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Journal of Virology, August 2000, p. 7651-7655, Vol. 74, No. 16
Department of Molecular and Cellular Biology,
Centro Nacional de Biotecnología, CSIC, Campus Universidad
Autónoma, E-28049 Madrid, Spain
Received 13 March 2000/Accepted 12 May 2000
A problem associated with the use of vaccinia virus recombinants as
vaccines is the existence of a large human population with preexisting
immunity to the vector. Here we showed that after a booster with
attenuated recombinant modified vaccinia virus Ankara (rMVA), higher
humoral and cellular immune responses to foreign antigens (human
immunodeficiency virus type 1 Env and Live-virus vector-based vaccines
have been proposed as excellent candidates for the design of novel
vaccines able to elicit long-term and protective humoral and
cell-mediated immune (CMI) responses, similar to the case during
natural infection with the corresponding pathogen. Certain properties
of poxvirus-derived vectors make them suitable for the development of
vaccines. Among properties of interest are the feasibility of
expressing in the same vector diverse and nonrelated antigens, the
potent and long-lasting immunity generated after immunization, and the
well-established protection raised upon vaccination with recombinant
vaccinia viruses (rVVs) in animal models and in field trials
(15). Low-cost production, temperature stability, and easy
administration are essential features when considering development of
vaccines for use worldwide, and poxvirus-derived vectors meet those
requirements (4). More recently, the beneficial
immunological effects when rVVs were administered during the booster in
bimodal immunization schemes have encouraged the use of poxvirus
vectors for human use against a variety of pathogens and malignancies
(22). However, safety concerns are present with the use of
live-virus-based vaccines, and particularly with rVV, as nondesired
side effects were demonstrated during the World Health
Organization-directed smallpox eradication program. Thus, the
development of strains of poxviruses that are highly safe for human use
are required for clinical trials. Modified vaccinia virus Ankara (MVA)
does not grow productively in cells of human origin, and in contrast to
the case for avirulent avipoxviruses, its beneficial potential was
evaluated in humans and showed no undesired effects, even in
immunocompromised individuals (14). Moreover, the levels of
expression of recombinant products from MVA in cultured cells are
higher than those from a wild-type strain, and vaccinated animals
developed protective immune responses to pathogens (16).
We have recently characterized in the mouse model the main
immunological features following infection with MVA by a systemic route
(19). We found that in comparison with a virulent laboratory strain (Western Reserve [WR]), the immune response elicited against the viral antigens was highly reduced, as neutralization antibodies were not significantly raised against MVA unless high viral doses were
employed. In addition, the specific cellular immune response against
MVA antigens was nearly five times lower than that to WR, irrespective
of the viral dose utilized. Significantly, the immune response elicited
against a foreign MVA-encoded antigen was at least as high, or even
higher at certain viral doses, than that with the virulent WR strain
(19). These data are of interest since in most systems
during repeated immunizations, a strong immune response against viral
vector proteins is associated with a lower effective immune response
against vector-expressed foreign antigens. In the case of VV this is
critical, as a large human population was vaccinated against smallpox
and still maintains a perdurable immunity against VV (2, 7).
Moreover, if recombinant poxvirus vectors are applied to humans,
preimmunized persons could not be recipients of further vaccines based
on the same vectors. This is an important issue considering that
several fatal diseases are endemic in third world and developing
countries, and for safety reasons, the immune status of the potential
vaccinees should thus be taken into account in the design of live-virus
vector-based vaccines.
The effects of preexisting immunity against VV upon secondary
inoculations with either the same vector encoding different exogenous
antigens or an rVV with distinct virulence have long been studied in
the context of the humoral immune response (6, 9, 18).
Briefly, it was demonstrated that immunity to virulent VV strains
influences negatively both the titer and duration of the antibody
response induced by a second distinct rVV (12) and that
prior immunization with an rVV reduces the protective immunity against
the pathogen achieved when the humoral immune response was raised after
a single or multiple inoculations of the rVV expressing the desired
antigen (21). However, a similar detailed analysis focusing
on the implications of the immunity elicited against the vector
antigens after priming, i.e., on the specific CMI response induced
against a new foreign antigen delivered by the vector following a
secondary immunization, has not been carried out. Our previous MVA
studies prompted us to analyze the implications of the fact that the
humoral and cellular immune responses elicited after priming against
MVA were low, while the CMI response against a foreign product
expressed from rMVA was similar to or even higher than that triggered
by rWR (19). Here, we compared the immune responses elicited
when either the virulent WR strain or MVA was employed in the first
immunization and the booster was given with rMVA.
