CN103987839A - 能够抑制幽门螺杆菌菌株粘附至上皮细胞的新德式乳杆菌保加利亚亚种菌株 - Google Patents
能够抑制幽门螺杆菌菌株粘附至上皮细胞的新德式乳杆菌保加利亚亚种菌株 Download PDFInfo
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Abstract
本发明涉及适合用于治疗或预防幽门螺杆菌感染的新德式乳杆菌保加利亚亚种菌株。
Description
本发明涉及益生菌领域。具体地说,本发明涉及用于治疗或预防幽门螺杆菌(Helicobacter pylori)感染的新德式乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp.bulgaricus)菌株。
根据美国国家酸奶协会(NYA)或国际生命科学会(ILSI)最近批准的定义,益生菌是在以充足量消化时发挥超出基本营养之外的健康益处的活微生物。在属于乳品工业常用的属乳杆菌属(Lactobacillus)、双歧杆菌属(Bifidobacterium)、链球菌属(Streptococcus)和乳球菌属(Lactococcus)的种类中已描述了益生菌。认为益生菌通过阻止致病微生物的发展和/或更直接地作用于免疫系统而在肠道微生物丛水平上进行干预。
幽门螺杆菌(H.pylori)是革兰氏阴性螺旋形细菌,其寄居在超过50%世界人口的人胃粘液层中。而被幽门螺杆菌感染的大多数个体是无症状的,虽然他们的胃上皮出现炎症体征,15%至20%的幽门螺杆菌感染个体出现疾病。实际上,幽门螺杆菌是慢性活动性胃炎、消化性溃疡疾病、萎缩、化生、发育异常、胃癌和胃粘膜相关淋巴样组织(MALT)淋巴瘤的主要病原体(参见综述Fox和Wang,2007以及Polk和Peek,2010)。
在感染期间,幽门螺杆菌特通过该细菌产生的多个粘附分子(粘附素)如BabA和SabA蛋白特异性地与内衬胃上皮的胃上皮细胞结合。粘附至胃上皮细胞使细菌免受流体流动、蠕动和粘液层的脱落。幽门螺杆菌粘附至胃粘膜诱导胃上皮细胞内的信号转导途径,通过氧化应激、细胞凋亡和/或自体吞噬机制造成胃上皮细胞损伤和萎缩。因此,幽门螺杆菌粘附至胃上皮细胞是建立胃粘膜感染的关键步骤。
对幽门螺杆菌感染患者的标准治疗是两种抗生素联合质子泵抑制剂(PPI),所谓的三联疗法。然而,由于抗生素耐药性或较差的顺应性,三联疗法治疗后的幽门螺杆菌的根除率在下降。此外,尽管有几次临床试验,目前尚无有效的疫苗上市。
由上述内容可见,需要对用于治疗或预防幽门螺杆菌感染的三联疗法的替代疗法或补充疗法。
已提出使用产生乳酸、细菌素和其它抗菌物质的益生乳酸菌(LAB)作为替代疗法。Boyanova等人(2009)已发现几种德式乳杆菌保加利亚亚种(L.bulgaricus)菌株,其以菌株依赖性方式在体外(使用琼脂孔扩散法)抑制幽门螺杆菌菌株包括耐抗生素菌株的生长。然而,所研究的德式乳杆菌保加利亚亚种菌株未被明确鉴定出来,且是公众不能获得的。Simova等人(2009)筛选了几种产生细菌素的乳酸菌。之后,他们通过采用两种体外方法(琼脂孔扩散法及Barefoot和Klaenhammer的临界稀释试验)来分析这些菌株的无细胞上清液(CFS)的抗微生物活性而测定了所述菌株的抗微生物活性。他们发现,由于其抗微生物活性,德式乳杆菌保加利亚亚种BB18菌株的CFS抑制了广谱致病微生物(细菌或酵母菌)包括幽门螺杆菌菌株的生长。
本申请发明人分离了一种新的德式乳杆菌保加利亚亚种菌株并发现该菌株在体外和体内抑制幽门螺杆菌菌株粘附至上皮细胞。这种德式乳杆菌保加利亚亚种菌株(命名为DN_100_0602或CNCM I-4428)因而能够用于通过靶向建立这一感染的主要步骤(即通过抑制幽门螺杆菌菌株粘附至上皮细胞)治疗或预防幽门螺杆菌感染。
