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Hum. Reprod. Advance Access published online on May 18, 2007
Human Reproduction, doi:10.1093/humrep/dem128
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Purification of antimicrobial factors from human cervical mucus

Li Ming1,2, Pan Xiaoling2, Li Yan3, Wang Lili2, Wu Qi2, Yu Xiyong1, Wang Boyao2 and Huang Ning2,4

1 Research Center of Medical Science, Guangdong Provincial People's Hospital, Guangzhou 510080, People's Republic of China 2 Research Unit of Infection and Immunity, West China Medical Center, Sichuan University, Chengdu 610041, People's Republic of China 3 College of Basic Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, People's Republic of China

4 Correspondence address. Tel: +86-28-85503159; Fax: +86-28-85501549; E-mail: lim940523{at}yahoo.com.cn

BACKGROUND: The aim of this study was to separate bactericidal proteins from healthy female cervical mucus.

METHODS: Cervical mucus was collected and dissolved in 1% acetic acid. The antimicrobial activity of acid-soluble extracts was detected by gel overlay assay against Escherichia coli ATCC 43827. The extracts showed considerable amount of antibacterial activity with a clearly visible band. The bactericidal band was purified by reverse-phase high performance liquid chromatography and the antibacterial activity of the eluate was examined using radial diffusion assay.

RESULTS: Two antimicrobial proteins were purified and were further characterized by Tricine sodium dodecyl sulphate-polyacrylamide gel electrophoresis, N-terminal sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The proteins were identified as high-mobility group nucleosomal-binding domain 2 (HMG N2) and secretory leukocyte peptidase inhibitor (SLPI). SLPI is an antimicrobial peptide already known in the cervical mucus while HMG N2 in the cervical mucus had not been previously reported. The expression of HMG N2 mRNA was detected in Hela cells and cervical epithelial cells by RT-PCR. Slit hybridization showed abundant amounts of the HMG N2 protein in the cervical mucus.

CONCLUSIONS: These results suggest that the expression of HMG N2 and SLPI in the healthy female cervical mucus may be relevant to their immune surveillance and defense against potential pathogens in human reproductive system.

Key words: innate immune/antimicrobial protein/cervical mucus/high-mobility group nucleosomal-binding domain 2/secretory leukocyte peptidase inhibitor

Submitted on January 29, 2007; resubmitted on April 8, 2007; accepted on April 17, 2007.


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