Isobe Naoki, Sugino Toshihisa, Taniguchi Kohzo, Moriya Naoko, Hosoda Kenji, Yoshimura Yukinori
Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
Vet Immunol Immunopathol. 2011 Jul 15;142(1-2):87-94. doi: 10.1016/j.vetimm.2011.03.020. Epub 2011 Apr 1.
Immunolocalization of lingual antimicrobial peptide (LAP), a member of the β-defensin family, in the digestive tract of calves was investigated to determine its distribution in the digestive tract of Holstein-Friesian calves. Various regions of the digestive tract were collected from slaughtered calves, and fixed in 10% formalin in PBS. Paraffin sections were stained with anti-LAP antibody, followed by visualization of immunoreactions products utilizing the avidin-biotin complex method. Expression of LAP mRNA was analyzed with reverse transcription-PCR. Immunoreactive LAP was localized in the stratum corneum of the stratified squamous epithelium of the tongue, esophagus, rumen, reticulum and omasum but not in their basal layer and lamina propria. In the gastric glands of the abomasum, only chief cells showed LAP positive reaction at the apical side of their cytoplasm. Lamina propria and Peyer's patch of the ileum had some leukocyte-like cells immunopositive for LAP. Weak immunoreaction of LAP was also detected in the mucosal epithelium of the intestinal gland of the cecum, colon and rectum. All regions of digestive tract showed LAP mRNA expression with PCR. These results indicate differential localization of LAP in the mucosal epithelium of digestive tracts in calves. The LAP expressed in stratum corneum of stratified squamous epithelium and chief cells of abomasum specifically may play role in the innate immune function in these tissues.
为了确定舌抗菌肽(LAP,β-防御素家族成员之一)在荷斯坦-弗里生犊牛消化道中的分布,对其在犊牛消化道中的免疫定位进行了研究。从屠宰的犊牛身上采集消化道的各个区域,并固定于含10%福尔马林的PBS中。石蜡切片用抗LAP抗体染色,随后利用抗生物素蛋白-生物素复合物法观察免疫反应产物。用逆转录PCR分析LAP mRNA的表达。免疫反应性LAP定位于舌、食管、瘤胃、网胃和瓣胃复层鳞状上皮的角质层,但不在其基底层和固有层。在皱胃的胃腺中,只有主细胞在其细胞质顶端显示LAP阳性反应。回肠的固有层和派伊尔结有一些对LAP呈免疫阳性的类白细胞。在盲肠、结肠和直肠肠腺的黏膜上皮中也检测到LAP的弱阳性反应。消化道的所有区域经PCR均显示LAP mRNA表达。这些结果表明LAP在犊牛消化道黏膜上皮中的定位存在差异。在复层鳞状上皮角质层和皱胃主细胞中表达的LAP可能在这些组织的固有免疫功能中发挥作用。