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. 2017 Nov 27;8:1649. doi: 10.3389/fimmu.2017.01649

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Copyright © 2017 Wu, Lu, Zhang, Mei, Feng, Peng, Hou, Kang, Liu and Tang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Serum-neutralization (SN) titers and hemagglutinin inhibition (HI) antibody titers in bronchoalveolar lavage (BAL) fluids. SN assays were performed in DF-1 cells. A fixed viral solution (200 TCID₅₀, ZJ, or DT viruses) was reacted with the same volume of twofold serial diluted serum solutions. The pooled (five pooled in one) sera samples (n = 5) were derived from each group of the specific pathogen-free chickens vaccinated with rHA or Re6 vaccine with or without adjuvant. Both DT (clade 7) and ZJ (subclade 2.3.4.6) viruses were isolated from chickens (A,B). The mucosal antibodies derived from BAL fluids (n = 3) were measured by HI assay with 4 HAU of Re6, Re7, Re8, and rHA at 3-week postvaccination (C). The statistical significance was analyzed with Student’s t-test or a one-way analysis of variance. **P < 0.01, ***P < 0.001.