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WO1999007869A1 - Vaccin vivant recombinant comprenant un virus ne se repliquant pas ou se repliquant sans efficacite - Google Patents

Vaccin vivant recombinant comprenant un virus ne se repliquant pas ou se repliquant sans efficacite Download PDF

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Publication number
WO1999007869A1
WO1999007869A1 PCT/US1997/013836 US9713836W WO9907869A1 WO 1999007869 A1 WO1999007869 A1 WO 1999007869A1 US 9713836 W US9713836 W US 9713836W WO 9907869 A1 WO9907869 A1 WO 9907869A1
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vaccine
virus
recombinant
mice
mva
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PCT/US1997/013836
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Parker A. Small, Jr.
Bradley Stephen Bender
Catherine Ann Meitin
Bernard Moss
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University Of Florida
The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services
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Priority to PCT/US1997/013836 priority Critical patent/WO1999007869A1/fr
Priority to AU39100/97A priority patent/AU3910097A/en
Publication of WO1999007869A1 publication Critical patent/WO1999007869A1/fr

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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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Definitions

  • Live Recombi nant vacci ne compri si ng i nefficiently or non-repl icati ng vi rus.
  • the subject invention described herein relates to the field of vaccines.
  • Each year millions of children die of vaccine-preventable diseases. It is estimated that at least one child in five, including many of the children in the United States under the age of 2. have not been fully vaccinated (Gibbons. 1994).
  • the Children's Vaccine Initiative proposed the development of a multivalent. heat-stable, inexpensive, orally- administered. safe and effective vaccine. Oral administration of vaccines may also help reduce children's fear of shots at the doctor's office.
  • Vaccinia is a non-oncogenic virus that reproduces entirely within the cytoplasm of a host eukaryotic cell.
  • Methods for inserting foreign DNA sequences into vaccinia are known in the art and at least 25 kb of heterologous DNA (representing about 10-20 genes) can be inserted into vaccinia virus genome (Smith et al.. 1983).
  • an immune response can be induced to the products of multiple heterologous genes expressed by a recombinant virus (Perkus et al.. 1985). Lyophilized vaccinia virus is very heat-stable, maintaining infectivity after two hours at 100°C (Arita. 1973).
  • Vaccinia vaccine is also inexpensive, having been produced for about $.03 to $.04/per dose during the smallpox eradication program (Fenner ei al.. 1988).
  • vaccinia virus can be readily grown in laboratory cultures and prepared in stable, freeze-dried forms.
  • vaccinia virus is rapidly destroyed by stomach acid or bile (Hochstein-Mintzel et al.. 1972), thereby eliminating intragastric immunization as a viable mode of administration.iHACi]
  • vaccinia can replicate in the eukaryotic cell cytoplasm, the potential for replication of a pathogenic virus vector cannot be completely ruled out when using standard vaccinia virus as a recombinant vector for vaccine purposes. It is for this reason that in a preferred embodiment of the present invention, a replication deficient virus is used as the vector.
  • the effectiveness of a particular vaccine may depend upon which cells or tissues of the immune system are activated by the vaccine.
  • the role of three arms of the immune system namely serum antibody, mucosal IgA antibody, and cytotoxic T-lymphocyte (CTL) activity, have been studied using an influenza viral challenge of mice.
  • secretory IgA antibody has been shown to prevent infection of the nose (Renegar et al., 1991a; Renegar et al.. 1991 b).
  • serum IgG antibody has been shown to prevent infection of the lung (Loosli et al. 1953).
  • anti- influenza CTL activity enhances recovery (Yap et al. 1978, Lin et al. 1981. and Wells et al, 1981).
  • Intradermal (scarification) administration of a recombinant vaccinia virus that expresses the influenza hemagglutinin protein (vac/Hi ) has been shown to induce both serum antibody that prevents viral pneumonia (Smith et al. 1983. Small et al. 1985. and Bennink et al, 1984) and CTL activ n that promotes recovery (Bennink et al. 1984. Andrew et al. 1987. and Bender et al. 1990).
  • this route of vaccine administration does not induce mucosal IgA (Small et al. 1985 and Meitin et al. 1993).
  • the subject invention concerns a novel recombinant vaccine composition for immunizing animals, including humans, against pathogenic disease organisms.
  • the vaccine comprises a live vaccinia, mutant vaccinia virus, such as a replication deficient or a highly attenuated vaccinia of which modified vaccinia virus Ankara (MVA) is a preferred virus, deletion or insertion mutants of vaccinia virus, or a canary pox virus, (each of which is referred to herein generally as “vaccinia " for ease of reference), that expresses a heterologous gene or genes.
  • VVA modified vaccinia virus Ankara
  • vaccinia canary pox virus
  • the recombinant virus is contained in an orally- administered, enteric-coated capsule or other suitable dosage form that will only dissolve and release virus when it reaches the host's small intestine. Once in the intestine, the recombinant virus induces a host immune response against the expression product of the heterologous genes.
  • an administration of the recombinant vaccine according to the subject invention is capable of inducing serum IgG, mucosal IgA and a cell-mediated immune response by the host animal.
  • a recombinant. replication deficient virus is provided in an intranasal dosage form.
  • a recombinant is provided in an intranasal dosage form.
  • replication deficient viral vector is used to induce immune responses in a geriatric mammal.
