DNA vaccination by immersion and ultrasound to trout viral haemorrhagic septicaemia virus

Fernandez‐Alonso, M.; Rocha, A.; Coll, Julio

doi:10.1016/s0264-410x(01)00046-9

Cited by 72 publications

(31 citation statements)

References 35 publications

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“…Among fish rhabdoviruses (novirhabdoviruses), the viral hemorrhagic septicemia virus (VHSV), which originated in Europe but has recently been found in America (26, 41), affects not only salmonids but also cod, turbot (38), sea bass, eels, flatfish, and shrimp (8,24,27). Despite many efforts, including successful DNA vaccines at the laboratory level, a commercial vaccine against VHSV is not yet available (1,18,25,30).The whole genome of VHSV has been sequenced (23), and the disulfide structure of its protein G (pG) has been elucidated (12). Reverse genetic methods have been developed for the VHSV-related rhabdovirus infectious hematopoietic necrosis virus (5-7), which could allow the design of new vaccines.…”

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“…Three of these mutations mapped around two pG locations, showing amino acid variations among VHSV isolates. Because 40% of the VHSV-immunized trout strongly recognized linear epitopes on these regions (15,17,35), the mutants described here might be useful in attempts to design attenuated VHSV strains to be used in DNA vaccines (2,3,18) or in vaccines obtained by reverse genetic methods (5,6).…”

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Conformation- and Fusion-Defective Mutations in the Hypothetical Phospholipid-Binding and Fusion Peptides of Viral Hemorrhagic Septicemia Salmonid Rhabdovirus Protein G

Rocha¹,

Ruiz²,

Tafalla³

et al. 2004

J Virol

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Fourteen single and two double point mutants in the highly conserved region (positions 56 to 159) of the G gene of viral hemorrhagic septicaemia virus (VHSV), a salmonid rhabdovirus, were selected and obtained in plasmids by site-directed mutagenesis. Fish cell monolayers transfected with the mutant plasmids were then assayed for protein G (pG) expression, conformation-dependent monoclonal antibody (MAb) reactivity, and cell-cell fusion. Some mutations located in the phospholipid-binding p2 peptide (positions 82 to 110; mutants P86A, A96E, G98A, and R107A) abolished both MAb recognition and fusion activity, while others (P79A, L85S, and R103A) abolished MAb recognition but retained fusion at similar or lower pHs compared to those for the wild type. Phospholipid-binding assays of p2-derived synthetic peptides suggested that phosphatidylserine binding was not affected by the mutations studied. On the other hand, three (P79A, L85S, and T135E) of the four mutants retaining fusion activity mapped around two locations showing amino acid variation in 22 VHSV isolates and in neutralizing MAb-resistant mutants described previously. Mutations located in the hypothetical fusion peptide (positions 142 to 159; mutants F147K, P148K, and W154K) abolished both MAb recognition and fusion activity. The existence of mutants with altered conformation and defective fusion in both p2 and fusion peptides provides further evidence in favor of the participation of these and adjacent regions in some of the steps of the VHSV fusion processes, as suggested by previous studies. In addition, because the studied region induced strong immunological responses in trout, some of the mutants described here might be used to design attenuated VHSV vaccines.Rhabdoviruses cause highly damaging diseases in fish reared by the worldwide salmon culture industry. Among fish rhabdoviruses (novirhabdoviruses), the viral hemorrhagic septicemia virus (VHSV), which originated in Europe but has recently been found in America (26, 41), affects not only salmonids but also cod, turbot (38), sea bass, eels, flatfish, and shrimp (8,24,27). Despite many efforts, including successful DNA vaccines at the laboratory level, a commercial vaccine against VHSV is not yet available (1,18,25,30).The whole genome of VHSV has been sequenced (23), and the disulfide structure of its protein G (pG) has been elucidated (12). Reverse genetic methods have been developed for the VHSV-related rhabdovirus infectious hematopoietic necrosis virus (5-7), which could allow the design of new vaccines. The study of rhabdovirus fusion in the VHSV model could be important in the design of alternative vaccines to fight these diseases.The pG mutants of vesicular stomatitis virus (VSV), a wellstudied mammalian rhabdovirus, with an altered extent of fusion and/or optimal fusion pH in vitro, had mutations located either in the fusion peptide or in carboxy-terminal regions affecting the low-pH conformational changes required for fusion (39,40,43). Alignment of the pG sequence of VSV with those of 14 o...

