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TWI402075B - 長雙歧桿菌ｎｃｃ２７０５（ｃｎｃｍ ｉ－２６１８）及免疫病症 - Google Patents

長雙歧桿菌ｎｃｃ２７０５（ｃｎｃｍ ｉ－２６１８）及免疫病症 Download PDF

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Publication number
TWI402075B
TWI402075B TW099115195A TW99115195A TWI402075B TW I402075 B TWI402075 B TW I402075B TW 099115195 A TW099115195 A TW 099115195A TW 99115195 A TW99115195 A TW 99115195A TW I402075 B TWI402075 B TW I402075B
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Taiwan
Prior art keywords
bifidobacterium longum
composition
replicating
longum ncc
cncm
Prior art date
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TW099115195A
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English (en)
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TW201043238A (en
Inventor
Valerie Petit
Clara Lucia Garcia-Rodenas
Monique Julita
Guenolee Prioult
Annick Mercenier
Sophie Helene Nutten
Original Assignee
Nestec Sa
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Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=42646295&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=TWI402075(B) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority claimed from EP09159925A external-priority patent/EP2251020A1/en
Priority claimed from EP09159929A external-priority patent/EP2251022A1/en
Application filed by Nestec Sa filed Critical Nestec Sa
Publication of TW201043238A publication Critical patent/TW201043238A/zh
Application granted granted Critical
Publication of TWI402075B publication Critical patent/TWI402075B/zh

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Description

長雙歧桿菌NCC2705(CNCM I-2618)及免疫病症
本發明大體上係關於預防及/或治療發炎性及感染性病症之領域，特定言之其係藉由加強內源性抗菌防禦。本發明之一項實施例為一種以長雙歧桿菌NCC2705(B. longum NCC2705)(寄存編號CNCM I-2618)於治療或預防與免疫系統相關之病症(包括感染)上之用途。
吾人之環境已被許多具潛在致病性之微生物污染。皮膚角質細胞，及在胃腸道、呼吸道、泌尿生殖道上之上皮細胞均提供物理性障壁，以抵抗微生物入侵體內。
此外，該等上皮組織有助於宿主防禦，其係藉由產生並分泌抗菌物質而限制細菌及其他微生物進入。該等抗菌性分子組成固有免疫基礎防禦線之關鍵組分。
防禦素為人類之其中一種最重要的抗菌性肽。防禦素係由肺、皮膚、口腔、泌尿生殖道、呼吸道及胃腸道之上皮細胞產生。其中，存在一種β-防禦素家族，包括防禦素1(hBD1)及2(hBD2)。
