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T Nagai,
Y Suzuki,
H Kiyohara,
E Susa,
T Kato,
T Nagamine,
Y Hagiwara,
S Tamura,
T Yabe, C Aizawa,
H Yamada
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ABSTRACT: Active substances from hot water extracts from 267 different Chinese and Japanese medicinal herbs were screened for mucosal adjuvant activity with influenza HA vaccine in mice. The extract from the root of Polygala tenuifolia was found to contain potent mucosal adjuvant activity. The active substances were purified and identified as onjisaponins A, E, F, and G. When each onjisaponin (10 microg) was intranasally (i.n.) inoculated with influenza vaccine (10 microg) in mice, serum hemagglutination-inhibiting (HI) antibody titers increased 3-14 times over control mice administered vaccine alone after 4 weeks. When each onjisaponin (10 microg) was i.n. inoculated with the vaccine (10 microg) followed by i.n. vaccination of the vaccine alone after 3 weeks, serum HI antibody titers increased 27-50 fold over those mice given i.n. vaccinations without onjisaponins. These same conditions also significantly increased nasal anti-influenza virus IgA antibody titers. Two inoculations with onjisaponin F (1 microg) and influenza HA vaccine (1 microg) at 3 weeks intervals, significantly increased serum HI antibody and nasal anti-influenza virus IgA and IgG antibody titers after only 1 week over mice given HA vaccine alone after the secondary vaccination. Intranasal vaccination with onjisaponin F inhibited proliferation of mouse adapted influenza virus A/PR/8/34 in bronchoalveolar lavages of infected mice. Separate intranasal vaccinations with onjisaponins A, E, F, and G (10 microg) each and diphtheria-pertussis-tetanus (DPT) vaccine (10 microg) of mice followed by i.n. vaccination with DPT vaccine alone after 4 weeks showed significant increases in serum IgG and nasal IgA antibody titers after 2 weeks following secondary vaccination over mice vaccinated with DPT vaccine alone. All onjisaponins showed little hemolytic activity at concentrations up to 100 microg/ml. The results of this study suggest that onjisaponins may provide safe and potent adjuvants for intranasal inoculation of influenza HA and DPT vaccines.
Vaccine 10/2001; 19(32):4824-34. · 3.77 Impact Factor
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ABSTRACT: Immune responses and protection against influenza virus infection were compared between young (2 months) and aged (18 months) BALB/c, C3H and C57BL/6 (B6) mice after intranasal vaccination. The mice were immunized with 2.5 microg protein of A/PR/8/34 (PR8) (H1N1) virus vaccine containing a cholera toxin adjuvant. In both the young and aged BALB/c mice, high levels of PR8-specific antibody-forming cell (AFC) responses were induced in the nasal-associated lymphoid tissue (NALT) 7 days after immunization. Nasal wash IgA and serum IgG antibody (Ab) responses to the PR8 haemagglutinin (HA) 4 weeks after immunization were slightly higher in the young mice than in the aged mice. The young mice showed complete protection against challenge infection, while the aged mice showed only a partial protection. In the C3H mice, NALT-AFC, and IgA and IgG Ab responses were higher in the young mice than those in the aged mice in parallel with the more efficient protection in the young mice than in the aged mice. Both the young and aged B6 mice showed no NALT-AFC responses, scarce IgA and IgG Ab responses and no protection. In the BALB/c mice, IgG1 and IgG2a levels were significantly lower in the aged mice. On the other hand, in the C3H mice, only IgG2a level was significantly lower in the aged mice. Similar results were obtained in terms of immune responses and protection between the young and aged mice of three different strains of mice after intra-nasal immunization with 0.1 microg of PR8 vaccine containing the adjuvant, two-times at 4-week intervals. In the B6 mice, the immune response was improved by immunization with a higher dose of the adjuvant-combined vaccine. These results suggest that local Ab responses, as well as systemic Ab responses, are downregulated in aged mice, although the degree of the downregulation of immune responses differs from strain to strain.
