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Recombinant Antigens E6 and E7 of Human Papilloma Virus Type 16: Preparation, Purification, and Assessment of the Immunogenicity and Antitumor Effects In Vivo

Authors
  • Subbotina, M. E.1, 2
  • Grunina, T. M.1, 2
  • Sergienko, O. V.2
  • Elkina, N. V.3
  • Elkin, D. S.3
  • Novikova, M. V.3
  • Kopnin, P. B.3
  • Vinokurova, S. V.3
  • 1 Centre for Strategic Planning of FMBA of Russia, Moscow, 119121, Russia , Moscow (Russia)
  • 2 All-Russia Research Institute of Agricultural Biotechnology, Moscow, 127550, Russia , Moscow (Russia)
  • 3 Blokhin Russian Cancer Research Center, Moscow, 115478, Russia , Moscow (Russia)
Type
Published Article
Journal
Applied Biochemistry and Microbiology
Publisher
Pleiades Publishing
Publication Date
May 01, 2021
Volume
57
Issue
3
Pages
392–402
Identifiers
DOI: 10.1134/S0003683821030121
Source
Springer Nature
Keywords
License
Yellow

Abstract

AbstractThe recombinant proteins E6-CBD and E7-CBD, which are antigens E6 and E7 of human papillomavirus type 16 (HPV16), were connected by a glycine-serine spacer with the cellulose-binding domain (CBD) of Anaerocellum thermophilum in preparations obtained via synthesis in a heterologous expression system in Escherichia coli. A simple, effective, one-step technology for the isolation and purification of recombinant proteins E6-CBD and E7-CBD has been developed. The presence of antigenic properties in the obtained protein preparations was confirmed via Western blot. Three variants of immunogenic compositions with the recombinant proteins E6-CBD and E7-CBD and a three-dose immunization scheme have been developed. The immunogenicity was assessed in the BALB/c mice, and the antitumor effect of immunogenic compositions was studied on an experimental model of BALB/c mice with HPV16-associated tumors. It is shown that E6-CBD and E7-CBD are highly immunogenic and have antitumor effects in vivo. Both proteins, E6-CBD and E7-CBD, must be present to achieve the highest efficiency as part of the immunogenic composition. The obtained HPV16 recombinant antigens E6-CBD and E7-CBD can be used in two ways: (a) in clinical practice as the main specific components of therapeutic vaccines for the treatment of HPV16-associated neoplastic lesions of various localizations (cervical cancer, carcinoma of the anogenital tract, oropharynx) and (b) in diagnostics for the development of diagnostic kits to detect antibodies to E6 and E7 oncoproteins in patients with HPV16-associated malignant neoplasms.

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