Leu Gln Tyr Arg Phe Glu Lys
65 70 75 80
Val Ile Ala Phe Ile Ile Ile Phe Gly Met Cys Lys Leu Ile Leu Tyr
85 90 95
Leu Val Val Val Thr Phe Asp Asn Ile Ile Thr Tyr Lys Lys Ile His
100 105 110
Leu Val Ser Arg Ile Ser Ser Val Val Leu Ser Ile Ile Ser Val Phe
115 120 125
Ile Tyr Leu Gln Ile Gly Leu Tyr Leu Leu Ser Leu Tyr Pro His Ser
130 135 140
Phe Ile Gln Tyr Gln Leu Ser Gln Ser Leu Val Ser Arg Val Val Ile
145 150 155 160
Glu Gln Ile Pro Tyr Leu Ser Gln Phe Ile Leu Asn Leu
165 170
<210> 53
<211> 1260
<212> DNA
<213> Staphylococcus epidermidis
<220>
<221> CDS
<222> (1)..(1260)
<400> 53
atg att ggt aga aaa aaa gaa acc ctt tta aaa aac gaa gtt att tct 48
Met Ile Gly Arg Lys Lys Glu Thr Leu Leu Lys Asn Glu Val Ile Ser
1 5 10 15
gcg ttt act act ttt ttt acc tgc agt tat ata ata att gtt aat ggt 96
Ala Phe Thr Thr Phe Phe Thr Cys Ser Tyr Ile Ile Ile Val Asn Gly
20 25 30
ata ttg tta cat caa gca gga atg tct ttg tta tgg acg att ata gct 144
Ile Leu Leu His Gln Ala Gly Met Ser Leu Leu Trp Thr Ile Ile Ala
35 40 45
act act cta gtt tgt tgc att agt tgc atc ctt ctt ggt ata tat gct 192
Thr Thr Leu Val Cys Cys Ile Ser Cys Ile Leu Leu Gly Ile Tyr Ala
50 55 60
aat gtt cca cta att att ata cca gga atc ggt gaa act att ttt ttt 240
Asn Val Pro Leu Ile Ile Ile Pro Gly Ile Gly Glu Thr Ile Phe Phe
65 70 75 80
act tat aca atc att aaa agt cat tac tat aat tat cat gaa gcg cta 288
Thr Tyr Thr Ile Ile Lys Ser His Tyr Tyr Asn Tyr His Glu Ala Leu
85 90 95
gct att gtt ttg att tca ggt ttg att ttc act ttt att gca tac aca 336
Ala Ile Val Leu Ile Ser Gly Leu Ile Phe Thr Phe Ile Ala Tyr Thr
100 105 110
ccg ttt gct aga gtt cta gac aag tcc ata cca aag aat tta aaa gaa 384
Pro Phe Ala Arg Val Leu Asp Lys Ser Ile Pro Lys Asn Leu Lys Glu
115 120 125
gga ata act att ggt ata ggt ctg ttt atg gcg ttt gtt gga cta caa 432
Gly Ile Thr Ile Gly Ile Gly Leu Phe Met Ala Phe Val Gly Leu Gln
130 135 140
aac agc aaa ata att ata cca aac agg caa agt att gtt gag cta aac 480
Asn Ser Lys Ile Ile Ile Pro Asn Arg Gln Ser Ile Val Glu Leu Asn
145 150 155 160
cac ata aac att tat agt ggg tta gcg ata cta cta cta tta ttt gca 528
His Ile Asn Ile Tyr Ser Gly Leu Ala Ile Leu Leu Leu Leu Phe Ala
165 170 175
att gtt ata ttt act tta ggg acc aag ttg gct ttc ttt tat aca gta 576
