抗过敏益生菌唾液乳杆菌PM-A0006调节过敏体质抗过敏

抗过敏益生菌唾液乳杆菌PM-A0006调节过敏体质抗过敏 J. Agric. Food Chem. 2007, 55, 11080–11086 Heat-Killed Cells of Lactobacilli Skew the Immune Response Toward T Helper 1 Polarization in Mouse Splenocytes and Dendritic Cell-Treated T Cells 论,热杀死抗过敏乳酸菌唾液乳杆菌PM-A0006调节过敏体质抗过敏 摘要 乳酸菌为人类及哺乳类肠胃道中的共生菌之一,已被认定为益生菌, 对于维持宿主健康扮演着重要的角色,包括了其对于免疫功能的调节。目 前大部分的研究皆比较着重于乳酸菌的活菌体 (viable cell)对于免疫调节 作用的影响,而本实验则主要是在于乳酸菌热杀死菌体的影响。我们藉由 将三株热杀死乳酸菌L. acidophilus A2、L. gasseri A5和L. salivarius A6与脾脏细胞共同培养,来探讨此三株热杀死乳酸杆菌对于免疫功能所产生 的影响。另一方面,我们亦将三株热杀死乳酸菌刺激树突细胞后,以观察T 细胞的免疫反应。由实验结果显示三株热杀死乳酸菌证实能刺激小鼠的脾 脏细胞增生和interleukin (IL)-10、IL-12 p70和interferon (IFN)-γ之分泌, 但对transforming growth factor (TGF)-β无影响。另外,三株热杀死乳酸菌 亦能刺激小鼠的树突细胞分泌高量的IL-12 p70,且由提高IFN-γ分泌,非 IL-5、IL-13及TGI-β之结果显示,T细胞的免疫反应趋向Th1。以上结果显 示乳酸杆菌 (lactobacilli) 对于调节免疫机能及过敏反应扮演了重要的角 色。 KEYWORDS: Lactobacillus; splenocytes; dendritic cells; T helper 1; cytokines 前言 益生菌被定义为“活菌,能在固有的营养条件下,增进健康”(1)。乳酸杆 菌是人和哺乳动物胃肠道的共生微生物的成员之一,通常被认定为益生菌 (2,3)。肠道乳酸菌一般被认为对于宿主的免疫反应之调节,扮演着一个重 要的角色(1,4-6)。乳酸菌具有影响肠周边免疫系统的功能(5,7)。在肠道中, 乳酸细菌细胞壁的组成成分在刺激免疫活性的细胞扮演一个重要的角色 (4,5,8,9)。 位于肠中的乳酸菌,包括各种乳酸杆菌,可交互调控肠中的一些细胞, 包括抗原呈现细胞(APCs)以及小肠上皮细胞(intestinal epithelial cells)(10-12)。近期的一些报告中也指出,乳酸菌可能可调控免疫路径Th1 与Th2之间的平衡,有助于减轻过敏反应1(5,13-15)。一般认为是靠一些 特殊的免疫细胞产生的细胞激素,例如,IL-10和TGF-β,来保持这样的平 衡(16,17)。而大部份的研究主要针对活的乳酸菌在免疫调节上之影响,而 the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 本篇研究则是研究经热杀死的乳酸菌。其拥有保存期限长、贮存方便、便于运输等优点。 图一, 热杀死乳酸杆菌对小鼠脾脏细胞增生之影响。柱状图分别代 表 关于同志近三年现实表现材料材料类招标技术评分表图表与交易pdf视力表打印pdf用图表说话 pdf 脾脏细胞对不同热杀死乳酸菌、PHA或无刺激物之结果,其中后两项为实验对照组。数据资料计算结果为平均数?标准差(个数=3,p<0.05) 由骨髓中衍生的树状细胞(Dendritic cells, DC),是引发适应性免疫对抗外来病源的专一性抗原呈现细胞(18)。未成熟的树状细胞藉由血液的流动进入非淋巴组织里,例如皮肤和黏膜,来捕捉和呈现抗原。然后移到淋巴腺器官,例如淋巴结、脾脏和黏膜伴生的淋巴组织(mucosal-associated lymphoid tissues, MALT),其中他们失去抗原加工活性并且成熟为强有力的免疫刺激细胞(18,19)。在肠黏膜中之树状细胞可能由乳酸菌调控,包括肠中原有的共生菌和经由口服(administered orally)入内的(20)。他们在Peyer氏斑间接通过M细胞(21),与乳酸菌交互作用,且绕过紧连的上皮中进入小肠内腔直接与乳酸菌作用(22)。而树状细胞藉由这些微生物的刺激而变成熟,包括上游调节共激分子(costimulatory molecule)以及cytokines和chemokines的分泌,以及提供各种T细胞的免疫反应。举例来说,树状细胞上游调节了共激分子CD80(B7-1)和CD86(B7-2) 及产生IL-12,造成Th1路径反应(23)。 他们也生产IL-4和IL-10,促进Th2路径或Treg反应(24-26)。 以人类及老鼠的试验中显示,乳酸菌能有效的刺激产生IL-12以及IFN-γ,并且调整免疫反应(13,14,27),它能调控树状细胞表面的分子表现和cytokine的分泌(28-30)。口服和原本在肠道中的乳酸杆菌,是十分相近于肠黏膜中之树突细胞,藉此产生Th1、Th2和Treg路径的免疫调控。在本文中将探讨三种经热杀的乳酸菌在小鼠脾脏细胞增生以及cytokine产生的the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 现象做探讨。同时我们也对于T细胞的影响做探讨。 