Vaccine 08/2001; 19(28-29):3981-9. · 3.77 Impact Factor
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ABSTRACT: Effects of intranasal administration of cholera toxin (CT) [or Escherichia coli heat-labile enterotoxin (LT)] B subunits supplemented with a trace amount of the holotoxin, CTB* or LTB*, on the brain were examined in BALB/c mice by comparing with those of the intracerebral injection. Intracerebral injection of CTB* at doses more than 10 microg/mouse caused significant body weight loss and dose-dependent death within 7 days, with localization of conjugates of horseradish peroxidase with CTB (HRP-CTB) in the ventricular system and in the perineural space of olfactory nerves of the nasal mucosa 3 h after injection. Intracerebral injection of CTB* at doses less than 3 microg/mouse (or LTB* at doses less than 22.7 microg/mouse) did not cause any significant body weight loss for 7 days, with localization of HRP-CTB in the brain but not in the nasal mucosa. On the other hand, intranasal administration of 10 microg of CTB* caused localization of HRP-CTB in the nasal mucosa but not in the brain 3 h after administration and caused body weight loss even after 30 administrations. Neither any histological changes of brain tissues nor marked changes in serum biochemical parameters were found in mice after the 30 administrations of CTB* or LTB*. These results suggest that 0.1 microg of CTB* or LTB*, which is known to be close to the minimal effective dose as an adjuvant for nasal influenza vaccine in mice and corresponds to 100 microg per person, can be used as a safe nasal adjuvant without adversely affecting the brain.
Vaccine 03/2001; 19(13-14):1652-60. · 3.77 Impact Factor
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Y Hagiwar,
T Tsuji,
T Iwasaki,
S Kadowaki,
H Asanuma,
Z Chen,
K Komase,
Y Suzuki, C Aizawa,
T Kurata,
S Tamura
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ABSTRACT: The effectiveness and safety of mutant Escherichia coli heat-labile enterotoxin, LT H44A (His to Arg substitution at position 44 from the N-terminus of the A1 fragment of the A subunit) as an adjuvant for nasal influenza vaccine were examined. (1) When 0.2 microg of LT H44A, together with 0.2 microg of influenza A/PR/8/34 virus (PR8, H1N1) vaccine, was administered intranasally into BALB/c mice (twice, 4 weeks apart), anti-PR8 hemagglutinin (HA) IgA and IgG antibody (Ab) responses were induced at levels that were sufficient to provide either complete protection against infection with a small volume of PR8 virus suspension or partial protection against infection with a lethal dose of the suspension. The dose of the mutant LT and vaccine used here (0.2 microg/ 20 g doses mouse) corresponded to the estimated dose per person, i.e. 0.1 mg/10 kg body weight. (2) Using these vaccination conditions, no additional total IgE Ab responses were induced. (3) The mutant was confirmed to be less toxic than the native LT when the toxicity was analyzed either using Y1 adrenal cells in vitro (1/483 EC(50)) or by an ileal loop test. (4) One hundred micrograms of the mutant, administered intranasally or intraperitoneally into guinea-pigs (Heartley strain, 0.3-0.4 kg), caused no body-weight changes 7 days after administration, although 100 microg of the native LT administered intraperitoneally caused death in all guinea-pigs due to diarrhea within 2 days. The intranasal administration of 100 microg of the mutant resulted in almost no pathological changes in the nasal mucosa 3 days after administration. These results suggest that LT H44A, which can be produced in high yields in an E. coli culture (about 5 mg/l), could be used as one of the effective and safe adjuvants for nasal influenza vaccine in humans.
Vaccine 03/2001; 19(15-16):2071-9. · 3.77 Impact Factor
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ABSTRACT: Electroporation for the transfer of plasmid DNA encoding influenza virus hemagglutinin (HA) into muscle or nasal mucosa was tried in BALB/c mice to examine the efficacy of this method for inducing anti-HA immune responses and providing protection against homologous A/PR/8/34 (PR8) virus infection. Mice were immunized by two injections, 3 weeks apart, of HA-DNA with electroporation into the muscle wherein a pair of electrode needles was inserted to deliver the electric pulses. One or 3 weeks after the immunization, the mice were infected with a lethal dose of the PR8 virus. Ten micrograms or more of HA-DNA/dose induced strong serum anti-HA IgG antibody (Ab) responses, in which both IgG1 and IgG2a were predominant, and weak cytotoxic T lymphocyte responses. These immune responses were sufficient to provide efficient protection against the lethal infection. In addition, mice were immunized by dropping HA-DNA (12 microg) three times, 2 weeks between each dose into nostrils where each of two electrode needles was placed on the right nostril or the palate. One week after the immunization, the mice were infected with a sublethal dose of the PR8 virus. The DNA immunization by electroporation provided reduced nasal virus titers, in parallel with a relatively high levels of serum anti-HA IgG Ab and a slight nasal anti-HA IgA Ab production. The intranasal administration of cholera toxin before HA-DNA immunization by electroporation enhanced the nasal IgA Ab production together with enhancement of the efficiency of protection. These results suggest that electroporation can be used as one of the efficient gene delivery systems for the transfer of influenza DNA-vaccine into muscle or nasal mucosa to provide protection against influenza virus infection.
