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Establishment of a Novel Autoimmune Experimental Model of Bladder Pain Syndrome/Interstitial Cystitis in C57BL/6 Mice

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Abstract

The aim of this study is to identify whether vaccinating twice with bladder homogenate can establish a new model of experimental autoimmune cystitis (EAC) in C57BL/6 strain mice. C57BL/6 mice were vaccinated with bladder homogenate in complete Freund’s adjuvant (CFA) and boost immunized with bladder homogenate in incomplete Freund’s adjuvant (IFA) after 2 weeks were used as the EAC model. Mice immunized with phosphate-buffered saline (PBS) in CFA or IFA were used as the control. Micturition habits and suprapubic–pelvic pain threshold were measured 4 weeks after primary immunization. Bladder to body weight ratios and expression of inflammatory cytokines and neurokinin 1 receptor (NK1R) were then examined. Histologic and immunohistochemical examination of the bladder was carried out, and IL-1β, IFN-γ, and TNF-α production by the kidneys, liver, and lungs was also tested. Double-immunized mice were extensively sensitive to pressure applied on the pelvic area (P < 0.001). Compared to single-immunized mice or controls, double-immunized mice showed more micturition frequency, lower urine output per micturition, higher bladder to body weight ratio, and significant elevation in the expression of inflammatory cytokines, including IL-1β, IL-4, IL-6, IL-10, IFN-γ, and TNF-α (all P < 0.05). NK1R gene expression was significantly increased in double-immunized mice compared to the other three groups (P < 0.001). A nonspecific immune response occurred in the liver but was much weaker than bladder inflammation. Our dual immunization EAC model in C57BL/6 mice can effectively mimic the symptoms and pathophysiologic characteristics of BPS/IC and thus can be widely used to investigate the pathogenesis and therapeutic strategies of BPS/IC.

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Abbreviations

BPS:

Bladder pain syndrome

CFA:

Complete Freund’s adjuvant

EAC:

Experimental autoimmune cystitis

IC:

Interstitial cystitis

IFA:

Incomplete Freund’s adjuvant

MHC:

Major histocompatibility complex

NK1R:

Neurokinin 1 receptor

SP:

Substance P

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Acknowledgments

This research was supported by the General Programs of the National Natural Science Foundation of China (No. 81270846) and Shanghai Municipal Commission of Health and Family Planning (No. 201540146).

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    Authors and Affiliations

    1. Department of Urology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, No. 197 Ruijin Er Road, Shanghai, 200025, China

      Xing-Wei Jin, Bo-Ke Liu, Xiang Zhang & Yuan Shao

    2. Department of Pathology, School of Basic Medical Sciences, Fudan University, Shanghai, China

      Zhong-Hua Zhao

    3. Department of Urology, Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine, Shanghai, China

      Yuan Shao

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    1. Xing-Wei Jin
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    3. Xiang Zhang
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    Corresponding author

    Correspondence to Yuan Shao.

    Ethics declarations

    All animal experiments were reviewed and approved by the Animal Care and Use Committee of Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine. The protocol was performed under the Animal Management Rule of the Ministry of Health, People’s Republic of China (documentation no. 55, 2001). All vaccinations were performed with mice under isoflurane anesthesia, and mouse sacrifice was carried out under an overdose of sodium pentobarbital.

    Competing Interests

    None.

    Authors’ Contributions

    XWJ: Data collection, analysis, and manuscript writing. BKL: Protocol development and data collection. XZ: Data collection. ZHZ: Pathologic evaluation. YS: Research design, data analysis, and manuscript editing.

    Additional information

    Xing-Wei Jin and Bo-Ke Liu contributed equally to this work.

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    Jin, XW., Liu, BK., Zhang, X. et al. Establishment of a Novel Autoimmune Experimental Model of Bladder Pain Syndrome/Interstitial Cystitis in C57BL/6 Mice. Inflammation 40, 861–870 (2017). https://doi.org/10.1007/s10753-017-0531-7

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    • DOI: https://doi.org/10.1007/s10753-017-0531-7

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