材料与 方法 快递客服问题件处理详细方法山木方法pdf计算方法pdf华与华方法下载八字理论方法下载 乳酸杆菌的处理,本研究所使用的三种乳酸杆菌由位于台南科学园区,东宇生物技术股份有限公司所提供,分别为经由热杀死的Lactobacillus acidophilus PM-A0002 (A2), Lactobacillus gasseri PM-A0005 (A5),以及 Lactobacillus salivarius PM-A0006 (A6)。在热处理之前,分别以MRS培养基,37?培养16小时,经离心(2000g 10min)后,使用去离子水洗涤两次,悬浮于phosphate-buffered saline(PBS; 0.85% NaCl, 2.68 mM KCl, 10 mM Na2HPO4, and 1.76 mM KH2PO4 at pH 7.7)溶液中,之后以100?煮沸15分钟杀死。 DC的备制,小鼠DC来源是由国家实验室动物中心(台北,台湾)购买的6至10星期成熟母鼠(BALB/c)骨髓中取得,经PBS洗涤两次后,将细胞数以RPMI-1640(含10%FBS、GM-CSF及IL-4)调整成1*106cell/mL,放入24-well 培养皿中培养6日,期间2~3日更换一次培养基。 脾脏细胞和T细胞的备制,来源是使用BALB/c母鼠,采断颈法牺牲后,在无菌环境取出脾脏,加入Ficoll-Hypaque后以400 g,离心30分钟。位于低密度的部分,约42.5~50%的介面,从其中分离出单一细胞悬浮于PBS中取出脾脏,加入Ficoll-Hypaque后以400 g,离心30分钟。位于低密度的部分,约42.5~50%的介面,从其中分离出单一细胞悬浮于PBS中。CD4+ T细胞的纯化采用Miltenyi Biotec.,Auburn, CA所制造的极性抗体和毫米磁珠系统纯化收集,之后将纯化的小鼠CD4+ T细胞培养在含有10% FBS的RPMI-1640培养基中,调整细胞数量成2*106 cell/mL,放入96-well培养皿中。 the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 乳酸杆菌刺激脾脏细胞,将细胞调整成4*105 cell/mL后,跟热杀死的 乳酸菌以1:3的比例,于37?高湿度、5% CO2的环境下共同培养24、48、 72小时和5日。以1 μg/mL PHA当作对照组。经24、48、72小时的培 养后,取上层液利用ELISA 分析 定性数据统计分析pdf销售业绩分析模板建筑结构震害分析销售进度分析表京东商城竞争战略分析 IL-10, IL-12 p70,IFN-γ, and TGF-β的分泌 量,而培养5日后,则使用MTT来分析脾脏细胞的增殖情况。 the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 乳酸杆菌刺激树状细胞,将细胞调整成1*106 cell/mL后,跟热杀死的 乳酸菌以1:10的比例,于37?高湿度5% CO2的环境下共同培养24、48、 72小时。以0.1 μg/mL LPS当作对照组。经24、48、72小时的培养后, 取上清液,以ELISA分析IL-4、IL-10及 IL-12 p70之分泌量。并以抗生素 (mitomycin C, 25 μg/mL),37?处理1小时,让DC停止生长。 the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 图四, 经由热杀死乳酸杆菌处理DC,对T细胞增生的影响。柱状图分别代表,树状细胞以热杀死处理之乳酸菌、LPS或无刺激时,与T细胞共培养24、48、72小时的结果。数据资料计算结果为平均数?标准差(个数=3,p<0.05)。A-C分别代表不同的经不同物质刺激后,所得到的数据在统计上是否归类为同一种类的表现,而X与Y分别代表不同的时间点,所得到的数据在统计上是否归类为同一种类的表现。 经乳酸杆菌所处理后的DC来刺激T细胞,停止增生的DC与T细胞以1:10的比例,于37?高湿度5% CO2的环境下共同培养2和5日。收集培养2日的细胞上层液,利用ELISA分析IL-5、IL-13、IFN-γ以及TGF-β之分泌量,而培养5日的细胞,则使用MTT来分析T细胞的增殖情况。 统计分析, 数据所代表的是平均数?标准差(mean?SD),样本数为3(n=3),统计的比较是藉由Duncan's multiple-range test之变异数来分析,p值少于0.05被认为统计上有差异性。 结果, 乳酸杆菌促进脾脏细胞的增生,三种经过热杀死的乳酸菌,包括L. acidophilus A2, L. gasseri A5, 和 L. salivarius A6,被测试用来引发脾脏细胞的增生能力(图一),PHA(1μg/mL)为此试验的正对照组。结果显示相对于负对照组,L. salivarius A6有效地诱导脾脏细胞之增生(约增加了11倍)。 诱导脾脏细胞来促进细胞激素的产生,为了确定这些热杀死的乳酸菌是否增进脾脏细胞分泌细胞激素,因此分析了脾脏细胞与热杀死乳酸菌共培养24、48、72小时之后,上层液所含IL-10、IL-12p70、IFN-γ、TGF-β的量(图二)。以PHA(1μg/mL)当作正对照组。结果显示三种菌都能诱导IL-10的分泌(图二A),但是远低于IL-12p70的量(图二B)。三种菌均对小鼠的脾脏细胞产生很强的刺激,分泌大量的IL-12p70,而在图2C显示,只有L. salivarius对于产生IFN-γ较显着(图三C)。L. acidophilus的试验组在24小时的培养下,能测得较高的IFN-γ生成量,L. gasseri和L. salivarius则以48小时的培养下得到较高。