The extent of preexisting immunity to VV antigens determines the
specific immune response against foreign antigens expressed upon
secondary immunization with rMVA.
The rVVs employed have the
inserted foreign genes in the thymidine kinase locus and have been
previously described (11, 20), except for MVAenv. They all
express
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Copyright © 2000, American Society for Microbiology. All rights reserved.
Attenuated Modified Vaccinia Virus Ankara Can Be
Used as an Immunizing Agent under Conditions of Preexisting
Immunity to the Vector
ABSTRACT
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Abstract
Text
References
-galactosidase) were found in
mice preimmunized with rMVA than in mice primed with the virulent
Western Reserve strain and boosted with rMVA. This enhancement
correlated with higher levels of expression of foreign antigens after
the booster.
TEXT
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Abstract
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References
-galactosidase (-Gal) under control of the p11 late
promoter. WRluc and MVAluc express luciferase under control of the p7.5
early-late promoter, WRenv expresses human immunodeficiency virus type
1 (HIV-1) IIIB gp160 under control of the p7.5 early-late promoter, as
does MVAenv, for which the levels of expression in baby hamster kidney
(BHK21) cells, as evaluated by Western blotting, were comparable to
those obtained with WRenv (data not shown). MVA derivatives were grown in BHK21 cells and plaque purified six times. WR derivatives were grown
in human HeLa cells. Sucrose cushion-purified viral stocks of rMVA and
rWR were titrated in BHK21 or African green monkey kidney BSC-40 cell
monolayers by immunostaining of fixed infected cultures with polyclonal
serum reactive against VV proteins (19) or by plaque assays, respectively.
-Gal gene and the luciferase gene (WRluc and
MVAluc, respectively). We first compared the humoral and cellular
immune responses after priming of mice with rMVA or rWR. Inoculated
animals were bled from the retro-orbital plexus, and immunoglobulin G
(IgG) antibodies to VV antigens at 14 days postinoculation were
determined by enzyme-linked immunosorbent assay (ELISA). Briefly,
96-well flat-bottom plates were coated with 100 µl of extracted
proteins from WR virions (1 µg/ml) (10) in carbonate
buffer at 4°C overnight. Antibodies were screened by plating serial
dilutions of sera, incubated (37°C, 1 h), and washed with
phosphate-buffered saline (PBS) plus 0.05% Tween 20 (PBS-T). After
incubation (37°C, 1 h) with peroxidase-conjugated goat
anti-mouse IgG specific antibodies (Southern Biotechnology, Birmingham,
Ala.), the color was developed with tetramethylbenzidine reagent
(Sigma, St. Louis, Mo.) for 5 to 10 min, and after the reaction was
stopped with 2 N SO4H2, absorbance was measured
at 450 nm on a Multiskan Plus plate reader (Labsystems, Chicago, Ill.).
The levels of antibodies to VV antigens are shown in Fig. 1A. As expected (19), at 14 days after the first immunization, MVAluc-inoculated mice had an IgG
response against VV that was sixfold lower than that of mice given
WRluc. Comparable results were obtained when ELISA plates were covered
with proteins extracted from purified MVA virions and reacted with sera
from the same group of animals (data not shown).
View larger version (21K):
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FIG. 1.
Humoral and CMI responses elicited against VV antigens
in mice primed with rMVA or rWR. Mice (four per group) were i.p.
inoculated with 107 PFU of MVAluc or WRluc and at 14 days
postinoculation pooled sera were tested for specific IgG antibodies
against VV by ELISA. (A) Averages of triplicate measurements of
absorbance at 450 nm of a 1/400 dilution of sera. (B) The number of
IFN- 
-secreting CD8+ T cells specific for VV antigens in
spleens was determined by ELISPOT assay; bars represent averages for
three pooled samples (± standard deviations) from triplicate cultures.
