scholarly journals Production of Human Papillomavirus Type 16 L1 Virus-Like Particles by Recombinant Lactobacillus casei Cells

2006 ◽  
Vol 72 (1) ◽  
pp. 745-752 ◽  
Author(s):  
Karina Araujo Aires ◽  
Aurora Marques Cianciarullo ◽  
Sylvia Mendes Carneiro ◽  
Luisa Lina Villa ◽  
Enrique Boccardo ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Lactobacillus Casei ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16 ◽  
Common Causes ◽  
Electron Microscopy Analysis ◽  
Microscopy Analysis ◽  
L1 Protein

ABSTRACT Infections with human papillomavirus type 16 (HPV-16) are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, the most promising vaccine against HPV-16 infection is based on the L1 major capsid protein, which self-assembles in virus-like particles (VLPs). In this work, we used a lactose-inducible system based on the Lactobacillus casei lactose operon promoter (plac) for expression of the HPV-16 L1 protein in L. casei. Expression was confirmed by Western blotting, and an electron microscopy analysis of L. casei expressing L1 showed that the protein was able to self-assemble into VLPs intracellularly. The presence of conformational epitopes on the L. casei-produced VLPs was confirmed by immunofluorescence using the anti-HPV-16 VLP conformational antibody H16.V5. Moreover, sera from mice that were subcutaneously immunized with L. casei expressing L1 reacted with Spodoptera frugiperda-produced HPV-16 L1 VLPs, as determined by an enzyme-linked immunosorbent assay. The production of L1 VLPs by Lactobacillus opens the possibility for development of new live mucosal prophylactic vaccines.

scholarly journals Nasal Immunization of Mice with Human Papillomavirus Type 16 (HPV-16) Virus-Like Particles or with the HPV-16 L1 Gene Elicits Specific Cytotoxic T Lymphocytes in Vaginal Draining Lymph Nodes

1999 ◽  
Vol 73 (11) ◽  
pp. 9063-9071 ◽  
Author(s):  
Catherine Dupuy ◽  
Dominique Buzoni-Gatel ◽  
Antoine Touzé ◽  
Daniel Bout ◽  
Pierre Coursaget
Keyword(s):  
T Cells ◽  
Human Papillomavirus ◽  
Immune Responses ◽  
Self Assembly ◽  
Genital Tract ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Mammalian Expression ◽  
Human Papillomavirus Type 16 ◽  
L1 Protein

ABSTRACT Human papillomavirus type 16 (HPV-16) infects the genital tract and is closely associated with the development of cervical cancer. HPV-16 initiates infection at the genital mucosal surface; thus, mucosal immune responses are likely to contribute to defense against HPV-16 infection. However, little information is available regarding the induction of immune responses in the genital tract mucosa. In this study, we evaluated the potential of intranasally administered papillomavirus vaccines to elicit both systemic and vaginal immune responses. HPV-16 virus-like particles (VLPs) produced by self-assembly of L1 protein and the HPV-16 L1 gene cloned into a mammalian expression vector were used as vaccines. Intranasally administered VLPs induced serum immunoglobulin G (IgG) and vaginal IgA secretory antibodies. Very weak serum IgG and vaginal IgA responses were found after DNA immunization. Both splenic and vaginal lymphocytes could be activated by intranasal immunization with VLPs and the HPV-16 L1 gene. Activated CD4+ Th1-like T cells were shown to synthesize gamma interferon, and activated CD8+ T cells were demonstrated to be cytotoxic.

scholarly journals The L1 Major Capsid Protein of Human Papillomavirus Type 16 Variants Affects Yield of Virus-Like Particles Produced in an Insect Cell Expression System

1998 ◽  
Vol 36 (7) ◽  
pp. 2046-2051 ◽  
Author(s):  
Antoine Touze ◽  
Slimane El Mehdaoui ◽  
Pierre-Yves Sizaret ◽  
Christine Mougin ◽  
Nubia Muñoz ◽  
...  
Keyword(s):  
Amino Acids ◽  
Human Papillomavirus ◽  
Expression System ◽  
The Philippines ◽  
Serological Tests ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16 ◽  
Amino Acid Region ◽  
L1 Protein

