Table of Contents

Volume 7, Supplemental Issue

  January/February 2011

  Pages 1 - 239

Special Issue: Vaccines, immunization and immunotherapy

Guest Editor: Professor Dr Edouard Kurstak

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 Issue Catalogue (MARC XML)

EDITOR'S CORNER

Open Access Article

Preface: Seventh World Congress on Vaccines, Immunisation and Immunotherapy

Edouard Kurstak
Page 1
http://dx.doi.org/10.4161/hv.7.0.14904
Abstract | PDF

This special issue of Human Vaccines is based on the Seventh World Congress on Vaccines, Immunisation and Immunotherapy (WCVII), which was organized from 26 to 28 of May 2010 by the Infections Control World Organization (ICWO) in Berlin, Germany. The venue of the Congress at the elegant Kaiserin Friedrich Haus (previously harboring the Academy of Arts of East Germany) was located in the historic city center. The full three days of Congress with morning plenary sessions, afternoon symposia, and poster presentations were attended, from all continents, by medical specialists in immunisation, immunotherapy and scientists devoted to developing new immunogenic and safe vaccines. The Congress was held under the auspices and with the scientific cooperation of Medical Faculties of the University of Montreal, Charité-Universitätsmedizin Berlin, University of Genoa, University of Barcelona, University of Florence, University of Milan, Griffith University, Gold Coast, Robert-Koch-Institute, Berlin, Max-Planck-Institute for Infection Biology, Berlin, Karolinska Institutet, Stockholm, Fraunhofer USA Center for Molecular Biotechnology, and Netherlands Vaccine Institute, among others. The opening lecture of the Congress on new vaccination strategies against tuberculosis was delivered by Professor Dr Stefan H.E. Kaufmann, Director of the Max-Planck-Institute.

Open Access Article

Letter from the Editor

Ronald W. Ellis and Eva Maria Riedmann
Pages 2 - 3
http://dx.doi.org/10.4161/hv.7.0.14905
Abstract | PDF








Part I. Towards new vaccines, design concepts, immunisation strategies and safety

REVIEWS

Open Access Article

Towards an AIDS vaccine: The transmembrane envelope protein as target for broadly neutralizing antibodies

Joachim Denner
Pages 4 - 9
http://dx.doi.org/10.4161/hv.7.0.14555
Abstract | PDF

Although the development of an effective vaccine is the main goal in the fight against AIDS, all attempts by numerous laboratories to develop a vaccine have failed so far. In addition, it is still unclear whether cytotoxic T cells or neutralizing antibodies or both should be induced. The major advantage of neutralizing antibodies is their ability to prevent infection and subsequent integration of the provirus into the cellular genome where it may persist in a form invisible to the immune system. Broadly neutralizing antibodies have been found in HIV infected individuals, including antibodies directed against a highly conserved region in the membrane proximal external region (MPER) of the transmembrane envelope (TM) protein gp41 of HIV-1. We successfully induced neutralizing antibodies against different gammaretroviruses by immunization with their respective TM proteins. These antibodies recognized epitopes not only in the MPER but also in the fusion peptide proximal region of the TM protein. In the case of feline leukaemia virus (FeLV), these antibodies protected cats from antigenemia following challenge. To understand the mechanism of neutralization, the interactions between neutralizing antibodies and their corresponding epitopes in the TM protein of gammaretroviruses and HIV-1 were analysed. These data may help to design antigens able to induce specific broadly neutralizing antibodies.

Open Access Article

Vaccines for Leishmaniasis: From proteome to vaccine candidates

Juliane Schroeder and Toni Aebischer
Pages 10 - 15
http://dx.doi.org/10.4161/hv.7.0.14556
Abstract | PDF

Leishmania spp. cause a wide spectrum of tropical diseases which are threatening an estimated 350 million people around the globe. While in most cases non-fatal, the disease is associated with high morbidity, social stigmata and poverty. However, the most severe form visceral leishmaniasis can be fatal if left untreated. Chemotherapeutics are available but show high toxicity, costs and are prone to resistance development due to prolonged treatment periods. Healing is associated with a life-long resistance to re-infection and this argues for the feasibility of vaccination. However, despite much effort, no such vaccine has become available yet. Here, the status of vaccine development in this field is briefly summarized before the focus is set on the promise of reverse vaccinology for anti-Leishmania vaccine development in the post-genomic era. We report on our own experience with this approach using an instructive example of successful candidate vaccine antigen identification.

Open Access Article

Arrested Plasmodium liver stages as experimental anti-malaria vaccines

Kai Matuschewski, Julius Clemence Hafalla, Steffen Borrmann and Johannes Friesen
Pages 16 - 21
http://dx.doi.org/10.4161/hv.7.0.14557
Abstract | PDF

Eukaryotic pathogens typically follow a complex life cycle, including host switch and morphologically distinct forms. Parasite stage conversion offers exceptional opportunities for whole organism vaccine development. In case of Plasmodium, the causative agent of malaria, disease is exclusively caused by asexual blood stages that invade and replicate within erythrocytes. Pathogenic blood stage infections are preceded by a silent parasite growth phase inside the liver. Two alternative experimental whole organisms vaccine strategies that lead to arrested Plasmodium liver stages elicit potent, lasting immunity against re-infection. Live irradiation- or genetically arrested parasites are metabolically active and correspond to classical attenuated vaccines. Specific antimalarial treatment during experimental natural sporozoite infections prevents a febrile malaria episode and, simultaneously, induces effective anti-liver stage immunity. Translation of these strategies into a safe, affordable, and accessible pediatric anti-malaria vaccine requires major bioengineering and pharmaceutical improvements, respectively, but holds promise for a truly effective immunization scheme against the most prevalent and fatal vector-borne disease.

Open Access Article

Piezoelectric permeabilization of mammalian dermal tissue for in vivo DNA delivery leads to enhanced protein expression and increased immunogenicity

Kate E. Broderick, Thomas Kardos, Jay R. McCoy, Michael P. Fons, Stephen Kemmerrer and Niranjan Y. Sardesai
Pages 22 - 28
http://dx.doi.org/10.4161/hv.7.0.14559
Abstract | PDF

Electropermeabilization of mammalian cells is a technique that has been used for the delivery of therapeutics, such as DNA plasmids or DNA vaccines. Typically, delivery via electropermeabilization occurs through injection of the substance into the tissue of interest followed by the insertion of electrodes at the site and the application of brief electrical pulses. Here we detail a novel and innovative contactless electropermeabilization method to deliver DNA plasmids to dermal tissue in vivo. This process has the advantage of eliminating the insertion of additional needles that serve as electrodes to facilitate the application of electric pulses in conventional electroporation processes. Plasmid encoding GFP was injected into guinea pig skin and pulsed with the novel contactless electropermeabilization method. Three days following treatment, robust GFP expression was observed on the skin of pulsed animals. Strong humoral immune responses were also achieved when a DNA vaccine expressing the influenza antigen NP was delivered and pulsed using the novel device in comparison to naked injection alone. This delivery method has the advantage of being contactless and suggests that gene transfer via this mode warrants further development.