於多種黏膜表面上表現hBD1，諸如口腔黏膜、唾液腺、胃部、小腸、結腸、肝臟及胰臟。於包括胃腸道之多種黏膜表面之上皮細胞中亦存在hBD2。此外，該兩種防禦素亦存在於唾液及呼吸道表面體液中(Cunliffe,R.N.及Mahida,Y.R. 2004,J Leukoc. Biol. 75:49-58)。
在正常組織中，所存在之hBD2量極低，且其表現係經細菌及促發炎性細胞激素向上調控。與hBD2相反，hBD1為組成性表現。從未顯示hBD1可由細菌或發炎組成性向上調控(Ou,G.,等人，2009,Scand. J Immunol 69:150-161)。
已熟知益生菌能夠加強多種腸道防禦線：免疫排除、免疫消除、及免疫調節。亦已知益生菌可刺激針對微生物病原體之非特異性宿主抵抗，且藉此有助於消除微生物病原體。
然而，儘管如此，已報導hBD1之組成性表現不受益生菌之影響(O'Neil,D.A.等人，J Immunol 163:6718-6724)，且受共生菌株(大腸桿菌(Escherichia coli))及病原性菌株(鼠傷寒沙門氏桿菌(Salmonella typhimurium))極溫和之向上調節影響(Ou,G.,等人，2009,Scand. J Immunol 69:150-161)。
當前，益生菌係應用於降低罹患與腸道障壁功能障礙相關之病症的風險(E. Isolauri等人，2002,Gut 2002; 50:iii 54-iii 59)。認為益生菌係透過在腸道中存活、具酸及膽汁安定性、且暫時定殖於腸道黏膜表面而發揮作用。
因此，大部份已報導之文獻係關於活益生菌。然而，一些研究已探討由非複製性細菌所傳遞之健康益處，且其中大部份指出，例如藉由熱處理使益生菌失活時，導致其喪失原本之健康益處(Rachmilewitz,D.,等人，2004,Gastroenterology 126:520-528；Castagliuolo等人，2005,FEMS Immunol. Med. Microbiol. 43:197-204; Gill,H. S. and K. J. Rutherfurd,2001,Br. J. Nutr. 86:285-289;Kaila,M.等人，1995,Arch. Dis. Child 72:51-53)。
然而，當前對食品中之存活細菌的處理存在一些缺點。存活細菌之抗壓性通常不強，且因此難以採用工業規模處理且同時保持存活率。更進一步，基於安全性考慮，一些產品種類可能不適於添加活的微生物至調配物中。
另一方面，含非複製性益生菌之食物可以讓例如粉末狀營養組合物進行熱復水，且同時保持對食用之患者之健康益處。
鑒於此，希望改用非複製性細菌替代其活的對應物進行操作，但關於該態樣之可獲得之研究並不樂觀。
文獻中已報導使用活益生菌作為治療或預防發炎性腸病的策略，且最近已概述於Dotan等人之文獻中(Dotan,I.及D. Rachmilewitz. 2005;Curr. Opin. Gastroenterol. 21:426-430)。例如，已顯示，八種活益生菌之高濃度混合物(VSL#3)可有效預防(Gionchetti,P.等人，2003,Gastroenterology 124:1202-1209)並治療人類之復發性或頑抗性結腸袋炎(Gionchetti,P.等人，2000,Gastroenterology 119:305-309；Mimura,T.等人，2004,Gut 53:108-114)。有趣的是，Rachmilewitz等人之文獻(Rachmilewitz,D.等人，2004,Gastroenterology 126:520-528)利用DSS誘發結腸炎之鼠類模型指出，以存活之VSL#3及經γ輻射之VSL#3處理，可保護防止罹患結腸病，而經熱滅活之VSL#3則不能。類似地，經熱滅活之鬈曲乳桿菌(L. crispatus)不能保護對抗DSS誘發之結腸病，而其存活之對應物可明顯減少體重減少量及腸道中MPO活性之下降程度(Castagliuolo等人，2005,FEMS Immunol. Med. Microbiol. 43:197-204)。該等研究說明，對於腸道發炎，活的益生菌比其非複製性對應物更有效。
已發現失活的羅伊氏乳桿菌(L. reuteri)(經熱滅活及經γ輻射)不能使由T84細胞產生的經TNFα-誘導之IL-8減少，而其活的對應物顯示出顯著之有益功效(Ma,D.等人，2004,Infect. Immun. 72:5308-5314)。
因此，相關技術中需要一種天然組合物，其在工業條件下易於處理，對投藥者安全且簡便，且特定言之藉由增強內源性抗菌性防禦而允許預防及/或治療發炎性及感染性病症。
天然組合物最佳應自益生菌培養物製得，特定言之自當今廣泛接受且由食用者認為可遞送健康益處之益生微生物製得。該組合物宜應含有非複製性細菌，且應比其活的對應物更有效。
本發明已滿足該種需求。