因此,本申请的保护主题是根据布达佩斯条约于2011年2月3日在CNCM(国家培养物和微生物保藏中心((Collection Nationale de Cultures deMicroorganismes),25rue du Docteur Roux,巴黎)以保藏号CNCM I-4428保藏的菌株德式乳杆菌保加利亚亚种。
该菌株CNCM I-4428具有以下特性:
-形态学:革兰氏阳性微生物,不动的、杆状的、具有颗粒的长杆菌,
-代谢:同型发酵(homofermentive),过氧化氢酶(-),
-糖发酵(在37℃下在MRS培养基中采用API50CH试验持续48h获得的结果):葡萄糖、果糖、甘露糖和乳糖。
此外,该菌株能够在体外和体内抑制幽门螺杆菌菌株粘附至上皮细胞。
如本文所用,术语“抑制幽门螺杆菌菌株粘附至上皮细胞”是指显著抑制(即全部或部分抑制)幽门螺杆菌菌株粘附至上皮细胞。可以如下面实施例1中所示在体外测量幽门螺杆菌菌株对上皮细胞粘附的抑制。
本发明还涵盖了突变菌株或由亲本菌株CNCM I-4428衍生的遗传转化菌株,前提条件是它们能够抑制幽门螺杆菌菌株粘附至上皮细胞。这些突变体或遗传转化菌株可以是其中亲本菌株的一种或多种内源性基因已经突变,例如以改变一些其代谢性质(例如其发酵糖的能力、其耐酸性、其在胃肠道转运的存活力、其酸化后性质或其代谢产物的生成)的菌株。它们也可以是由亲本菌株CNCM I-4428通过一种或多种目标基因进行遗传转化得到的菌株,例如为了赋予所述遗传转化的菌株额外的生理特征,或允许其表达具有治疗或疫苗益处的蛋白,人们希望通过所述菌株给予这样的蛋白。这些菌株可以由CNCMI-4428菌株通过用于乳酸菌属随机诱变或定点诱变以及遗传转化的常规技术,如Gury等人(2004)或Perea Vélez等人(2007)描述的那些,或者通过被称为“基因组改组”的技术(Patnaik等人,2002和Wang等人,2007)获得。
本发明的保护主题也是用于获得能够抑制幽门螺杆菌菌株粘附至上皮细胞的德式乳杆菌保加利亚亚种菌株的方法,该方法包括诱变或遗传转化菌株CNCM I-4428的步骤。
本发明的保护主题也是细胞组分(cell fractions),其可以由如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428获得,前提条件是它们能够抑制幽门螺杆菌菌株粘附至上皮细胞。它们特别是由所述菌株的培养物获得的DNA制剂或细菌细胞壁制剂。它们还可以是培养物上清或这些上清的部分。举例来说,菌株CNCM I-4428的无细胞上清(CFS)可以通过使用用于从Simova等人(2009)公开的另一种德式乳杆菌保加利亚亚种菌株获得CFS的方法而获得。
本发明的保护主题也是用于获得能够抑制幽门螺杆菌菌株粘附至上皮细胞的细胞组分的方法,该方法包括以下步骤:
a)培养如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCMI-4428,以及
b)由步骤a)中的培养物获得细胞组分。
本发明的保护主题也是包含根据本发明的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或由根据本发明的所述菌株获得的细胞组分的组合物。
在本发明的组合物中,所述菌株可以以全菌形式使用,所述全菌可以是活的或死的。可选地,所述菌株可以以细菌裂解物的形式或以细菌成分的形式使用;例如,可以通过测试它们在幽门螺杆菌对上皮细胞的粘附方面的性质来选择适合这种用途的细菌成分。优选细菌细胞以活的、有生命力的细胞存在。
本发明的组合物可以是任何适合施用特别是经口施用的形式。这包括例如固体、半固体、液体和粉末。液体组合物由于更易施用,例如作为饮料,而是普遍优选的。
组合物可以包含每克如上所述的至少一种细菌菌株干重至少105个cfu、优选至少106个cfu。
组合物还可以包含除了根据本发明的菌株以外的其他细菌菌株,特别是益生菌菌株,如乳杆菌属、双歧杆菌属、链球菌属或乳球菌属菌株。