(C) In an independent experiment, groups of four mice were inoculated
as described above, and neutralizing antibodies 40 days after the virus
inoculation were determined by measuring the inhibition of virus plaque
formation on BSC-40 cells using the wild-type WR strain. The percentage
of neutralization was calculated as (number of PFU with immune
serum/number of PFU with naive serum) × 100. Averages of two
independent measurements are shown.
)-secreting T cells
(10). Briefly, 96-well nitrocellulose plates (Millipore
Corporation, Bedford, Mass.) were coated with 8 µg of anti-mouse
IFN- monoclonal antibody R4-6A2 (PharMingen, San Diego, Calif.) per
ml in 75 µl of PBS. After overnight incubation at room temperature,
wells were washed with RPMI complete medium and blocked with RPMI plus
10% fetal calf serum during 1 h at 37°C. Afterwards,
triplicates of erythrocyte-depleted spleen cells were plated in twofold
dilutions starting with 5 × 105 cells/well. P815
cells (a mastocytoma cell line that expresses only major
histocompatibility complex class I molecules) were used as
antigen-presenting cells (105 cells/well), after infection
with 5 PFU of WRluc per cell and treatment with mitomycin C (30 µg/ml) (Sigma) at 4.5 h postinfection. The control was
mock-infected P815 cells. Plates were incubated in a humidified
atmosphere (37°C, 24 h), washed extensively with PBS-T, and
incubated (room temperature, 2 h) with a solution in PBS-T of
biotinylated anti-mouse IFN- monoclonal antibody XMG1.2 (2 µg/ml)
(PharMingen). The plates were washed with PBS-T, and 100 µl of
peroxidase-labeled avidin (1/800 dilution in PBS-T) (Sigma) was added
to each well and incubated at room temperature for 1 h. After
washing, spots were developed by adding the substrate 3,3'-diaminobenzidine tetrahydrochloride (1 µg/ml) (Sigma) in 50 mM
Tris-HCl (pH 7.5) containing 0.015% hydrogen peroxide and counted
using an MZ122 APO and Imaging System QWIN software (Leica, Cambridge,
United Kingdom). As depicted in Fig. 1B, 14 days after primary
inoculation, the number of anti-VV CD8+ IFN--secreting T
cells was approximately four times lower in mice inoculated with MVAluc
than in mice inoculated with WRluc. The results in Fig. 1A and B show
that the humoral as well as the cellular immune response induced
against VV antigens was quantitatively different in animals primed with
rMVA versus rWR.
Neutralizing antibody titers in heat-inactivated (56°C, 30 min) sera
from immunized and control mice were evaluated. Serial dilutions of
sera in PBS plus 2% fetal calf serum were incubated with 200 PFU of
wild-type WR (37°C, 1 h); confluent BSC-40 cell monolayers were
infected in triplicate, and plaques were visualized at 48 h
postinfection by crystal violet staining and counted. As a negative
control, sera from mock-infected mice were used. The number of plaques
obtained with each serum was referred to the control value and used to
calculate the neutralization titers (Fig. 1C). Clearly, the
neutralization titers were considerably lower in sera from mice primed
with MVA than in those from mice primed with WR.
Next, we compared the immune responses with a prime-boost combination
of vectors. Mice primed as for Fig. 1 were boosted 40 days later with
5 × 107 PFU of MVAenv expressing the HIV-1 Env gene
of clade B (IIIB). Ten days after the second immunization with MVAenv,
the specific cellular immune response against an epitope on the V3 loop
was evaluated by ELISPOT assay as described for Fig. 1B, except that P815 cells were pulsed with a 10
6 M concentration of the
10-amino-acid specific peptide RGPGRAFVTI (24). Control mice
received only one inoculation with either MVAenv or the similar
WR-based virus (WRenv). A single immunization with MVAenv or WRenv
induces a specific anti-Env CD8+-T-cell response of similar
magnitudes for both vectors (Fig. 2A,
insert). When mice were preimmunized with either MVAluc or WRluc, the
total amount of anti-Env CD8+ IFN--secreting T cells
diminished after the booster (Fig. 2A). However, nearly fourfold more
specific CD8+ T cells were present after immunization with
MVAenv in mice preimmunized with MVA compared to mice preimmunized with
WR. Thus, it appears that the lower immune response raised against the
MVA antigens compared to WR antigens after a primary inoculation
allowed a more potent response to an MVA-delivered antigen to which
mice were naive (the HIV-1 Env). The low titer of anti-gp160 antibodies generated limited our ability to measure the antibody response in
primed mice to a newly presented antigen delivered by immunization with
a second recombinant virus (data not shown). The limitation of
preexisting immunity to VV antigens in mounting of a humoral immune
response to a foreign antigen delivered from a second recombinant VV
vector has been extensively studied by other authors (12, 20).