Open Access Article

Adjuvants and alternative routes of administration towards the development of the ideal influenza vaccine

Paolo Durando, Rocco Iudici, Cristiano Alicino, Marisa Alberti, Daniela de Florentis, Filippo Ansaldi and Giancarlo Icardi
Pages 29 - 40
http://dx.doi.org/10.4161/hv.7.0.14560
Abstract | PDF

Vaccination is universally considered as the principal measure for the control of influenza, which represents a significant burden worldwide, both from a health-care and a socio-economic viewpoint. Conventional non-adjuvanted trivalent influenza vaccines (TIVs) have been recognized as having some deficiencies, such as suboptimal immunogenicity particularly in the elderly, in patients with severe chronic diseases and immunocompromized, indeed, those groups of the population at higher risk of developing severe complications following influenza infection, when compared to healthy adults. Moreover, the protection offered by conventional vaccines may be reduced by periodic antigenic drifts, resulting in a mismatch between the circulating and vaccinal viral strains. Another gap regarding currently available vaccines is related to the egg-based manufacturing system for their production: not only the length of time involved with the latter but also the limited capacity of this platform technology represent a major limitation for the active prevention of influenza, which is particularly important in the case of a new pandemic strain. New technologies used in vaccine composition, administration and manufacture have led to major advances during the last few years, and clinical researchers have continued to work hard, investigating several different strategies to improve the performance of influenza vaccines: namely, the addition of different adjuvants (i.e., MF59- and AS03-vaccines, virosomal formulations), the use of alternative routes of administration or manufacture (i.e., intradermal, nasal and oral vaccines and cell culture- and reverse genetic-based vaccines) or of high doses of antigen, and the development of DNA-vaccines, or the use of conserved viral epitopes (i.e., the extracellular portion of the M2 protein, the nucleoprotein and some domains of the hemagglutinin), in the attempt to produce a “universal target” antigen vaccine. The knowledge acquired represents a fundamental challenge for the control of influenza. An overview of the most recent and interesting results, some of which gained from our own research experience, particularly concerning two successful approaches, of those outlined above, namely the use of: (i) the oil-in-water MF59-adjuvant, and (ii) the intradermal (ID) route for vaccine administration, through a novel microinjection system, will be reported and discussed, together with the possible implications and perspectives to optimize immunization policies against influenza in the near future.

Open Access Article

Plant-based rapid production of recombinant subunit hemagglutinin vaccines targeting H1N1 and H5N1 influenza

Yoko Shoji, Jessica A. Chichester, Mark Jones, Slobodanka D. Manceva, Emily Damon, Vadim Mett, Konstantin Musiychuk, Hong Bi, Christine Farrance, Moneim Shamloul, Natasha Kushnir, Satish Sharma and Vidadi Yusibov
Pages 41 - 50
http://dx.doi.org/10.4161/hv.7.0.14561
Abstract | PDF

In 2009, a novel H1N1 swine influenza virus was isolated from infected humans in Mexico and the United States, and rapidly spread around the world. Another virus, a highly pathogenic avian influenza virus of the H5N1 subtype, identified by the World Health Organization as a potential pandemic threat in 1997, continues to be a significant risk. While vaccination is the preferred strategy for the prevention and control of influenza infections, the traditional egg-based approach to producing influenza vaccines does not provide sufficient capacity and adequate speed to satisfy global needs to combat newly emerging strains, seasonal or potentially pandemic. Significant efforts are underway to develop and implement new cell substrates with improved efficiency for influenza vaccine development and manufacturing. In recent years, plants have been used to produce recombinant proteins including subunit vaccines and antibodies. The main advantages of using plant systems for the production of vaccine antigens against influenza are their independence from pathogenic viruses, and cost and time efficiency. Here, we describe the large-scale production of recombinant hemagglutinin proteins from A/California/04/09 (H1N1) and A/Indonesia/05/05 (H5N1) strains of influenza virus in Nicotiana benthamiana plants, and their immunogenicity (serum hemagglutination inhibition and virus neutralizing antibodies), and safety in animal models. These results support the testing of these candidate vaccines in human volunteers and also the utility of our plant expression system for large-scale recombinant influenza vaccine production.

Open Access Article

Challenges for the evaluation of Staphylococcus aureus protein based vaccines: Monitoring antigenic diversity

Ellen Murphy, Shuo L. Lin, Lorna Nunez, Lubomira Andrew, Pamela S. Fink, Deborah A. Dilts, Susan K. Hoiseth, Kathrin U. Jansen and Annaliesa S. Anderson
Pages 51 - 59
http://dx.doi.org/10.4161/hv.7.0.14562
Abstract | PDF

Clumping factors A (ClfA) and B (ClfB) are Staphylococcus aureus virulence proteins that are displayed on the cell surface of the organism and have potential as vaccine antigens for the prevention of S. aureus disease. Here we evaluate the phylogeny of S. aureus in the context of antigenic variation of these two surface proteins. ClfA and ClfB gene sequences, along with epidemiological markers (MLST, spa and capsule genotype) were obtained for 224 S. aureus isolates including both historical strains and a collection representative of current MRSA isolates from the United States. Variation within ClfA and ClfB was consistent with the established population biology of S. aureus, namely, that S. aureus strains belong to a relatively small number of clonal lineages, with evolution proceeding mainly by mutation and with little to no recombination between clades. Thus most variation in ClfA and ClfB occurs between but not within lineages, and particular groups of ClfA and ClfB variants are closely linked. This has important implications for vaccine development and assessment as it suggests that a relatively small survey of strains will be representative of the total population variation, whereas for species that evolve mainly by recombination, such as Neisseria meningitidis, analysis of a much larger number of strains is needed to accomplish the same purpose. Our study also revealed evidence for the de-evolution of ClfB and therefore its reduced suitability as a target for vaccine development compared to ClfA.

Open Access Article

Novel therapeutic vaccine: Granulysin and new DNA vaccine against Tuberculosis

Masaji Okada and 22 others
Pages 60 - 67
http://dx.doi.org/10.4161/hv.7.0.14563
Abstract | PDF

[Purpose] Multi-drug resistant (MDR) Mycobacterium Tuberculosis (M.TB) is a big problem in the world. We have developed novel TB therapeutic vaccines. [Methods and Results] DNA vaccine expressing mycobacterial heat shock protein 65 and IL-12 was delivered by the hemagglutinating virus of Japan (HVJ)-envelope. M. TB, MDR-TB or extremenly drug resistant (XDR-TB) was injected i.v. into DBA/1 mice, and treated with the vaccine three times. This HVJ-E/Hsp65DNA+IL-12DNA vaccine provided strong therapeutic efficacy against MDR-TB and XDR-TB (prolongation of survival time and the decrease in the number of TB) in mice. Therapeutic effect of this vaccine on TB infection was also demonstrated in chronic TB infection murine model using aerosol infection intratracheally. On the other hand, granulysin protein produced from CTL has lethal activity against TB. Granulysin protein vaccine also exerted strong therapeutic effect. Furthermore, we extended our studies to monkey model, which is currently the best animal model of human TB. Hsp65DNA+IL-12 DNA vaccine exerted strong therapeutic efficacy (100% survival and augmentation of immune responses) in the TB-infected monkeys. In contrast, the survival of the saline control group was 60% at 16 week post-challenge. HVJ-Envelope/HSP65 DNA+IL-12 DNA vaccine increased the body weight of TB-infected monkeys, improved the erythrocyte sedimentation rate, and augmentated the immune responses (proliferation of PBL and IL-2 production). The enhancement of IL-2 production from monkeys treated with this vaccine was correlated with the therapeutic efficacy of the vaccine. [Conclusion] These data indicate that novel vaccines might be useful against TB including XDR-TB and MDR-TB for human therapeutic clinical trials.