Ile Val Ile Phe Thr Leu Gly Thr Lys Leu Ala Phe Phe Tyr Thr Val
180 185 190
att att ggt atc att ata tct ttt tta gct ggg att ata aag gtg aaa 624
Ile Ile Gly Ile Ile Ile Ser Phe Leu Ala Gly Ile Ile Lys Val Lys
195 200 205
tat cat ttt tat aat ttt agt ttg cga tca ata gta agc gag aat aat 672
Tyr His Phe Tyr Asn Phe Ser Leu Arg Ser Ile Val Ser Glu Asn Asn
210 215 220
att ttt agt tac agt ttt gat aaa ata ggt cat ttt tct ttt tgg tct 720
Ile Phe Ser Tyr Ser Phe Asp Lys Ile Gly His Phe Ser Phe Trp Ser
225 230 235 240
tta gtg ttc tca ctt act att ttg tta ctg ttt caa aat tta ggt aca 768
Leu Val Phe Ser Leu Thr Ile Leu Leu Leu Phe Gln Asn Leu Gly Thr
245 250 255
tta cat gga ttg aaa att aat gat aaa gta aaa ttg tca aga att ttt 816
Leu His Gly Leu Lys Ile Asn Asp Lys Val Lys Leu Ser Arg Ile Phe
260 265 270
aaa atg gtc ggt att act aac ata att tca agc tta ttt ggt gtg agt 864
Lys Met Val Gly Ile Thr Asn Ile Ile Ser Ser Leu Phe Gly Val Ser
275 280 285
tct aca gtt att gca gtc gaa agt tct act gca act cat tca gga gct 912
Ser Thr Val Ile Ala Val Glu Ser Ser Thr Ala Thr His Ser Gly Ala
290 295 300
aaa aca gga aaa gta tct att ttt gta ggt ata atg ttt ctt tta tct 960
Lys Thr Gly Lys Val Ser Ile Phe Val Gly Ile Met Phe Leu Leu Ser
305 310 315 320
ttg ttg ata atg ccc gtt att ata gca ata cct agt tta gtt gta tca 1008
Leu Leu Ile Met Pro Val Ile Ile Ala Ile Pro Ser Leu Val Val Ser
325 330 335
cct atc tta ata att gtt ggc ggt tta atg ttt act aat att aaa gaa 1056
Pro Ile Leu Ile Ile Val Gly Gly Leu Met Phe Thr Asn Ile Lys Glu
340 345 350
tta gat ttt aat gat atg act gaa ttt att cct tgt tat ata aca att 1104
Leu Asp Phe Asn Asp Met Thr Glu Phe Ile Pro Cys Tyr Ile Thr Ile
355 360 365
ata atg ata cca ctt act ttt gat att gca act gga atg gga ttt gga 1152
Ile Met Ile Pro Leu Thr Phe Asp Ile Ala Thr Gly Met Gly Phe Gly
370 375 380
ttt att tca tat gtt cta att aat ttt gta tgc aaa aaa acc gaa cgt 1200
Phe Ile Ser Tyr Val Leu Ile Asn Phe Val Cys Lys Lys Thr Glu Arg
385 390 395 400
tta aat cca att tta ata att att gct tta ctt ttt aca ata aat tta 1248
Leu Asn Pro Ile Leu Ile Ile Ile Ala Leu Leu Phe Thr Ile Asn Leu
405 410 415
gtt tta caa taa 1260
Val Leu Gln
<210> 54
<211> 419
<212> PRT
<213> Staphylococcus epidermidis
<400> 54