因此，本發明之標的為改善當前技術，並提供一種天然組合物，該種天然組合物特定言之係藉由加強內源性抗菌性防禦而可以預防及/或治療發炎性及感染性病症，並滿足上述需求。
本發明人驚奇發現，可藉由獨立專利申請範圍中之標的達成本發明之目的。附屬專利申請範圍進一步定義本發明之較佳實施例。
本發明之標的可藉由投與一種含有諸如非複製性微生物之微生物(例如經熱處理之微生物)而加強哺乳動物之內源性抗菌防禦。
本發明人闡述，長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)具有比彼等先前於文獻中定義及闡述者更佳之誘發抗菌性肽表現的功效。
已發現，例如：
-　長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)可比其他微生物及8種細菌之組合更強力誘發hBD2表現(Mondel,M. 等人，2009,Mucosal. Immunol 2:166-172；Schlee,M.等人，2007,Infect. Immun. 75:2399-2407)，且
-　經熱處理之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)可比其活的對應物更強力向上調節hBD2，且另外還向上調節hBD1。
hBD1及hBD2顯示出抗細菌活性，其可抵抗廣譜之細菌，包括大腸桿菌(E. coli)及綠膿桿菌(Pseudomonas aeruginosa)、幽門螺旋桿菌(H. pylori)(Nuding,S.等人，2009,Microbes. Infect. 11:384-393)；其亦對抗酵母菌，諸如白色念珠菌(Candida albicans)(O'Neil,D.A. 2003,Mol. Immunol 40:445-450)；及病毒(人類免疫缺陷病毒(human immunodeficiency virus))(Kota,S.等人，2008,J. Biol. Chem 283:22417-22429)。因此，該等抗菌性肽可加強黏膜障壁，且因此限制細菌之黏附及入侵。
有越來越多證據顯示，於某些病理生理條件下之防禦素含量會減少，且此係病理學上及諸如以下感染性及發炎性病症的併發症之危險因子(Doss,M.等人，2010,J Leukoc. Biol 87:79-92；Rivas-Santiago,B.等人，2009,Infect. Immun. 77:4690-4695)：
-　於呼吸道中：囊腫性纖維化、反應性呼吸道疾病、吸菸引起之肺部感染、哮喘、肺炎、鼻炎、耳炎、竇炎、結核病
-　於胃腸道中：克隆氏病(Crohn's disease)(結腸及回腸)、潰瘍性結腸炎、由幽門螺旋桿菌感染所誘發之胃炎及胃潰瘍、感染性腹瀉、壞死性腸炎、與抗生素相關之腹瀉、腸不成熟症。
-　於泌尿生殖道中：細菌性陰道炎、HIV、單純疱疹病毒、尿道感染
-　於皮膚中：異位性皮膚炎、慢性潰瘍、癌瘤、異位性濕疹、燒傷
-　於口腔中：HIV患者、扁桃腺炎、齒齦炎、齲齒
-　於眼中之角膜炎。
文中所示結果說明，長雙歧桿菌NCC 2705(CNCM I-2618)加強內源性抗菌性防禦的能力比過去鑑定之益生菌更強，且因此可更有效預防及治療SIBO(小腸細菌過度生長症)、發炎性及感染性病症。此外，本發明者之數據顯示，與文獻中所述相反，熱處理非但不降低，而且進一步增加長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之強力抗菌功效。
因此，本發明之一項實施例為一種包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物，其係用於治療或預防與免疫系統相關之病症(包括感染)。
根據本發明，可藉由增加內源性hBD1及/或hBD2表現而治療或預防與免疫系統相關之病症。
本發明亦關於一種包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物，其係用於治療或預防與hBD1表現減少相關之病症(諸如細菌感染)。
本發明亦關於一種以長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)於製備組合物上之用途，該組合物係用於治療或預防與免疫系統相關之病症。
可使用至少部份呈非複製性之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。非複製性(特定言之，經熱處理之)長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)所具有之優勢為比其活的對應物更有效。
使用非複製性微生物(諸如經熱處理之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618))，而並非其活的對應物，具有如下之其他優勢：
