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Generation and characterization of a bivalent protein boost for future clinical trials: HIV-1 subtypes CR01_AE and B gp120 antigens with a potent adjuvant
- Source :
- PLoS ONE, Vol 13, Iss 4, p e0194266 (2018), PLoS ONE
- Publication Year :
- 2018
- Publisher :
- Public Library of Science (PLoS), 2018.
-
Abstract
- The RV144 Phase III clinical trial with ALVAC-HIV prime and AIDSVAX B/E subtypes CRF01_AE (A244) and B (MN) gp120 boost vaccine regime in Thailand provided a foundation for the future development of improved vaccine strategies that may afford protection against the human immunodeficiency virus type 1 (HIV-1). Results from this trial showed that immune responses directed against specific regions V1V2 of the viral envelope (Env) glycoprotein gp120 of HIV-1, were inversely correlated to the risk of HIV-1 infection. Due to the low production of gp120 proteins in CHO cells (2-20 mg/L), cleavage sites in V1V2 loops (A244) and V3 loop (MN) causing heterogeneous antigen products, it was an urgent need to generate CHO cells harboring A244 gp120 with high production yields and an additional, homogenous and uncleaved subtype B gp120 protein to replace MN used in RV144 for the future clinical trials. Here we describe the generation of Chinese Hamster Ovary (CHO) cell lines stably expressing vaccine HIV-1 Env antigens for these purposes: one expressing an HIV-1 subtype CRF01_AE A244 Env gp120 protein (A244.AE) and one expressing an HIV-1 subtype B 6240 Env gp120 protein (6240.B) suitable for possible future manufacturing of Phase I clinical trial materials with cell culture expression levels of over 100 mg/L. The antigenic profiles of the molecules were elucidated by comprehensive approaches including analysis with a panel of well-characterized monoclonal antibodies recognizing critical epitopes using Biacore and ELISA, and glycosylation analysis by mass spectrometry, which confirmed previously identified glycosylation sites and revealed unknown sites of O-linked and N-linked glycosylations at non-consensus motifs. Overall, the vaccines given with MF59 adjuvant induced higher and more rapid antibody (Ab) responses as well as higher Ab avidity than groups given with aluminum hydroxide. Also, bivalent proteins (A244.AE and 6240.B) formulated with MF59 elicited distinct V2-specific Abs to the epitope previously shown to correlate with decreased risk of HIV-1 infection in the RV144 trial. All together, these results provide critical information allowing the consideration of these candidate gp120 proteins for future clinical evaluations in combination with a potent adjuvant.
- Subjects :
- RNA viruses
Glycosylation
Physiology
HIV Antigens
Glycobiology
Polysorbates
lcsh:Medicine
HIV Infections
HIV Antibodies
HIV Envelope Protein gp120
Pathology and Laboratory Medicine
Biochemistry
Antigen-Antibody Reactions
Immunologic Adjuvants
Epitopes
Immunodeficiency Viruses
Immune Physiology
Cricetinae
Medicine and Health Sciences
Public and Occupational Health
Post-Translational Modification
lcsh:Science
Immune Response
Mammals
AIDS Vaccines
Vaccines
Immune System Proteins
Eukaryota
virus diseases
CHO cells
Animal Models
Vaccination and Immunization
Recombinant Proteins
Infectious Diseases
Experimental Organism Systems
Medical Microbiology
Viral Pathogens
Viruses
Vertebrates
Cell lines
Female
Pathogens
Biological cultures
Research Article
Squalene
Infectious Disease Control
Immunology
Guinea Pigs
Microbiology
Rodents
Cricetulus
Adjuvants, Immunologic
Retroviruses
Animals
Humans
Antigens
Microbial Pathogens
Lentivirus
lcsh:R
Organisms
Biology and Life Sciences
HIV
Proteins
Antibodies, Neutralizing
Research and analysis methods
Amniotes
HIV-1
lcsh:Q
Preventive Medicine
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 13
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.pmid.dedup....e88ba7add8390f3b27311973dd84bdcc