  • a recombinant. replication deficient viral vector is used to induce mucosal. serum and cellular immune responses against an antigen in large mammals, including horses, pigs, goats, sheep, cows, and primates, including humans.
  • the vaccine of the subject invention is capable of providing multiple levels of immune protection against pathogenic infections in a form that is inexpensive, environmentally stable, easily administered, safe and effective.
  • the subject invention further concerns a method of inducing a protective immune response by immunization with an orally administered live recombinant vaccinia virus.
  • the induced serum IgG, mucosal IgA and cell-mediated responses are directed against the heterologous gene product(s) expressed by the recombinant ⁇ irus.
  • the multi-level immune response induced by the subject vaccine confers protective immunity on a host from targeted pathogens.
  • the enteric administration of a recombinant vaccinia virus that expresses the influenza hemagglutinin gene (this recombinant virus is referred to herein as vac/Hi) induced mucosal IgA and serum IgG anti-Hl antibody, in addition to inducing CTL activity in mice.
  • This immune response provided protection of both the nose and lungs of the mice from a subsequent viral challenge with influenza.
  • Enteric immunization with MVA and recombinant MVA is very safe; neither MVA nor MVA containing the influenza virus HI and NP genes (MVA HA-NP) killed any SCID (B- and T-cell deficient) mice.
  • MVA(H1+NP) induce high levels of immunity in a mouse model.
  • the titers persisted for at least 52 weeks (half the life-span of the mouse) and the noses and lungs of the immunized mice were still protected from an influenza challenge at 52 weeks.
  • 3 doses of intragastric MVA(H1+NP) induced anti-influenza serum IgG antibody and mucosal IgA antibody in aged (22-24-month-old) mice, even though the levels were about 5-fold lower in the aged animals.
  • the vaccine induced neutralizing antibody in both groups of animals.
  • MVA (Hl+NP) given intranasally in an equine model induces serum and mucosal influenza neutralizing antibody, and blood from immunized horses gives approximately a 3 log reduction in the in vitro growth of MVA(H1+NP).
  • MVA(H1 +NP) cannot be isolated from the blood of inoculated horses.
  • MVA(H1+NP) cannot be isolated from horse feces following intrajejunal inoculation.
  • MVA(H1+NP) cannot be recovered from nasal secretions following intranasal inoculation.
  • Figure 1 shows influenza virus titers from the nose (upper graph) and lungs (lower graph) of research animals. Titers are reported as the logio of the 50% egg infectious doses
  • FIG. 1 shows the induction of anti-Hl influenza virus specific serum antibodies by various vaccines or control. Mice were given 10 pfu of MVA HA-NP by two intragastric (i.g.) inoculations (C). Control mice were inoculated intranasally (i.n.) with influenza A/Puerto Rico/8/34 (h); i.m.
  • Serum was obtain from tail vein and vaginal wash fluid was obtained by flushing the vagina 6-8 times with the same 80 :1 of PBS (Meitin. et al, 1994) during weeks 2. 4. and 8.
  • the ELISA to measure anti-Hl serum antibodies was performed as previously described (Meitin et al, 1991) and data points are plotted as X 10°. Vaginal wash samples were frozen at -20E C and later tested in an ELISA for anti-Hl IgA antibodies. To dissolve mucous strands, an equal quantity of 0.01 M dithiothreitol (Sigma. St. Louis.
  • Titers are expressed as the highest dilution for which the OD of the positive (antigen-containing) well divided by the OD of the respective negative (control) well gave a ratio greater than or equal to 2.
  • Figure 3 shows nasal (A) and pulmonary (B) influenza virus titers in vivo following virulent homologous influenza virus challenge of animals pretreated with various vaccines or control. Twenty four days after the second inoculation with MVA HA-NP. mice were challenged with H1N1 and sacrificed one (closed symbols) and three (open symbols) days later. The data are a summary of two experiments challenging mice with using 10 7 0 TCID 50 (•) and 10 4 ' TCID 50 ⁇ Z.. O) of influenza virus. Virus titers were virtually identical for naive or i.m. MVA mice and are not shown.
  • Day one nasal viral titers were significantly lower than those of control mice (naive and MVA i.m.) for MVA HA-NP i.g. (PO.001. Exp. #1 : p ⁇ 0.05, Exp. #2) and H1N1 recovered (p ⁇ .001. Exp. #1 : p ⁇ 0.05. Exp. #2). but not for MVA HA-NP i.m. (p>0.05. Exp. #1 and #2).
  • Day one pulmonary viral titers were significantly lower than control for MVA HA-NP i.g. (p ⁇ .001. Exp. #1 and #2), H 1N 1 recovered (p ⁇ 0.001. Exp. #1 and #2). and MVA HA-NP i.m. (p ⁇ 0.001. Exp. #1 and #2).
  • Data were analyzed using InStat 2.00 (GraphPadSoftware. San Diego. CA) and a Power Macintosh 6100/66 computer. A one way ANOVA was followed by Student-Newman-Keuls multiple
  • Figure 4 shows mean nasal (A) and pulmonary (B) viral titer in vivo 5 days following virulent heterologous influenza virus (H3N2) challenge of animals pre-treated with various vaccines or control.