Vaccine 07/2000; 18(25):2779-88. · 3.77 Impact Factor
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ABSTRACT: Protection against influenza virus infection and antibody responses in mice vaccinated with plasmid DNAs encoding haemagglutinin (HA), neuraminidase (NA) and nucleoprotein (NP) were compared among BALB/c (H-2d), B10 (H-2b) and C3H (H-2k) mice. Mice were inoculated with each DNA construct twice, 3 weeks apart, at a dose of 1 microg per mouse by particle-mediated DNA transfer (gene gun) to the epidermis. They were challenged with a lethal dose of the homologous virus 7 days after the second vaccination. NA-DNA provided significant protection in all strains of mouse, whereas HA-DNA afforded significant protection only in BALB/c mice. The serum antibody titres against NA or HA molecules in BALB/c, C3H and B10 mice were high, intermediate and low, respectively. NP-DNA failed to provide protection in any strain of mouse, and elicited low titres of anti-NP antibodies. These results suggest that NA-DNA can be used as a vaccine component to provide effective protection against influenza virus infection in various strains of mouse.
Journal of General Virology 11/1999; 80 ( Pt 10):2559-64. · 3.36 Impact Factor
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ABSTRACT: The effectiveness and safety of mutants of cholera toxin (CT) as an adjuvant for nasal influenza vaccine was examined. Four CT mutants, called CT7 K (Arg to Lys), CT61F (Arg to Phe), CT112 K (Glu to Lys), and CT118E (Glu to Gln), were produced by the replacement of one amino acid at the A1-subunit using site-directed mutagenesis. All these mutants were confirmed to be less toxic than native CT when the toxicity was analysed by using Y1 adrenal cells in vitro. When high (1 microg) and low (0.1 microg) doses of these CT mutants, together with high (1 microg) and low (0.1 microg) doses of influenza A/PR/8/34 virus (H1N1) vaccine, respectively, were administered intranasally into BALB/c mice in a two dose regimen (twice, 4 weeks apart), they produced both anti-PR8 hemagglutinin (HA) IgA and IgG antibody (Ab) responses roughly in a dose-dependent manner. The relatively low level of anti-HA Ab responses, induced by the low dose CT mutants, were enough to provide complete protection against the homologous virus infection. Under these vaccination conditions, no anti-CTB IgE Ab responses were induced. The mutant CT112 K, which showed a relatively high adjuvant activity, the lowest toxicity and relatively high yields in a bacterial culture, seems to be the most effective and safest adjuvant for nasal influenza vaccine among those examined. The low dose of CT derivatives or vaccine used in the mouse model (0.1 microg/20 g mouse) corresponded to 100 microg/20 kg, the estimated dose per person. A tentative plan for safety standards for human use of CT (or LT) derivatives as an adjuvant of nasal influenza vaccine is discussed.