当细菌是活细菌形式时,组合物可以通常包含每克组合物干重105至1013个菌落形成单位(cfu),优选至少106个cfu,更优选至少107个cfu,更优选至少108飞cfu以及最优选至少109个cfu。在液体组合物的情况中,这一般对应于104至1012个菌落形成单位(cfu),优选至少105个cfu,更优选至少106个cfu,更优选至少107个cfu,以及最优选至少109个cfu/ml。
组合物可以是营养组合物,包括食品、食品补充剂和功能性食品。"食品补充剂"是指由一般在食品中使用的化合物制成的产品,但其是以片剂、粉剂、胶囊剂、饮剂的形式或一般与食物不相关的任何其它形式,且其对人们的健康是有益处的。"功能性食品"是对人们的健康也有益处的食物。具体地说,食品补充剂和功能性食品能够对疾病例如慢性疾病具有保护性或治疗性的生理效应。
根据本发明的营养组合物也包括婴儿食品、婴儿乳配方或婴儿后续配方(infant follow-on formula)。优选地,本发明组合物是营养品或药物产品、营养补充剂或医疗食品。
组合物可以是乳制品,优选发酵乳制品。发酵产品可以以液体形式存在或者以通过干燥发酵的液体获得的干粉形式存在。乳制品的实例包括凝固、搅拌或可饮用形式的发酵乳和/或发酵乳清、乳酪和酸乳酪。
发酵产品也可以是发酵的植物,如凝固、搅拌或可饮用形式的发酵的大豆、谷物和/或水果。
在优选的实施方案中,发酵的产品是新鲜产品。未经过剧烈的热处理步骤的新鲜产品具有如下优点:细菌菌株以活体形式存在。
本发明的保护主题也是用作药物的如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或者如上面所定义的组合物。
本发明的保护主题也是用作用于治疗或预防幽门螺杆菌感染的药物的如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或者如上面所定义的组合物。
本发明的保护主题也是如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或者如上面所定义的组合物作为药物优选用于治疗或预防幽门螺杆菌感染的药物的用途。
本发明的保护主题也是如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或者如上面所定义的组合物在制备药物优选用于治疗或预防幽门螺杆菌感染的药物中的用途。
本发明的保护主题也是用于治疗或预防有需要的受试者中幽门螺杆菌感染的方法,所述方法包括向所述受试者施用治疗有效量的如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或如上面所定义的组合物。
治疗有效量的确定对于本领域技术人员是公知的,尤其是考虑到本文所提供的详细公开内容后。
本发明的保护主题也是用于生产药物,优选用于治疗或预防幽门螺杆菌感染的药物的方法,所述方法包括将如上面所定义的德式乳杆菌保加利亚亚种菌株,优选菌株CNCM I-4428,或者如上面所定义的组合物并入到至少一种药学上可接受的稀释剂、载体或赋形剂中。
如本文所用,治疗或预防由其涵盖:预防感染、稳定幽门螺杆菌的载量和/或降低幽门螺杆菌的载量。治疗或预防还涵盖缓解下面所述的与幽门螺杆菌相关的至少一种症状。
用于诊断幽门螺杆菌感染的方法是本领域中已知的。举例来说,可以通过血液抗体试验、粪便抗原试验或碳尿素呼气试验来进行幽门螺杆菌感染的诊断。也可以通过内窥镜下活组织检查之后进行尿素酶试验、组织学检查或微生物培养来进行幽门螺杆菌感染的诊断。
与幽门螺杆菌感染相关的症状或疾病是胃痛、胃灼热、酸反流、腹痛、反胃、呕吐、嗳气、胃气胀、恶心、胃炎如慢性活动性胃炎、消化性溃疡疾病、萎缩、化生、发育异常、胃癌和胃粘膜相关淋巴样组织(MALT)淋巴瘤。
根据下面的进一步描述可以更清楚地理解本发明,即示出CNCM I-4428菌株的抗感染性质的实施例以及随附的附图。