然而与负对照组的比较中发现,此三种热the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 杀死乳酸菌对脾脏细胞产生TGF-β则没有明显的作用(图二D)。 促进树状细胞产生细胞激素,为了确定这些热杀死的乳酸菌是否增进树状细胞分泌细胞激素,因此分析了树状细胞与热杀死乳酸菌共培养24、48、72小时后,上层液所含IL-4、IL-10、IL-12p70的量(图三)。以LPS当作正对照组。结果显示,培养48小时的条件下,任合一组所测试出的IL-4量均低于负对照组(图三A),但在24和72小时均未侦测到(p<0.05)。而IL-10的变化,在三个时间点观察下,各组差异均不大(图三B),相对的,IL-12p70则都有极高的表现量(图三C)。 菌处理的树状细胞刺激T细胞增殖,为了测试成熟的DC是否可活化T细胞, DCs先经这些热杀死的乳酸菌刺激后,再与T细胞共同培养5天。结果显示(图四),在共同培养24小时的条件下,T细胞的数量的确增加许多(增加约2~3倍)。在48小时或72小时后的观察下发现,除了经由L. gasseri处理的试验组外,其余均比负对照组高出约1.5倍。 菌处理的树状细胞诱导T细胞分泌细胞激素,进一步确定T细胞所分泌细胞激素的种类,我们收取培养2日后的上层液,分析IL-5、IL-13、IFN-γ以及TGF-β的生成量(图五)。结果发现IL-5的量没有明显的改变(图五A),而IL-13的量也远低于负对照组(图五B)。然而,经过乳酸杆菌刺激后的DC与T细胞共同培养,却可刺激T细胞产生IFN-γ(图五C),当T细胞以L. acidophilus或L. salivarius处理的树状细胞刺激24小时及L. gasseri处理的树状细胞刺激48小时下,有高,的IFN-γ被产生。至于TGF-β,除了L. gasseri的组别在24小时观察下,量略低以外,其余组别之间在统计上无明显差异(图五D)。 the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 讨论, 包含新型抗过敏乳酸菌菌株组合物,康敏元益生菌,,目前已知道对于健康有很多益处,例如,免疫调节,改善过敏体质。多数研究集中于探讨抗过敏乳酸菌或其细胞壁萃取物对于免疫的调节作用,然而,本篇在于探讨热杀死乳酸菌的研究。 过敏反应是由于Th1/Th2间之平衡被破坏使反应趋向Th2免疫反应(31),巨噬细胞(Macrophage)的IL-12,与T细胞衍生的IFN-γ为Th1路径下所产生的cytokines,它可以增进细胞免疫反应的调节。IL-12刺激T和NK细胞产生IFN-γ,并且促进未成熟CD4+ T细胞进入Th1的分化路径(32,33)。IFN-γ亦增加IL-12的产生,并减低Th2路径细胞的增殖与活化,提高体液免疫能力(34)。IL-10和TGF-β为免疫抑制的cytokines,能抑制作用T细胞(effector T cell)的活性与能力,例如Th1和Th2细胞。 他们在Treg细胞族群的分化和作用也扮演一个重要角色(16,17)。本研究结果显示,根据IFN-γ在脾脏细胞的产生结果,L. salivarius可以促进小鼠脾脏细胞的大幅度增生,但是L. acidophilus和L. gasseri只有小幅度增加。然而,三the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine 种乳酸杆菌刺激脾脏细胞所得到的结果显示,均产生高量IL-12p70,但IL-10的表现量相对较低,TGF-β甚至侦测不到表现,这样显示乳酸菌强烈刺激脾细胞偏向Th1的路径反应(35)。利用L. casei Shirota也能刺激脾脏细胞产生IL-12和IFN-γ(27)。也有报告指出,定时摄取乳酸杆菌可以增加鼠类脾脏细胞产生IFN-γ,但对于IL-4或是IL-5则没有影响(36,37)。 所以肠道中的乳酸菌可以透过肠中的DC来调节免疫功能。DC如果产生IL-12,则造成Th1反应(23),同样的如果产生IL-4和IL-10,则促进Th2或Treg反应(24-26)。由Th2细胞所分泌的IL-5和IL-13来恢复粒状效应细胞,例如嗜酸性、硷性及柱状细胞(mast cell) 在过敏发炎部位的作用(38,39)。 在这项研究中,三种热杀死乳酸菌能引起小鼠DC分泌IL-12p70,但是IL-10及IL-4则较无影响。以乳酸菌处,的树状细胞有促进T细胞的增殖,及增加IFN-γ的表现量,此代表成熟的DC经乳酸杆菌的刺激,将T细胞免疫反应趋向Th1路径。Mohamadzadeh等人指出(29),L. johnsonii, L. reuteri, 和 L. gasseri可刺激人类单核球树状细胞(monocyte derived DCs)产生IL-12,而非IL-10,显示将免疫路径导向Th1。但有文献指出L. casei 与 L. plantarum Lb1刺激小鼠DC强烈引起IL-12的生成,而L. reuteri DSM12246却是引起IL-10的生成(28)。近来也有报导指出L. acidophilus 与 L. paracasei能强烈刺激人类单核球树状细胞产生IL-12(30)。所以到底是么机制造成各种乳酸杆菌在树状细胞上产生不同的反应,目前仍未知,但在免疫调节的反应上造成的差异,可以推断于几个原因,例如使用的不同的乳酸细菌和人或老鼠免疫细胞。根据早先观察和我们的结果,推断乳酸菌所诱导产生之细胞激素与乳酸菌菌种有关。 本研究发现,经热杀死的乳酸杆菌在体外试验中有免疫调控能力。