  • H3N2 virulent heterologous influenza virus
  • mice were challenged with H3N2 and sacrificed one and five days later.
  • Day one titers (not shown) varied from 1.8 to 2.5 TCID 50 in the nose and 3.0 to 3.6 TCID50 in the lungs and were not significantly different between groups.
  • the data are a combination of two separate experiments using IO 6 3 (•) and 10 4 6 ( I) TCLD 50 as challenge inoculation.
  • Day 5 nasal viral titers were significantly lower than control (naive and MVA i.m.) for MVA HA-NP i.g. ( ⁇ 0.05, Exp. #1 and #2) and H1N1 recovered (p ⁇ .01. Exp. #1 and #2). but not MVA HA- NP i.m. (p>0.05, Exp. #1 and #2).
  • Day 5 pulmonary viral titers were significantly lower than control for MVA HA-NP i.g. (p ⁇ 0.05. Exp. #1 and #2 XI or X2 or both). H1N1 recovered (p ⁇ .001, Exp #1 and #2). and MVA HA-NP i.m. (p ⁇ 0.05. Exp. #1 p ⁇ 0.05. Exp. #2).
  • Figure 5 shows the results of inoculation of nude mice with recombinant vaccinia viruses is safer by the intragastric than parenteral route.
  • Figure 6 shows that MVA, H 1 +NP is not lethal in SCID mice (no deaths post inoculation, top panel; no weight loss, bottom panel), while vaccinia recombinants carrying the influenza HA gene are lethal.
  • Figure 7 shows that three doses of MVA HA-NP induced long-term serum and mucosal anti-influenza ( serum IgG. top panel: mucosal IgA, bottom panel), and since an increased response is achieved between the second and third doses, elicitation of anti-vector (MVA) antibodies does not appear to limit booster of desired immune responses.
  • MVA anti-vector
  • Figure 8 shows the comparative elicitation of serum (top panels) and mucosal (bottom panels) immune responses in young (left hand panels) and aged (right hand panels) of mice, showing a reduced but still clearly significant immune response in aged mice.
  • Figure 9 shows the induction of influenza neutralizing antibodies in young and aged mice.
  • Figure 10 shows protection of mice challenged with H1N1 virus after no treatment (negative control, in convalescent mice, in mice that received standard vaccine, or MVA-HA- NP orally or intranasally). with the vaccine of this invention providing the greatest protection against weight loss in the challenged animals.
  • the subject invention concerns novel recombinant vaccinia, mutant vaccinia virus, such as a replication deficient or a highly attenuated vaccinia of which modified vaccinia virus Ankara (MVA) is a preferred virus, deletion or insertion mutants of vaccinia virus, or a canary pox virus, (each of which is referred to herein generally as "vaccinia" for ease of reference), compositions and methods of inducing a broad immune response in a host organism using this vaccine.
  • VVA modified vaccinia virus Ankara
  • the subject invention concerns a live recombinant vaccinia virus that can express the products of one or more heterologous genes.
  • the host animal When expressed by the vaccinia virus in a host animal these gene products induce a multi- component immune response in the host.
  • the immune response includes serum IgG antibody, mucosal IgA antibody, and cell-mediated responses directed against the heterologous gene products.
  • the host animal is either protected from infection or at least primed to mount a fully protective secondary immune response upon exposure to any pathogen expressing the heterologous gene products to which the host has been sensitized. Accordingly, the host animal vaccinated with the vaccine of the subject invention can be effectively immunized against a wide variety of pathogens including bacteria, viruses, fungi, and parasites.
  • the live recombinant vaccinia virus is administered enterically to a host animal.
  • a mutant form of a recombinant vaccinia virus such as a strain that is replication deficient in mammalian cells, can be used according to the subject invention.
  • the recombinant virus is orally administered in a form that releases the virus only in the intestine of the host animal.
  • Techniques for preparing live vaccine in entericallv-coated dosage forms are known in the art (see. for example. Stickl, A.H.. British Patent No. 1 -333-512).
  • the term "intestine" is meant to include both the large and small intestinal tracts.
  • immunization with the subject vaccine occurs primarily in the small intestine.
  • IgA precursor B cells in lymphoid tissue of the intestine, such as Peyer's Patches, to the expressed heterologous gene products. These IgA precursor B cells can then migrate to mucosal tissues, such as the respiratory tract, where they may differentiate into mature IgA- secreting plasma cells. In addition. Ig secreting B cells in other lymphoid tissues, and various components of the cellular immune system, such as T cells and macrophage. are stimulated upon exposure to the heterologous gene products.
  • higher mammals are inoculated intranasally with a replication deficient recombinant viral vector, and neutralizing immune responses are thereby induced.
  • the recombinant vaccinia virus of the subject invention contains multiple heterologous genes that encode polypeptide antigens which are expressed after introduction into the host system.
  • a polyvalent vaccine according to the subject invention can be used to immunize a host animal against multiple diseases produced by a variety of pathogenic organisms.
  • such a polyvalent vaccine can be used to induce a broad immune response against a single type of pathogen, particularly those pathogens that express various forms of antigens or that express different antigens at different times during their life- cycle.
  • the recombinant vaccinia virus of the subject invention can be used with a variety of heterologous genes or gene fragments.
  • genes from pathogens that cause influenza, measles, hepatitis B. diphtheria, tetanus, mumps, rubella, and others can be inserted into the recombinant vaccinia.
