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Stronger In Vitro Phagocytosis by Monocytes-Macrophages Is Indicative of Greater Pathogen Clearance and Antibody Levels In Vivo

S. F. Sun^*,,, Q. Z. Pan^,, X. Hui, B. L. Zhang^*,, H. M. Wu^#, H. Li^,, W. Xu, Q. Zhang^,, J. Y. Li, X. M. Deng^,, J. W. Chen^||, Z. X. Lian^*,,,1 and N. Li^*,

^* National Key Laboratory of Agricultural Biotechnology, Beijing 100094, China; College of Animal Science and Technology, China Agricultural University, Beijing 100094, China; Laboratory of Animal Genetics and Breeding, Ministry of Agriculture, Beijing 100094, China; National Laboratory for Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; ^# Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100094, China; and ^|| University of Pennsylvania School of Veterinary Medicine, 382 West Street Road, Kennett Square, PA 19348

¹ Corresponding author: lianzhx{at}cau.edu.cn or ninglcau{at}cau.edu.cn

Monocytes-macrophages are crucial players in specific and nonspecific immune responses to protect organisms from invasion of bacteria or viruses. In this study, monocytes in circulation from 2 lines of Silky and Starbro chickens with different disease resistance were separated and cultured in vitro. After identification with acridine orange (AO), Giemsa staining, and CD14 immunostaining, monocytes-macrophages were used for adherence and phagocytosis test. The overall percentages of adherence of Silky monocytes was 1.5 times greater than that of Starbro (P < 0.01), which were 26.85% ± 8.24% and 18.34% ± 8.15%, respectively (mean ± SD). The monocytes-macrophages phagocytic index, phagocytic product, and percentage of phagocytosis in Silkies were greater than in Star-bros, respectively. The difference of phagocytic index was significant (P < 0.05), that is, 3.70 ± 1.75 and 1.97 ± 0.31, respectively (mean ± SD). Then, 20 Silkies were divided into 2 groups according to phagocytic index: high phagocytic index (HPI) group and low phagocytic index (LPI) group, to study the relationship between phagocytic activity in vitro and pathogen clearance. After being challenged against Salmonella Pullorum C79-13, the Silky birds with HPI produced a 3-fold greater level of specific antibodies compared with those with LPI (P < 0.01), 50.21 ± 6.67 and 16.85 ± 4.52, respectively (mean ± SD). In contrast to LPI birds, HPI birds shed less Salmonella Pullorum bacteria (P < 0.05), that is, 168.98 x 10⁸ ± 294.74 x 10⁸ compared to 385.40 x 10⁸ ± 399.94 x 10⁸ (mean ± SD), and the shedding peak of Salmonella Pullorum in the test span appeared 4 d earlier. These results indicated that phagocytosis of monocytes-macrophages had strong effects on antibody titer and bacteria shedding postchallenge, which could be used to predict the disease resistance in animals.

Key Words: monocyte • macrophage • phagocytosis • pathogen clearance • Salmonella Pullorum