|
-Gal antigen
(a protein that must be expressed after viral infection to be exposed
to the immune system) delivered by rMVA and rWR was also analyzed. The
levels of IgG antibodies against the -Gal antigen were evaluated by
ELISA as described above, except that plates were coated with 5 µg of
a solution of commercial -Gal (Sigma) per ml. Anti--Gal
antibodies in pooled serum were measured 14 days after priming with
either WRluc or MVAluc and also 14 days after the booster with MVAenv.
While after priming, low antibody titers to -Gal are produced (data
not shown) (19), after the booster, specific IgG levels were
nearly three times higher in animals preimmunized with MVAluc than in
those animals preimmunized with WRluc (Fig. 2B, left panel). As
expected, the anamnestic response against VV envelope antigens (which
are exposed on the virus particles and do not need to be expressed to
affect the immune system) was different. While anti-WR antibody titers
were high after a booster with MVAenv, these levels were significantly lower in mice preimmunized with MVAluc and boosted with MVAenv (Fig.
2B, right panel), in correlation with the primary anti-VV antibody
response observed in WR-primed mice with respect to those preimmunized
with MVAluc (Fig. 1A).
The outcome of the immune response in animals primed with VV is
determined by the levels of expression of the recombinant antigen
during the booster.
We next investigated whether differences in
the anti-Env and anti--Gal immune responses in mice primed with
WRluc or MVAluc and boosted with MVAenv could correlate with a
different expression of the antigen product upon secondary MVA
inoculation. To circumvent difficulties associated with the
quantitation of expressed Env protein in animals, we primed mice with
1 × 107 PFU of WRenv or MVAenv and then carried out a
secondary inoculation with 5 × 107 PFU of MVAluc
(Fig. 3). Since MVAluc expresses the
luciferase gene under control of the same early-late promoter as the
rVV expressing the HIV-1 Env gene, luciferase levels at short times postinoculation indicate the extent of virus gene expression in target
tissues. Luciferase activity in spleens and ovaries of infected mice
was measured at various times postinoculation from cleared tissue
homogenates in extraction buffer (300 µl/spleen and 100 µl/ovary)
(Promega Corp., Madison, Wis.) in the presence of luciferin and ATP
using a Lumat LB 9501 Berthold luminometer (Berthold, Nashua, N.H.) and
was expressed as relative luciferase units per milligram of protein.
Protein content in tissue extracts was measured with the bicinchoninic
acid protein assay reagent kit (Pierce Co., Rockford, Ill.). As shown
in Fig. 3, after the booster with MVAluc there is a clear difference in
levels of luciferase expression between WRenv-primed mice (with a
strong anti-VV immune response) and animals primed with MVAenv (with a
low anti-VV immune response). At 6 h postbooster, when MVA reaches
maximum values of gene expression in vivo (19), the
differences between groups are 2.2 log units in ovaries and about 1 log
unit in spleen. These findings highlight the differences in gene
expression when animals are primed with WR or MVA and then boosted with
rMVA, which are related to the virulence of the virus strain used for
priming.
|
-Gal) than in an rWR
prime-rMVA boost combination. Although here we have not characterized
the boosting capacity of MVA against HIV-1 Env, it has been
demonstrated (23) that anti-simian immunodeficiency virus
Gag/Pol cytotoxic-T-lymphocyte responses can be boosted with the same
MVA recombinant upon intramuscular injection of rhesus monkeys. By
contrast, Belyakov et al. (1) showed in mice that there was
no increase in the CMI elicited upon repeated subcutaneous injections
with MVA expressing the HIV-89.6 Env protein (rMVA89.6) in comparison
with a single vector injection; moreover, in mice preimmunized with
WR-based recombinant virus, there was no response to HIV-89.6 Env upon
booster with rMVA89.6 (1). In this investigation we showed
that although prior immunity to the MVA vector causes a significant
inhibitory effect on the CMI response against the HIV-1 Env, this
immune response is measurable and is nearly fourfold higher than that
observed in mice preimmunized with the WR vector (Fig. 2). Differences
due to the inoculation routes (i.p. versus subcutaneous) and viral
doses used in the two studies might explain these discrepancies, as the
virus input has a relevant effect on the extent of the specific humoral
and CMI responses against MVA antigens (19). Moreover,
Belyakov et al. evaluated the specific cytotoxic-T-lymphocyte response after a longer period of time following the booster than we did in our
study, and also they used an assay less sensitive than the ELISPOT assay.
