Structural Mimicry and Enhanced Immunogenicity of Peptide Epitopes Displayed on Filamentous Bacteriophage: The V3 Loop of HIV-1 gp120

doi:10.1006/jmbi.1994.1643

Journal of Molecular Biology

Volume 243, Issue 2, 20 October 1994, Pages 167-172

https://doi.org/10.1006/jmbi.1994.1643 Get rights and content

Abstract

The principal neutralizing determinant of the human immunodeficiency virus type 1 (HIV-1) is an intra-chain disulphide-bridged loop, designated V3, in the third hypervariable region of the surface glycoprotein gp 120. Peptide sequences from the V3 loop of gp120 from HIV-1 strain MN (HIV-1_MN) were engineered into the N-terminal region of the major coat protein of filamentous bacteriophage fd, leading to their display in multiple copies on the surface of the bacteriophage virion. Peptides displayed in this way were shown to be remarkably effective structural mimics of the natural epitope. They were recognised by human HIV antisera and evoked high titres of antibodies in mice, which cross-reacted with other strains of HIV and were capable of neutralizing the virus. In addition, antibody production could be stimulated by simultaneous inoculation with T-cell epitopes similarly displayed on filamentous bacteriophage. The bacteriophage display system offers a powerful means of studying the immunological recognition of proteins and is a promising vaccine model.

References (0)

Cited by (74)

