應用免疫組織化學染色法及電子顯微鏡技術研究豬迴腸培耶氏斑圓頂上皮M細胞之來源、分佈及細胞形態

Immunohistochemical and Transmission Electron Microscopical Studies on the Distribution, Origin and Morphology of M Cell in Peyer's Patches of Porcine Ileum

周國政(Gwo-Jeng Jou) ； 劉瑞生(Michael RS Liu) ； 劉振軒(Chen-Hsuan Liu) ； 朱瑞民(Redman M Chu)

《中華民國獸醫學會雜誌》 22卷1期 (1996/02) Pp. 35-48

https://doi.org/10.7009/JTCSVS.199602.0035

摘要

本研究之目的是藉免疫組織化學染色法及穿透式電子顯微鏡技術瞭解豬迴腸培耶氏斑圓頂上皮M細胞之來源、分佈及細胞形態。首先以10％中性福馬林或絕對酒精固定的豬迴腸組織石蠟切片進行卵白－生物素酵素複合體（Avidin-Biotin Enzyme Complex, ABC），6種不同檢查包括Vimentin，Desmin，S-100（P），MAK-6（P），Factor 8（P）與Cytokeratin 18（M）免疫組織化學染色，結果顯示M細胞內不具Vimentin及Desmin這2種Intermediate filaments，也不顯現S-100蛋白質成分，同時也不表現出Factor-8 相關抗原，但MAK-6染色結果則顯示，幾乎所有迴腸上皮細胞及腺窩均表現出強烈陽性呈色。同時Intermediate filament-Cytokeratin 18亦在M細胞處表現出強烈特異性呈色。其次以Cytokeratin 18染色與光學顯微鏡檢查比較在不同年齡（1日齡到20週齡）及不同飼養環境下，M細胞於豬迴腸中數量及分佈位置之差異。顯示組織以10％中性福馬林固定者，幾乎完全無陽性呈色，而以絕對酒精固定者，則有強烈陽性呈色。M細胞之分佈主要在培耶氏斑圓頂上皮區域及近腺窩上皮。有關M細胞在傳統不同年齡健康豬迴腸中之平均數量，在16週齡之前，隨著年齡的漸增而增多，但在16週齡之後，則反是隨著年齡的漸增而減少。此外，圓頂上皮中，M細胞與非M細胞之比例，則是隨著年齡的漸增而降低。另外以穿透式電子顯微鏡觀察豬迴腸M細胞之顯微構造。結果可在圓頂上皮中見到具典型細胞型態的M細胞，M細胞之形態包括腸腔表面具短、稀疏而不規則的微細絨毛，細胞質頂端有大量胞飲小囊泡，上皮內淋巴球等。同時在近腺窩部則常可見到細胞形態與M細胞相似卻不含上皮內淋巴球的細胞。由它們分佈位置的相關性與細胞形態的相似性來推判，M細胞底是由腺窩部細胞往上移行而來的。

關鍵字

免疫組織化學染色法 ； 培耶氏斑 ； M細胞

並列摘要

The purpose of our study is to detect the distribution, origin and morphology of M cells in dome epithelium of porcine ileal Peyer's patches by immunohistochemistry and electron microscopy. First, the paraffin sections of porcine ileal Peyer's patches, fixed in buffered formaldehyde or absolute alcohol, were stained by the peroxidase-anti-peroxidase method. The results reveal that there were no vimentin or desmin in the cytoplasm of M cells and there were also no appearances of S-100 protein or Factor 8 related antigen in M cells. But the results of MAK-6 stain reveals that almost all the ileal epitheIium were intensely labelled by the polyclonal antibody and M cells were also intensely labelled by the anti-cytokeratin 18 antibody. In the second, detection of the differences of amount and distribution of M cells in porcine ileum between different ages and circumstances was studies by cytokeratin 18 stain. The result reveals that the ileal tissues, fixed in absolute alcohol, were intensely labelled but were poorly labelled of the buffered formaldehyde fixed tissues. Microscopic observation reveals that the distribution of M cells were mainly in the dome epithelium of Peyer's patches and near the crypt. According to the observation, we propose that M cells must be the epithelial origin, especially the cryptal stem cells. About the average amount of M cells in conventional healthy porcine ileum, the amount increased according the age before the age of 16 weeks but decreased according the age after the age of 16 weeks. In addition, the ratio of M cells to non-M cells in the dome epithelium decreased according the age. Finally, we can observe typical M cells in the dome epithelium under the transmission electron microscope: irregular, short and sparse microvilli in the surface of the intestinal lumen, endocytoplasmic vesicles and intraepithelial lymphocytes. We can also observe some cells near the crypt similar to the M cells in morphology but without intraepithelial lymphocytes in their cytoplasm. Because of the relationship of distribution and the similarity of the morphology between M cells and those M-cell-like cells, we think that M cells must migrate from the crypt.

並列關鍵字

Immunohistochemistry ； Peyer's patches ； M cell