China Journal of Oral and Maxillofacial Surgery ›› 2019, Vol. 17 ›› Issue (3): 216-220.doi: 10.19438/j.cjoms.2019.03.005

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Application of double stained immunohistochemistry technology in lymphoepithelial lesions of salivary gland

GU-Ting, HU Yu-hua, XIA Rong-hui, TIAN Zhen, WANG Li-zhen, ZHANG Chun-ye, LI Jiang   

  1. Department of Oral Pathology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine;National Clinical Research Center for Oral Disease;Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology.Shanghai 200011, China
  • Received:2018-10-17 Revised:2019-01-16 Online:2019-05-20 Published:2019-06-21

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Abstract: PURPOSE: Depending on the sites of positive antibody expression (nucleus, cytoplasm and membrane), this study was aimed to investigate the best method of applying double stained immunohistochemistry technology in lymphoepithelial lesions of salivary gland. METHODS: The expression of AE1/AE3，Ki-67 and CD20cy was examined by single stained immunohistochemistry and double stained immunohistochemistry respectively in 20 benign lymphoepithelial lesions (BLEL), 20 lymphoepithelial carcinomas (LEC) and 20 extranodal marginal zone lymphomas of mucosa-associated lymphoid tissue (MALT) . Depending on the different sites of antibody expression (nucleus, cytoplasm and membrane), the sequence of staining and chromogenic system, three methods of double stained immunohistochemistry were performed: ① Ki-67(DAB chromogen)，AE1/AE3 or CD20cy(AEC chromogen); ②AE1/AE3 or CD20cy(DAB chromogen)，Ki-67(AEC chromogen); ③ Ki-67(AEC chromogen)，AE1/AE3 or CD20cy(BCIP/NBT chromogen). The results of double stained immunohistochemistry were compared with single stained immunohistochemistry. The data were analyzed by using SPSS 17.0 software package. RESULTS: The results of immunohistochemistry demonstrated specificity and accuracy of the immunostaining procedures in all three methods, no significant difference of the positive strength (P=0.765) and positive ratio of antibody (P>0.05) was found in method 1. Otherwise, there were significant differences of the positive strength and positive ratio of antibody in method 2 and 3 (P<0.05). CONCLUSIONS: The best method is that nucleus positive antibody should be chosen first (Ki-67), followed by DAB chromogen, cytoplasm and membrane positive antibody should be chosen second(AE1/AE3 or CD20cy), combined with AEC chromogen for double stained immunohistochemistry in lymphoepithelial lesions of salivary gland.

Key words: Double stained immunohistochemistry technology, Benign lymphoepithelial lesion, Lymphoepithelial carcinoma, Extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue, Strength of positivity, Positive ratio

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