Anti-V2 antibody deficiency in individuals infected with HIV-1 in Cameroon
2019, Virology
Citation Excerpt :
We analyzed 3–9 gp120 sequences per subject. Sequence analysis and alignment was performed using DNASTAR and Clustal W (di Marzo Veronese et al., 1994). The gp120 nucleotide sequences of 12 plasma viruses from HIV-1 infected individuals with deficient and cross-reactive anti-V2 Abs are available from GenBank with the accession numbers: MH632751– MH632762.
The results of the RV144 vaccine clinical trial showed a correlation between high level of anti-V1V2 antibodies (Abs) and a decreased risk of acquiring HIV-1 infection. This turned the focus of HIV vaccine design to the induction of elevated levels of anti-V2 Abs to increase vaccine efficacy. In plasma samples from HIV-1 infected Cameroonian individuals, we observed broad variations in levels of anti-V2 Abs, and 6 of the 79 plasma samples tested longitudinally displayed substantial deficiency of V2 Abs. Sequence analysis of the V2 region from plasma viruses and multivariate analyses of V2 characteristics showed a significant difference in several features between V2-deficient and V2-reactive plasma Abs. These results suggest that HIV vaccine immunogens containing a shorter V2 region with fewer glycosylation sites and higher electrostatic charges can be beneficial for induction of a higher level of anti-V2 Abs and thus contribute to HIV vaccine efficacy.
Phagemid vectors for phage display: Properties, characteristics and construction
2012, Journal of Molecular Biology
Citation Excerpt :
Only when there is no gene III in the genome of helper phage would five copies be present in the progeny phages.89 However, the progenies of type VIII phagemid can append much more than five copies of foreign proteins,90 even hundreds or thousands of copies, which are determined by the genome size of phagemids when the gene VIII of helper phage is missing. With the increase in the copies of displayed foreign proteins in this displaying system, the length of displayed peptides would decrease drastically to a few or a dozen amino acids.91
Phagemids are filamentous-phage-derived vectors containing the replication origin of a plasmid. Phagemids usually encode no or only one kind of coat proteins. Other structural and functional proteins necessary to accomplish the life cycle of phagemid are provided by the helper phage. In addition, other elements such as molecular tags and selective markers are introduced into the phagemids to facilitate the subsequent operations, such as gene manipulation and protein purification. This review summarizes the elements of the phagemids and their corresponding functions. Finally, the possible trends and future direction to improve the characteristics of the phagemids are highlighted.
Serum anti-p53 antibody detection in carcinomas and the predictive values of serum p53 antibodies, carcino-embryonic antigen and carbohydrate antigen 12-5 in the neoadjuvant chemotherapy treatment for III stage non-small cell lung cancer patients
2011, Clinica Chimica Acta
Citation Excerpt :
Phage display technology was once used for tumor-associated antigens and autoantibodies' screening during biopanning [18]. Portefaix et al. and di Marzo Veronese et al. [19,20] compared the immunogenicity of phage-displayed peptides and the synthetic peptides in ELISA; the outcome indicated that phage-displayed peptides had advantage over synthetic peptides inasmuch as they effectively mimicked the natural epitope structure and were correctly recognized by antibodies. With the purpose of increasing detection sensitivity and making a full use of hybrid phages' advantage, we designed the p53-ELISA and phage-ELISA double detection system to measure the titer of s-p53 Abs in cancers.
The serum p53 antibody (s-p53 Ab) is a valuable prognostic factor for carcinomas, but its common detection method, based on enzyme linked immunosorbent assay (ELISA), needs to be improved due to low sensitivity. Although neoadjuvant chemotherapy (NACT) is widely used in the treatment of non-small cell lung cancer (NSCLC) in China, forecasting chemoresistance is still a pressing problem.
Hybrid phage and wild-type p53 protein (wt p53 protein) were produced before the establishment of phage-ELISA and p53-ELISA. S-p53 Abs of 829 patients with various types of cancer was detected by a double ELISA system. 47 ΙΙΙ stage NSCLC patients treated with mitomycin, vindesine and cisplatin (MCV)-based NACT were chosen for s-p53 Abs, carcino-embryonic antigen (CEA) and carbohydrate antigen (CA) 12-5 predictive value analysis.
Through the combination of p53-ELISA and phage-ELISA (p53-phage ELISA), the sensitivity of s-p53 Abs in lung, breast, colorectal, gastric, esophageal, liver and ovarian cancer increased to 39.0%, 33.3%, 41.7%, 32.1%, 30.9%, 23.1% and 43.2% respectively. S-p53 Abs proved to correlate with nodal involvement, TNM stage, histological type (in lung cancer) or tumor size (in gastric cancer). As for the 47 ΙΙΙ stage NSCLC treated with NACT, s-p53 Abs and CA12-5 remarkably decreased after NACT treatment (P = 0.034 and P = 0.007) and pre-NACT low s-p53 Abs correlated with high objective chemoresponse rate (P = 0.016).
p53-phage ELISA system has an edge over single p53-ELISA. S-p53 Abs level correlates with cancer patients' clinicalpathological parameters and can predict the chemoresponse of ΙΙΙ stage NSCLC patients during MCV-based NACT treatment.
Antigenic properties of HCMV peptides displayed by filamentous bacteriophages vs. synthetic peptides
2008, Immunology Letters
