Screening of Anti-Hepatitis B Virus Polypeptides

Screening of Anti-Hepatitis B Virus Polypeptides

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Author(s)

Author(s): Hui Wang, Wenqiong Wang, Jiejie Yang, Lin Hou, Ning Li

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DOI: 10.18483/ijSci.1881 43 118 65-69 Volume 7 - Dec 2018

Abstract

Objective: HepG2.215 cells are used as an experimental model, the central interaction region (CID) of MxA protein is divided into different regions according to its structure, and different plasmids are constructed to find the most powerful functional polypeptide against hepatitis B virus. Methods: Using full-length MxA plasmid as a template and pCMV-tag-3A as a vector, we constructed plasmids A1, A1A2, and A2A3 according to the CID. The plasmids were sequenced and transfected into HepG2.2.15 cells respectively. 24 hours later, the expression of the plasmid was detected by qRT-PCR. And the expression of HBsAg and HBeAg were detected by enzyme-linked immunosorbent assay (ELISA). The relative DNA expression of HBV DNA was detected by qRT-PCR. Combined with the peptide structure and the screening results, the segment was again divided into different short peptides, which were constructed into different plasmids and transfected into cells. And we detected the corresponding indicators again. Thereby repeat in this cycle until the shortest functional polypeptide is found. Result: The constructed plasmids were correct and their expression was normal. The expression of HBV DNA, HBsAg and HBeAg in the A1 group were significantly lower than that in the other two groups, and it was equivalent to the CID group. Separating the A1 component into A1N, A1C, the results of the test showed that the expression levels of HBV DNA, HBsAg and HBeAg in the two groups were higher than those in the A1 group and the CID group, and there was no difference between the two groups and the blank group.

Keywords

HBV, DNA, Peptide, Plasmid Construction

References

  1. K. N. D. kim k H, seong B L, "Discovery and development of anti-HBV agents and their resistance," Molecules, vol. 15, p. 30, 2010.
  2. N. M. Lozano R, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, "Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010_ a systematic analysis for the Global Burden of Disease Study 2010," Lancet, vol. 380, p. 33, 2012.
  3. M. B. Lok AS, "AASLD Practice Guidelines.Chronic hepatitis B update of therapeutic guidelines," Gastroentero, vol. 12, p. 5, 2004.
  4. Senko Tsukuda, 2 Koichi Watashi,1,3,4 Taichi Hojima,5 Masanori Isogawa,6 Masashi Iwamoto,1,3 Katsumi Omagari,6, H. A. Ryosuke Suzuki, 1 Soichi Kojima,2 Masaya Sugiyama,7 Akiko Saito,5 Yasuhito Tanaka,6 Masashi Mizokami,7, and a. T. W. Camille Sureau, "A New Class of Hepatitis B and D Virus Entry Inhibitors, Proanthocyanidin and Its Analogs, That Directly Act on the Viral Large Surface Proteins," HEPATOLOGY, vol. 65, p. 13, 2017.
  5. C. Peltekian, E. Gordien, F. Garreau, V. Meas-Yedid, P. Soussan, V. Willams, et al., "Human MxA protein participates to the interferon-related inhibition of hepatitis B virus replication in female transgenic mice," J Hepatol, vol. 43, pp. 965-72, Dec 2005.
  6. N. Li, L. Zhang, L. Chen, W. Feng, Y. Xu, F. Chen, et al., "MxA inhibits hepatitis B virus replication by interaction with hepatitis B core antigen," Hepatology, vol. 56, pp. 803-11, Sep 2012.
  7. S. C. Deres K, Paessens A, Goldmann S, Hacker HJ, Weber O, Krämer T, Niewöhner, P. U. U, Stoltefuss J, Graef E, Koletzki D, Masantschek RN,Reimann A, Jaeger R,, and B. B. Gross R, Schlemmer KH, Haebich D, R ü bsamen-Waigmann H., "Inhibition of hepatitis B virus replication by drug-induced depletion of nucleocapsids," Science, vol. 299, p. 4, 2003.
  8. M. Lutgehetmann, T. Bornscheuer, T. Volz, L. Allweiss, J. H. Bockmann, J. M. Pollok, et al., "Hepatitis B virus limits response of human hepatocytes to interferon-alpha in chimeric mice," Gastroenterology, vol. 140, p. 12, Jun 2011.
  9. M. Fernandez, J. A. Quiroga, and V. Carreno, "Hepatitis B virus downregulates the human interferon-inducible MxA promoter through direct interaction of precore/core proteins," J Gen Virol, vol. 84, pp. 2073-82, Aug 2003.
  10. H. Xiao, M. J. Killip, P. Staeheli, R. E. Randall, and D. Jackson, "The human interferon-induced MxA protein inhibits early stages of influenza A virus infection by retaining the incoming viral genome in the cytoplasm," J Virol, vol. 87, pp. 13053-8, Dec 2013.
  11. M. Wadhwa, M. Subramanyam, S. Goelz, J. Goyal, V. Jethwa, W. Jones, et al., "Use of a standardized MxA protein measurement-based assay for validation of assays for the assessment of neutralizing antibodies against interferon-beta," J Interferon Cytokine Res, vol. 33, pp. 660-71, Nov 2013.
  12. N. I. Maria, Z. Brkic, M. Waris, C. G. van Helden-Meeuwsen, K. Heezen, J. P. van de Merwe, et al., "MxA as a clinically applicable biomarker for identifying systemic interferon type I in primary Sjogren's syndrome," Ann Rheum Dis, vol. 73, pp. 1052-9, Jun 2014.
  13. E. Gordien, O. Rosmorduc, C. Peltekian, F. Garreau, C. Brechot, and D. Kremsdorf, "Inhibition of hepatitis B virus replication by the interferon-inducible MxA protein," J Virol, vol. 75, pp. 2684-91, Mar 2001.
  14. G. Wu, B. Liu, Y. Zhang, J. Li, A. Arzumanyan, M. M. Clayton, et al., "Preclinical characterization of GLS4, an inhibitor of hepatitis B virus core particle assembly," Antimicrob Agents Chemother, vol. 57, pp. 5344-54, Nov 2013.
  15. M. R. Campagna, F. Liu, R. Mao, C. Mills, D. Cai, F. Guo, et al., "Sulfamoylbenzamide derivatives inhibit the assembly of hepatitis B virus nucleocapsids," J Virol, vol. 87, pp. 6931-42, Jun 2013.
  16. M. H. Cho, J. S. Song, H. J. Kim, S. G. Park, and G. Jung, "Structure-based design and biochemical evaluation of sulfanilamide derivatives as hepatitis B virus capsid assembly inhibitors," J Enzyme Inhib Med Chem, vol. 28, pp. 916-25, Oct 2013.

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International Journal of Sciences is Open Access Journal.
This article is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) License.
Author(s) retain the copyrights of this article, though, publication rights are with Alkhaer Publications.

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