  • the subject invention can be used with a wide variety of gene inserts.
  • the subject invention can be used to prevent diseases, such as pertussis, tuberculosis, cholera, and immune deficiency conditions induced by infection, upon insertion into the vector of appropriate gene inserts.
  • the subject invention can be readily used in other areas of vaccine technology, such as in cancer prevention or fertility control, upon insertion into the vector of appropriate antigens.
  • the subject invention also contemplates the use of chimeric genes that express a fusion product comprising the expression products of a portion of two or more heterologous genes.
  • the subject invention further contemplates the use of genes that encode protein structures that mimic polysaccharide antigens.
  • the subject invention can be used in vaccinating both animals and humans against pathogenic organisms.
  • the vaccine can be administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutical ly effective and immunogenic.
  • the quantity to be administered can depend upon the subject to be treated, the immunogenicitv of the expressed gene products, and the degree of protection desired. Dosage parameters can be readily determined by those skilled in the art using the disclosure provided herein.
  • intrajejunal immunization with a recombinant vaccinia expressing the influenza hemagglutinin protein (vac/Hi ) in a influenza mouse model system consistently induced nasal, gut, and vaginal wash IgA antibody as well as serum IgG antibody and CTL activity in the host mouse that was directed against the hemagglutinin.
  • the nasal IgA antibody was responsible for a significant reduction in nasal virus following challenge of enterically immunized mice (Renegar et al, 1991a.b).
  • the serum IgG antibody was responsible for a significant reduction in lung virus following challenge of immunized mice.
  • mice immunized intrajejunally multiple times with the recombinant vaccinia vac/Hi developed an immune response to the hemagglutinin in spite of having been immunized parenterally fifty-three days earlier with a wild-type (WR) strain of vaccinia.
  • a vaccine must be safe for both the vaccinated person and that person's contacts.
  • Mice immunized with the Vac/Hi recombinant showed no ill effects from enteric immunization. The mice had no change in bowel habits or ruffled fur. The enterically immunized mice did not shed vaccinia virus in their feces.
  • replication deficient mutant forms of vaccinia strains such as MVA (Sutter et al. 1994) or a canary pox virus (Cadoz et al. 1992) can be used.
  • MVA modified vaccinia virus Ankara
  • MVA poxvirus host-restricted mutants
  • mice injected intranasally, subcutaneously, intramuscularly or intravenously with a recombinant MVA that expresses the genes of influenza hemagglutinin (HA) and nucleoprotein (NP) developed anti-Hl serum IgG antibodies and CTL activity and were protected from a lethal challenge with a homologous influenza virus (Sutter et al. 1994). There was no evidence from this report relating to oral immunization using recombinant MVA. and induction of IgA is not demonstrated.
  • Non-replicating or replication defective vectors can also include a gene for the expression of interleukin-2 in order to supplement the induced immune response (Flexner et al, 1987).
  • oral administration of an enteric coated recombinant vaccinia or mutant forms of recombinant vaccinia can provide a safe and effective mode of immunization.
  • the development of multivalent vaccinia recombinants makes the subject invention even more effective.
  • a replication- deficient viral vector exemplified herein by a recombinant MVA. is capable of inducing significant immune response which in appropriate animal models exhibit significant indicia of efficacy in man and other vertebrates.
  • replication defective means that the virus cannot replicate, or replicates inefficiently, absolute absence of any replication being difficult to confirm.
  • HA-NP as an oral, enteric or intranasal vaccine.
  • One oral dose of the vaccine induced low levels of both mucosal and serum antibodies.
  • the biological effectiveness of these immunizations was shown in a series of challenge experiments. The mice that received one i.g. dose of MVA HA-NP were partially protected from a homotypic challenge and all of those that received two doses were fully protected from pulmonary infection. Most (10 to 17 mice) were also protected from nasal infection. One i.m.
  • MVA HA-NP completely protected the lungs, but not the noses.
  • These data serve to emphasize the importance of mucosal versus serum antibodies in the prevention of infection of the upper and lower respiratory tract, respectively (Renegar et al. 1991a; Renegar et al, 1991b; Barber et al, 1978; Clements et al. 1986).
  • Recovery was evaluated and demonstrated herein by data generated in experiments with heterologous. H3N2 influenza virus. These showed that viral clearance of both the lung and the nose was enhanced with i.g. MVA HA-NP. while i.m. MVA HA-NP enhanced recovery of only the lungs ( Figure 4).
  • MVA replication-deficient recombinant vaccinia virus
  • this method is not only effective in inducing immune response in all three arms of the immune system)serum antibody, mucosal IgA antibody, and cell-mediated immunity)it is also efficacious.
  • Use of an oral or intranasal rather than a parenteral v accine may enhance patient (and/or parental) acceptance and would obviate the need for ringes and needles.
  • MVA is an extremely safe vector that has undergone extensiv e safety testing in humans and animals (Hochstein-Mintzel, et al . 1972: Werner et al . 1980; ayr.
  • multivalent recombinant MVA can be constructed. Even if it proves to be too difficult to transfer more than a few genes into MVA or have them expressed in appropriate concentrations, a cocktail of recombinant MVA viruses, each containing several genes, would be expected to work based on the instant disclosure.