图1显示了对接受对照产品的被幽门螺杆菌SS1感染的小鼠或被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-4428治疗的小鼠或被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-1632(Lb130)治疗的小鼠在治疗前(第一棒)、治疗后3周(第二棒)及处死前(第三棒)测量的体重变化(以克计)。
图2显示了通过免疫组织化学使用抗-幽门螺杆菌抗体在(i)未被幽门螺杆菌感染的小鼠、(ii)接受对照产品(非发酵乳品)的被幽门螺杆菌SS1感染的小鼠、(iii)被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-1632(Lb130)治疗的小鼠以及(iv)被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-4428治疗的小鼠中获得的感染得分。得分的定义:0:无感染腺;1:极少感染腺,2:25%感染腺,3:25至50%感染腺,4:>50%感染腺。
图3显示了通过实时PCR在(i)未被幽门螺杆菌感染的小鼠、(ii)被幽门螺杆菌SS1感染但接受对照产品(非发酵乳品)的小鼠、(iii)被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-4428治疗的小鼠以及(iv)被幽门螺杆菌SS1感染且用德式乳杆菌保加利亚亚种CNCM I-1632(Lb130)治疗的小鼠中获得的幽门螺杆菌SS1DNA的量化。
实施例1:选择德式乳杆菌保加利亚亚种菌株评估其对幽门螺杆菌对胃上
皮细胞的粘附的作用
使用经过改良的Oleastro等人(2008)描述的方法来评估两种德式乳杆菌保加利亚亚种菌株(CNCM I-4428和于1995年10月25日在CNCM保藏的CNCMI-1632)对幽门螺杆菌对上皮细胞的粘附性质的作用。
1.1材料&方法
测试了两种幽门螺杆菌菌株:菌株J99(BabA2;ATCC号:700824)和菌株09.193(BabA1,由Yoshio Yamaoka在日本分离得到,来自Oita大学)。采用50的MOI(感染复数)对这些菌株进行测试。将幽门螺杆菌细胞在幽门琼脂(Biomérieux,法国)中培养24h之后转移到PBS缓冲液中。将两种幽门螺杆菌菌株的细菌悬浮液的浓度分别标准化至2.108cfu/mL(对应于OD600nm=1)。
采用50的MOI对这两种德式乳杆菌保加利亚亚种悬浮液进行测试(对应于每孔25μL标准化的德式乳杆菌保加利亚亚种的悬浮液)。
在感染实验前一天在500μL补充有10%胎牛血清的DMEM培养基F12中接种100000个上皮AGS细胞(ATCC号:CRL-1739)。
幽门螺杆菌悬浮液的染色
在10000g下将1mL幽门螺杆菌悬浮液离心10分钟。将沉淀物(pellet)重悬浮于1mL稀释液A(Diluent A)(来自PKH2绿色荧光细胞连接试剂盒(PKH2Green Fluorescent Cell Linker Kit),Sigma Aldrich,Saint-Louis),并加入2.5μL PKH2(Sigma Aldrich,Saint-Louis)。将悬浮液在室温下孵育2分钟30秒。通过加入2mL胎牛血清停止染色。然后加入4mL DMEM/F12培养基,之后在10000g下离心悬浮液10分钟。将沉淀物重悬浮于4mL培养基中并在10000g下离心10分钟。将该洗涤步骤重复两次以除去多余的荧光染料。
粘附的测量
用PBS洗涤3次以除去未粘附的细菌之后,采用胰蛋白酶遵循标准条件解离上皮AGS细胞。通过流式细胞术分析荧光发射。使用FACSCalibur流式细胞仪(Becton Dickinson)测量荧光。以三重重复进行每种条件。
在两种条件下进行幽门螺杆菌感染
-在感染前(pre)条件下:在用幽门螺杆菌感染前1h30将待测试的德式乳杆菌保加利亚亚种悬浮液加入到上皮AGS细胞。在感染后1h30(孵育时间共计3h)通过流式细胞术分析细胞;