Vaccine 08/1999; 17(22):2918-26. · 3.77 Impact Factor
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ABSTRACT: The ability of plasmid DNA encoding hemagglutinin (HA), neuraminidase (NA) or matrix protein (M1) from influenza virus A/PR/8/34 (PR8) (H1N1), and mixtures of these plasmid DNAs (HA + NA and HA + NA + M1) to protect against homologous or heterologous virus infection was examined in BALB/c mice. Each DNA was inoculated twice, 3 weeks apart, or four times, 2 weeks apart, at a dose of 1 microg of each component per mouse by particle-mediated DNA transfer to the epidermis (gene gun). Seven days after the last immunization, mice were challenged with a lethal homologous or heterologous virus and the ability of each DNA to protect the mice from influenza was evaluated by observing lung virus titers and survival rates. The administration of a plasmid DNA mixture of either (HA + NA) or (HA + NA + M1) provided almost complete protection against the PR8 virus challenge, and this protection was accompanied by high levels of specific antibody responses to the respective components. The degree of protection afforded in these groups is significantly higher than that in mice given either HA- or NA-expressing DNA alone, which provided only a partial protection against PR8 challenge or that in mice given M1-expressing DNA, which failed to provide any protection. In addition, both of the plasmid DNA mixtures (HA + NA) and (HA + NA + M1) showed a slight tendency to provide cross-protection against an A/Yamagata/120/86 (H1N1) virus challenge, and this was accompanied by a relatively high level of cross-reacting antibodies. Thus, there was no clear difference between the ability of the HA + NA and HA + NA + M1 plasmid DNA mixtures in providing protection against either a PR8 or heterologous virus challenge. These results suggest that in mice immunized by gene gun, a mixture of plasmid DNAs encoding HA and NA can provide the most effective protection against the virus challenge. The addition of the M -expressing plasmid DNA to this mixture does not enhance the degree of protection afforded.
Vaccine 03/1999; 17(7-8):653-9. · 3.77 Impact Factor
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ABSTRACT: The ability of plasmid DNA encoding various influenza viral proteins from the A/PR/8/34 (H1N1) virus to protect against influenza was compared in BALB/c mice. The plasmid DNA encoded hemagglutinin (HA), neuraminidase (NA), matrix protein (M1), nucleoprotein (NP) or nonstructural protein (NS1) in a chicken beta-actin-based expression vector (pCAGGS). Each DNA was inoculated twice 3 weeks apart at a dose of 1 microgram per mouse by particle-mediated DNA transfer to the epidermis (gene gun). Seven days after a second immunization, mice were challenged with the homologous virus and the ability of each DNA to protect mice from influenza was evaluated by decreased lung virus titers and increased survival. Mice, given HA- or NA-expressing DNA, induced a high level of specific antibody response and protected well against the challenge virus. On the other hand, mice given M1-, NP-, or NS1-DNA failed to provide protection, although M1- and NP-DNAs did induce detectable antibody responses. These results indicate that both HA- and NA-expressing DNAs for the surface glycoproteins are most protective against influenza from among the various viral protein-expressing DNAs used here.
Vaccine 11/1998; 16(16):1544-9. · 3.77 Impact Factor
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ABSTRACT: Effects of a single intranasal (i.n.), subcutaneous (s.c.) or intravenous (i.v.) vaccination and their combined vaccination of priming and boosting on a primary and a secondary IgA antibody forming cell (AFC) response were examined in the nasal associated lymphoid tissue (NALT), spleen and popliteal lymph nodes (pLNs) of BALB/c mice. Mice were primed with the vaccine prepared from A/Yamagata/120/86 (H1N1) together with a cholera toxin-adjuvant and boosted with the same vaccine 3 weeks later. Three days after boosting, IgA-AFC responses in each lymphoid tissue were measured as an index of the immunological memory that mediates a secondary IgA-AFC response. Single i.n. vaccination induced a greater primary IgA-AFC response in the NALT not only than that in the spleen or pLNs, but also than that induced by single i.v. or s.c. vaccination. The combination of i.n. priming and i.n. boosting afforded a greater anamnestic IgA-AFC response in the NALT not only than that in the spleen or pLNs, but also than that induced by any other combinations of priming and boosting (i.n.-i.v., i.n.-s.c., s.c.-i.n., s.c.-i.v., and s.c.-s.c.). These results showed that i.n. priming induced a greater primary IgA-AFC response in the NALT and simultaneously induced the immunological memory that mediated a greater secondary-type AFC response following i.n. boosting in the NALT.