Several efforts have been invested in the identification of CTL and Th epitopes, as well as in the characterization of their immunodominance and MHC restriction, for the generation of a peptide-based HCMV vaccine. Small synthetic peptides are, however, poor antigens and carrier proteins are important for improving the efficacy of synthetic peptide vaccines. Recombinant bacteriophages appear as promising tools in the design of subunit vaccines.
To investigate the antigenicity of peptides carried by recombinant bacteriophages we displayed different HCMV MHCII restricted peptides on the capsid of filamentous bacteriophage (fd) and found that hybrid bacteriophages are processed by human APC and activate HCMV-specific CD4 T-cells. Furthermore we constructed a reporter T-cell hybridoma expressing a chimeric TCR comprising murine αβ constant regions and human variable regions specific for the HLA-A2 restricted immunodominant NLV peptide of HCMV. Using the filamentous bacteriophage as an epitope carrier, we detected a more robust and long lasting response of the reporter T-cell hybridoma compared to peptide stimulation. Our results show a general enhancement of T-cell responses when antigenic peptides are carried by phages.
Fc receptor-mediated, antibody-dependent enhancement of bacteriophage lambda-mediated gene transfer in mammalian cells
2008, Virology
Citation Excerpt :
Since gene expression is enhanced by 10- to 50-fold by disruption of the microtubule network, it appears likely that most of the internalized phage-immune complexes are ultimately destined for degradation, under normal circumstances. The ability of phage-immune complexes to efficiently transduce FcγR-bearing mammalian cells has important implications for phage-mediated gene transfer (Eguchi et al., 2001; Hajitou et al., 2006; Ivanenkov et al., 1999; Kassner et al., 1999; Larocca et al., 1999; Sathaliyawala et al., 2006; Zanghi et al., 2007), and for applications in which phage vectors are being used to elicit immune responses to encoded or displayed antigens (Chen et al., 2001; Clark and March, 2004; di Marzo Veronese et al., 1994; Irving et al., 2001; Jepson and March, 2004; Minenkova et al., 1993; Sathaliyawala et al., 2006). For example, in vaccine applications, it may be beneficial to generate phage-immune complexes in vitro with the goal of targeting vector to FcγR-expressing antigen presenting cells (APC) in vivo.
Lambda phage vectors mediate gene transfer in cultured mammalian cells and in live mice, and in vivo phage-mediated gene expression is increased when mice are pre-immunized with bacteriophage lambda. We now show that, like eukaryotic viruses, bacteriophage vectors are subject to Fc receptor-mediated, antibody-dependent enhancement of infection in mammalian cells. Antibody-dependent enhancement of phage gene transfer required FcγRI, but not its associated γ-chain, and was not supported by other FcγR family members (FcγRIIA, FcγRIIB, and FcγRIII). Studies using chlorpromazine and latrunculin A revealed an important role for clathrin-mediated endocytosis (chlorpromazine) and actin filaments (latrunculin A) in antibody-enhanced phage gene transfer. This was confirmed by experiments using inhibitors of endosomal acidification (bafilomycin A1, monensin) and by immunocytochemical colocalization of internalized phage particles with early endosome-associated protein-1 (EAA1). In contrast, microtubule-targeting agents (nocodazole, taxol) increased the efficiency of antibody-enhanced phage gene transfer. These results reveal an unexpected antibody-dependent, FcγRI-mediated enhancement of phage transduction in mammalian cells, and suggest new approaches to improve bacteriophage-mediated gene transfer.
The detection of serum anti-p53 antibodies from patients with gastric carcinoma in China
2007, Cancer Detection and Prevention
Background: Gastric carcinoma is one of the most frequently occurring cancers. The aim of this research was to increase the detection efficiency of anti-p53 antibodies in the sera of patients with gastric carcinoma and to improve the diagnosis for patients with gastric carcinoma. Methods: We prepared phage-displayed peptide DO7 and established an enzyme-linked immunosorbent assay method to detect the anti-p53 antibodies. We detected the anti-p53 antibodies of 61 patients with gastric carcinoma using the method and our previous ELISA method assisted by the recombinant wild-type human p53 protein to detect the anti-p53 antibodies. We studied the correlation between the anti-p53 antibodies and the clinicopathological data including sex, age, carcinoembryonic antigen, tumor size, tumor TNM staging, and lymph-node status. Results: The anti-p53 antibodies positive rate for patients with gastric carcinoma was increased (31.1%, 19/61) through the combination of p53-ELISA and phage-ELISA. We found that the positive anti-p53 antibodies correlated significantly with tumor size (P = 0.047). The combination of the anti-p53 antibodies and carcinoembryonic antigen could improve the diagnosis for patients with gastric carcinoma. Conclusions: This approach indicated an increased anti-p53 antibodies positive rate for patients with gastric carcinoma and provided a useful marker for clinical diagnosis for patients with gastric carcinoma.

View all citing articles on Scopus

View full text

Journal of Molecular Biology

CommunicationStructural Mimicry and Enhanced Immunogenicity of Peptide Epitopes Displayed on Filamentous Bacteriophage: The V3 Loop of HIV-1 gp120

Abstract

Communication
Structural Mimicry and Enhanced Immunogenicity of Peptide Epitopes Displayed on Filamentous Bacteriophage: The V3 Loop of HIV-1 gp120