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Active immunization against tetanus in man. II. Combined active and passive prophylaxis with human tetanus immune globulin.

19 Persons were actively immunized with adsorbed tetanus toxoid and were simultaneously given tetanus immune globulin of human origin, TIG(H), in doses of 500-1500 IU. Their antitoxin titres were followed for 1 year. Seven persons were given only TIG(H), 500 IU and 1500 IU and their antitoxin titres were followed for 3 months to 1 year. For comparison, 30 military recruits were actively immunized with adsorbed tetanus toxiod according to common practice. Their antitoxin titres were followed for 1 year. The response to complete active immunization could not be demonstrated to be impaired by passive immunization, when 500 IU or 1500 IU OF TIG(H) were given simultaneously with toxoid. The titres were in accordance with those achieved by active immunization of the recruits.

Adult↗

Immune processes in the course of infection with dysentery bacilli. III. Protective activity of spleen cells and serum from mice immunized with killed dysentery bacilli.

Killed dysentery bacilli induce immunity in mice which can be transferred to other mice with serum. Spleen cells do not transfer this immunity, in contrast to spleen cells from mice immunized with live dysentery bacilli. The results of this and previous studies 5,17 suggest that immunity in mice infected with dysentery bacilli depends on two coexisting effector mechanisms--cell-mediated and humoral immunity. Live bacilli induce both, however killed bacilli stimulated humoral immunity and protect mice equally effectively against lethal infection with dysentery bacilli.

Animals↗

Cytokine and hepatitis B virus DNA co-immunizations enhance cellular and humoral immune responses to the middle but not to the large hepatitis B virus surface antigen in mice.

Genetic immunization is a potentially useful strategy to prevent or treat hepatitis B virus (HBV) infection. We have previously shown that HBV envelope proteins are highly immunogenic using this technique. The large envelope protein (LHBs), however, induced significantly weaker humoral and cellular immune responses when compared with the middle envelope protein (MHBs). We studied the effect of co-immunizations with cytokine DNA expression constructs encoding for interleukin (IL)-2 and (GM-CSF) on the immunogenicity of LHBs at the B-and T-cell level. Co-immunizations of mice with plasmids encoding for MHBs and IL-2 or GM-CSF increased anti-HBs responses, helper T-cell proliferative activity, and cytotoxic T lymphocyte (CTL) killing. In contrast, co-immunizations of plasmids encoding for LHBs and IL-2 or GM-CSF had no effect on humoral and cellular immune responses. LHBs did not inhibit the production or secretion of IL-2 and GM-CSF. In addition, IL-2, tumor necrosis factor alfa (TNF-alpha), and interferon gamma (IFN-gamma) had no suppressive effect on HBV envelope protein expression in vitro. Based on these data, MHBs, but not LHBs, genetic immunization can be augmented by IL-2 or GM-CSF cytokines.

Animals↗

Update on the mechanisms of immune suppression of injury and immune modulation.

Major trauma results in massive impairment of immunologic reactivity, the clinical consequence of which consists in the high susceptibility of the traumatized individual toward serious infection. Whereas parts of the immune system are stimulated within a systemic, nondiscriminant, excessive whole-body inflammation, other functions within the complex of cell-mediated immunity (CMI) are dramatically paralyzed. Immune abnormalities in the aftermath of trauma occur in a sequence of states of cellular activation and within a complex order of events that is not yet well understood. Traumatic stress is causing disintegration of the intact monocyte (Mphi)-T cell interaction, which is associated with profound changes in Mphi forward-regulatory capacities and substantial depression of T cell function. Extensive tissue destruction results in the generation of numerous stimuli, such as phagocytosis, immune complexes, complement split products, and endo- and exotoxins, all of which contribute to excessive Mphi activation. Mphi then rapidly produce and release prostaglandin E2 (PGE2), a powerful endogenous immune suppressant. PGE2 is an inhibitor of T cell mitogenesis, interleukin 2 (IL-2) production, and IL-2 receptor expression; and it has a massive impact on the quality of B cell antibody synthesis. Most importantly, PGE2 represents an important cofactor for the induction of T-helper lymphocyte (TH) activity toward the TH2 direction. TH2 cells are associated with the synthesis of immunosuppressive cytokines, such as IL-4 and IL-10. Although immunosuppressive substrates are inhibitory for TH1 cells-the functional carriers of CMI-they support TH2 activity, which predisposes the host to develop infection. The endogenous ability of the organism to survive overwhelming trauma is insufficient and requires major exogenous support. Immune modulatory interventions, depending on the immune abnormalities seen in the traumatized host, should be started as early as possible after trauma in a preventive fashion to protect against organ tissue destruction. Ideally, it should protect all cellular host defense compartments from hyperactivation as well as from exhaustion. We do believe that only a combination of drugs can effectively control the posttraumatic dyshomeostasis of the various cell systems.

Adjuvants, Immunologic↗

Characterization of the mucosal immune response in breast milk after peroral immunization of chimpanzees (Pan troglodytes) with Streptococcus mutans.