Met Ile Gly Arg Lys Lys Glu Thr Leu Leu Lys Asn Glu Val Ile Ser
1 5 10 15
Ala Phe Thr Thr Phe Phe Thr Cys Ser Tyr Ile Ile Ile Val Asn Gly
20 25 30
Ile Leu Leu His Gln Ala Gly Met Ser Leu Leu Trp Thr Ile Ile Ala
35 40 45
Thr Thr Leu Val Cys Cys Ile Ser Cys Ile Leu Leu Gly Ile Tyr Ala
50 55 60
Asn Val Pro Leu Ile Ile Ile Pro Gly Ile Gly Glu Thr Ile Phe Phe
65 70 75 80
Thr Tyr Thr Ile Ile Lys Ser His Tyr Tyr Asn Tyr His Glu Ala Leu
85 90 95
Ala Ile Val Leu Ile Ser Gly Leu Ile Phe Thr Phe Ile Ala Tyr Thr
100 105 110
Pro Phe Ala Arg Val Leu Asp Lys Ser Ile Pro Lys Asn Leu Lys Glu
115 120 125
Gly Ile Thr Ile Gly Ile Gly Leu Phe Met Ala Phe Val Gly Leu Gln
130 135 140
Asn Ser Lys Ile Ile Ile Pro Asn Arg Gln Ser Ile Val Glu Leu Asn
145 150 155 160
His Ile Asn Ile Tyr Ser Gly Leu Ala Ile Leu Leu Leu Leu Phe Ala
165 170 175
Ile Val Ile Phe Thr Leu Gly Thr Lys Leu Ala Phe Phe Tyr Thr Val
180 185 190
Ile Ile Gly Ile Ile Ile Ser Phe Leu Ala Gly Ile Ile Lys Val Lys
195 200 205
Tyr His Phe Tyr Asn Phe Ser Leu Arg Ser Ile Val Ser Glu Asn Asn
210 215 220
Ile Phe Ser Tyr Ser Phe Asp Lys Ile Gly His Phe Ser Phe Trp Ser
225 230 235 240
Leu Val Phe Ser Leu Thr Ile Leu Leu Leu Phe Gln Asn Leu Gly Thr
245 250 255
Leu His Gly Leu Lys Ile Asn Asp Lys Val Lys Leu Ser Arg Ile Phe
260 265 270
Lys Met Val Gly Ile Thr Asn Ile Ile Ser Ser Leu Phe Gly Val Ser
275 280 285
Ser Thr Val Ile Ala Val Glu Ser Ser Thr Ala Thr His Ser Gly Ala
290 295 300
Lys Thr Gly Lys Val Ser Ile Phe Val Gly Ile Met Phe Leu Leu Ser
305 310 315 320
Leu Leu Ile Met Pro Val Ile Ile Ala Ile Pro Ser Leu Val Val Ser
325 330 335
Pro Ile Leu Ile Ile Val Gly Gly Leu Met Phe Thr Asn Ile Lys Glu
340 345 350
Leu Asp Phe Asn Asp Met Thr Glu Phe Ile Pro Cys Tyr Ile Thr Ile
355 360 365
Ile Met Ile Pro Leu Thr Phe Asp Ile Ala Thr Gly Met Gly Phe Gly
370 375 380
Phe Ile Ser Tyr Val Leu Ile Asn Phe Val Cys Lys Lys Thr Glu Arg
385 390 395 400
Leu Asn Pro Ile Leu Ile Ile Ile Ala Leu Leu Phe Thr Ile Asn Leu
405 410 415
Val Leu Gln
<210> 55
<211> 10
<212> PRT
<213> Staphylococcus epidermidis
<220>
<221> MISC_FEATURE
<222> (10)..(10)
<223> Xaa at 10 is V or L
<400> 55
Lys Asn Gly Ala Tyr Lys Ala Gln Gly Xaa
1 5 10
<210> 56
<211> 30
<212> PRT
<213> Staphylococcus hominis
<400> 56