The stronger response to foreign antigens elicited by MVA after a
booster in mice primed with MVA versus mice primed with WR is, in part,
due to levels of expression of the antigen. Indeed, higher levels of
luciferase are produced in mice with preexisting immunity against MVA
antigens than in those with immunity against WR (Fig. 3). This is
probably because virus-neutralizing antibodies are lower in MVA-primed
mice than in WR-primed mice (Fig. 1C). We found that those differences
were linked to the ability of MVA administered in the second dose to
infect target cells, which has also implications for the anamnestic
antibody responses against an antigen expressed from rVV. Indeed, those
reasons were pointed out by several authors (6, 9, 21) but
were not demonstrated. The preexisting immunity to VV proteins,
revealed by both neutralizing antibody titers and anti-VV
CD8+ T cells present in mice at the time of the boost (Fig.
1), will limit the virus input that can reach and infect target tissues during booster and, hence, decrease the expression of virus-encoded genes.
Our findings establish benefits, besides safety concerns, of the use of
MVA as a live-virus-based vaccine, due to the ability to elicit
specific secondary immune responses upon repeated inoculations with
MVA. This in conjunction with the proven efficient boosting capacity of
MVA in bimodal immunization protocols (22) makes MVA a
strain with potential use as a VV-based vaccine vector. Obviously, the
intrinsic properties of MVA as an immunogen can be improved further by
varying the time in the immunization scheme, increasing the strength of
the VV promoter used to express the antigens, or coexpressing
immunomodulatory molecules.
| |
ACKNOWLEDGMENTS |
|---|
Juan C. Ramírez and M. Magdalena Gherardi contributed equally to this work.
The excellent technical assistance of M. Victoria Jiménez is acknowledged.
This work was supported by grants 08.6/0020/97 from Comunidad Autónoma de Madrid (CAM), SAF98-0056 from Comision Interministerial de Ciencia y Tecnología (CICYT), Spain, and BIO4-CT98-0456 from the European Union. J.C.R. is a recipient of postdoctoral fellowship from the CAM, Spain. M.M.G. is a researcher from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
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FOOTNOTES |
|---|
* Corresponding author. Mailing address: Centro Nacional Biotecnologia (CSIC), Campus Cantoblanco, 28049 Madrid, Spain. Phone: 34-91-585-4503. Fax: 34-91-585-4506. E-mail: mesteban{at}cnb.uam.es.
Present address: Department of Applied Microbiology, Centro de
Estudios Farmacológicos y Botánicos (CEFYBO-CONICET),
Buenos Aires, Argentina.
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REFERENCES |
|---|
|
Top
Abstract
Text
References
|
|---|
| 1. |
Belyakov, I. M.,
B. Moss,
W. Strober, and J. A. Berzofsky.
1999.
Mucosal vaccination overcomes the barrier to recombinant vaccinia immunization caused by preexisting poxvirus immunity.
Proc. Natl. Acad. Sci. USA
96:4512-4517 |
| 2. | Cooney, E. L., A. C. Collier, P. D. Greenberg, R. W. Coombs, J. Zarling, D. E. Arditti, M. C. Hoffman, S. L. Hu, and L. Corey. 1991. Safety of and immunological response to a recombinant vaccinia virus vaccine expresing HIV envelope glycoprotein. Lancet 337:567-572[CrossRef][Medline]. |
| 3. |
Cooney, E. L.,
M. J. McElrath,
L. Corey,
S. Hu,
A. C. Collier,
D. Arditti,
M. Hoffman,
R. W. Coombs,
G. E. Smith, and P. D. Greenberg.
1993.