Part II. Progress in infectious diseases vaccines, immunisation and immunotherapy

REVIEWS

Open Access Article

Preclinical evidence for the potential of a bivalent fHBP vaccine to prevent Neisseria meningitidis Serogroup C Disease

Shannon L. Harris, Duzhang Zhu, Ellen Murphy, Lisa K. McNeil, Xin Wang, Leonard W. Mayer, Lee H. Harrison, Kathrin U. Jansen and Annaliesa S. Anderson
Pages 68 - 74
http://dx.doi.org/10.4161/hv.7.0.14564
Abstract | PDF

A bivalent factor H binding protein (fHBP) vaccine for the prevention of disease caused by Neisseria meningitidis serogroup B is currently in clinical development. Since fHBP is also expressed by other meningococcal serogroups, anti-fHBP antibodies may have bactericidal activity against meningococci independent of serogroup. To begin examining the susceptibility of other meningococcal serogroups to anti-fHBP antibodies, meningococcal serogroup C invasive isolates (n = 116) were collected from the Centers for Disease Control and Prevention’s Active Bacterial Core surveillance (ABCs) sites during 2000–2001. These isolates were analyzed for the presence of the fhbp gene. All serogroup C isolates contained the gene, and sequence analysis grouped the proteins into two subfamilies, A and B. Flow cytometry analysis demonstrated that fHBP was expressed on the surface of ~70% of isolates in vitro with varying levels of expression. fHBP was accessible to antibodies on the cell surface even in the presence of the polysaccharide capsule. Nine isolates from different geographic regions were identified which harboured an identical single nucleotide deletion that could result in a truncated subfamily B fHBP. Analysis by flow cytometry using a polyclonal fHBP antibody preparation revealed that a subpopulation of each of these isolates expressed fHBP. Rabbit and non-human primate immune sera generated with bivalent fHBP vaccine were tested for bactericidal activity against a panel of diverse serogroup C clinical isolates using human complement. Sera from both species demonstrated serum bactericidal antibody activity against the serogroup C isolates tested. These promising findings suggest that a bivalent fHBP vaccine may be capable of providing protection against meningococcal disease caused by both serogroup C and B.

Open Access Article

13-valent pneumococcal conjugate vaccine immune sera protects against pneumococcal serotype 1, 3, and 5 bacteremia in a neonatal rat challenge model

Philip Fernsten, Kathryn W. Mason, Xinhong Yu, Donna Tummolo, Kelly A. Belanger, How Tsao, Duzhang Zhu, David Cooper, Michael Hagen and Kathrin U. Jansen
Pages 75 - 84
http://dx.doi.org/10.4161/hv.7.0.14566
Abstract | PDF

PCV7 was first licensed in the United States in 2000 based on clinical efficacy studies. Since the introduction, PCV7 has demonstrated protective effectiveness for each of the vaccine serotypes. More recently, PCV13 has been licensed in more than 60 countries based on serological noninferiority to PCV7 for the shared serotypes and noninferiority to the least immunogenic serotypes of PCV7 for the additional 6 serotypes in PCV13. To evaluate whether the functional antibody responses to serotypes 1, 3, and 5 were sufficient to protect animals challenged with virulent strains of these serotypes, rhesus macaques were immunized with three clinical doses of PCV13. The macaques mounted robust anti-capsular polysaccharide IgG and opsonophagocytic killing (OPA) responses to each serotype contained in the vaccine. Pooled pre-immunization sera and post-immunization serum pools were tested in a neonatal rat bacteremia model. Passive transfer of pooled post-immunization sera, but not pre-immunization sera, protected neonatal rats from lethal IP challenge with serotype 1, 3, or 5 strains. The functional activity of PCV13 immune sera against a virulent type 3 strain was further evaluated using sera from human children immunized with 4 doses of PCV7 or PCV13. Pooled sera from children immunized with PCV13, but not pooled sera from children immunized with PCV7, which does not contain the serotype 3 polysaccharide conjugate, protected neonatal rats from lethal IP challenge with a highly encapsulated and virulent serotype 3 strain. These data suggest that PCV13 will provide protection against pneumococcal serotype 1, 3, and 5 disease in human populations, even at relatively low OPA titers.

Open Access Article

Toll-like receptor-mediated adjuvanticity and immunomodulation in dendritic cells: Implications for peptide vaccines

Daniel Mitchell, Michelle Yong, Jyothy Raju, Nicole Willemsen, Matthew Black, Amanda Trent, Matthew Tirrell and Colleen Olive
Pages 85 - 93
http://dx.doi.org/10.4161/hv.7.0.14567
Abstract | PDF

Considerable success has been made with many peptide antigen formulations and it appears that peptide-based vaccines are emerging as the next generation of prophylactic and remedial immunotherapy. However, peptides are typically poorly immunogenic and rely on delivery with potent immunostimulatory adjuvants that activate the innate and adaptive arms of the immune system. Our research aims to develop novel peptide antigen delivery systems that incorporate multiple pattern-recognition receptor (PRR) agonists and is focused on those designed to stimulate Toll-like receptors (TLRs) on dendritic cells (DCs). The cytokine (IL-4, IL-6, IL-10, IL-12 and IL-23) profiles of DCs induced by individual TLR agonists have been evaluated. From this data we predicted which TLR agonists may influence a particular T helper cell (Th) response. Using purified DCs that were derived from precursor cells in murine bone marrow and then stimulated simultaneously with multiple TLR agonists, we have shown synergy between various TLR agonist pairs leading to enhanced cytokine production. Using various mitogen-activated protein kinase (MAPK) inhibitors (c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 MAPK) we have demonstrated the importance of p38 MAPK and ERK signaling pathways in IL-12p70 and IL-12p40 production in DCs induced by TLR stimulation, whereas the JNK pathway appeared to have a negative regulatory role on cytokine production in DCs stimulated with certain TLR agonists. An important role for nuclear factor-kappa B and phosphoinositol-3-kinase as positive regulators of TLR signaling in DCs leading to cytokine production was also demonstrated. The significance of this research lies not only in improving potency, but by understanding the immunological mechanisms of adjuvanticity, in being able to tailor peptide vaccines to generate specific types of Th responses required for immunity against various types of pathogens.