Lys Cys Ser Trp Trp Asn Ala Ser Cys His Leu Gly Asn Asn Gly Lys
1 5 10 15
Ile Cys Thr Val Ser His Glu Cys Ala Ala Gly Cys Asn Leu
20 25 30
<210> 57
<211> 38
<212> PRT
<213> Staphylococcus hominis
<400> 57
Ala Thr Pro Thr Ile Thr Thr Ser Ser Ala Thr Cys Gly Gly Ile Ile
1 5 10 15
Val Ala Ala Ser Ala Ala Gln Cys Pro Thr Leu Ala Cys Ser Ser Arg
20 25 30
Cys Gly Lys Arg Lys Lys
35
本開示は下記の実施形態を包含する。
[1] 1種以上のプロバイオティック細菌株の局所濃厚製剤、並びに場合によりプレバイオティック化合物、保護剤、湿潤剤、皮膚軟化剤、研磨剤、塩及び/又は界面活性剤を含む組成物であって、
前記1種以上のプロバイオティック細菌株は、スタフィロコッカス属の1種以上の細菌株を含み;及び
前記組成物は、皮膚、頭皮、又は粘膜のディスバイオシスの障害の局所治療用に製剤化されている、組成物。
[2] 前記1種以上のプロバイオティック細菌株が、表皮ブドウ球菌(Staphylococcus epidermidis)、スタフィロコッカス・ホミニス(Staphylococcus hominis)、又は表皮ブドウ球菌とスタフィロコッカス・ホミニスの組み合わせを含む、実施形態1に記載の組成物。
[3] 前記1種以上のプロバイオティック細菌株が、表皮ブドウ球菌株MO34、MO38、A11、AMT1、AMT5-C5及び/又はAMT5-G6を含む、実施形態2に記載の組成物。
[4] 前記1種以上のプロバイオティック細菌株が、スタフィロコッカス・ホミニス菌株A9、C2、AMT2、AMT3、AMT4-C2、AMT4-G1及び/又はAMT4-D12を含む、実施形態2に記載の組成物。
[5] 各プロバイオティック細菌株が、図11、図12、図13、図14、図15、図16、図17、図18、又は図19のいずれかに示されるものの1つに対応する脂肪酸メチルエステルプロファイルを示す、実施形態2に記載の組成物。
[6] 前記1種以上のプロバイオティック細菌株が、配列番号2、4、28、30、32、34、36、38、40、42、44、46、48、50、52、54及び55、並びにそれらの任意の組み合わせからなる群から選択される配列を有するペプチドを産生し、このようなペプチドは、場合により翻訳後修飾されている、実施形態2に記載の組成物。
[7] 前記1種以上のプロバイオティック細菌株が生存形態で提供される、実施形態1に記載の組成物。
[8] 前記1種以上のプロバイオティック細菌株が、凍結乾燥形態又は噴霧乾燥形態で提供される、実施形態1に記載の組成物。
[9] 前記プロバイオティック細菌を生存形態に再構成することができる、実施形態8に記載の組成物。
[10] 有効量の実施形態1~9のいずれかに記載の組成物をそれを必要とする対象に対し皮膚又は粘膜に適用することによって、ヒト若しくは他の哺乳動物における皮膚又は粘膜感染、アトピー性皮膚炎、乾癬、乳腺炎、座瘡、又は皮膚ディスバイオシスに関連する他の障害を治療する方法。
[11] 前記組成物が局所的に適用される、実施形態10に記載の方法。
[12] 前記組成物が、クリーム、軟膏、噴霧剤、散剤、油剤、濃厚製剤又は湿布剤として製剤化されている、実施形態10に記載の方法。
[13] ホゴシジンペプチド、誘導体若しくはバリアント、SH-ランチビオティックペプチド、SH-抗菌剤、SE-ランチビオティックペプチド、及び/又はSE抗菌剤のうちの1つ以上を含み;1つ以上の増粘剤、溶媒、乳化剤、又は薬学的に許容される担体若しくは賦形剤をさらに含む組成物。
[14] カテリシジンペプチド、誘導体又はバリアントをさらに含む、実施形態13に記載の組成物。
[15] 前記ペプチドが、1つ以上のD-アミノ酸又は非天然アミノ酸を含む、実施形態13又は14に記載の組成物。
[16] 前記ホゴシジンペプチド、SH-ランチビオティックペプチド、SH-抗菌剤、SE-ランチビオティックペプチド、又はSE抗菌剤が、インサイチュで、スタフィロコッカス・ホミニス菌株A9、スタフィロコッカス・ホミニス菌株C2、スタフィロコッカス・ホミニス菌株AMT2、スタフィロコッカス・ホミニス菌株AMT3、スタフィロコッカス・ホミニス菌株AMT4-C2、スタフィロコッカス・ホミニス菌株AMT4-G1、スタフィロコッカス・ホミニス菌株AMT4-D12、表皮ブドウ球菌株AMT1、表皮ブドウ球菌株SE-A11、表皮ブドウ球菌株AMT5-C5、表皮ブドウ球菌株AMT5-G6及び表皮ブドウ球菌株MO34のうちの1つ以上によって産生される、実施形態13に記載の組成物。
[17] 前記ペプチドが局所投与用に製剤化されている、実施形態13、14又は15に記載の組成物。
[18] 前記製剤が、ローション、軟膏クリーム、粉末、軟膏、油剤又は噴霧剤を含む、実施形態17に記載の組成物。