Enhanced immunity to HIV envelope elicited by a combined vaccine regimen consisting of priming with a vaccinia recombinant expressing HIV envelope and boosting with gp160 protein.
Proc. Natl. Acad. Sci. USA
90:1882-1886 |
| 4. | Cox, W. I., J. Tartaglia, and E. Paoletti. 1992. Poxvirus recombinants as live vaccines, p. 123-162. In M. M. Binns, and G. L. Smith (ed.), Recombinant poxviruses. CRC Press, Boca Raton, Fla. |
| 5. |
Cremer, K.,
M. Mackett,
C. Wohlenberg,
A. L. Notkins, and B. Moss.
1985.
Vaccinia virus recombinants expressing herpes simplex type 1 glycoprotein D prevents latent herpes in mice.
Science
228:737-740 |
| 6. | Dallo, S., J.-S. Maa, J. R. Rodriguez, D. Rodriguez, and M. Esteban. 1989. Humoral immune response elicited by highly attenuated variants of vaccinia virus and by attenuated recombinant expressing HIV-1 envelope protein. Virology 173:323-329[CrossRef][Medline]. |
| 7. |
Erickson, A. L., and C. M. Walker.
1993.
Class I major histocompatibility complex-restricted cytotoxic T cell responses to vaccinia virus in humans.
J. Gen. Virol
74:751-754 |
| 8. | Etlinger, H., and W. Altenburger. 1991. Overcoming inhibition of antibody responses to a malaria recombinant vaccinia virus caused by prior exposure to wild type virus. Vaccine 9:470-472[CrossRef][Medline]. |
| 9. | Flexner, C., B. R. Murphy, J. F. Rooney, C. Wohlenberg, V. Yuferov, A. L. Notkins, and B. Moss. 1988. Succesful vaccination with a polyvalent live vector despite existing immunity to an expressed antigen. Nature 335:259-262[CrossRef][Medline]. |
| 10. | Gherardi, M. M., and M. Esteban. 1999. Mucosal and systemic immune responses induced after oral delivery of vaccinia virus recombinants. Vaccine 17:1074-1083[CrossRef][Medline]. |
| 11. |
Gherardi, M. M.,
J. C. Ramirez,
D. Rodriguez,
J. R. Rodriguez,
G. Sano,
F. Zavala, and M. Esteban.
1999.
IL-12 delivery from recombinant vaccinia virus attenuates the vector and enhances the cellular immune response against HIV-1 env in a dose-dependent manner.
J. Immunol.
162:6724-6733 |
| 12. | Kündig, T. M., C. P. Kalberer, H. Hengartner, and R. M. Zinkernagel. 1993. Vaccination with two different vaccinia recombinants viruses: long-term inhibition of secondary vaccination. Vaccine 11:1154-1158[CrossRef][Medline]. |
| 13. |
Kutinovä, L.,
V. Ludvíkova,
L. Maresová,
J. Broucek,
P. Hainz, and V. Vonka.
1999.
Effect of virulence on immunogenicity of single and double vaccinia virus recombinants expressing differently immunogenic antigens: antibody-response inhibition induced by immunization with a mixture of recombinants differing in virulence.
J. Gen. Virol.
80:2901-2908 |
| 14. | Mayr, A., H. Stickl, H. K. Muller, K. Denner, and H. Singer. 1978. The smallpox vaccination strain MVA: marker, genetic structure, experience gained with the parenteral vaccination and behaviour in organisms with a debilitated defence mechanism. Zentbl. Backteriol. Hyg. B 167:375-390. |
| 15. |
Moss, B.
1996.
Genetically engineered poxviruses for recombinant gene expression, vaccination and safety.
Proc. Natl. Acad. Sci. USA
93:11341-11348 |
| 16. | Moss, B., M. W. Carroll, L. S. Wyatt, J. R. Bennink, V. Hirsch, S. Golstein, W. R. Elkins, T. R. Fuerst, J. D. Lifson, M. Piatak, N. P. Restifo, W. Overwijk, R. Chamberlain, S. A. Rosenberg, and G. Sutter. 1996. Host range restricted non-replicating vaccinia virus vectors as vaccine candidates. Adv. Exp. Med. Biol. 397:7-13[Medline]. |
| 17. |
Paoletti, E.,
B. R. Lipinskas,
C. Samsonoff,
S. Mercer, and D. Panicalli.
1984.