Open Access Article

Aging and impaired immunity to influenza viruses: Implications for vaccine development

Wai Ming Liu, Bernard A.M. van der Zeijst, Claire J.P. Boog and Ernst C. Soethout
Pages 94 - 98
http://dx.doi.org/10.4161/hv.7.0.14568
Abstract | PDF

Influenza infections are responsible for significant morbidity and mortality each year, with the highest infection rates found in the elderly population. The main strategy to reduce the impact of influenza infections in the elderly population is vaccination. However, the efficacy of influenza vaccines that are licensed for use in the elderly is relatively low (17-53%). The complex age-related changes that occur in both innate and adaptive immunity are thought to hamper the immune response to influenza immunization and to reduce protection against infection in the elderly. For the development of improved vaccines that overcome the limitations of an aged immune system, it is crucial to understand the mechanisms that lead to immune dysfunction. Here, we review the recent progress in unravelling the mechanisms behind the age-related immune dysfunction in elderly, as well as the recent developments in improving influenza vaccines and identification of new correlates of protection.

RESEARCH PAPER

Open Access Article

Effectiveness of engineering the nontypeable Haemophilus influenzae antigen Omp26 as an S-layer fusion in bacterial ghosts as a mucosal vaccine delivery

Eva M. Riedmann, Werner Lubitz, John McGrath, Jennelle M. Kyd and Allan W. Cripps
Pages 99 - 107
http://dx.doi.org/10.4161/hv.7.0.14569
Abstract | PDF

The potential of empty bacterial cell envelopes (ghosts) as a delivery system for mucosal immunization was assessed in a rat model and different routes of immunization were evaluated. Animals were mucosally immunized targeting either gut only or gut and lung mucosal sites with Escherichia coli ghosts harbouring the nontypeable Haemophilus influenzae (NTHi) antigen Omp26. Omp26 was expressed as either a part of an S-layer fusion or as a soluble protein in the periplasm. In the gut/lung regime two initial gut targeted inoculations with the ghosts were followed by an intratracheal (IT) boost with purified Omp26. The gut only immunization regime showed a moderate enhancement of bacterial clearance following pulmonary challenge whereas the gut/lung immunization regime resulted in significantly enhanced pulmonary clearance of NTHi. Both immunization regimes induced high levels of Omp26 specific antibodies in the serum of immunized rats, with higher levels in the groups that received the IT boost with purified Omp26. Analysis of IgG isotypes present in serum suggest that the immune response was predominantly of a T-helper1 type. Additionally, immunization induced a significant cellular immune response with lymphocytes from animals vaccinated using the gut/lung regime responding significantly to Omp26 when compared to control groups. The results of this study show that mucosal immunization with recombinant Omp26 in E. coli ghosts followed by a boost with purified Omp26 can induce a specific and protective immune response in a rodent model of acute lung infection.

REVIEW

Open Access Article

Development of therapeutic and prophylactic vaccine against Tuberculosis using monkey and transgenic mice models

Yoko Kita and 21 others
Pages 108 - 114
http://dx.doi.org/10.4161/hv.7.0.14571
Abstract | PDF

[Purpose] BCG is not efficacious against M. tuberculosis (TB) in adult. Therefore, novel TB vaccines were established by using three kinds of animal models (cynomolgus monkey model which is the best animal model of human TB, IL-2R knock out SCID mice as a human immune model, and granulysin transgenic mouse). [Methods and Results] DNA vaccine expressing TB Hsp65 and IL-12 was delivered by the hemagglutinating virus of Japan (HVJ)-envelope. The BCG prime followed by Hsp65+IL-12/HVJ vaccine boost showed a synergistic effect in the TB-infected cynomolgus monkey (100% survival). In contrast, 33% of monkeys were alive in BCG alone group. Furthermore, the prolongation of survival period of the monkey was observed by the combination of BCG and DNA vaccine even when the boost was performed after long-term period (4month) from prime. This combination also improved the erythrocyte sedimentation rate (ESR), increased the body weight, and augmented the proliferation of PBL and IL-12 production at higher levels than BCG alone or saline. Furthermore, this vaccine exerted therapeutic efficacy in IL-2R knock out SCID-PBL/hu mice, which were transplanted with human T cells. Granulysin is an important defensive molecule expressed by human T cells and NK cells and has a cytolytic activity against microbes including Mycobacterium tuberculosis (TB) and tumors. Expression of 15kD (15K) granulysin protein and mRNA in CD8 positive T cells in the patients infected with drug sensitive (TB) or multi-drug resistant (MDR-TB) M. tuberculosis were lower than that in the healthy volunteers, suggesting that granulysin treatment might improve the tuberculous disease in human. Therefore, we established two kinds of granulysin transgenic mice (15K granulysin transgenic mice and 9K granulysin transgenic mice). It was demonstrated that 15K granulysin transgenic mice as well as 9K granulysin transgenic mice exerted in vivo anti-TB effect, including the decrease of the number of TB and augmentation of the CTL activity. These are the first findings which demonstrate in vivo effects of 15K granulysin and 9K granulysin against TB infection. Moreover, DNA vaccine expressing 15K granulysin showed a therapeutic activity against TB in mice. [Conclusion] These data indicate that monkey, IL-2R gene-knock out SCID-PBL/hu and granulysin transgenic mice models provide useful tools for the development of novel vaccines (HVJ-Envelope/Hsp65 DNA + IL-12 DNA vaccine and granulysin vaccine) against TB.




Part III. Perspectives in immune therapy and anti-cancer vaccines development

REVIEWS

Open Access Article

Therapeutic vaccination for cancer immunotherapy: Antigen selection and clinical responses

Astrid Geldmacher, Anja Freier, Florian O. Losch and Peter Walden
Pages 115 - 119
http://dx.doi.org/10.4161/hv.7.0.14573
Abstract | PDF

Background: Because of its high specificity and low toxicity therapeutic vaccination is considered a desirable treatment for cancer. So far, however, the results of cancer vaccination trials have been disappointing, which is often attributed to the problem identifying appropriate vaccine antigens. Tumorassociated antigens are mostly autoantigens and therefore expected to be subject to immunosuppressive mechanisms. Cancer-testis antigens are the most prominent exception as, still being self, they are physiologically only expressed in immunopriviledged tissues and should therefore not induce autotolerance. This leads to the widely accepted hypothesis that cancer-testis antigens should be more efficient inducers of anti-tumor cellular immune responses than differentiation antigens. Aim of the study was to test this hypothesis by evaluating the published reports on clinical therapeutic vaccination trials for the objective clinical response rates to vaccination with cancer testis antigen vs. differentiation antigens. Approach: The results of vaccination clinical trials with cancer testis and/or differentiation antigens published in literature and databanks were analyzed for clinical outcome versus vaccine antigens. 21 publications on cancer testis antigen-based trials in which clinical outcome was reported according to WHO or RECIST were identified and analyzed. Results: The rate of objective responses to cancer testis antigen vaccines in 239 patients was 3.8% and for the 235 patients vaccinated with cancer testis plus 3 differentiation antigens 4.3% compared to 2.6% for the 496 patients vaccinated with differentiation antigens alone. Conclusions: Cancer testis antigen-based vaccines seem slightly superior over vaccines based on differentiation antigens providing support for the hypothesis.