在大鼠身上,经热杀死处理之L. plantarum L-137会诱导IL-12和IFN-γ的产生,并抑制因IL-4之诱导而生成的IgE,进而压抑Th2免疫反应 (40)。而且,使用热杀死的L. casei Shirota经由体外试验刺激脾脏细胞或口服,均能诱导IL-12和IFN-γ及Th1路径的细胞激素产生(27,41)。从本试验可得知,经热杀死的乳酸菌对于免疫调节也具有功效,并非活菌才能引发免疫反应。 总之,研究所使用的三种热杀死的乳酸菌,包括L. acidophilus A2, L. gasseri A5, L.salivarius A6,能活化老鼠脾脏细胞和DC,驱使T细胞往Th1免疫反应。 从此结果可得知,经热杀死的特定乳酸菌菌株可运用在免疫调节上并扮演着重要的角色。经热杀死的乳酸菌有着长产品贮藏期限、更容易保存和运输等优点。便于出外旅游携带,也许可以扮演免疫预防的目的。 LITERATURE CITED the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine (1) Guarner, F.; Schaafsma, G. J. Probiotics. Int. J. Food Microbiol.1998, 39, 237–238. (2) Molin, G.; Jeppsson, B.; Johansson, M. L.; Ahrne, S.; Nobaek,S.; Stahl, M.; Bengmark, S. Numerical taxonomy of Lactobacillus spp. associated with healthy and diseased mucosa of the human intestines. J. Appl. Bacteriol. 1993, 74, 314–323. (3) Ahrne, S.; Nobaek, B.; Jeppsson, I; Adlerberth, A.; Wold, E.; Molin, G. The normal Lactobacillus flora of healthy human rectal and oral mucosa. J. Appl. Microbiol. 1998, 85, 88–94. (4) Erickson, K. L.; Hubbard, N. E. Probiotic immunomodulation in health and disease. J. Nutr. 2000, 130, 403. (5) Cross, M. L.; Stevenson, L. M.; Gill, H. S. Anti-allergy properties of fermented foods: An important immunoregulatory mechanism of lactic acid bacteria. Int. Immunopharmacol. 2001, 1, 891–901. (6) Macpherson, A. J.; Harris, N. L. Interactions between commensal intestinal bacteria and the immune system. Nat. ReV. Immunol.2004, 4, 478–485. (7) Elson, C. O.; Cong, Y.; Iqbal, N.; Weaver, C. T. Immuno-bacterial homeostasis in the gut: New insights into an old enigma. Semin. Immunol. 2001, 13, 187–194. (8) Kankaanpää, P.; Sütas, Y.; Salminen, S.; Isolauri, E. Homogenates derived from probiotic bacteria provide down-regulatory signals for peripheral blood mononuclear cells. Food Chem. 2003, 83, 269–277. (9) Amrouche, T.; Boutin, Y.; Fliss, I. Effects of bifidobacterial cytoplasm peptide and protein fractions on mouse lymphocyte proliferation and cytokine production. Food Agric. Immunol. 2006, 17, 29–42. (10) Kagnoff, M. F.; Eckmann, L. Epithelial cells as sensors for microbial infection. J. Clin. InVest. 1997, 100, 6–10. (11) Rescigno, M.; Urbano, M.; Valzasina, B.; rancolini, M.; Rotta, G.; Bonasio, R.; Granucci, F.; Kraehenbuhl, J. P.; Ricciardi- Castagnoli, P. Dendritic cells express tight junction proteins and penetrate gut epithelial monolayers to sample bacteria. Nat. Immunol. 