  • lyophilized vaccinia is extremely heat-stable. Heating to 100E C for two hours lead to a loss of only one log of infectivity. After storage at 45E C for two years, it was still 100% successful in vaccination of volunteers (Cross, 1957).
  • both of these pre-treatments may be acceptable.
  • pre-treatment with a cholecystokinin antagonist such as are well known in the art. may be more clinically acceptable. This pre-treatment would ensure that bile is not released when the vaccine is en-route.
  • These pre-treatments may be less important with an efficiently enteric-coated vaccine, but may be of assistance. For the purposes of the present studies, these pre-treatments provided for efficient oral immunization.
  • a recombinant vaccine according to this invention harboring a gene for any given antigen may be used to induce serum, cellular and mucosal immune responses upon appropriate administration thereof either to the intestinal or nasal mucosa.
  • mice Female BALB/c mice. 6-8 weeks old were given a single immunization either by scarification of the tail, intravenous, intranasal. intragastric, or intrajejunal administration of 10 8 pfu of a vaccinia-influenza recombinant containing the hemagglutinin gene from H lN l PR8 influenza (this recombinant is referred to herein as vac/Hi ) which was constructed as previously described (Flexner et al, 1987). All mice were fasted overnight before immunization. Intragastric administration was accomplished by use of an oral gastric tube. Intrajejunal administration was done by injection during laparotomy.
  • mice were anesthetized with 0.1 -0.2 ml of sodium pentobarbital (0.09 g/ml) and the peritoneal cavity was entered. The stomach and small bowel were identified and the vaccine was placed into the lumen of the jejunum with a 26 gauge needle. The incision was then closed.
  • the three control groups consisted of naive mice, mice injected intraperitoneally (i.p.) two times with inactivated HlNl influenza vaccine, and mice infected intranasally while awake with live HlNl PR8 influenza virus.
  • mice Six weeks following inoculation, the mice were sacrificed. Splenic lymphocytes were collected for measurement of anti-influenza cytotoxic T lymphocyte (CTL) activity. Serum, nasal wash (Renegar et al. 1991b). gut wash, and vaginal wash (Wu et al. 1993) were collected for anti-influenza antibody determination. These were later assayed by ELISA for serum IgG as well as nasal and gut wash IgA anti-influenza antibody, and the titers were then calculated by comparison with monoclonal controls (Meitin et al. 1991 ). Data were analyzed using a one factor ANOVA. Student-Newman-Keuls post hoc test was used to analyze differences between groups.
  • CTL cytotoxic T lymphocyte
  • Spleens were obtained from BALB/c (H-2 d ) mice 6- 8 weeks post-intrajejunal immunization with 10 8 pfu of vac/Hi .
  • Spleens were also obtained from HlNl -infected mice, wild-type vaccinia-immunized mice, and naive mice. Spleen cells were then cultured for seven days with HlNl -sensitized autologous splenocytes and then tested in a 6 fir 51 Cr release assay against vac/Hi-, vaccinia containing the nucleoprotein gene (vac NP-).
  • HlNl -sensitized P815 (H-2 d ) mastocytoma cells (Bender et al. 1991 ). Percent (%) specific lysis was determined as: (experimental release-spontaneous release/total release- spontaneous release) x 100. Multiplicity of infection was 100 pfu per cell for vaccinia targets and 10 TCID 50 per cell for HlNl targets. Spontaneous release was less than 15% for all groups. The effector cell/target cell ratio was 30: 1.
  • mice were immunized as described in the methods section above and tested for the presence of IgG and IgA antibodies to HI . Titers of detected antibodies from each immunization group are shown in Table 1. Naive mice had no detectable anti-Hl antibody. Convalescent mice developed high levels of serum IgG, nasal IgA. and gut IgA anti-Hl antibody. Control mice immunized intraperitoneally with inactivated vaccine developed the highest titers of anti-influenza serum IgG antibody.
  • mice immunized with the vac/Hl recombinant by either scarification or intravenous injection developed high titers of anti- influenza serum IgG antibody, similar to those seen in convalescent mice, although not quite as high as seen in mice immunized with standard vaccine.
  • Mice immunized intranasally and intrajejunally developed lower but readily detectable levels of serum IgG antibody.
  • Intragastric immunization produced inconsistent results: four of six mice responded similarly to the intranasal and intrajejunal groups, but two were unresponsive.
  • mice tails scratched with the vac/H l developed low titers of both nasal and gut wash IgA. This is most likely due to auto- or cross-immunization from grooming (Meitin et l . 199 )
  • mice that were immunized by the intragastric route were divided into two groups.
  • Statistically significant differences were not seen due to the low numbers of animals in the subdivided intragastric groups, i.e.. 4 and 2.
  • mice were immunized by either the intrajejunal or intragastric route described in the methods section. Six weeks later, vaginal wash fluid was obtained (Wu et al, 1993). No detectable IgA anti-Hl antibody was present in the vaginal wash of naive mice.
  • Memory CTL activity was determined for six of the intrajejunally immunized mice in two separate experiments as shown in Table 2. Mice immunized with wild-type vaccinia or naive mice had minimal activity. Mice convalescent from an influenza HlNl infection yielded 32% lysis against vac/Hl -sensitized targets. 49% against vaccinia containing the nucleoprotein gene (vac/NP)-sensitized targets, and 24% of lysis against HlNl -sensitized targets in the first experiment and 58% lysis against vac/Hl targets and 45% against vac NP targets in the second experiment.