Vaccine 09/1998; 16(13):1257-62. · 3.77 Impact Factor
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ABSTRACT: Antibody-forming cell (AFC) responses in the nasal-associated lymphoid tissue (NALT) of BALB/c mice were examined following intranasal infection, mainly of the upper respiratory tract, with a small volume of influenza virus. The infection induced significant accumulation of T and B cells in NALT, peaking around day 7 post-infection. Virus-specific IgA, IgG and IgM AFC responses were induced, developing from day 5 and peaking at day 7; responses were predominantly IgA and IgG, followed by IgM. At peak, NALT contained the greatest number of IgA AFCs per total cells of the lymphoid tissues examined in the upper respiratory tract. The IgM AFC responses were induced in NALT cell cultures from uninfected mice following in vitro culture with influenza virus, indicating that at least a part of the AFCs in infected mice may have originated from specific B cell precursors in NALT. In parallel with the detection of AFCs in infected mice, virus-specific IgA antibodies appeared in the nasal wash and their appearance correlated well with virus clearance from the nasal area. These results suggest that virus-specific IgA antibodies, produced by IgA AFCs in NALT, play an important role in recovery from infection.
Journal of General Virology 03/1998; 79 ( Pt 2):291-9. · 3.36 Impact Factor
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ABSTRACT: In previous papers, we have shown that Escherichia coli heat-labile enterotoxin B subunit, supplemented with a trace amount of the holotoxin (LTB*) could be used as a potent adjuvant for a nasal influenza HA (haemagglutinin) vaccine in humans. The present study was designed to determine whether the effectiveness of a combined LTB*-HA vaccine could be limited by preexisting immunity to LTB and how many times the adjuvant-combined vaccine could be administered intranasally without reducing its protective efficacy in BALB/c, C3H and B10 mice. The magnitude of both nasal and serum Ab responses to HA vaccine was correlated with the degree of protection against virus infection. Higher doses of LTB*-combined vaccine were required for inducing high enough levels of anti-HA Ab responses to provide complete protection in low responder mice. Repeated pretreatments with LTB* alone (more than six times), which provided high levels of preexisting Abs to LTB, inhibited the induction of anti-HA Ab responses and reduced the protective efficacy of the adjuvant-combined vaccine. However, the LTB*-combined vaccine could be given repeatedly (about ten times) to mice without reducing the effectiveness of the adjuvant-combined vaccine. These results suggest that the LTB*-combined nasal influenza vaccine can be given to humans once every few years when an epidemic of influenza may occur.
Vaccine 12/1997; 15(16):1784-90. · 3.77 Impact Factor
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ABSTRACT: A method for isolation of mouse nasal-associated lymphoid tissue (NALT), which is a principal mucosal lymphoid tissue of the respiratory tract in rodents, was developed. The paired lymphoid organs could be separated from the upper jaw by peeling away the palate where NALT was localized bilaterally on the posterior side. About 3 x 10(5) lymphocytes could be obtained from one NALT fragment. The NALT lymphocyte fraction from normal BALB/c mice contained T- and B-cells in about equal numbers, and contained about 4 times as many CD4+ T-cells as CD8+ T-cells when analyzed with a FACScan fluorescence analyzer. The composition of the NALT lymphocytes was similar to that of the lymphocytes from the portion of the nasal cavity remaining after isolation of the NALT. The NALT lymphocyte fraction from mice infected 7 days previously with influenza virus was also characterized. The numbers of NALT T- and B-cells from the infected mice were approximately 2 and 3 times higher than those of non-infected mice, respectively. In parallel with the cell increase, NALT lymphocytes produced IFN-gamma when cultured for 24 h and contained cells secreting influenza virus-specific IgA and IgG antibodies. The results suggest that this method can be successfully used for investigating cellular dynamics of mucosal immunology in the upper respiratory tract.
Journal of Immunological Methods 04/1997; 202(2):123-31. · 2.20 Impact Factor
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ABSTRACT: Oral administration of a small amount of antigen conjugated cholera toxin B subunit is known to induce tolerance to the antigen. In the present experiments, whether nasal administration of allergen conjugated to Escherichia coli heat-labile toxin B subunit (LTB) induced tolerance was examined in BALB/c mice. A single administration of a small amount of LTB-coupled ovalbumin (OVA) suppressed the induction of delayed-type hypersensitivity and IgE antibody responses to OVA which was administered parenterally after nasal administration of LTB-coupled OVA. The antigen-specific suppression was abrogated by the addition of the holotoxin to LTB-coupled OVA. The suppression, induced by nasal administration with a small amount of allergen conjugated to a mucosa-binding molecule, may be applicable for preventing the development of allergy.
