Development of a flow cytometry method for detection of bovine multi-cytokines.

Zhaocheng Zhu; Aihong Xia; Zhaoli Cao; Xin LI; Xiang Chen; Zhengzhong XU; Xin An Jiao

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Development of a flow cytometry method for detection of bovine multi-cytokines.

Author: Zhaocheng ZHU ¹ ; Aihong XIA ¹ ; Zhaoli CAO ² ; Xin LI ² ; Xiang CHEN ¹ ; Zhengzhong XU ¹ ; Xin An JIAO ¹
Author Information

1. Jiangsu Key Laboratory of Zoonosis/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, Jiangsu, China.
2. Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture and Rural Affairs, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Publication Type:Journal Article
Keywords: bovine cytokine; cellular immune response; disease diagnosis; flow cytometry; intracellular staining
MeSH: Cattle; Animals; Cytokines; BCG Vaccine/metabolism*; Tumor Necrosis Factor-alpha/metabolism*; Interleukin-2; Flow Cytometry/methods*; Chemokine CXCL10/metabolism*; Leukocytes, Mononuclear; CD4-Positive T-Lymphocytes/metabolism*; Tuberculosis; Antibodies, Monoclonal/metabolism*
From: Chinese Journal of Biotechnology 2023;39(1):347-358
CountryChina
Language:Chinese
Abstract: This study aims to develop a method to detect bovine multi-cytokines based on flow cytometry. Previously we have prepared and screened monoclonal antibodies against bovine cytokines IFN-γ, IL-2, TNF-α, IP-10 and MCP-1. These bovine cytokine monoclonal antibodies were fluorescently labeled, and the combination of antibody and cell surface molecules were used to develop the method for detecting bovine multi-cytokines. Subsequently, the developed method was used to determine the cytokine expression profile of Mycobacterium bovis BCG infected bovine peripheral blood mononuclear cells in vitro, and evaluate the cytokine expression level of peripheral blood CD4⁺ T cells of tuberculosis-positive cattle. The bovine multi-cytokine flow cytometry detection method can effectively determine the cytokine expression of BCG-infected bovine peripheral blood T lymphocytes. Among them, the expression levels of IFN-γ, IL-2, and TNF-α continue to increase after 40 hours of infection, while the expression levels of IP-10 and MCP-1 decreased. The combined detection of IFN-γ, IL-2, and TNF-α on CD4⁺ T lymphocytes in peripheral blood of cattle can effectively distinguish tuberculosis-positive and tuberculosis-negative samples. This method may facilitate evaluating the level of cellular immune response after bovine pathogen infection and vaccine injection.