  • vac/NP nucleoprotein gene
  • mice immunized intrajejunally with vac/Hl had 32% lysis against vac/Hl- sensitized targets. 1 1% lysis against vac/NP-sensitized targets, and 18% against H1N1- sensitized targets in the first experiment and 42% lysis against vac/Hl and 16% lysis against vac/NP in the second experiment.
  • lysis of H3N2-sensitized P815 cells by splenocytes from vac/Hl -immunized mice was less than 25% compared to 86% for lysis from a HIN l-infected mouse.
  • most of the CTL activity was directed against the hemagglutinin and not to the vaccinia.
  • the low anti-vaccinia CTL activity was undoubtedly due to the secondary in vitro stimulation with HlN l -sensitized stimulator cells rather than vaccinia-sensitized stimulator cells.
  • mice that were immunized as described in the methods section above were challenged six weeks later with 20 MID 50 of live influenza A PR8 (HlNl) administered intranasally while anesthetized (Yetter et al, 1980).
  • Mice were sacrificed 1, 3, 5. and 7 days after challenge and virus titers determined for nasal and lung tissues (Figure 1).
  • the mice convalescent from a previous influenza infection had no virus in their noses.
  • Naive mice had high titers of virus in their noses throughout the 7 days.
  • Significantly lower levels of nasal virus were found in the three groups receiving successful mucosal immunization, i.e.. vac/H l intranasal. vac/Hl intrajejunal.
  • mice had Day 1 nasal virus titers that were statistically indistinguishable from naive mice. Viral clearance after Day 1 was more rapid in mice that had received vac/Hl than either the naive mice, the non-responders, or those given standard vaccine i.p. Because respiratory viruses are primarily cleared by cytotoxic T-cells. this more rapid clearance is most likely due to the higher levels of anti- influenza CTL activity induced by vac/Hl than by standard vaccine (Bender et al. 1990).
  • Virus titers in the lungs are also shown in Figure 1 .
  • Convalescent mice were solidly immune as evidenced by the lack of virus.
  • Naive mice had high titers of virus throughout the seven days studied.
  • the gastric non-responders were not statistically different from the naive mice in regard to virus shedding. All the remaining groups had statistically significantly (p ⁇ 0.05) lower amounts of virus than the naive controls.
  • Live recombinant vaccinia virus is prepared containing heterologous DNA sequences that encode various HIV antigens.
  • the HIV gene sequences can include those that encode GP160, GP120. or subunits of these or other HIV proteins.
  • the live recombinants are then prepared in an entericallv-coated dosage form.
  • the host animal or human ingests the vaccine which dissolves upon reaching the host's intestine. In the intestine, the free virus replicates and induces a host immune response against the HIV expression products. Subsequent administrations of vaccine can be determined and administered when necessary.
  • Live recombinant vaccinia virus is prepared containing heterologous DNA sequences that encode hepatitis B antigens.
  • the live recombinants are then prepared in an enterically- coated dosage form.
  • the animal or human to be vaccinated orally ingests the vaccine preparation, which dissolves upon reaching the host's intestine and releases free virus.
  • the free recombinant virus replicates and induces a host immune response against the hepatitis B surface antigen expression products of the heterologous genes.
  • the immunized host Upon subsequent exposure to hepatitis B virus, the immunized host can produce a strong, multi- level immune response against hepatitis B which protects the host from infection with the virus.
  • Optimal dosage levels and subsequent administration of the recombinant vaccine can be determined by those skilled in the art.
  • mice Female BALB/c mice were immunized intragastrically (i.g.) with MVA HA-NP one hour after co-administration of cimetidine to inhibit gastric acid secretion and cholecystokinin to induce emptying of the gallbladder prior to vaccine administration.
  • Positive control mice were convalescent from an HlNl (influenza A/PR/8/34) infection or given MVA HA-NP i.m.
  • Negative control mice were naive animals or mice given wild-type MVA i.m.
  • each mouse received a combination of 3 mg of cimetidine HCI (SmithKline Beecham, Philadelphia, PA) and 0.02 Fg of sincalide (the C-terminal octapeptide of cholecystokinin) (Squibb, Princeton. NJ) in 100 FI of PBS i.p.
  • Mice received 200 FI of MVA HA-NP in 50 mM HEPES buffer (Mediatech. Washington. D.C.) containing 10 8 p.f.u. via a 1 " (2.5 cm) feeding needle. In some mice, i.g. inoculation was repeated five weeks later. For i.m. inoculations, 100 FI of wild-type MVA or MVA HA-NP.
  • mice were anesthetized with sodium pentobarbital and inoculated i.n. with HlNl .
  • Serum anti-Hl antibody was found in all mice receiving MVA HA-NP (either i.m. or i.g.) or HlN l influenza virus (Figure 2A), though levels were lower in the i.g. immunized mice.
  • Mucosal anti-Hl IgA antibody was detected in the vaginal secretions of 31 of 33 mice that received MVA HA- NP i.g. and in 9 of 9 positive controls convalescent from an H l N l infection.