Vaccine 03/1997; 15(2):225-9. · 3.77 Impact Factor
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ABSTRACT: The protective roles of influenza viral nucleoprotein (NP), together with the cellular mechanism of the protection in the nasal site, were examined in BALB/c mice immunized intranasally with an adjuvant (cholera toxin B subunit containing 0.2% of the whole toxin)-combined A or B virus recombinant NP. The NP-immune mice, when challenged intranasally with a sublethal dose of the virus 3 wk after immunization, had accelerated virus clearance from the nasal site in both an influenza type-specific and a nonspecific manner, as shown by the protection from high morbidity from the second day after challenge. Both type-specific and nonspecific acceleration of recovery was confirmed by the increased survival rate after challenge with a lethal dose of virus in mice immunized and boosted with adjuvant-combined NP. The acceleration of nasal virus clearance was accompanied with acceleration of type-specific systemic delayed-type hypersensitivity (DTH) and with IFN-gamma production by nasal lymphocytes. The nasal lymphocytes from the immunized and challenged mice generated a significantly high level of DTH when transferred locally, but no class I MHC-restricted CTL response. Moreover, nasal CD4+ T cells, induced by NP immunization and increased in number by the subsequent challenge, were involved in the accelerated IFN-gamma production. These results suggest that nasal Th1 cells, capable of producing IFN-gamma and mediating DTH, are involved in the type-specific acceleration of recovery from influenza after challenge in mice immunized intranasally with adjuvant-combined NP, although the nonspecific mechanism of accelerated recovery remains to be solved.
The Journal of Immunology 06/1996; 156(10):3892-900. · 5.79 Impact Factor
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K Hashigucci,
H Ogawa,
T Ishidate,
R Yamashita,
H Kamiya,
K Watanabe,
N Hattori,
T Sato,
Y Suzuki,
T Nagamine, C Aizawa,
S Tamura,
T Kurata,
A Oya
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ABSTRACT: Evaluation of the efficacy of nasal influenza vaccine combined with Escherichia coli heat-labile enterotoxin B subunit (LTB) containing a trace amount of the holotoxin (LT) in inducing antibody responses among volunteers, which was conducted during the winter season of 1993-1994, is reported. A trivalent inactivated vaccine, composed of A/Yamagata/32/89 (H1N1), A/Kitakyusyu/159/93 (H3N2) and B/Bangkok/163/90 influenza virus strains, was used alone or together with the adjuvant, recombinant LTB supplemented with 0.5% recombinant LT (LTB*). The volunteers were divided into two groups: 73 volunteers (mean age 35.0 +/- 12.0 years) inoculated intranasally (i.n.) with LTB*-combined vaccine and 49 volunteers (37.9 +/- 11.3) inoculated i.n. with the vaccine alone. Vaccination was done twice 4 weeks apart. Salivary secretory IgA and serum hemagglutination-inhibiting (HI) antibodies were measured before and 8 weeks after the primary vaccination. For the sake of convenience, more than a 1.4-fold rise in IgA antibody response (units of specific IgA antibody per microgram of total IgA) and a fourfold or greater rise in HI antibody titer after vaccination were regarded as a positive antibody response. Thirty-seven (50.3%) and 36 (49.3%) of the 73 vaccinees, respectively, given the nasal LTB*-combined vaccine showed positive IgA and HI antibody responses to one or more of the three vaccine strains. In comparison, positive antibody responses in the group given vaccine alone were 32.7% for IgA and 30.6% for HI antibody. There was a significant difference between these two groups. These results suggest that the nasal LTB*-combined vaccine could enhance the production of higher levels not only of serum HI antibody but IgA antibodies in the respiratory tract than do the nasal vaccine alone.
Vaccine 03/1996; 14(2):113-9. · 3.77 Impact Factor
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ABSTRACT: A reproducible method for isolation of mouse nasal lymphocytes was developed. The cells were released from tissue fragments of dissected mouse nose by enzyme extraction with collagenase and separated by a stepwise Percoll gradient centrifugation. The partially purified nasal lymphocyte fraction from normal BALB/c mice contained CD4+ T cells (18-23%), CD8+ T cells (7-10%) and B cells (20-38%), when analysed with a FACScan fluorescence analyser. The ratio of T to B cells and that of CD4+ to CD8+ T cells in the nasal cell fraction were about twice as high as those in Peyer's path lymphocytes. The nasal lymphocyte fraction from the mice infected with influenza virus was then characterized. The nasal lymphocytes contained a twice larger number of CD4+ and a three times larger number of CD8+ T cells than those of normal mice 7 days after infection. They produced IFN-gamma which increased after infection. They contained the cells secreting influenza virus-specific IgA and IgG antibodies 4 weeks after infection. Moreover, the nasal lymphocytes from infected C3H mice lysed the virus infected-target cells. These results suggest that this method can successfully be used for investigating cellular dynamics of mucosal immunity in the upper respiratory tract of experimental animals.