[19] 前記ホゴシジンペプチド、誘導体又はバリアントが、配列番号2又は配列番号4から選択される配列を含む、実施形態13に記載の組成物。
[20] ホゴシジンペプチド、誘導体又はバリアント、SH-ランチビオティックペプチド、SH-抗菌剤、SE-ランチビオティックペプチド、SE抗菌剤、及びカテリシジンペプチド、誘導体又はバリアントのうちの1つ以上が、スタフィロコッカス・ホミニス菌株A9、スタフィロコッカス・ホミニス菌株C2、スタフィロコッカス・ホミニス菌株AMT2、スタフィロコッカス・ホミニス菌株AMT3、スタフィロコッカス・ホミニス菌株AMT4-C2、スタフィロコッカス・ホミニス菌株AMT4-G1、スタフィロコッカス・ホミニス菌株AMT4-D12、表皮ブドウ球菌株AMT1、表皮ブドウ球菌株SE-A11、表皮ブドウ球菌株AMT5-C5、表皮ブドウ球菌株AMT5-G6及び表皮ブドウ球菌株MO34の抽出物又は溶解物として提供される、実施形態13に記載の組成物。
[21] 対象をホゴシジンペプチド、誘導体又はバリアント、SH-ランチビオティックペプチド、SH-抗菌剤、SE-ランチビオティックペプチド、及び場合によりカテリシジンペプチド、誘導体又はバリアントのうちの1つ以上を含む有効量の組成物と接触させることを含む、対象における皮膚若しくは粘膜感染又はアトピー性皮膚炎を治療するための方法。
[22] 前記接触が、局所投与による、又は場合により、前記対象をSH-ランチビオティック又はバクテリオシン産生スタフィロコッカス・ホミニス菌株A9、C2、AMT2、AMT3、AMT4-C2、AMT4-G1、AMT4-D12及び表皮ブドウ球菌株AMT5-G6及びMO34のうち1つ以上と接触させることによるものである、実施形態21に記載の方法。
[23] 配列番号2若しくは4と少なくとも95%同一であるポリペプチド、又は抗菌活性を有するその生物学的に活性な断片をコードするポリヌクレオチドを含む組換えベクター。
[24] 前記ベクターが、配列番号2又は4のポリペプチドをコードするポリヌクレオチドを含む、実施形態23に記載の組換えベクター。
[25] 前記ベクターが、配列番号2のアミノ酸約32~アミノ酸約61のポリペプチドをコードするポリヌクレオチドを含む、実施形態23に記載の組換えベクター。
[26] 前記ベクターが、配列番号4のアミノ酸約29~アミノ酸約66のポリペプチドをコードするポリヌクレオチドを含む、実施形態23に記載の組換えベクター。
[27] 前記ベクターが、配列番号1又は3と少なくとも95%同一であり、それぞれ配列番号2又は4のポリペプチドをコードするポリヌクレオチドを含む、実施形態23に記載の組換えベクター。
[28] 前記ベクターが配列番号1又は3の断片を含む、実施形態25又は26に記載の組換えベクター。
[29] 実施形態23~28のいずれかに記載の組換えベクターを発現するように遺伝子操作された宿主細胞。
[30] 前記宿主細胞が、非病原性弱毒化宿主細胞である、実施形態29に記載の宿主細胞。
[31] 実施形態29に記載の宿主細胞によって産生された組換えポリペプチド。
[32] 実施形態29に記載の宿主細胞を含む組成物。
[33] 実施形態30に記載の宿主細胞を含む組成物。
[34] 実施形態31に記載のポリペプチドを含む組成物。
Claims (34)
前記1種以上のプロバイオティック細菌株は、スタフィロコッカス属の1種以上の細菌株を含み;及び
前記組成物は、皮膚、頭皮、又は粘膜のディスバイオシスの障害の局所治療用に製剤化されている、組成物。
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CN113730648B (zh) * | 2021-09-06 | 2022-11-01 | 温州瑞司特生物科技有限公司 | 结合表皮葡萄球菌的水凝胶及其在治疗创面中的应用 |
CZ2021461A3 (cs) * | 2021-09-30 | 2023-02-15 | BiomServ s.r.o. | Přípravky na ochranu mikrobiomu kůže a sliznic proti patogenním mikrobům a virům |
CA3237750A1 (en) * | 2021-11-11 | 2023-05-19 | Concerto Biosciences, Inc. | Microbial compositions for the treatment of skin diseases |
WO2023119287A1 (en) * | 2021-12-22 | 2023-06-29 | The State Of Israel, Ministry Of Agriculture & Rural Development, Agricultural Research Organization (Aro) (Volcani Institute) | Nomadic bacteria and uses thereof |