The characteristics of the mucosal immune response to Streptococcus mutans cells, antigen A, antigen B, glucosyltransferases and glucan-binding proteins were examined in four pregnant chimpanzees that had been immunized perorally with Strep. mutans. Six pregnant chimpanzees served as non-immunized controls. None of the chimpanzees harbored S. mutans. Samples of milk were collected from all animals throughout the experiment. Peroral immunization resulted in an overall 17-fold median increase in SIgA in milk. Although SIgA1 comprised almost two-thirds of milk SIgA, Strep. mutans whole-cell antibody activity was contained predominantly in the SIgA2 subclass. The difference between the specific activities of anti-Strep. mutans SIgA1 and SIgA2 antibodies compared over time reached the borderline of statistical significance (p = 0.08). The avidity of anti-Strep. mutans antibodies was low in three of four chimpanzees and there was no evidence of affinity maturation. SIgA antibodies from the milk of all four immunized chimpanzees recognized antigen A. In three animals these antibodies were restricted to the SIgA1 subclass and, in one animal, anti-A antibodies were confined to SIgA2. Antibodies from all of the immunized chimpanzees recognized degradation products of antigen B in both the SIgA1 and the SIgA2 subclasses. Only two of four immunized chimpanzees responded to glucosyltransferases and these antibodies were restricted to the SIgA1 subclass. None of the chimpanzees responded to the 74-kDa glucan-binding protein. However, three animals produced SIgA1 antibodies against the 59-kDa glucan-binding protein and two of these also produced SIgA2 antibodies against this protein.

Analysis of Variance↗

Adoptive transfer of natural antibodies to non-immunized chickens affects subsequent antigen-specific humoral and cellular immune responses.

To determine a regulatory function of natural antibodies in the immune response of chickens, pooled plasma obtained from non-immunized (naïve) 15 months old hens was subjected to keyhole limpet hemocyanin (KLH) antigen-affinity chromatography. Purified KLH-binding antibodies were adoptively transferred intravenously to 5 weeks-old cocks that were subsequently immunized subcutaneously 24 h later with KLH. Control groups consisted of birds that were either adoptively transferred with KLH-binding antibodies purified from plasma of KLH-immunized chickens, or PBS, or a salt precipitated total immunoglobulin fraction obtained from the corresponding pooled nai;ve chicken plasma, respectively.Total, IgM and IgY antibody titers to KLH in the plasma of recipients adoptively transferred with KLH-NAb, but not in the plasma of the groups transferred with salt precipitate or KLH-binding specific antibodies, were significantly enhanced as compared to the non-treated, KLH immunized group. Titers of IgA antibodies binding KLH were decreased in the plasma of the group that received specific KLH-binding antibodies, but not in the plasma of the other groups. Proliferation from peripheral blood leucocytes in whole blood from the KLH-NAb treated group, the group treated with KLH-binding specific antibodies and the group treated with salt precipitate, respectively, to both concanavalin A and KLH were significantly decreased as compared to the group receiving PBS. Our data show that antigen-specific antibodies can be isolated from plasma obtained from non-immunized chickens. Such antibodies that resemble natural antibodies as described in mammals may perform an important role in the enhancement of subsequent antigen-specific antibody responses or the maturation of the immune system, which may differ from the role of specific antibodies.

Animals↗

Induction of mucosal immune responses following enteric immunization with antigen delivered in alginate microspheres.

Oral immunization is the most effective way of inducing immune responses in the intestinal tract. Biodegradable microspheres have been used extensively for the delivery of antigens to the Peyer's patches (PPs) within the gut-associated lymphoid tissue (GALT). We evaluated various formulations of alginate microspheres for their capacity to induce mucosal immune responses in vivo. Multiple intestinal "loops" each containing a single PP, were surgically prepared in lambs. We have previously showed that PP in individual intestinal loops function as independent sites for the induction of immune responses. This animal model provides a system for directly comparing different antigen formulations within the same animal. Individual intestinal loops were injected with a model antigen, porcine serum albumin (PSA) encapsulated in three different formulations of alginate micropsheres. Three weeks after immunization, PSA-specific immune responses were assayed with antibody secreting cell (ASC) ELISPOT, lymphocyte proliferative responses (LPRs), IFN-gamma production and antibody secreted into intestinal loops. PSA encapsulated in alginate micropsheres or in saline induced humoral immune responses as indicated by the presence of numerous ASC. However, PSA-specific T-cell responses (LPR and IFN-gamma production) were not induced.

Alginates↗

Intranasal immunization with liposome-formulated Yersinia pestis vaccine enhances mucosal immune responses.