Construction of live vaccines using genetically engineered poxviruses: biological activity of vaccinia virus recombinants expressing the hepatitis B virus surface antigen and the herpes simplex virus glycoprotein D.
Proc. Natl. Acad. Sci. USA
81:193-197 |
| 18. |
Perkus, M. E.,
A. Piccini,
B. R. Lipinskas, and E. Paoletti.
1985.
Recombinant vaccinia virus: immunization against multiple pathogens.
Science
229:981-984 |
| 19. |
Ramírez, J. C.,
M. M. Gherardi, and M. Esteban.
2000.
The biology of attenuated modified vaccinia virus Ankara recombinant vector in mice: fate and activation of B- and T-cell immune responses in comparison with the Western Reserve strain and advantages as a vaccine.
J. Virol.
74:923-933 |
| 20. |
Rodríguez, J. F.,
D. Rodríguez,
J. R. Rodríguez,
E. B. McGowan, and M. Esteban.
1988.
Expression of the firefly luciferase gene in vaccinia virus: a highly sensitive gene marker to follow virus dissemination in tissues of infected animals.
Proc. Natl. Acad. Sci. USA
85:1667-1671 |
| 21. |
Rooney, J. F.,
C. Wohlenberg,
K. J. Cremer,
B. Moss, and A. L. Notkins.
1988.
Immunization with a vaccinia virus recombinant expressing herpes simplex virustype 1 glycoprotein D: long-term protection and effect of revaccination.
J. Virol.
62:1530-1534 |
| 22. | Schneider, J., S. C. Gilbert, C. M. Hannan, P. Degano, E. prieur, E. G. Sheu, M. Plebanski, and A. V. S. Hill. 1999. Induction of CD8+ T cells using heterologous prime-boost immunisation strategies. Immunol. Rev. 170:29-38[CrossRef][Medline]. |
| 23. |
Seth, A.,
I. Ourmanov,
M. J. Kuroda,
J. E. Schmitz,
M. W. Carroll,
L. S. Wyatt,
B. Moss,
M. A. Forman,
V. M. Hirsch, and N. L. Letvin.
1998.
Recombinant modified vaccinia virus Ankara-simian immunodeficiency virus gag pol elicits cytotoxic T lymphocytes in rhesus monkeys detected by a major histocompatibility complex class I/peptide tetramer.
Proc. Natl. Acad. Sci. USA
95:10112-10116 |
| 24. | Takeshita, T., H. Takahashi, S. Kozlowski, J. D. Ahlers, c. D. Pendleton, R. L. Moore, Y. Nakagawa, K. Yokomuro, B. S. Fox, D. H. Margulies, and J. A. Berzofsky. 1995. Molecular analysis of the same HIV peptide functionally binding to both a class I and a class II MHC molecule. J. Immunol. 154:1973-1986[Abstract]. |
Journal of Virology, August 2000, p. 7651-7655, Vol. 74, No. 16
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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[Abstract] [Full Text] -
Stober, C. B., Lange, U. G., Roberts, M. T. M., Alcami, A., Blackwell, J. M.
(2007). Heterologous Priming-Boosting with DNA and Modified Vaccinia Virus Ankara Expressing Tryparedoxin Peroxidase Promotes Long-Term Memory against Leishmania major in Susceptible BALB/c Mice. Infect. Immun.
75: 852-860
[Abstract] [Full Text] -
Walther, M., Thompson, F. M., Dunachie, S., Keating, S., Todryk, S., Berthoud, T., Andrews, L., Andersen, R. F., Moore, A., Gilbert, S. C., Poulton, I., Dubovsky, F., Tierney, E., Correa, S., Huntcooke, A., Butcher, G., Williams, J., Sinden, R. E., Hill, A. V. S.
(2006). Safety, Immunogenicity, and Efficacy of Prime-Boost Immunization with Recombinant Poxvirus FP9 and Modified Vaccinia Virus Ankara Encoding the Full-Length Plasmodium falciparum Circumsporozoite Protein.. Infect. Immun.
74: 2706-2716
[Abstract] [Full Text] -
Palena, C., Foon, K. A., Panicali, D., Yafal, A. G., Chinsangaram, J., Hodge, J. W., Schlom, J., Tsang, K. Y.