The L1 major capsid proteins of six human papillomavirus type 16 (HPV-16) strains were expressed in insect cells by using recombinant baculoviruses. Virus-like particles (VLPs) which appeared similar to empty virions were identified by electron microscopy for all HPV strains investigated. However, the yield of VLPs produced varied in a range from 1 to 79 depending on the HPV-16 strain. The L1 proteins of these strains differed by up to 15 amino acids from the L1 protein of the prototype HPV-16 strain. Mutations in the amino acid region from residues 83 to 97 seemed to affect the level of expression of the L1 protein. These results are important when considering the development of HPV vaccines and serological tests. They indicate that strains inducing high levels of VLP production must be selected for the development of vaccines. Moreover, the L1 proteins of all strains investigated were able to bind with DNA. We also investigated the seroreactivities of VLPs derived from three different HPV-16 strains from Algeria, Senegal, and the Philippines by testing sera from women from 11 countries in immunoglobulin G-specific enzyme-linked immunosorbent assays. We observed a strong correlation between the reactivities of the three different VLP variants, independent of the geographical origin of the sera investigated. These results indicate that the three strains investigated are serologically cross-reactive despite the fact that their L1 proteins differ in 14 amino acids and suggest that VLPs derived from only one HPV-16 strain could be sufficient for the development of an HPV-16 vaccine and anti-HPV-16 tests.

scholarly journals Nasal Immunization of Mice with Human Papillomavirus Type 16 Virus-Like Particles Elicits Neutralizing Antibodies in Mucosal Secretions

1998 ◽  
Vol 72 (10) ◽  
pp. 8220-8229 ◽  
Author(s):  
Carole Balmelli ◽  
Richard Roden ◽  
Alexandra Potts ◽  
John Schiller ◽  
Pierre De Grandi ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Antibody Response ◽  
Neutralizing Antibodies ◽  
Secretory Iga ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16 ◽  
Specific Igg ◽  
Nasal Immunization ◽  
Mucosal Secretions

ABSTRACT To specifically induce a mucosal antibody response to purified human papillomavirus type 16 (HPV16) virus-like particles (VLP), we immunized female BALB/c mice orally, intranasally, and/or parenterally and evaluated cholera toxin (CT) as a mucosal adjuvant. Anti-HPV16 VLP immunoglobulin G (IgG) and IgA titers in serum, saliva, and genital secretions were measured by enzyme-linked immunosorbent assay (ELISA). Systemic immunizations alone induced HPV16 VLP-specific IgG in serum and, to a lesser extent, in genital secretions but no secretory IgA. Oral immunization, even in the presence of CT, was inefficient. However, three nasal immunizations with 5 μg of VLP given at weekly intervals to anesthetized mice induced high (>104) and long-lasting (>15 weeks) titers of anti-HPV16 VLP antibodies in all samples, including IgA and IgG in saliva and genital secretions. CT enhanced the VLP-specific antibody response 10-fold in serum and to a lesser extent in saliva and genital secretions. Nasal immunization of conscious mice compared to anesthetized mice was inefficient and correlated with the absence of uptake of a marker into the lung. However, a 1-μg VLP systemic priming followed by two 5-μg VLP intranasal boosts in conscious mice induced both HPV16 VLP-specific IgG and IgA in secretions, although the titers were lower than in anesthetized mice given three intranasal immunizations. Antibodies in serum, saliva, and genital secretions of immunized mice were strongly neutralizing in vitro (50% neutralization with ELISA titers of 65 to 125). The mucosal and systemic/mucosal HPV16 VLP immunization protocols that induced significant titers of neutralizing IgG and secretory IgA in mucosal secretions in mice may be relevant to genital HPV VLP-based human vaccine trials.

scholarly journals Mucosal but Not Parenteral Immunization with Purified Human Papillomavirus Type 16 Virus-Like Particles Induces Neutralizing Titers of Antibodies throughout the Estrous Cycle of Mice

