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Chu S, Zhu X, You N, et al. The Fab Fragment of a Human Anti-Siglec-9 Monoclonal Antibody Suppresses LPS-Induced Inflammatory Responses in Human Macrophages. Front Immunol. 2016;7:649doi: 10.3389/fimmu.2016.00649.
Chu, S., Zhu, X., You, N., Zhang, W., Zheng, F., Cai, B., Zhou, T., Wang, Y., Sun, Q., Yang, Z., Zhang, X., Wang, C., Nie, S., Zhu, J., & Wang, M. (2016). The Fab Fragment of a Human Anti-Siglec-9 Monoclonal Antibody Suppresses LPS-Induced Inflammatory Responses in Human Macrophages. Frontiers in immunology, 7649. https://doi.org/10.3389/fimmu.2016.00649
Chu, Sasa, et al. "The Fab Fragment of a Human Anti-Siglec-9 Monoclonal Antibody Suppresses LPS-Induced Inflammatory Responses in Human Macrophages." Frontiers in immunology vol. 7 (2016): 649. doi: https://doi.org/10.3389/fimmu.2016.00649
Chu S, Zhu X, You N, Zhang W, Zheng F, Cai B, Zhou T, Wang Y, Sun Q, Yang Z, Zhang X, Wang C, Nie S, Zhu J, Wang M. The Fab Fragment of a Human Anti-Siglec-9 Monoclonal Antibody Suppresses LPS-Induced Inflammatory Responses in Human Macrophages. Front Immunol. 2016 Dec 26;7:649. doi: 10.3389/fimmu.2016.00649. eCollection 2016. PMID: 28082984; PMCID: PMC5183739.
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Front Immunol. 2016 Dec 26;7:649. doi: 10.3389/fimmu.2016.00649. eCollection 2016.

The Fab Fragment of a Human Anti-Siglec-9 Monoclonal Antibody Suppresses LPS-Induced Inflammatory Responses in Human Macrophages.

Frontiers in immunology

Sasa Chu, Xuhui Zhu, Na You, Wei Zhang, Feng Zheng, Binggang Cai, Tingting Zhou, Yiwen Wang, Qiannan Sun, Zhiguo Yang, Xin Zhang, Changjun Wang, Shinan Nie, Jin Zhu, Maorong Wang

Affiliations

  1. Department of Infectious Disease, Anhui Medical University Affiliated with Bayi Clinical College, Hefei, China; Institute of Liver Disease, Nanjing Jingdu Hospital, Nanjing, China.
  2. Huadong Medical Institute of Biotechniques, Nanjing, China; Department of Emergency Medicine, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.
  3. Department of Emergency Medicine, Jinling Hospital, Medical School of Nanjing University , Nanjing , China.
  4. Huadong Medical Institute of Biotechniques , Nanjing , China.
  5. Institute of Liver Disease, Nanjing Jingdu Hospital , Nanjing , China.
  6. Department of Traditional Chinese Pharmacology, Chinese Pharmaceutical University , Nanjing , China.
  7. Huadong Medical Institute of Biotechniques, Nanjing, China; Department of Pathology, Key Laboratory of Antibody Technique of the Ministry of Health, NJMU, Nanjing, China.

PMID: 28082984 PMCID: PMC5183739 DOI: 10.3389/fimmu.2016.00649

Abstract

Sepsis is a major cause of death for hospitalized patients and is characterized by massive overreaction of immune responses to invading pathogens which is mediated by cytokines. For decades, there has been no effective treatment for sepsis. Sialic acid-binding, Ig-like lectin-9 (Siglec-9), is an immunomodulatory receptor expressed primarily on hematopoietic cells which is involved in various aspects of inflammatory responses and is a potential target for treatment of sepsis. The aim of the present study was to develop a human anti-Siglec-9 Fab fragment, which was named hS9-Fab03 and investigate its immune activity in human macrophages. We began by constructing the hS9-Fab03 prokaryotic expression vector from human antibody library and phage display. Then, we utilized a multitude of assays, including SDS-PAGE, Western blotting, ELISA, affinity, and kinetics assay to evaluate the binding affinity and specificity of hS9-Fab03. Results demonstrated that hS9-Fab03 specifically bind to Siglec-9 antigen with high affinity, and pretreatment with hS9-Fab03 could attenuate lipopolysaccharide (LPS)-induced TNF-α, IL-6, IL-1β, IL-8, and IFN-β production in human PBMC-derived macrophages, but slightly increased IL-10 production in an early time point. We also observed similar results in human THP-1-differentiated macrophages. Collectively, we prepared the hS9-Fab03 with efficient activity for blocking LPS-induced pro-inflammatory cytokines production in human macrophages. These results indicated that ligation of Siglec-9 with hS9-Fab03 might be a novel anti-inflammatory therapeutic strategy for sepsis.

Keywords: Fab fragment; LPS; Siglec-9; TLR4; human anti-Siglec-9 antibody; human macrophages; sepsis

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