-在同时感染(co)条件下:同时将待测试的德式乳杆菌保加利亚亚种悬浮液和幽门螺杆菌菌株加入到上皮AGS细胞,并在孵育1h30后通过流式细胞术分析细胞。
对照
采用与幽门螺杆菌菌株一起孵育1h30的上皮AGS细胞获得荧光强度的对照值。
相对于未暴露于德式乳杆菌保加利亚亚种的幽门螺杆菌对上皮AGS细胞的粘附(其被设定为100%)给出粘附百分比值。
根据三重重复获得的平均值分析结果,并且当根据Student's检验结果是显著不同(P<0.05)时对其进行评分。
1.2结果
结果在下文表1中示出。
表1:用2种德式乳杆菌保加利亚亚种菌株获得的它们对幽门螺杆菌对上皮AGS细胞的粘附的作用结果。
得分定义:-3:粘附显著增加,0:对粘附无显著性作用,1:对粘附的显著抑制为1至10%,2:对粘附的显著抑制为11至20%,3:对粘附的显著抑制为21至30%,,4:对粘附的显著抑制超过30%。
结果表明菌株CNCM I-4428的细菌悬浮液在两种不同的条件(孵育前和同时孵育)下抑制两种幽门螺杆菌菌株(J99和09.193)对上皮AGS细胞的粘附,但是菌株CNCM I-1632的细菌悬浮液不具有这种性质。
实施例2:菌株CNCM I-4428对小鼠模型中幽门螺杆菌的载量的作用
2.1材料&方法
幽门螺杆菌
使用对小鼠胃粘膜具有非常好的定殖能力的幽门螺杆菌菌株SS1(Lee等人,1997)。通过测定基因glm和hspA和vacA的序列来核实菌株的正身。
德式乳杆菌保加利亚亚种
按如下方法制备采用2种不同的德式乳杆菌保加利亚亚种菌株(CNCMI-4428或CNCM I-1632)发酵的乳制品:由冷冻菌株在MRSn中制备第一培养物并在37℃下孵育17h。在富含酵母菌提取物(2g/L)的脱脂乳中通过1%接种第一培养物来制备第二培养物,并在37℃下孵育17h。在富含酵母菌提取物(2g/L)的脱脂乳中通过1%接种第二培养物来制备第三培养物,并在37℃下孵育直到达到pH4.7。最后通过用第三培养物以1%接种富含酵母菌提取物(2g/L)的乳直到达到pH4.7来制备产品。将产品储存在-80℃下。孵育48h后在MRSn中进行细菌计数。对于菌株CNCM I-4428和CNCM I-1632,细菌计数分别为6.108和1.3.109cfu/mL。
小鼠
将55只5周龄并检测为SPF(《无特定病原体》)的雌性BALB/cBy/J小鼠(Charles River,法国)进行分组:使3组15只小鼠被感染,使用1组10只小鼠作为未感染对照。用缺少维生素的食物饲喂小鼠以增加幽门螺杆菌诱导的损害发展。
感染(8周)
6周龄小鼠接受含水饮食(hydric diet)1天,然后在次日早晨强行饲喂250μL菌株幽门螺杆菌SS1的浓缩悬浮液(对于5只小鼠1至2培养皿的幽门螺杆菌)。将小鼠放在提供正常饮食的笼子中。然后,小鼠在晚上再次接受含水饮食。将该方案重复3天。
治疗(6周)
在小鼠感染后8周,将小鼠用含有德式乳杆菌保加利亚亚种菌株的乳制品治疗6周。在饲喂瓶中给予每天每笼120g乳制品,而不是水。每天更换饲料瓶。为了评估每只动物摄取的产品的量,对饲料瓶进行称重。此外,在治疗之前、治疗后3周和处死前对小鼠进行称重(结果在图1中显示)。
小鼠对照组接受富含酵母菌提取物(2g/L)的乳品(即,不含有任何德式乳杆菌保加利亚亚种菌株)。
处死
通过颈椎脱臼法处死小鼠。实施剖腹术。分离出胃,并在生理血清(serum)中清洗胃粘膜。
从食道中间到十二指肠切割胃。对于右半侧胃,清除贲门,然后将该半侧胃放入生理血清中以用于分子研究。将左半侧胃用于组织学。
组织学
将左半侧胃在3.7%福尔马林中固定1夜并用70%乙醇清洗,然后石蜡包埋和切片,厚度为3μm。
采用抗幽门螺杆菌抗原的抗体实施免疫组织化学:一抗:抗幽门螺杆菌抗体(Dako,Ref.B0471);二抗和DAB:Dako EnVision+System-HRP(DAB)(Dako,Ref.K4011)。
分子研究(q RT-PCR)
采用Potter-Elvehjem(在含胃组织和不含胃组织的情况下对管进行称重以获知组织的重量)将右侧胃在0.2ml生理血清中匀浆(分散)。