2001, 2, 361–367. (12) Sansonetti, P. J. War and peace at mucosal surfaces. Nat. ReV.Immunol. 2004, 4, 953–964. (13) Pochard, P.; Gosset, P.; Grangette, C.; Andre, C.; Tonnel, A. B.; Pestel, J.; Mercenier, A. Lactic acid bacteria inhibit TH2 cytokine the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine production by mononuclear cells from allergic patients. J. Allergy Clin. Immunol. 2002, 110, 617–623. (14) Fujiwara, D.; Inoue, S.; Wakabayashi, H.; Fujii, T. The antiallergic effects of lactic acid bacteria are strain dependent and mediated by effects on both Th1/Th2 cytokine expression and balance. Int. Arch. Allergy Immunol. 2004, 135, 205–215. (15) Peng, S.; Lin, J. Y.; Lin, M. Y. Antiallergic effect of milk fermented with lactic acid bacteria in a murine animal model. J. Agric. Food Chem. 2007, 55, 5092–5096. (16) Groux, H.; O’Garra, A.; Bigler, M.; Rouleau, M.; Antonenko, S.; de Vries, J. E.; Roncarolo, M. G. A CD4+ T-cell subset inhibits antigen-specific T-cell responses and prevents colitis. Nature 1997, 389, 737–742. (17) von der Weid, T.; Bulliard, C.; Schiffrin, E. J. Induction by a lactic acid bacterium of a population of CD4+ T cells with low proliferative capacity that produce transforming growth factor _ and interleukin-10. Clin. Diagn. Lab. Immunol. 2001, 8, 695–701. (18) Banchereau, J.; Steinman, R. M. Dendritic cells and the control of immunity. Nature 1998, 392, 245–252. (19) Cella, M.; Sallusto, F.; Lanzavecchia, A. Origin, maturation and antigen presenting function of dendritic cells. Curr. Opin. Immunol.1997, 9, 10–16. (20) Stagg, A. J.; Hart, A. L.; Knight, S. C.; Kamm, M. A. Microbial-gut interactions in health and disease. Interactions between dendritic cells and bacteria in the regulation of intestinal immunity. Best Pract. Res., Clin. Gastroenterol. 2004, 18, 255–270. (21) Iwasaki, A.; Kelsall, B. L. I. Mucosal dendritic cells: Their specialized role in initiating T cell responses. Am. J. Physiol. 1999, 276, G1074– G1078. (22) Niess, J. H.; Brand, S.; Gu, X.; Landsman, L.; Jung, S.; McCormick, B. A.; Vyas, J. M.; Boes, M.; Ploegh, H. L.; Fox, J. G.; Littman, D. R.; Reinecker, H. C. CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance. Science 2005, 307, 254–258. (23) Macatonia, S. E.; Hosken, N. A.; Litton, M.; Vieira, P.; Hsieh,C. S.; Culpepper, J. A.; Wysocka, M.; Trinchieri, G.; Murphy, K. M.; O’Garra, A. Dendritic cells produce IL-12 and direct the development of Th1 cells from the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine naive CD4+ T cells. J. Immunol. 