1999 ◽  
Vol 73 (11) ◽  
pp. 9609-9613 ◽  
Author(s):  
Denise Nardelli-Haefliger ◽  
Richard Roden ◽  
Carole Balmelli ◽  
Alexandra Potts ◽  
John Schiller ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Estrous Cycle ◽  
Neutralizing Antibodies ◽  
Female Genital Tract ◽  
Neutralization Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16 ◽  
Parenteral Immunization

ABSTRACT We have recently shown that nasal immunization of anesthetized mice with human papillomavirus type 16 (HPV16) virus-like particles (VLPs) is highly effective at inducing both neutralizing immunoglobulin A (IgA) and IgG in genital secretions, while parenteral immunization induced only neutralizing IgG. Our data also demonstrated that both isotypes are similarly neutralizing according to an in vitro pseudotyped neutralization assay. However, it is known that various amounts of IgA and IgG are produced in genital secretions along the estrous cycle. Therefore, we have investigated how this variation influences the amount of HPV16 neutralizing antibodies induced after immunization with VLPs. We have compared parenteral and nasal protocols of vaccination with daily samplings of genital secretions of mice. Enzyme-linked immunosorbent assay analysis showed that total IgA and IgG inversely varied along the estrous cycle, with the largest amounts of IgA in proestrus-estrus and the largest amount of IgG in diestrus. This resulted in HPV16 neutralizing titers of IgG only being achieved during diestrus upon parenteral immunization. In contrast, nasal vaccination induced neutralizing titers of IgA plus IgG throughout the estrous cycle, as confirmed by in vitro pseudotyped neutralization assays. Our data suggest that mucosal immunization might be more efficient than parenteral immunization at inducing continuous protection of the female genital tract.

scholarly journals Influence of Codon Bias on Heterologous Production of Human Papillomavirus Type 16 Major Structural Protein L1 in Yeast

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Milda Norkiene ◽  
Alma Gedvilaite
Keyword(s):  
Gene Expression ◽  
Human Papillomavirus ◽  
Codon Bias ◽  
Heterologous Gene Expression ◽  
Heterologous Production ◽  
Structural Protein ◽  
Hpv 16 ◽  
Major Structural Protein ◽  
Human Papillomavirus Type 16 ◽  
L1 Protein

Heterologous gene expression is dependent on multistep processes involving regulation at the level of transcription, mRNA turnover, protein translation, and posttranslational modifications. Codon bias has a significant influence on protein yields. However, sometimes it is not clear which parameter causes observed differences in heterologous gene expression as codon adaptation typically optimizes many sequence properties at once. In the current study, we evaluated the influence of codon bias on heterologous production of human papillomavirus type 16 (HPV-16) major structural protein L1 in yeast by expressing five variants of codon-modified open reading frames (OFRs) encoding HPV-16 L1 protein. Our results showed that despite the high toleration of various codons used throughout the length of the sequence of heterologously expressed genes in transformed yeast, there was a significant positive correlation between the gene's expression level and the degree of its codon bias towards the favorable codon usage. The HPV-16 L1 protein expression in yeast can be optimized by adjusting codon composition towards the most preferred codon adaptation, and this effect most probably is dependent on the improved translational elongation.

Genetic variability in the major capsid L1 protein of human papillomavirus type 16 (HPV-16) and 18 (HPV-18)

2011 ◽  
Vol 11 (8) ◽  
pp. 2119-2124 ◽  
Author(s):  
E. Frati ◽  
S. Bianchi ◽  
D. Colzani ◽  
A. Zappa ◽  
G. Orlando ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Genetic Variability ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
L1 Protein ◽  
Hpv 18

Chimeric Virus-like Particles of the Human Papillomavirus Type 16 (HPV 16) as a Prophylactic and Therapeutic Vaccine

1999 ◽  
Vol 30 (4) ◽  
pp. 269-274 ◽  
Author(s):  
Ingrid Jochmus ◽  
Klaus Schäfer ◽  
Stefan Faath ◽  
Martin Müller ◽  
Lutz Gissmann
Keyword(s):  
Human Papillomavirus ◽  
Therapeutic Vaccine ◽  
Chimeric Virus ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16