采用Arrow Stool DNA试剂盒(NorDiag,Norvège)按照供应商的推荐从破碎的胃提取总DNA。对于每个破碎的胃,将总DNA重悬浮在180μL TRIS缓冲液(10mM)中。
通过实时PCR量化在DNA提取物中存在的幽门螺杆菌的DNA。遵循Oleastro等人(2003)描述的方法采用靶向在两拷贝幽门螺杆菌中存在的23SrRNA的引物进行扩增。对于20μl20μM的的混合物(MgCl225mM,Ménard等人(2002)描述的引物HPY-A和HPY-S20μM,5’标记有LC-Red640和3’磷酸化的传感器(sensor)探针和3’标记有荧光素的锚定(anchor)探针(这两种探针均是由Oleastro等人(2003)描述的),含有酶的缓冲液(10X,试剂盒FastStartDNA Master杂交探针(kit FastStart DNA Master Hybridization Probes),RocheDiagnostics),加入5μl200ng/μl DNA以在Light Cycler ROCHE中使用以下程序进行扩增:
2.2结果
通过免疫组织化学对德式乳杆菌保加利亚亚种菌株CNCM I-4428和CNCM I-1632(Lb130)获得的感染得分在图2中示出。这些结果表明,对被幽门螺杆菌感染的小鼠施用用菌株CNCM I-4428发酵的乳制品显著降低了感染得分,但是用菌株CNCM I-1632发酵的乳制品进行处理未降低幽门螺杆菌的载量。
通过实时PCR对德式乳杆菌保加利亚亚种菌株CNCM I-4428和CNCMI-1632(Lb130)获得的结果在图3中示出。这些结果表明,在小鼠中,用菌株CNCM I-4428发酵的乳制品进行处理显著降低了幽门螺杆菌的载量,但是用菌株CNCM I-1632发酵的乳品进行处理未降低幽门螺杆菌的载量。
参考文献
-Boyanova L et al.,Lett Appl Microbiol.2009;48:579-84.
-Fox JG and Wang TC.,J Clin Invest.2007;117:60-9.
-Gury J et al.,Arch Microbiol.2004;182:337-45.
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PCT/RO/134表
Claims (11)
1.一种德式乳杆菌保加利亚亚种菌株,特征在于其以编号I-4428保藏在CNCM。
2.一种用于获得能够抑制幽门螺杆菌粘附至上皮细胞的德式乳杆菌保加利亚亚种菌株的方法,所述方法包括诱变或遗传转化权利要求1的德式乳杆菌保加利亚亚种菌株的步骤。
3.一种德式乳杆菌保加利亚亚种菌株,特征在于其能够通过根据权利要求2所述的方法获得,以及其能够抑制幽门螺杆菌粘附至上皮细胞。
4.一种用于获得能够抑制幽门螺杆菌粘附至上皮细胞的细胞组分的方法,所述方法包括以下步骤:
a)培养根据权利要求1或3所述的德式乳杆菌保加利亚亚种菌株,和
b)从步骤a)的培养物中获得细胞组分。
5.一种细胞组分,特征在于其能够通过权利要求4所述的方法获得,以及其能够抑制幽门螺杆菌粘附至上皮细胞。
6.一种组合物,其包含根据权利要求1或3所述的德式乳杆菌保加利亚亚种菌株或根据权利要求5所述的细胞组分。
7.根据权利要求6所述的组合物,特征在于其包含每克根据权利要求1或3所述的德式乳杆菌保加利亚亚种菌株干重至少105个cfu,优选至少106个cfu。
8.根据权利要求6或7所述的组合物,特征在于其是营养组合物。
9.根据权利要求8所述的组合物,特征在于其是乳制品。
10.根据权利要求1或3所述的菌株或根据权利要求6或7所述的组合物,其用作药物。
11.根据权利要求10所述的菌株或组合物,特征在于所述药物用于治疗或预防幽门螺杆菌感染。
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