1995, 154, 5071–5079. (24) Iwasaki, A.; Kelsall, B. L. Freshly isolated Peyer’s patch, but not spleen, dendritic cells produce interleukin 10 and induce the differentiation of T helper type 2 cells. J. Exp. Med. 1999, 190, 229–240. (25) Kelleher, P.; Maroof, A.; Knight, S. C. Retrovirally induced switch from production of IL-12 to IL-4 in dendritic cells. Eur.J. Immunol. 1999, 29, 2309–2318. (26) Roncarolo, M.; Levings, M. K.; Traversari, C. Differentiation of T regulatory cells by immature dendritic cells. J. Exp. Med. 2001, 193, F5– F9. (27) Kato, I.; Tanaka, K.; Yokokura, T. Lactic acid bacterium potently induces the production of interleukin-12 and interferon-γ by mouse splenocytes. Int. J. Immunopharmacol. 1999, 21, 121–131. (28) Christensen, H. R.; Frokiar, H.; Pestka, J. J. Lactobacilli differentially modulate expression of cytokines and maturation surface markers in murine dendritic cells. J. Immunol. 2002, 168, 171–178. (29) Mohamadzadeh, M.; Olson, S.; Kalina, W. V.; Ruthel, G.; Demmin, G. L.; Warfield, K. L.; Bavari, S.; Klaenhammer, T. R. Lactobacilli activate human dendritic cells that skew T cells toward T helper 1 polarization. Proc. Natl. Acad. Sci. U.S.A. 2005, 102,2880–2885. (30) Zeuthen, L. H.; Christensen, H. R.; Frokiaer, H. Lactic acid bacteria inducing a weak interleukin-12 and tumor necrosis factor R response in human dendritic cells inhibit strongly stimulating lactic acid bacteria but act synergistically with Gram-negative bacteria. Clin. Vaccine Immunol. 2006, 13, 365–375. (31) Robinson, D. S.; Kay, A. B. Role of Th1 and Th2 cells in human allergic disorders. Chem. Immunol. 1996, 63, 187–203. (32) Abbas, A. K.; Murphy, K. M.; Sher, A. Functional diversity of helper T lymphocytes. Nature 1996, 383, 787–793. (33) Trinchieri, G. Interleukin-12: A proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu. ReV. Immunol. 1995,13, 251– 276. (34) Gajewski, T. F.; Fitch, F. W. Anti-proliferative effect of IFN-γ in immune regulation. I. IFN-γ inhibits the proliferation of Th2 but not Th1 murine helper T lymphocyte clones. J. Immunol. 