alexa Replication and Translation of Sperm-introduced Hepatit
E- ISSN: 2320 - 3528
P- ISSN: 2347 - 2286

Research & Reviews: Journal of Microbiology and Biotechnology
Open Access

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Research Article

Replication and Translation of Sperm-introduced Hepatitis B Virus Genes in Embryonic Cells

Qiu-Ju Zhang1#, Ying Zhong2#, Ji-Hua Huang2, Qing-Dong Xie1, Xiao-Fang Tan1#, Chen-Yao Zhong3, Lan Xu1, Tian-Hua Huang2*

1Research Center for Reproductive Medicine, Shantou University Medical College, Shantou 515041, PR China

2Jinxin Research Institute for Reproductive Medicine and Genetics, Chengdu Jinjiang Hospital for Maternal and Child Health Care, Chengdu 610066, PR China

3 Faculty of Science and Engineering, University Paul Sabatier, Toulouse 31062, France

#Both authors contributed equally to this work

*Corresponding Author:
Tian-Hua Huang
Jinxin Research Institute for Reproductive Medicine and Genetics
Chengdu Jinjiang Hospital for Maternal and Child Health Care
Chengdu 610066, PR China
Tel: +86 28 6507 8734
Fax: +86 28 6507 8734
E-mail: [email protected]

Received 01/03/2016 Accepted 13/05/2016 Published 23/05/2016

 

Abstract

Objective: Given that hepatitis B virus (HBV) can infect human sperm cells and HBV genes introduced by sperm can be transcribed in developing embryos, we sought to confirm the presence of mature HBV antigens and to assess the mechanism of viral gene replication in embryonic cells. Methods: Human sperm were transfected with recombinant plasmids pIRES2-EGFP-HBs or pIRES2-EGFP-HBc and were then used to fertilize zona-free golden hamster ova. Two-celled embryos with and without green fluorescence were collected for detection of HBV DNA or protein by fluorescence in situ hybridization (FISH), immunofluorescence assay (IA), and enzyme-linked immunosorbent assay (ELISA). Results: 1. Positive FISH signals for HBs- and HBc-DNA were observed in the sperm nuclei and in the nuclei of the two-celled embryos in the test samples but were not detected in the controls. 2. Positive IA signals for HBsAg or HBcAg were detected within the cytoplasm of the two-celled embryos in the test samples but not in the controls. 3. Using ELISA, the level of HBsAg in a single two-celled embryo was 0.064 ng/ ml. Likewise, HBcAg was detected in the five-embryo experimental group. Conclusion: The sperm-introduced HBV S or C genes were able to replicate concomitantly host embryo genome via a semiconservative mode of DNA replication and were able to express viral proteins in the embryonic cells.

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