Human Papillomavirus Type 16 Exists in Bacteria Isolated from Cervical Cancer Biopsies

2009 ◽  
Vol 37 (4) ◽  
pp. 1065-1074 ◽  
Author(s):  
Zhenghai Ma ◽  
Lihong Liu ◽  
Fuchun Zhang ◽  
Meng Yu ◽  
Kai Wang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
Reverse Transcription Pcr ◽  
Bacterial Rna ◽  
L1 Protein

This study investigated the association between infectious microbes and persistent infection with human papillomavirus type 16 (HPV-16) in cervical cancer. Bacterial strains (identified as Enterococcus, Staphylococcus, Bacillus and Corynebacterium, based on their partial 16S rDNA sequence) were HPV-16 positive from 12 out of 14 cervical cancer biopsies. Total DNA was isolated from the four bacterial strains, and HPV-16 genes and genome were detected using polymerase chain reaction (PCR) and Southern blotting. RNA transcripts for HPV-16 E6 and L1 genes were detected in total bacterial RNA samples using reverse transcription-PCR, and HPV-16 L1 protein expression was detected in bacterial cells by Western blotting and immunocolloidal gold electron microscopy. The presence of virus particles in bacterial cells was demonstrated by transmission electron microscopy. The results suggest that bacteria carrying HPV-16 could provide a potential explanation for how infectious microbes contribute to the progression from HPV-16 infection to cervical cancer.

scholarly journals Type-specific and cross-reactive antibodies induced by human papillomavirus 31 L1/L2 virus-like particles

2007 ◽  
Vol 56 (7) ◽  
pp. 907-913 ◽  
Author(s):  
Yufei Xu ◽  
Qingyong Wang ◽  
Yehua Han ◽  
Guoxing Song ◽  
Xuemei Xu
Keyword(s):  
Human Papillomavirus ◽  
Western Blotting ◽  
Neutralizing Antibodies ◽  
Cross Reaction ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Hpv Types ◽  
Cross Neutralization ◽  
The Cross ◽  
L1 Protein

The aim of this study was to determine whether antibodies induced by human papillomavirus (HPV) type 31 L1/L2 virus-like particles (VLPs) could cross-react with VLPs of the closely related HPV-16 and distantly related HPV-11, and to investigate the potential role of the L2 protein in L1/L2 VLPs in inducing cross-neutralizing antibodies. Antisera were prepared from rabbits immunized with intact or denatured HPV-31 L1/L2 VLPs. Cross-reaction and cross-neutralization were analysed by Western blotting and ELISA, and by haemagglutination inhibition, respectively. Western blotting results showed that H31 L1/L2 (D) antiserum (antiserum from a rabbit immunized with denatured HPV-31 L1/L2 VLPs) could cross-react with the L1 protein of HPV-11 and -16. HPV-31 L1/L2 VLP antiserum showed strong cross-reaction with and cross-neutralization of HPV-16 VLPs, but this was significantly less with HPV-11 VLPs. In addition, the cross-neutralizing activity against HPV-16 L1/L2 VLPs was slightly higher than that against HPV-16 L1 VLPs, although the difference was not statistically significant. Epitope-blocking ELISA showed that mAb H16.V5 could partially inhibit the cross-reaction of HPV-31 L1/L2 VLP antiserum with HPV-16 L1/L2 VLPs. These results suggested that (i) H31 L1/L2 (D) antiserum could cross-react with L1 protein from both closely related and distantly related HPV types, but HPV-31 L1/L2 VLP antiserum could only cross-neutralize closely related HPV types, (ii) surface-exposed epitopes of the L2 protein in L1/L2 VLPs may induce only low titres of cross-neutralizing antibodies and (iii) certain epitopes that cross-reacted with HPV-31 L1/L2 VLP antiserum are located close to the epitopes recognized by mAb H16.V5. These findings may provide suggestions for the design of multivalent VLP vaccines.

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