-　在敏感之目標人群中，降低可能罹患與活益生菌相關之敗血症的風險，
-　代表對免疫受損之患者更安全之替代物，及
-　加工限制減少，可併入存放期長之具存放穩定性之液態產品中。
因此，於本發明之一項實施例中，至少90%，例如至少95%，較佳至少98%，最佳至少99%，最理想至少99.9%、或全部之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性。
本發明亦關於一種包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物，其中至少95%，較佳至少98%，最佳至少99%，最理想至少99.9%或100%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性。
因此，本發明亦關於一種具生物活性之非複製性(例如經熱處理)長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。
「非複製性」長雙歧桿菌NCC 2705包括經熱處理之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。其包括非活性、死亡、非存活及/或呈諸如DNA片段、代謝物、細胞質化合物及/或細胞壁材料存在的長雙歧桿菌NCC 2705。
「非複製性」意指，由傳統之平板培養方法測得沒有存活的細胞及/或菌落形成單位存在。該等傳統之平板培養方法概述於微生物學書籍中：James Monroe Jay、Martin J. Loessner、David A. Golden. 2005. Modern food microbiology.第7版，Springer Science,New York,N.Y.第790頁。通常可由以下現象顯示沒有存活的細胞存在：在接種不同濃度之細菌製劑(「非複製性」樣本)，並於適宜條件(有氧及/或無氧氛圍中，至少24 h)下培養之後，於瓊脂平板上沒有可見之菌落，或液態生長培養基中之渾濁度不會增加。
可藉由熱滅活使長雙歧桿菌NCC 2705呈非複製性。可於至少約70℃下進行熱滅活。
可使用任一種熱處理使益生菌失活，只要其進行時間足以使其失活。例如，該種熱處理可進行至少10秒鐘。
通常，高溫所需之加熱時間短，而較低溫度所需之加熱時間長。
例如，可於110℃至140℃下熱處理1至30秒鐘，例如10至20秒鐘或於75℃至95℃下熱處理10至30分鐘，使長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
所示之時間範圍係指長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)處於所示溫度下之時間。注意：取決於提供長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物之性質及數量，且取決於所使用加熱裝置的構造，加熱時間可能不同。
可於常壓下進行溫度處理，但亦可於高壓下進行。通常之壓力為1至50 bar，較佳自1至10 bar，更佳自2至5 bar。所施用之理想壓力取決於添加微生物之組合物的性質，且取決於所採用之溫度。
已添加長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物於例如封裝及配送之前，若依任何方式熱處理，則較佳為採用這種熱處理步驟，使長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)失活。
處理含有長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物的方法通常為高溫短時(HTST)處理、瞬間巴斯德滅菌處理或超高溫(UHT)處理。
UHT處理為超高溫處理或超熱處理(二者之縮寫皆為UHT)，及包括藉由於超過135℃(275℉)之溫度下(用於殺死牛奶中細菌孢子時所需之溫度)，短時(約1至10秒鐘)處理組合物，使至少部份組合物無菌。例如，採用135℃以上之溫度，依該方式處理牛奶一段必要且能夠進行連續流動操作之保持時間(2至5秒)，使得細菌含量減少。
有兩種主要類型之UHT系統：直接及間接系統。於直接系統中，藉由注射蒸汽或輸注蒸汽處理產品；而於間接系統中，係利用平板式熱交換器、管式熱交換器或刮板式熱交換器熱處理產品。可於製備產品過程中的任一步驟或多個步驟中應用UHT系統之組合。
HTST處理之定義如下(高溫/短時)：巴斯德滅菌法之設計在於可減少5-log，且可殺死牛奶中存活微生物數量之99,9999%。認為其足以殺死幾乎所有酵母菌、黴菌及常見腐敗細菌，且亦確保足以殺死常見致病性耐熱微生物。於HTST方法中，將牛奶加熱至71.7℃(161℉)，並保持15至20秒鐘。