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confidence: 99%

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Conformation- and Fusion-Defective Mutations in the Hypothetical Phospholipid-Binding and Fusion Peptides of Viral Hemorrhagic Septicemia Salmonid Rhabdovirus Protein G

Rocha¹,

Ruiz²,

Tafalla³

et al. 2004

J Virol

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“…However, most changes in gene expression that occur with resistance mechanisms in short-term and long-term immunity are not fully understood (Goetz & MacKenzie, 2008). Furthermore, more basic knowledge on mucosal immunity is required to move rhabdoviral DNA vaccines from the laboratory into the field, as existing vaccines still require either intramuscular injection in individual fish or stronger (adjuvanted) immune responses to facilitate mass delivery methods, such as those using oral- (delasHeras et al, 2010;Tian et al, 2008) or ultrasound-aided (Fernandez-Alonso et al, 2001) immunization. Studies using microarrays could greatly contribute to furthering this basic knowledge (Secombes, 2008).…”

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confidence: 99%

Use of Microarray Technology to Improve DNA Vaccines in Fish Aquaculture - The Rhabdoviral Model

Encinas¹,

Gómez-Casado²,

Estepa³

et al. 2012

Health and Environment in Aquaculture

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“…While the literature on ultrasound-induced transdermal drug delivery is substantial, that on ultrasound-assisted transcutaneous immunisation is scant and includes a few reports on ultrasound-assisted vaccination in fish, for application in fish farming (Fernandez-Alonso et al, 2001;Zhou et al, 2002a-b;Navot et al, 2004), a report on the role of the macrophage system in the mediation of the immunostimulating effects of low frequency ultrasound (Khodareva & Sizyakina 1991) and a paper on transcutaneous vaccination in mice (Tezel et al, 2005). In the latter study, the authors showed enhanced levels of serum antibody titres, following topical application of antigen on to skin pre-treated with low-frequency ultrasound in the presence of sodium dodecyl sulphate (SDS).…”

Section: Introductionmentioning

confidence: 99%

Transcutaneous immunisation assisted by low-frequency ultrasound

Dahlan

Alpar

Stickings

et al. 2009

International Journal of Pharmaceutics

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Low-frequency ultrasound application is known to increase the skin's permeability to large molecules such as vaccines, and to enable transcutaneous immunisation. Sodium dodecyl sulphate (SDS) -a skin irritant -is often included in the coupling medium at 1% w/v, as this has been found to enhance skin permeability. In this paper we show, for the first time, the feasibility of low-frequency ultrasound-assisted transcutaneous immunisation in the absence of SDS. Antibody titres were strongly influenced by experimental conditions. SDS presence in the coupling medium increased antibody titres, though a lower concentration of 0.5% w/v generated much higher titres than the commonly used 1%w/v, despite causing less skin damage. A lower ultrasound duty cycle of 10% generated higher antibody titres than a duty cycle of 20%, also despite causing lower skin damage. Such lack of correlation between skin damage and immune responses indicates that enhancement of skin permeability to topically-applied antigen (as indicated by changes in skin integrity) was not the main mechanism of low-frequency ultrasound-assisted skin immunisation.

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DNA vaccination by immersion and ultrasound to trout viral haemorrhagic septicaemia virus

Cited by 72 publications

References 35 publications

Conformation- and Fusion-Defective Mutations in the Hypothetical Phospholipid-Binding and Fusion Peptides of Viral Hemorrhagic Septicemia Salmonid Rhabdovirus Protein G

Conformation- and Fusion-Defective Mutations in the Hypothetical Phospholipid-Binding and Fusion Peptides of Viral Hemorrhagic Septicemia Salmonid Rhabdovirus Protein G

Use of Microarray Technology to Improve DNA Vaccines in Fish Aquaculture - The Rhabdoviral Model

Transcutaneous immunisation assisted by low-frequency ultrasound

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