  • vaginal IgA anti-Hl titers of mice which received two does of MVA HA-NP were about half the titer of mice convalescent from influenza (Figure 2B).
  • mice with a wide range of antibody titers were chosen for challenge with HlNl influenza virus. Following this challenge, influenza virus reached a peak titer on day 3 post challenge of JO 4 TCID 50 in the noses of naive healthy mice ( Figure 3 A). The noses were protected best in those mice receiving two i.g. doses of MVA HA-NP ( Figure 3 A). Five of the 12 mice receiving two i.g. doses of MVA HA-NP shed no virus from their noses on day 1 post-challenge and the mean titer of the total group (0.65"0.2) was significantly lower than naive (1.9"0.4), MVA HA-NP i.m.
  • Example 8 For determination of anti-influenza CTL activity, spleens were removed and cultured in vitro with H3N2-infected autologous splenocytes for 7 days and cytotoxicity was assayed against H3N2-sensitized P815 cells (Bender et al . 1991. Bender et al. 1993). Table 3 demonstrates that two i.g. doses of MVA HA-NP induced lower levels of anti-influenza CTL activity than did nasal HlNl influenza virus. As a measure of the efficacy of this cell- mediated immune response. MVA HA-NP immunized mice were challenged with influenza A/Port Chalmers/1/73 (H3N2).
  • p eens were o a ne c - m e s pos u a on p u o - , PR8 infection, or wild-type MVA; cultured for 7 days with H3N2-sensitized autologous splenocytes: and tested in a 6- hr 5l Cr release assay against H3N2-sensitized P8 I 5 ( H-2d) cells [Bender et ai , 1991 : Bender et ai . 1993]. Percent specific lysis was determined as [(experimental release-spontaneous release (/(total release-spontaneous release)] X 100. Spontaneous release was ⁇ 10% of total.
  • inoculation is safer than i.p. inoculation. Death is delayed by approximately five days with i.g. inoculation with VAC and does not occur with i.g. inoculation with VAC/HA, perhaps due to some viral inactivation in the gut. This observation also may be related to the fact that even nude mice have ⁇ - ⁇ T-cells lining their gut.
  • HA high titers (10'to more than ⁇ 0 pfu) of virus were isolated from brain, lung, liver, and spleen.
  • Example 10 Experiments to establish the dose-response and duration of intragastric administration of MVA/Hl+NP.
  • mice were immunized i.g. with various doses of MVA H1+NP at three-week intervals and blood and vaginal wash obtained for determination of anti-influenza serum IgG antibody and mucosal IgA antibody. Mean titers and numbers of responding mice are shown in Table 4 and Figure 7. (Mice were immunized at weeks 0, 3, and 6. Comparable mean values for six HlNl convalescent mice were serum ELISA titer of 7,400 and IgA of 32; na ⁇ ve or MVA immunized mice had values of ⁇ 0.1 and ⁇ 2). Table 4. Enhanced serum and mucosal antibody response with three i.g. doses of MVA/H l+NP.
  • Inoculum 1 IgG Serum IgA Vaginal IgG Serum IgA Vaginal
  • Vaccines 85 (Lerner. R.A.. Chanock. R.M.. Brown. F.. eds.), Cold Spring Harbor Laboratory. Cold Spring Harbor. NY, pp. 175-177.

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Abstract

La présente invention concerne un vaccin du virus de la vaccine recombinant et servant à immuniser des animaux et des hommes contre une maladie. Ce vaccin comprend un virus de la vaccine vivant ou une réplication du virus de la vaccine mitante déficient, mais capable d'exprimer un ou plusieurs gènes ou fragments de gènes hétérologues. Selon une réalisation préférée, le virus recombinant est contenu dans une enveloppe d'administration orale qui ne se dissout que dans l'intestin de l'animal hôte. L'invention concerne également un procédé permettant d'induire une large réponse immunitaire protectrice par administration par voie orale, intranasale ou autrement muqueuse, du vaccin du virus de la vaccine recombinant.