瞬間巴斯德滅菌法為一種使諸如果汁及蔬菜汁、啤酒及奶產品之飲料進行加熱巴斯德滅菌的方法。其係於充填至容器之前進行，以殺死腐敗性微生物，使得產品更安全並延長存放期。使液體在控制下連續流動，同時施加71.5℃(160℉)至74℃(165℉)之溫度約15至30秒鐘。
為本發明之目的，術語「短時高溫處理」包括例如高溫短時(HTST)處理、UHT處理、及低溫長時之瞬間巴斯德滅菌處理。
本發明組合物可包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)，其數量足以至少部份治療感染、及與免疫系統相關之病症、及/或其併發症。足以達成該目的的數量定義為「治療有效劑量」。有效達成該目的之數量取決於彼等熟習此項技術者所知之數種因素，諸如病症之嚴重程度、及食用者之體重及一般健康狀態，且取決於食物成份之功效。
於預防性應用中，向易於罹患或處於罹患與免疫系統相關之病症之風險中之食用者投與數量足以至少部份降低發展出該等病症之風險的根據本發明之組合物。該種數量定義為「預防有效劑量」。同樣地，確切數量取決於數種患者特異性因素，諸如患者之健康狀態及體重，並取決於食物成份之功效。
彼等熟習此項技術者能夠適宜地調整治療有效劑量及/或預防有效劑量。
本發明組合物通常含有治療有效劑量及/或預防有效劑量之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。
治療有效劑量及/或預防有效劑量之範圍通常為每份日劑量含約0.005 mg至1000 mg Lal。
就數量而言，於組合物中長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)所占之數量對應於每公克乾組合物含104 至1012 當量cfu。顯然，非複製性微生物不形成菌落，因此，該術語應理解為獲自104 至1012 cfu/g複製性細菌之非複製性微生物數量。其包括失活、非存活或死亡之微生物，或呈諸如DNA之片段或細胞壁或細胞質化合物存在。換言之，組合物所含之微生物數量係以該數量之微生物的菌落形成能力(cfu)表示，就如同所有微生物均存活，而不管其事實上是否呈非複製性(諸如失活或死亡、呈片段或該等狀態中任一種或全部之混合物)。
例如，根據本發明組合物之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)含量相當於每份日劑量含約104 至1012 cfu。
本發明組合物每份日劑量可含有約0.005 mg至1000 mg之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。
本發明組合物可為任一種組合物。該組合物之投與方式可為例如經口、經腸、非經腸(經皮下或經肌內)、經外部或經眼、藉由吸入、經直腸內及經陰道內。
因此，本發明組合物可選自以下組成之群中：食物組合物、食品(包括寵物食物、飲料、用於完全營養之調配物、營養補充物、營養藥劑、食品添加劑、醫藥組合物、化妝品組合物、外用組合物及醫藥物)。
可添加益菌助生質。益菌助生質可支持益生菌在呈非複製性之前生長。益菌助生質亦可與存在於組合物中及/或可能添加之活益生菌協同作用。
與免疫系統相關之病症可選自以下組成之群中：感染，特定言之細菌、病毒、真菌及/或寄生蟲感染；發炎；吞噬細胞功能不足症；上皮障壁功能障礙或免疫系統不成熟症、SIBO及其組合。
於本發明之一項實施例中，包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物可用於治療或預防微生物感染，諸如病毒、真菌及/或寄生蟲感染。
與免疫系統相關之病症亦可選自與防禦素含量減少相關之病症所組成之群中，特定言之hBD2。
此外，若使用非複製性長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)，則與免疫系統相關之病症亦可選自與hBD1含量減少有關之病症所組成之群中。
該等病症可選自以下組成之群中：囊腫性纖維化、反應性呼吸道疾病、由於吸菸導致之肺部感染、哮喘、肺炎、鼻炎、耳炎、竇炎、結核病、克隆氏病(Crohn's disease)(結腸及回腸)、潰瘍性結腸炎、腸不成熟症、由幽門螺旋桿菌感染所誘發之胃炎及胃潰瘍、感染性腹瀉、壞死性腸炎、與抗生素相關之腹瀉、細菌性陰道炎、HIV、單純疱疹病毒、尿道感染、異位性皮膚炎、慢性潰瘍、癌瘤、異位性濕疹、燒傷、扁桃腺炎、齒齦炎、齲齒、眼部角膜炎、及其組合。
本發明組合物亦可用於加強內源性抗菌性防禦。
其機轉可為例如藉由加強內源性hBD2表現，且若使用非複製性長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)，則另外亦藉由加強hBD1表現。