Open Access Article

Co-delivery of PSA and PSMA DNA vaccines with electroporation induces potent immune responses

Bernadette Ferraro, Neil J. Cisper, Kendra T. Talbott, Lindsey Philipson-Weiner, Colleen E. Lucke, Amir S. Khan, Niranjan Y. Sardesai and David B. Weiner
Pages 120 - 127
http://dx.doi.org/10.4161/hv.7.0.14574
Abstract | PDF

Prostate cancer (PCa) remains a significant public health problem. Current treatment modalities for PCa can be useful, but may be accompanied by deleterious side effects and often do not confer long-term control. Accordingly, additional modalities, such as immunotherapy, may represent an important approach for PCa treatment. The identification of tissue-specific antigens engenders PCa an attractive target for immunotherapeutic approaches. Delivery of DNA vaccines with electroporation has shown promising results for prophylactic and therapeutic targets in a variety of species including humans. Application of this technology for PCa immunotherapy strategies has been limited to single antigen and epitope targets. We sought to test the hypothesis that a broader collection of antigens would improve the breadth and effectiveness of a PCa immune therapy approach. We therefore developed highly optimized DNA vaccines encoding prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA) as a dual antigen approach to immune therapy of PCa. PSA-and PSMA-specific cellular immunogenicity was evaluated in a mouse model for co-delivery and single antigen vaccination. Mice received 2 immunizations spaced 2 weeks apart and immunogenicity was evaluated 1 week after the second vaccination. Both the PSA and PSMA vaccines induced robust antigen-specific IFNγ responses by ELISpot. Further characterization of cellular immunogenicity by flow cytometry indicated strong antigen-specific TNFα production by CD4+ T cells and IFNγ and IL-2 secretion by both CD4+ and CD8+ T cells. There was also a strong humoral response as determined by PSA-specific seroconversion. These data support further study of this novel approach to immune therapy of PCa.

Open Access Article

An overview on the implementation of HPV vaccination in Europe

Paolo Bonanni, Miriam Levi, Nina B. Latham, Angela Bechini, Emilia Tiscione, Piero Lai, Donatella Panatto, Roberto Gasparini and Sara Boccalini
Pages 128 - 135
http://dx.doi.org/10.4161/hv.7.0.14575
Abstract | PDF

The discovery that the Human PapillomaVirus (HPV) is the necessary cause of cervical cancer has led to the development of prophylactic vaccines. Cervical cancer is the second most common cause of death from cancer among young women in Europe: mortality is still high, despite its important reduction due to screening programs for early detection. Besides cervical cancer, HPV is responsible for a significant proportion of other anogenital cancers and an increasing number of oropharyngeal cancers, representing together an at least equal burden compared to cervical cancer. HPV is also responsible for conditions such as condyloma acuminata (genital warts) and recurrent respiratory papillomatosis. Organized vaccination programs against HPV have the potential to prevent about 70% of cervical cancers and the vast majority of the other HPV-related conditions. Recommendations for HPV vaccination of at least one cohort of females have been issued in nearly all western European countries, and national/regional publicly funded vaccination programs have been introduced in most of them. Different approaches have been chosen for the implementation of HPV vaccination, based on the organization of each country's health care system. A brief outline of these programs in Europe is presented. As for all preventive public health interventions, high coverage of the target population with HPV vaccines pre-exposure is essential to achieve maximum reduction of cases: therefore, in order to obtain the maximum and most equitable coverage and future benefit, programs targeting adolescents before exposure to HPV should be preferred and population-based. Catch-up programs should also be implemented wherever possible, in order to deliver more and even earlier benefits, and effective communication strategies need to be adopted.

Open Access Article

Safety and tolerability of bivalent HPV vaccine: An Italian post-licensure study

Roberto Gasparini, Paolo Bonanni, Miriam Levi, Angela Bechini, Sara Boccalini, Emilia Tiscione, Daniela Amicizia, Piero Luigi Lai, Klodiana Sulaj, Antonio Giuseppe Patria and Donatella Panatto
Pages 136 - 146
http://dx.doi.org/10.4161/hv.7.0.14576
Abstract | PDF

One of the most important scientific discoveries of the last century was that persistent infection by some types of HPV is a precondition for the development of cervical cancer. The oncogenic types of HPV are also associated with other tumours (vaginal, vulvar and anal carcinomas, tumours of the head and neck, urethra and penis). Two preventive vaccines are currently available (Cervarix® and Gardasil®). Both have shown very good efficacy, safety and tolerability profiles. Nonetheless, extensive vaccination requires long-term monitoring of safety and tolerability. The aim of our study was to evaluate the safety and tolerability of the bivalent vaccine Cervarix® in Italy. Every participant in the study completed a questionnaire after each dose of vaccine received, with a view to recording adverse events during the first 7 days after vaccination. We registered local (pain, redness, swelling) and systemic symptoms (fever, headache, myalgia, fatigue, arthralgia, itching, gastrointestinal disorders, rash and urticaria). A total of 4,643 subjects were recruited. In all, 7,107 questionnaires were collected: 3,064 after the first dose, 2,367 after the second and 1,676 after the third. No serious adverse events were observed. The most frequent local symptom was pain at the injection site, while fatigue, headache and myalgia were the most common systemic reactions. Pain was reported more frequently after the first dose than after the others, while all the other local and general symptoms were reported most frequently after the third dose. Almost all of the local and general reactions proved to be of negligible intensity and duration and required no medical intervention. Our results show better tolerability of the vaccine in comparison with the data from some controlled clinical studies and from other surveillance programmes conducted internationally. That tolerability proved to be better than in clinical studies could be explained by the absence of the typical apprehension felt by subjects involved in clinical experimentation.