1988,140, 4245–4252. the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine (35) Morita, H.; He, F.; Fuse, T.; Ouwehand, A. C.; Hashimoto, H.; Hosoda, M.; Mizumachi, K.; Kurisaki, J. Cytokine production by the murine macrophage cell line J774.1 after exposure to lactobacilli. Biosci., Biotechnol., Biochem. 2002, 66, 1963–1966. (36) Gill, H. Stimulation of the immune system by lactic cultures. Int. Dairy J. 1998, 8, 535. (37) Gill, H. S.; Rutherfurd, K. J.; Prasad, J.; Gopal, P. K. Enhancement of natural and acquired immunity by Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019). Br. J. Nutr. 2000, 83, 167–176. (38) Djukanovic, R.; Wilson, J. W.; Britten, K. M.; Wilson, S. J.; Walls, A. F.; Roche, W. R.; Howarth, P. H.; Holgate, S. T. Quantitation of mast cells and eosinophils in the bronchial mucosa of symptomatic atopic asthmatics and healthy control subjects using immunohistochemistry. Am. ReV. Respir. Dis. 1990, 142, 863–871. (39) Hamelmann, E.; Gelfand, E. W. Role of IL-5 in the development of allergen-induced airway hyperresponsiveness. Int. Arch. Allergy Immunol. 1999, 120, 8–16. (40) Murosaki, S.; Yamamoto, Y.; Ito, K.; Inokuchi, T.; Kusaka, H.; Ikeda, H.; Yoshikai, Y. Heat-killed Lactobacillus plantarum L-137 suppresses naturally fed antigen-specific IgE production by stimulation of IL-12 production in mice. J. Allergy Clin. Immunol. 1998, 102, 57–64. (41) Matsuzaki, T.; Chin, J. Modulating immune responses with probiotic bacteria. Immunol. Cell Biol. 2000, 78, 67–73. Received for review June 18, 2007. Revised manuscript received October 28, 2007. Accepted November 1, 2007. This study was supported in part by grants from the Taichung Veterans General Hospital and the National Chung Hsing University (TCVGH-NCHU-977606), Taichung, Taiwan. the needs of future rehabilitation centre. facilitates future expansion of the hospital-wide voice communications, reducing costs due to expansion. Taking into account the special nature of public hospital, from a security point of view, the internal network of centres for the rehabilitation of the project system and Internet construction independently. to protect data within medical rehabilitation center from influence of external network, ensure safe and reliable medical information. Intranet for the hospital's internal network and not the hospital external link external network and an external network and data Association, two independent sets of network equipment. Medical network to four-room for 2 large Gigabit switches at the core, double-machine