本發明者已發現長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)強力誘發組成性hBD2表現；且例如經熱處理之非複製性長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)比其活的對應物更能向上調節hBD2表現；且經熱處理之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)亦向上調節hBD1表現。
因此，本發明之標的亦包括一種提高長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)於治療或預防與免疫系統相關之病症上之效力的方法，包括使長雙歧桿菌NCC 2705呈非複製性之步驟，例如藉由熱處理。
該與免疫系統相關之病症可為例如上述任一種病症。
特定言之，病症可為與β-防禦素含量減少相關之病症。
於本發明之一項實施例中，該方法包括至少於70℃下熱處理至少約10秒鐘之步驟。
彼等熟習此項技術者咸瞭解，在不偏離所揭示之本發明之範圍下，其可自由組合文中所述之本發明之所有特徵。特定言之，所闡述之本發明組合物之特徵可應用於本發明之用途及/或方法中，反之亦然。
自以下實例及圖示中可見本發明之其他優勢及特徵。
圖1顯示，經於120℃下熱處理15秒鐘之長雙歧桿菌(寄存編號CNCM I-2618)能夠比其他經熱處理之菌株更強力於活體外誘發腸上皮細胞中之hBD2 mRNA。由T84細胞與經熱處理之菌株一起培養4小時。由即時定量PCR分析hBD2之基因表現量。數據代表經過未經刺激之細胞中基線表現量校正之平均值±sem。
圖2顯示，經於85℃下熱處理20分鐘之長雙歧桿菌(寄存編號CNCM I-2618)可最強力誘發hBD2(A)及hBD1(B)。以活的及經於120℃下熱處理15秒鐘，或於85℃下熱處理20分鐘的長雙歧桿菌(寄存編號CNCM I-2618)刺激T84細胞4小時。由即時定量PCR分析hBD2之基因表現量。數據代表經過未經刺激之細胞中基線表現量校正之平均值±sem。
實例： 實驗方法：
使用第30至40繼代之T84細胞，並培養於含有5%胎牛血清(FCS)(Amined BioConcept)及2 mM穀胺醯胺之杜氏改良最基本培養基(Dulbecco's modified essential medium)/F-12(Sigma D 6421)中。依2×106 個細胞/孔將細胞接種於6孔培養平板中，並使其於37℃下，於5% CO2 -95%空氣氛圍中呈單層生長。在細胞生長至匯合1週後，與不含血清及抗生素之培養基一起培養細胞至少12小時。需要該步驟，以消除由血清誘發之防禦素表現，並防止抗生素對益生菌及細胞免疫反應之任何影響。再與益生菌或經熱處理之菌株一起培養細胞4小時。在結束培養時，根據供應商之說明，以PBS洗滌細胞，並利用TriPureTM 分離試劑收集。由定量PCR分析經如此處理之細胞中之人類hBD1及hBD2基因表現量。
該實驗中所採用之細菌菌株為長雙歧桿菌NCC2705(寄存編號CNCM I-2618)、乳雙歧桿菌(B. lactis)(NCC 2818，寄存編號CNCM I-3446)、約氏乳桿菌(L. johnsonii)(Lal、NCC 533，寄存編號CNCM I-1225)、副乾酪乳桿菌(L. paracasei)(ST11、NCC 2461、寄存編號CNCM I-2116)。所測試該等菌株為活菌，或經於120℃下熱處理15秒鐘或經於85℃下熱處理20分鐘。
結果：
所有研究之經熱處理(120℃，15秒鐘)之菌株均向上調節hBD2 mRNA之表現，但長雙歧桿菌(NCC2705)(寄存編號CNCM I-2618)之誘發功效比其他經熱處理之菌株更強(圖1)。
此外，所示數據顯示，熱處理長雙歧桿菌NCC2705(寄存編號CNCM I-2618)可增進NCC2705對hBD2 mRNA表現之效力。事實上，經於120℃下熱處理15秒鐘，或經於85℃下熱處理20分鐘之長雙歧桿菌NCC2705(寄存編號CNCM I-2618)比活菌株更有效(圖2)。
更進一步，於低溫(85℃)下，長時間(20分鐘)處理長雙歧桿菌NCC2705(寄存編號CNCM I-2618)以使其失活，可強烈增加hBD2及hBD1 mRNA表現(圖2A及B)。
圖1顯示，經於120℃下熱處理15秒鐘之長雙歧桿菌(寄存編號CNCM I-2618)能夠比其他經熱處理之菌株更強力於活體外誘發腸上皮細胞中之hBD2 mRNA；及
圖2顯示，經於85℃下熱處理20分鐘之長雙歧桿菌(寄存編號CNCM I-2618)可最強力誘發hBD2(A)及hBD1(B)。
(無元件符號說明)

Claims (18)