Open Access Article

Mutants of plant genes for developing cancer vaccines

Silvia Massa, Francesca Paolini, Laura Spanò, Rosella Franconi and Aldo Venuti
Pages 147 - 155
http://dx.doi.org/10.4161/hv.7.0.14577
Abstract | PDF

Preventive Human Papillomavirus (HPV) vaccination is an expensive practice and it may be an insufficient tool to tackle cervical cancer worldwide. Therapeutic intervention is seeking for safe/effective vaccines inducing the activation of CD8+ cytotoxic T lymphocytes (CTLs) that is required to clear the tumor. Linking a tumor-specific antigen (i.e. the E7 oncoprotein of the ‘high risk’ HPVs) to molecules able to increase its immune ‘visibility’ represents a strategy to force the immune system to fight cancer. We focused on plants as sources of innovative immunostimulatory sequences. We have already shown the anti-cancer activity obtained by fusing E7GGG (a mutagenized E7 gene from the high risk HPV type 16) to the coat protein of a plant virus, the Potato Virus X. We are now investigating plant-derived carriers, such as the ‘Ribosome inactivating proteins’ (RIPs), so far used to develop immunotoxins for targeted cancer therapy. Beside toxicity, RIPs have other features (i.e. immunogenicity, ability to modulate immune functions and apoptosis induction) that could be useful tools to use in tumor immunotherapy. A non toxic mutant of saporin (SAP-KQ) was used as a carrier for the E7GGG gene in the context of a DNA-based vaccine. We show here that fusion constructs of SAP-KQ with E7GGG can induce E7-specific Immunoglobulins (IgGs), CTLs and Delayed-Type Hypersensitivity (DTH) affecting the growth of E7-expressing tumors in mice. These data demonstrate that mutant plant genes hold promise to improve the poor immunogenicity of tumor-associated cancer antigens and could contribute to the evolution of new cancer immunotherapy.

Open Access Article

Glycan mediated immune responses to tumor cells

Anastas D. Pashov, Behjatolah Monzavi-Karbassi and Thomas Kieber-Emmons
Pages 156 - 165
http://dx.doi.org/10.4161/hv.7.0.14578
Abstract | PDF

Preclinical animal studies convincingly demonstrate that tumor immunity to self-antigens can be actively induced and can translate into effective anti-tumor responses. Among the most challenging of clinical targets for cancer immunotherapy is Tumor Associated Carbohydrate Antigens (TACA). The molecular characterization of TACA suggest that these glycans are both altered and self-antigens. A new appreciation of the interaction of glycans with immune effector cells that will benefit immunotherapy strategies is emerging as more information on the nature of molecular interactions of glycan recognition molecules is obtained. Carbohydrate recognition affects more or less every aspect of the innate and adaptive immune response and their role in immunotherapy of cancer should be considered beyond the existing paradigm of traditional TACA based-vaccines.

RESEARCH PAPER

Open Access Article

Evaluation of adjuvants for a candidate conjugate vaccine against benzo[a]pyrene

Mario T. Schellenberger, Sophie Farinelle, Stéphanie Willième and Claude P. Muller
Pages 166 - 173
http://dx.doi.org/10.4161/hv.7.0.14579
Abstract | PDF

We have recently developed an experimental vaccine based on benzo[a]pyrene (B[a]P) conjugated to tetanus toxoid as a carrier protein. In combination with Freund adjuvant, this vaccine induces high levels of B[a]P-specific antibodies to protect against detrimental effects of this carcinogen. Here we evaluate this conjugate vaccine by replacing Freund adjuvant by adjuvants that are potentially compatible with their use in humans. We showed that all adjuvants tested induced specific antibodies against B[a]P and 7,8-diol-B[a]P, its carcinogenic metabolite. The best antibody levels were obtained with Quil A, MF-59 and Alum. Biological activity in terms of enhanced retention of B[a]P was confirmed in mice immunised with Quil A, Montanide, Alum and MF-59. Our findings demonstrate that a vaccination against B[a]P is feasible in combination with adjuvants licensed in humans.




Part IV. Advances in immunisation research and epidemiological analysis

REVIEWS

Open Access Article

Cross-protective immunity against influenza virus infections induced by intranasal vaccination together with a TLR3-mucosal adjuvant

Akira Ainai, Masato Tashiro and Hideki Hasegawa
Pages 174 - 182
http://dx.doi.org/10.4161/hv.7.0.14584
Abstract | PDF

A new pandemic of influenza virus could result from the emergence of an unpredictable viral strain in an unexpected fashion. Thus, developing methods to protect the population from the spread of a new influenza virus is an urgent and important public health concern. Although vaccines can induce protective and prophylactic immune responses, the immunity induced by the current parenteral inactivated vaccine preparation is less effective in preventing heterologous virus infection. The induction of cross-protective mucosal immunity in the respiratory tract, the initial site of infection, is the most effective method for defending against heterologous influenza virus infection. Secretory immunoglobulin A plays a critical role in cross-protective mucosal immunity. Such cross-protective immunity can be induced by the intranasal administration of a vaccine together with an appropriate adjuvant that can mimic natural influenza virus infection. In this review, we describe the development of mucosal vaccines against influenza viruses and discuss their advantages. In addition, we describe data indicating that synthetic double-stranded RNAs, agonists of Toll-like receptor 3, are effective mucosal adjuvants for intranasally administered inactivated influenza virus vaccines.

Open Access Article

A non-glycosylated, plant-produced human monoclonal antibody against anthrax protective antigen protects mice and non-human primates from B. anthracis spore challenge

Vadim Mett, Jessica A. Chichester, Michelle L. Stewart, Konstantin Musiychuk, Hong Bi, Carolyn J. Reifsnyder, Anna K. Hull, Mark T. Albrecht, Stanley Goldman, Les W. J. Baillie and Vidadi Yusibov
Pages 183 - 190
http://dx.doi.org/10.4161/hv.7.0.14586
Abstract | PDF

The health and economic burden of infectious diseases in general and bioterrorism in particular necessitate the development of medical countermeasures. One proven approach to reduce the disease burden and spread of pathogen is treatment with monoclonal antibodies (mAb). mAbs can prevent or reduce severity of the disease by variety of mechanisms, including neutralizing pathogen growth, limiting its spread from infected to adjacent cells, or by inhibiting biological activity of toxins, such as anthrax lethal toxin. Here, we report the production of glycosylated (pp-mAb^PA) and non-glycosylated (pp-mAb^PANG) versions of a plant-derived mAb directed against protective antigen (PA) of Bacillus anthracis in Nicotiana benthamiana plants using agroinfiltration. Both forms of the antibody were able to neutralize anthrax lethal toxin activity in vitro and protect mice against an intraperitoneal challenge with spores of B. anthracis Sterne strain. A single 180 µg intraperitoneal dose of pp-mAb^PA or pp-mAb^PANG provided 90% and 100% survival, respectively. When tested in non-human primates, pp-mAb^PANG was demonstrated to be superior to pp-mAb^PA in that it had a significantly longer terminal half-life and conferred 100% protection against a lethal dose of aerosolized anthrax spore challenge after a single 5 mg/kg intravenous dose compared to a 40% survival rate conferred by pp-mAb^PA. This study demonstrates the potential of a plant-produced non-glycosylated antibody as a useful tool for the treatment of inhalation anthrax.