WO2023223993A1 (ja) * | 2022-05-16 | 2023-11-23 | マルホ株式会社 | 白癬治療用組成物 |
CN117343856A (zh) * | 2022-06-28 | 2024-01-05 | 上海菌济健康科技有限公司 | 具有促进毛发生长的人体共生菌及其应用 |
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US5925556A (en) * | 1996-10-02 | 1999-07-20 | Agency Of Industrial Science And Technology | Method of degrading polylactic acid resin using staphylococcus hominis and staphylococcus epidermidis |
WO2007114510A1 (en) * | 2006-03-31 | 2007-10-11 | Canon Kabushiki Kaisha | Probe, probe set, probe-immobilized carrier, and genetic testing method |
US8148352B2 (en) * | 2006-05-12 | 2012-04-03 | The Regents Of The University Of California | Antimicrobial therapy for bacterial infections |
US20080026999A1 (en) * | 2006-07-28 | 2008-01-31 | Van Der Donk Wilfred A | Two-component bacillus lantibiotic and methods for producing and using the same |
WO2008103751A2 (en) * | 2007-02-20 | 2008-08-28 | The Regents Of The University Of California | Antimicrobial and anti-inflammatory therapies and compositions |
US20090041727A1 (en) * | 2007-08-08 | 2009-02-12 | Conjugon, Inc. | Compositions and Methods for Microbe Storage and Delivery |
US9073990B2 (en) * | 2010-04-05 | 2015-07-07 | Bar-Llan University | Protease-activatable pore-forming polypeptides |
WO2012112548A2 (en) * | 2011-02-15 | 2012-08-23 | The Regents Of The University Of California | Firmocidin, an antimicrobial molecule produced by staphylococcus epidermidis |
DK3035802T3 (da) * | 2013-08-19 | 2021-10-11 | Syngulon Sa | Kontrolleret vækst af mikroorganismer |
AU2016256882B2 (en) * | 2015-05-05 | 2022-06-02 | The Regents Of The University Of California | Antimicrobial therapy |
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BR112017023707A2 (pt) | 2018-07-31 |
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WO2016179440A3 (en) | 2017-02-09 |
KR102624665B1 (ko) | 2024-01-12 |
JP2022028833A (ja) | 2022-02-16 |
BR112017023707B1 (pt) | 2022-03-29 |
WO2016179440A2 (en) | 2016-11-10 |
HK1250639A1 (zh) | 2019-01-11 |
US20210228652A1 (en) | 2021-07-29 |
CN107735098A (zh) | 2018-02-23 |
EP3291824A4 (en) | 2019-01-23 |
US20180289751A1 (en) | 2018-10-11 |
AU2016256882A1 (en) | 2017-11-23 |
CN107735098B (zh) | 2022-07-29 |
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