(2005). Potential approach to immunotherapy of chronic lymphocytic leukemia (CLL): enhanced immunogenicity of CLL cells via infection with vectors encoding for multiple costimulatory molecules. Blood
106: 3515-3523
[Abstract] [Full Text] -
Wang, Z., La Rosa, C., Mekhoubad, S., Lacey, S. F., Villacres, M. C., Markel, S., Longmate, J., Ellenhorn, J. D. I., Siliciano, R. F., Buck, C., Britt, W. J., Diamond, D. J.
(2004). Attenuated poxviruses generate clinically relevant frequencies of CMV-specific T cells. Blood
104: 847-856
[Abstract] [Full Text] -
Gherardi, M. M., Perez-Jimenez, E., Najera, J. L., Esteban, M.
(2004). Induction of HIV Immunity in the Genital Tract After Intranasal Delivery of a MVA Vector: Enhanced Immunogenicity After DNA Prime-Modified Vaccinia Virus Ankara Boost Immunization Schedule. J. Immunol.
172: 6209-6220
[Abstract] [Full Text] -
Wang, Z., La Rosa, C., Maas, R., Ly, H., Brewer, J., Mekhoubad, S., Daftarian, P., Longmate, J., Britt, W. J., Diamond, D. J.
(2004). Recombinant Modified Vaccinia Virus Ankara Expressing a Soluble Form of Glycoprotein B Causes Durable Immunity and Neutralizing Antibodies against Multiple Strains of Human Cytomegalovirus. J. Virol.
78: 3965-3976
[Abstract] [Full Text] -
Cosma, A., Buhler, S., Nagaraj, R., Staib, C., Hammarin, A.-L., Wahren, B., Goebel, F. D., Erfle, V., Sutter, G.
(2004). Neutralization Assay Using a Modified Vaccinia Virus Ankara Vector Expressing the Green Fluorescent Protein Is a High-Throughput Method To Monitor the Humoral Immune Response against Vaccinia Virus. CVI
11: 406-410
[Abstract] [Full Text] -
Hodge, J. W., Poole, D. J., Aarts, W. M., Gomez Yafal, A., Gritz, L., Schlom, J.
(2003). Modified Vaccinia Virus Ankara Recombinants Are as Potent as Vaccinia Recombinants in Diversified Prime and Boost Vaccine Regimens to Elicit Therapeutic Antitumor Responses. Cancer Res.
63: 7942-7949
[Abstract] [Full Text] -
Hornemann, S., Harlin, O., Staib, C., Kisling, S., Erfle, V., Kaspers, B., Hacker, G., Sutter, G.
(2003). Replication of Modified Vaccinia Virus Ankara in Primary Chicken Embryo Fibroblasts Requires Expression of the Interferon Resistance Gene E3L. J. Virol.
77: 8394-8407
[Abstract] [Full Text] -
Gherardi, M. M., Najera, J. L., Perez-Jimenez, E., Guerra, S., Garcia-Sastre, A., Esteban, M.
(2003). Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes. J. Virol.
77: 7048-7057
[Abstract] [Full Text] -
Espenschied, J., Lamont, J., Longmate, J., Pendas, S., Wang, Z., Diamond, D. J., Ellenhorn, J. D. I.
(2003). CTLA-4 Blockade Enhances the Therapeutic Effect of an Attenuated Poxvirus Vaccine Targeting p53 in an Established Murine Tumor Model. J. Immunol.
170: 3401-3407
[Abstract] [Full Text] -
Harrington, L. E., Most, R. v. d., Whitton, J. L., Ahmed, R.
(2002). Recombinant Vaccinia Virus-Induced T-Cell Immunity: Quantitation of the Response to the Virus Vector and the Foreign Epitope. J. Virol.
76: 3329-3337
[Abstract] [Full Text] -
Sharpe, S., Polyanskaya, N., Dennis, M., Sutter, G., Hanke, T., Erfle, V., Hirsch, V., Cranage, M.
(2001). Induction of simian immunodeficiency virus (SIV)-specific CTL in rhesus macaques by vaccination with modified vaccinia virus Ankara expressing SIV transgenes: influence of pre-existing anti-vector immunity. J. Gen. Virol.
82: 2215-2223
[Abstract] [Full Text]
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