Open Access Article

Antibodies to plant-produced Plasmodium falciparum sexual stage protein Pfs25 exhibit transmission blocking activity

Christine E. Farrance, Jessica A. Chichester, Konstantin Musiychuk, Moneim Shamloul, Amy Rhee, Slobodanka D. Manceva, R. Mark Jones, Tarlan Mamedov, Satish Sharma, Vadim Mett, Stephen J. Streatfield, Will Roeffen, Marga van de Vegte-Bolmer, Robert W. Sauerwein, Yimin Wu, Olga Muratova, Louis Miller, Patrick Duffy, Robert Sinden and Vidadi Yusibov
Pages 191 - 198
http://dx.doi.org/10.4161/hv.7.0.14588
Abstract | PDF

Malaria is a serious and sometimes fatal mosquito-borne disease caused by a protozoan parasite. Each year, it is estimated that over one million people are killed by malaria, yet the disease is preventable and treatable. Developing vaccines against the parasite is a critical component in the fight against malaria and these vaccines can target different stages of the pathogen’s life cycle. We are targeting sexual stage proteins of P. falciparum which are found on the surface of the parasite reproductive cells present in the mosquito gut. Antibodies against these proteins block the progression of the parasite’s life cycle in the mosquito, and thus block transmission to the next human host. Transmission blocking vaccines are essential to the malaria eradication program to ease the disease burden at the population level. We have successfully produced multiple versions of the Pfs25 antigen in a plant virus-based transient expression system and have evaluated these vaccine candidates in an animal model. The targets are expressed in plants at a high level, are soluble and most importantly, generate strong transmission blocking activity as determined by a standard membrane feeding assay. These data demonstrate the feasibility of expressing Plasmodium antigens in a plant-based system for the economic production of a transmission blocking vaccine against malaria.

Open Access Article

An influenza N1 neuraminidase-specific monoclonal antibody with broad neuraminidase inhibition activity against H5N1 HPAI viruses

Yoko Shoji, Jessica A. Chichester, Gene A. Palmer, Christine E. Farrance, Robert Stevens, Michelle Stewart, Lauren Goldschmidt, Varough Deyde, Larisa Gubareva, Alexander Klimov, Vadim Mett and Vidadi Yusibov
Pages 199 - 204
http://dx.doi.org/10.4161/hv.7.0.14595
Abstract | PDF

H5N1 avian influenza continues to be a potential pandemic threat. Several vaccine candidates based on potentially pandemic influenza strains and antiviral drugs have been tested in preclinical and clinical studies. The data obtained so far have shown some promise, but have also revealed some shortcomings with both of these approaches. We have identified and characterized an H5N1 neuraminidase-specific monoclonal antibody which specifically inhibits N1 neuraminidase activity of highly pathogenic avian influenza (HPAI) strains from clades 1 and 2. We have also shown the protective efficacy of this antibody in animal challenge models using homologous virus. Specific and effective inhibition of N1 NA could make this mAb a useful therapeutic tool in the treatment of human infection, in particular with oseltamivir- and zanamivir-resistant strains of HPAI.

Open Access Article

Hepatitis A outbreaks in the vaccination era in Catalonia, Spain

Ana Martínez, Sonia Broner, Nuria Torner, Pere Godoy, Joan Batalla, Josep Álvarez, Irene Barrabeig, Neus Camps, Gloria Carmona, Sofía Minguell, Rosa Sala, Joan Caylà and Angela Domínguez
Pages 205 - 210
http://dx.doi.org/10.4161/hv.7.0.14598
Abstract | PDF

Hepatitis A outbreaks have a major impact on public health services and involve case investigation and intervention measures to susceptible contacts. At the end of 1998 a universal vaccination program with a combined hepatitis A+B vaccine was started in Catalonia (Spain) in 12-years-old preadolescents. The objective of this study was to compare the characteristics of hepatitis A outbreaks in the periods before and after the introduction of the preadolescent vaccination program and to estimate the preventable fraction of cases associated to outbreaks. The incidence rates of outbreaks, cases and hospitalization associated with each outbreak were calculated. Two periods were considered: before (1991-1998) and after (2000-2007) the introduction of mass vaccination. The preventable fraction and 95% confidence intervals (CI) of cases associated with outbreaks was calculated. The rate of associated cases with outbreaks was higher in the period before the vaccination program than in the post vaccination period (1.53 per 100,000 person-year vs 1.12 ; p<0.001), but the rate of hospitalization was greater in the period after the introduction of vaccination program than in the period previous to vaccination (0.70 per million persons-year vs 0.08; p<0.001). The preventable fraction of cases associated to outbreaks was 19.6%(95%CI 6.7-32.5) in the 0-4 years group and 16.7% (95% CI 6.0-27.5) in the 5-14 years group, but the highest figure (38.6%; 95%CI 21.3-55.9) was observed in the 15-24 years age group. The estimated proportion of cases associated with outbreaks that would theoretically have been prevented with the vaccination program suggests that substantial benefits have been obtained in Catalonia in people aged less than 25 years.

Open Access Article

Epidemiological changes after PCV7 implementation in Italy: Perspective for new vaccines

Filippo Ansaldi, Daniela de Florentis, Paola Canepa, Roberto Bandettini, Maria Cristina Diana, Mariano Martini, Paolo Durando and Giancarlo Icardi
Pages 211 - 216
http://dx.doi.org/10.4161/hv.7.0.14602
Abstract | PDF

The emergence of serotypes not included in 7-valent pneumococcal conjugate vaccine (PCV7), so-called “replacement”, could erode the great advantage offered by vaccination. The effect of the replacement phenomena is very variable in different Countries and depends on one or more of several factors i.e. serotype before PCV7 implementation, PCV7 coverage, period from PCV implementation, vaccination schedule, antibiotic use, etc. The long-term epidemiological picture in Europe regarding serotype change is limited because of the relatively recent introduction of PCV7 - the majority of European Countries have only experienced PCV implementation since 2006 - and the difficulty in reaching very high vaccination coverage. In May 2003, a large-scale program of vaccination against Streptococcus pneumoniae with PCV7 was started in Liguria achieving coverage higher than 90%. The results of this program anticipate the results likely to be achieved when high coverage has been reached in Europe for a period of several years. The project included the immunogenicity evaluation of the co-administration of PCV7 and exavalent vaccine using the 3-5-11 month schedule, effectiveness evaluation and the active and passive surveillance system of invasive pneumococcal disease (IPD) and non-IPD. Since the beginning of PCV7 implementation, the proportion of PCV7 serotype has declined and in 2009-10 it accounted for about 10% of all Streptocuccus pn responsible for IPD and non-IPD. The new 13-valent pneumococcal conjugate vaccine, available since July 2010, will offer a significant added benefit covering about 90%, 100% of IPD and more than 40% and 60% of non-IPD detected in pre-school and school children, respectively, after PCV7 introduction.