PCT/US1997/013836 1997-08-05 1997-08-05 Vaccin vivant recombinant comprenant un virus ne se repliquant pas ou se repliquant sans efficacite WO1999007869A1 (fr)

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Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6960345B2 (en) * 2003-03-06 2005-11-01 Incell Corporation, Llc Oral vaccinia formulation
US7189536B2 (en) 2000-11-23 2007-03-13 Bavarian Nordic A/S Modified vaccinia ankara virus variant
EP1465658B1 (fr) * 2002-01-15 2007-05-02 Bio Life Science Forschungs- und Entwicklungsges.m.b.H. VACCINATION ORALE AVEC LE MIMOTOPE D'ANTIGENE Gln-Met-Trp-Ala-Pro-Gln-Trp-Gly-Pro-Asp
EP1327450B1 (fr) * 2002-01-15 2007-08-15 Bio Life Science Forschungs- und Entwicklungsges.m.b.H. Vaccination orale par des mimotopes d'antigene tumoraux dénudés
US7445924B2 (en) 2000-11-23 2008-11-04 Bavarian Nordic A/S Modified Vaccinia Ankara virus variant and cultivation method
US7628980B2 (en) 2000-11-23 2009-12-08 Bavarian Nordic A/S Modified vaccinia virus ankara for the vaccination of neonates
US7695939B2 (en) 2002-09-05 2010-04-13 Bavarian Nordic A/S Method for the cultivation of primary cells and for the amplification of viruses under serum free conditions
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US7897156B2 (en) 2001-11-22 2011-03-01 Bavarian Nordic A/S Modified vaccinia virus ankara for the vaccination of neonates
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US8128922B2 (en) 1999-10-20 2012-03-06 Johns Hopkins University Superior molecular vaccine linking the translocation domain of a bacterial toxin to an antigen
US9011866B2 (en) 2005-01-06 2015-04-21 The Johns Hopkins University RNA interference that blocks expression of pro-apoptotic proteins potentiates immunity induced by DNA and transfected dendritic cell vaccines
US9085638B2 (en) 2007-03-07 2015-07-21 The Johns Hopkins University DNA vaccine enhancement with MHC class II activators
US9701725B2 (en) 2003-05-05 2017-07-11 The Johns Hopkins University Anti-cancer DNA vaccine employing plasmids encoding signal sequence, mutant oncoprotein antigen, and heat shock protein
EP3333265A1 (fr) * 2010-05-14 2018-06-13 Oregon Health & Science University Vecteurs hcmv et rhcmv recombinants et utilisations associées
US10532099B2 (en) 2016-10-18 2020-01-14 Oregon Health & Science University Cytomegalovirus vectors eliciting T cells restricted by major histocompatibility complex E molecules
US10688164B2 (en) 2015-11-20 2020-06-23 Oregon Health & Science University CMV vectors comprising microRNA recognition elements
US10760097B2 (en) 2011-06-10 2020-09-01 Oregon Health & Science University CMV glycoproteins and recombinant vectors
US10995121B2 (en) 2014-07-16 2021-05-04 Oregon Health & Science University Human cytomegalovirus comprising exogenous antigens
US11091779B2 (en) 2015-02-10 2021-08-17 Oregon Health & Science University Methods and compositions useful in generating non canonical CD8+ T cell responses

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1333512A (en) * 1971-06-19 1973-10-10 Stickl A H Vaccine composition for oral smallpox inoculation
WO1989003429A1 (fr) * 1987-08-28 1989-04-20 Health Research Inc. Virus recombinant de la variole avicole
WO1992016616A1 (fr) * 1991-03-20 1992-10-01 Virogenetics Corporation Vaccin contre le poxvirus de recombinaison de la malaria

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1333512A (en) * 1971-06-19 1973-10-10 Stickl A H Vaccine composition for oral smallpox inoculation
WO1989003429A1 (fr) * 1987-08-28 1989-04-20 Health Research Inc. Virus recombinant de la variole avicole
WO1992016616A1 (fr) * 1991-03-20 1992-10-01 Virogenetics Corporation Vaccin contre le poxvirus de recombinaison de la malaria

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BENDER ET AL.: "Cytotoxic T lymphocyte activity induced by influenza vaccination in young and aged mice", VACCINES90, 1990, pages 69 - 73, XP002055083 *
BENDER ET AL.: "Oral immunization with a replication-deficient recombinant vaccinia virus protects mice against Influenza", JOURNAL OF VIROLOGY, vol. 70, no. 9, September 1996 (1996-09-01), pages 6418 - 6424, XP002055082 *
BLANCOU ET AL.: "Oral Vaccination of the fox against rabies using a live recombinant vaccinia virus", NATURE, vol. 322, 1986, pages 373 - 375, XP002055040 *
MEITIN ET AL.: "Enteric immunization of mice against influenza with recombinant vaccinia", PROC. NATL. ACAD. SCIE. USA, vol. 91, November 1994 (1994-11-01), pages 11187 - 11191, XP002055039 *
SUTTER ET AL.: "A recombinant vector derived from the host range-restricted and highly attenuated MVA strain of vaccinia virus stimulates protective immunity in mice to inflenza virus", VACCINE, vol. 12, no. 11, 1994, pages 1032-1040, XP002019163 *

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EP3333265A1 (fr) * 2010-05-14 2018-06-13 Oregon Health & Science University Vecteurs hcmv et rhcmv recombinants et utilisations associées
US11266732B2 (en) 2010-05-14 2022-03-08 Oregon Health & Science University Recombinant HCMV and RHCMV vectors and uses thereof
US10760097B2 (en) 2011-06-10 2020-09-01 Oregon Health & Science University CMV glycoproteins and recombinant vectors
US10995121B2 (en) 2014-07-16 2021-05-04 Oregon Health & Science University Human cytomegalovirus comprising exogenous antigens
US11692012B2 (en) 2014-07-16 2023-07-04 Oregon Health & Science University Human cytomegalovirus comprising exogenous antigens
US11091779B2 (en) 2015-02-10 2021-08-17 Oregon Health & Science University Methods and compositions useful in generating non canonical CD8+ T cell responses
US10688164B2 (en) 2015-11-20 2020-06-23 Oregon Health & Science University CMV vectors comprising microRNA recognition elements
US10532099B2 (en) 2016-10-18 2020-01-14 Oregon Health & Science University Cytomegalovirus vectors eliciting T cells restricted by major histocompatibility complex E molecules
US11305015B2 (en) 2016-10-18 2022-04-19 Oregon Health & Science University Cytomegalovirus vectors eliciting T cells restricted by major histocompatibility complex E molecules

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