Journal of Immunological Methods 12/1995; 187(1):41-51. · 2.20 Impact Factor
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ABSTRACT: Cholera toxin B subunit (CTB) (1 microgram) and a trace amount of cholera toxin (CT) (0.1-10 ng), when inoculated intranasally into Balb/c mice together with influenza vaccine, induced synergistically a greater delayed-type hypersensitivity (DTH) response to the vaccine than did a trace amount of CT alone. In parallel with the in vivo response, normal peritoneal macrophages that were incubated in vitro with the vaccine and the CT-containing CTB, induced a higher adenylate cyclase activity and a greater ability to transfer DTH response into naive recipient mice than did the macrophages incubated with the vaccine and CT. The treatment of macrophages with the vaccine and CTB failed to induce either adenylate cyclase or DTH response. From these results, the mechanism by which CTB and a trace amount of CT enhance immune responses synergistically could be explained by the enhancement of the CT action on macrophages or by the efficient binding of a trace amount of CT to antigen-presenting cells in the presence of a relatively large amount of CTB, resulting in enhanced cyclic AMP formation followed by enhanced antigen presentation.
Vaccine 04/1995; 13(4):339-41. · 3.77 Impact Factor
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ABSTRACT: The effects of a combination of intranasal (i.n.) and subcutaneous (s.c.) administration of inactivated influenza vaccine for priming and boosting on the cross-protection against antigenically drifted virus challenge were examined in Balb/c mice. Mice were primed through the i.n. or s.c. route with a CTB*-A/Kumamoto/37/79 (H1N1) combined vaccine (CTB*: cholera toxin B subunit supplemented with 0.2% of the holotoxin) and boosted through the i.n. or s.c. route with another drift virus vaccine, A/Bangkok/10/83 (H1N1), 4 weeks later. Two weeks after boosting, the mice were challenged with a third drift virus, A/Yamagata/120/86 (H1N1). The combination of i.n. priming and i.n. boosting afforded the highest cross-protection, while combinations of s.c. priming and i.n. or s.c. boosting afforded little cross-protection. In parallel with the protective activity, anti-A/Yamagata haemagglutinin-reactive IgA and IgG antibodies were detected in nasal and bronchoalveolar wash specimens. These results suggest that cross-protection against a variant virus challenge is most favourably provided by i.n. priming with the CTB* combined vaccine and i.n. boosting with the vaccine, which optimally induces cross-protective IgA and IgG antibodies.
Vaccine 02/1995; 13(1):3-5. · 3.77 Impact Factor
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ABSTRACT: Effects of cholera toxin B subunits supplemented with 0.1% cholera toxin (CTB*) on systemic IgE antibody responses to ovalbumin (OVA) were examined in BDFI (H-2b/d), Balb/c (H-2d) and C3H (H-2k) mice given OVA intragastrically or intranasally. Two successive doses of OVA intragastrically administered to Balb/c and C3H mice induced little IgE response and resulted in almost complete unresponsiveness to subsequent intraperitoneal challenge with OVA in Al(OH)3, while the intragastric administration to BDF1 mice induced high IgE response and resulted in abrogation of the unresponsiveness to the subsequent challenge. The intranasal administration of OVA induced little IgE response and suppressed response to the subsequent challenge in any strain of mice. On the other hand, two successive doses of intragastric or intranasal OVA together with CTB* enhanced IgE response in all three strains and the subsequent challenge with OVA in Al(OH)3 induced higher IgE responses. These results suggest that CTB* augments IgE response to OVA and abrogates the unresponsiveness when administered orally or intranasally into mice together with OVA, regardless of the H-2 haplotype of the mice.
Vaccine 11/1994; 12(13):1238-40. · 3.77 Impact Factor