  1. 一種包括長雙歧桿菌(B.longum)NCC 2705(寄存編號CNCM I-2618)的組合物，其係用於治療或預防感染及與免疫系統相關之疾病(包括感染)，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  2. 如請求項1之組合物，其中進行該熱處理至少10秒鐘。
  3. 如請求項1或2之組合物，其中於110℃至140℃下熱處理10至20秒鐘，或於75℃至95℃下熱處理10至30分鐘使長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  4. 如請求項1或2之組合物，其中該組合物所含之非複製性長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之數量相當於每份日劑量含約104 至1012 cfu。
  5. 如請求項1或2之組合物，其中該組合物每份日劑量含有約0.005 mg至1000 mg非複製性長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)。
  6. 如請求項1或2之組合物，其中至少95%長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  7. 如請求項6之組合物，其中所有長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  8. 如請求項1或2之組合物，其中該組合物係自以下組成之群中選出：食物組合物、食品，包括寵物食物、飲料、營養調配物、餵養調配物、營養藥劑、食品添加劑、醫 藥組合物、化妝品組合物、外用組合物及醫藥物。
  9. 如請求項1或2之組合物，其中該與免疫系統相關之病症係自以下組成之群中選出：感染，特定言之細菌、病毒、真菌及/或寄生蟲感染；吞噬細胞功能不足症；上皮障壁功能障礙或免疫系統不成熟症、小腸細菌過度生長症(SIBO)及其組合。
  10. 如請求項1或2之組合物，其中該與免疫系統相關之病症係選自由與防禦素(特定言之hBD2)含量減少相關之病症所組成之群。
  11. 如請求項10之組合物，其中該與hBD2含量減少相關之病症係自以下組成之群中選出：囊腫性纖維化、反應性呼吸道疾病、吸菸引起之肺部感染、哮喘、肺炎、鼻炎、耳炎、竇炎、結核病、克隆氏病(Crohn's disease)(結腸及回腸)、潰瘍性結腸炎、腸不成熟症、由幽門螺旋桿菌(Helicobacter pylori)感染所誘發之胃炎及胃潰瘍、感染性腹瀉、壞死性腸炎、與抗生素相關之腹瀉、細菌性陰道炎、HIV、單純疱疹病毒、尿道感染、異位性皮膚炎、慢性潰瘍、癌瘤、異位性濕疹、燒傷、扁桃腺炎、齒齦炎、齲齒、眼部角膜炎。
  12. 如請求項1或2之組合物，其係用於加強內源性抗菌性防禦、及/或內源性hBD2及hBD1表現。
  13. 一種提高長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)於治療或預防與免疫系統相關之病症上之功效之方法，包括藉由於至少約75℃下熱處理至少約10秒鐘使長雙歧 桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性之步驟。
  14. 一種包括長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)之組合物，其中至少95%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  15. 如請求項14之組合物，其中至少98%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  16. 如請求項14之組合物，其中至少99%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  17. 如請求項14之組合物，其中至少99.9%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
  18. 如請求項14之組合物，其中100%之長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)係呈非複製性，其中藉由熱滅活，藉由於至少約70℃下處理，使至少部份長雙歧桿菌NCC 2705(寄存編號CNCM I-2618)呈非複製性。
TW099115195A 2009-05-11 2010-05-11 長雙歧桿菌ｎｃｃ２７０５（ｃｎｃｍ ｉ－２６１８）及免疫病症 TWI402075B (zh)

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