Open Access Article

Burden of the 1999-2008 seasonal influenza epidemics in Italy: Comparison with the H1N1v (A/California/07/09) pandemic

Piero Luigi Lai, Donatella Panatto, Filippo Ansaldi, Paola Canepa, Daniela Amicizia, Antonio Giuseppe Patria and Roberto Gasparini
Pages 217 - 225
http://dx.doi.org/10.4161/hv.7.0.14607
Abstract | PDF

Despite preventive efforts, seasonal influenza epidemics are responsible for substantial morbidity and mortality every year worldwide, including developed countries. The A/H1N1v pandemic imposed a considerable healthcare and economic burden. In order to obtain an accurate estimate of the economic burden of influenza, and hence to guide policy-makers effectively, systematic studies are necessary. To this end, data from epidemiological surveillance are essential. To estimate the impact of the 1999-2008 seasonal influenza epidemics and the H1N1v pandemic, we analyzed data from the Italian Influenza Surveillance System (CIRINET). In the period 1999-2008, the Italian surveillance network consisted of sentinel general practitioners and pediatricians, who reported cases of Influenza-Like Illness (ILI) and Acute Respiratory Infections (ARI) observed during their clinical practice from mid-October to late April each year; reports were sent to the Center for Research on Influenza and other Viral Infections (CIRI-IV). CIRI-IV receives data from 9 of the 20 Italian Regions: Liguria, Abruzzo, Calabria, Friuli Venezia Giulia, Lombardy, Puglia, Sicily, Tuscany and Umbria. Previous estimates of influenza case costs were used in economic evaluations. Clinical-epidemiological and Virological surveillance of the seasonal epidemics from 1999-2008 showed that the highest epidemic period was 2004-2005, when a new variant of the H3N2 influenza virus subtype emerged (A/California/07/04). Indeed, the highest peak of morbidity in the decade occurred in February 2005 (12.6 per 1,000 inhabitants). In 1999-2008, H1N1 subtype strains circulated, and co-circulated with strains belonging to the H3N2 subtype and B type. Regarding B viruses in 2001-02, viruses belonged to the B/Victoria/02/07 lineage re-emerged, and in subsequent years co-circulated with viruses belonging to the B/Yamagata/lineage. The estimated costs of seasonal epidemics from 1999-2008 in Italy ranged from €15 to €20 billion, and the costs of the H1N1v pandemic ranged from €1.3 to €2.3 billion. This Italian study yields interesting conclusions: the results of Influenza surveillance in several developed countries vary markedly; influenza imposes a considerable social, healthcare and economic burden; most cases that occurred during the pandemic involved subjects under 14 years of age, and, although the clinical course of H1N1v influenza was usually mild, the related economic burden was heavy.

Open Access Article

Epidemiological analysis of severe hospitalized 2009 pandemic influenza A (H1N1) cases in Catalonia, Spain

Neus Cardeñosa, Anna Rodés, Núria Follia, Elsa Plasencia, Sarah Lafuente, César Arias, Roser Torra, Sofia Minguell, Glòria Ferrús, Irene Barrabeig and Pere Godoy
Pages 226 - 229
http://dx.doi.org/10.4161/hv.7.0.14609
Abstract | PDF

From 25th April 2009 to 20th January 2010, 771 laboratory confirmed cases of 2009 pandemic influenza A (H1N1) were admitted to intensive care units (ICU) or hospitalized in medical wards with clinical criteria of severe disease (hospitalized pneumonias, multiorganic failure, septic shock or admitted to ICU or death while hospitalized). 82% of cases were hospitalized between epidemiological week 43 and 48 (25th October – 5th December 2009). Median age of patients was 40 years (range 0-89 years) and 56% were males. 38.7% cases were in the 15-44 year age-group, 29.4% in the 45-64 and 21.8% were children under 15 years of age. 36.8% were admitted to an ICU and 48 died. Underlying conditions were absent in 29% of patients (up to 38% among those under 15 years old). Differences in the prevalence of underlying conditions were found between children and adults. In children less than 15 years old, asthma (16.2%), other respiratory diseases (12.7%), cognitive disorders (10.2%), epilepsy (8.7%) and neuromuscular disorders (7.1%) were the most frequent. In adults, chronic obstructive pulmonary disease (14.8%), cardiovascular diseases (12.6%), asthma (11.5%), diabetes (11.2%) and morbid obesity (10.6%) were the most frequent. Further surveillance is needed to better characterize the epidemiology of this pandemic.

Open Access Article

Differences in sentinel influenza confirmed incidence rates and clinical presentation of influenza virus: 2008-09 seasonal vs 2009-10 pandemic influenza

Núria Torner, Maite Morteruel, Ana Martinez, Luca Basile, Ricard Isanta and Tomas Pumarola
Pages 230 - 233
http://dx.doi.org/10.4161/hv.7.0.14612
Abstract | PDF

The Influenza sentinel surveillance network of Catalonia (PIDIRAC) allows for the study of circulating influenza virus (IV). The aim of this work was to assess differences between two influenza seasons, the 2008-2009 A(H3N2) season and the 2009-2010 season with predominance of pandemic influenza virus circulation. Incidence rate (IR) of confirmed influenza illness were calculated for both periods and age group. Clinical presentation features by age group (0-4,5-14,15-64 and >64 y.o.) were studied and compared for both seasons. Statistical significance of proportion differences assessed by statistic z and Mantel Hanzel’s Woolf test. The level of statistical significance was established at α=0.05. In both seasons studied, the 5-14 y.o. age group presented the highest confirmed influenza IR and highest presentation of cephalalgia as a symptom in the pandemic A(H1N1)2009 season. In conclusion, the significant burden of influenza, both seasonal and pandemic , on children should encourage upgrading vaccination coverage in these age groups and especially for those included in risk groups for whom yearly vaccination is recommended.

Open Access Article

Consultancy and surveillance of post-immunisation adverse events in the Veneto region of Italy for 1992-2008

Fabiola Micheletti, Ugo Moretti, Giuseppe Tridente and Giovanna Zanoni
Pages 234 - 239
http://dx.doi.org/10.4161/hv.7.0.14613
Abstract | PDF

Prevention and control of adverse events following immunization (AEFI) are fundamental activities of successful immunisation programs. AEFI reporting, investigation and analysis, integrated by consultancy for subjects needing a specialized evaluation, represent an ideal model for vaccine safety surveillance. In the Veneto Region of Italy the Green Channel Centre has been created by the local Public Health authority, to offer a consultancy activity for vaccinations at risk of adverse events and to ensure an efficient AEFI surveillance system with regular feedback data for vaccine personnel. This report updates the overall activity provided by the Green Channel between 1992 and 2008, concerning consultations for previous AEFI and contraindications to vaccinations and analysis of AEFI reports. After 1280 consultancy cases, 998 (78%) subjects were found eligible for vaccination, with personalized precautions suggested in 42% of cases. Of a total of 724 patients actually vaccinated as per the Green Channel instructions, only 55 subjects (7.6%) reported mild symptoms and one (0.3%) a moderate allergic reaction. Since 1993, a total of 5,006 AEFI reports have been collected and evaluated by the Green Channel against more than 20 millions of vaccine doses administered with an estimate mean AEFI rate of 2.3 x 10.000 doses per year. The majority of them (94%) were found in causal relationship with vaccines; of these, 267 reports (5,6% - 0.1/10,000 doses) were serious and 9 of these subjects, affected by a neurological event, were not recovered or were still on therapy at follow up. This regional activity has proven efficacious in evaluating and managing individual cases at potential risk of AEFI and integrating the national passive surveillance system.