Selective induction of Th2 cells in murine Peyer's patches by oral immunization.
Selective induction of Th2 cells in murine Peyer's patches by oral immunization.
复制标题
通过口服免疫选择性诱导小鼠派尔氏淋巴集结中的 Th2 细胞。
DOI：
10.1093/intimm/4.4.433
复制
发表时间：
1992
影响因子：
4.4
通讯作者：
McGhee,JR
中科院分区：
文献类型：
--
作者：
Xu-Amano,J;Aicher,WK;Taguchi,T;Kiyono,H;McGhee,JR
我们使用了三种不同的方法来测定小鼠Peyer’s patches （PP）口服绵羊红细胞（SRBC）免疫后辅助性T细胞（Th）的反应，包括Th1和Th2型。这些包括：(1)使用细胞因子特异性（IFN-γ和IL-5），单细胞检测分别估计Th1和Th2细胞的频率，（II）细胞因子特异性mRNA- cdna点印迹和Northern凝胶杂交检测特异性mRNA的水平，以及（II） T细胞克隆技术确定Th1和Th2克隆的频率。小鼠分别通过口服和口服途径免疫SRBC。分离PP和脾（SP） CD3+和CD3+CD4+T细胞亚群，并与抗原、饲养细胞和IL-2一起培养，并在不同的时间间隔（0,1,3和6天）使用酶联免疫斑点（ELISPOT）方法评估产生IFN-γ或IL-5的T细胞数量。口服SRBC小鼠PP或SP培养的T细胞中，IL-5斑点形成细胞（SFC）的频率较高，IFN-γSFC的数量较少。然而，免疫小鼠脾脏的CD3+T细胞和CD3+CD4+Th细胞主要表现出IFN-γSFC， IL-5 SFC数量较少，mRNA分析支持了这种独特的细胞因子产生模式，口服诱导小鼠PP T细胞培养物中IL-5特异性mRNA水平较高，而免疫小鼠SP CD3+T细胞和CD3+CD4+Th细胞培养物中IFN-γmRNA含量较高。对经口腔或系统免疫小鼠克隆的Th细胞中IFN-γ或IL-5 SFC的频率进行评估时，经SRBC口腔免疫小鼠的PP克隆的Th细胞中有74%为IL-5产生者（Th2型），而经ipp途径免疫小鼠的SP克隆的Th细胞中有67%为IFN-γ产生者（Th1型）。我们的研究表明，通过全身途径给予抗原的小鼠SP中产生IFN-γ的th1型细胞频率较高，而口服免疫导致PP中主要产生IL-5的th2型细胞。
We have used three different methods to determine the T helper (Th) cell response, Including Th1 and Th2 types, in murine Peyer's patches (PP) following oral immunization with sheep red blood cells (SRBC). These Include: (I) use of cytokine-spectfic (IFN-γand IL-5), single cell assays to estimate the frequencies of Th1 and Th2 cells respectively, (II) cytokine-speciflc mRNA-cDNA dot blot and Northern gel hybridizations to detect levels of specific mRNA, and (ill) T cell cloning techniques to determine the frequency of Th1 and Th2 clones. Mice were immunized with SRBC by either the oral or i.p. route. The PP and splenic (SP) CD3+and CD3+CD4+T cell subsets were Isolated and cultured with antigen, feeder cells, and IL-2, and were assessed at various intervals (days 0, 1, 3, and 6) for numbers of T cells producing either IFN-γor IL-5 by use of an enzyme-linked immunospot (ELISPOT) procedure. Cultures of T cells from PP or SP of mice given SRBC by the oral route had a high frequency of IL-5 spot forming cells (SFC), with lower numbers of IFN-γSFC. However, cultures of CD3+T cells and CD3+CD4+Th cells from spleens of i.p. immunized mice exhibited predominantly IFN-γSFC, with smaller but significant numbers of IL-5 SFC. This distinct pattern of cytokine production was supported by mRNA analysis where high IL-5 specific mRNA levels were noted in PP T cell cultures of orally primed mice, while IFN-γmRNA was predominant In the SP CD3+T cell and CD3+CD4+Th cell cultures from i.p. immunized mice. When the frequencies of IFN-γor IL-5 SFC were assessed among cloned Th cells from orally- or systemically-lmmunlzed mice, 74% of Th cell clones from PP of mice orally immunized with SRBC were IL-5 producers (Th2 type), while 67% of Th cell clones from SP of mice Immunized by the i.p. route were IFN-γproducers (Th1 type). Our studies show that higher frequencies of IFN-γproducing Th1-type cells occur in SP of mice given antigen by the systemic route, while oral immunization results in predominantly IL-5 producing, Th2-type cells in PP.