The induction of mucosal immune responses by a liposome-formulated Y. pestis vaccine (formaldehyde-killed whole cell vaccine; KWC) was evaluated. We demonstrated that intranasal immunization of mice with Y. pestis KWC vaccine, formulated with liposomes, significantly enhanced mucosal immune responses in the lung when compared to the responses induced with KWC vaccine alone. These immune responses were characterized by increased titres of specific IgA and IgG in mucosal secretions (lung and nasal washes), and an increased frequency of specific antibody-secreting cells in the lungs. In addition, antigen-specific proliferative responses and IFN-gamma-secreting cells were also significantly enhanced in the spleens of mice immunized with the KWC vaccine formulated in liposomes. Animals that were immunized intranasally with the KWC vaccine showed significant protection against an intranasal challenge with Y. pestis. These results highlight the importance of mucosal administration of vaccine antigens to stimulate immunity in the respiratory tract and demonstrate that liposome formulations can improve the effectiveness of conventional vaccines.

Administration, Intranasal↗

Transcutaneous immunization: T cell responses and boosting of existing immunity.

Transcutaneous immunization (TCI) is a novel immunization strategy by which antigen and adjuvant are applied topically to intact, hydrated skin to induce potent antibody and cell-mediated immune responses specific for both the antigen and the adjuvant. Using tetanus toxoid as a model antigen, we examined the T cell response to tetanus toxoid after topical immunization with a variety of adjuvants. TCI readily induced systemic antigen specific T cell responses with a mixed Th1/Th2 phenotype but with a Th2 bias. We also investigated whether priming by the intramuscular route, which is known to induce T cell memory, could be followed by a boosting immunization on the skin to induce secondary responses. TCI could augment existing immunity, but interestingly, this strategy induced potent responses only if the antibody titer was low at the time of TCI boosting. These and previous observations suggest that TCI follows known immunological principles that govern other routes of vaccine delivery. Furthermore, booster immunization using tetanus toxoid may provide a useful model for further development of important patch and formulation concepts for TCI, and act as an early candidate for validating product feasibility of TCI in humans.

Adjuvants, Immunologic↗

The effect of pre-existing adenovirus-specific immunity on immune responses induced by recombinant adenovirus expressing glycoprotein D of bovine herpesvirus type 1.

We investigated whether pre-existing adenovirus-specific immunity influenced the development of immunity to a foreign antigen expressed by recombinant adenovirus. Active adenovirus-specific immunity was induced in cotton rats by i.n. administration of wild type human adenovirus type 5 (HAd5) two weeks before immunisation with a HAd5 vector expressing the glycoprotein D (gD) of bovine herpesvirus type 1 (gD-dE3 recombinant adenovirus). Active adenovirus-specific immunity inhibited gD-specific immune responses, following either i.n. or gastrointestinal immunisation with gD-dE3. An inhibitory effect was present even if infection with HAd5 and immunisation with gD-dE3 were 13 weeks apart. Passive transfer of adenovirus specific antibodies to cotton rats one day before immunisation, however, did not significantly inhibit gD-specific immune responses induced by i.n. immunisation with gD-dE3. Repeated administration of an adenovirus vector, therefore, may have a limited ability to deliver antigen, while passive immunity to adenovirus may not interfere with the success of immunisation.

Adenoviruses, Human↗

Immunity to influenza in ferrets. V. Immunization with inactivated virus in adjuvant 65.

Ferrets infected with influenza virus A2/Hong Kong/3/68 responded with a febrile reaction; the temperature was elevated by 1.0 degrees C. or greater to a level of 40 degrees C. or more. In addition, relatively high titres of virus were recovered from nasal washings taken 3 days after virus infection, serum antibody was produced, increased nasal protein was detected and nasal washings contained both HI and neutralizing antibody. Of four ferrets immunized with 400 CCA units of inactivated influenza virus A2/Aichi/2/68 in saline, only one produced detectable serum HI antibody, and none produced detectable nasal antibody. These ferrets were subsequently found to be susceptible to intranasal infection with influenza virus A2/Hong Kong/3/68. Thus, the temperature response, the titre of virus recovered from nasal washings and the serum HI antibody response found after virus infection was similar to that found after infection of non-immunized ferrets. However, the increase in protein concentration and the titre of HI and neutralizing antibody found in nasal washings after virus infection was detectably less than that found after virus infection of non-immunized ferrets.Four ferrets were immunized with 400 CCA units of inactivated A2/Aichi/2/68 virus in adjuvant 65, and these ferrets produced relatively high titres of serum HI antibody but no detectable nasal antibody. After subsequent virus infection with influenza virus A2/Hong Kong/3/68, these ferrets showed a modified temperature response, reduced titres of virus in nasal washings compared to that found in nasal washings from non-immunized ferrets, no increase in nasal protein and no detectable nasal HI antibody. Thus, immunization with inactivated virus in adjuvant 65 resulted in a significant modification of the response of ferrets to challenge virus; however, the immunity was not complete, and appreciably less than that found after infection with live homologous virus.

Adjuvants, Immunologic↗

[Active immunization against hepatitis A. Comparison of various immunization schedules].

Three different immunization schedules were compared in 144 young, healthy adults (81 men, 63 women; mean age 28.5 years). They were randomly assigned to one of three groups: group 1, immunization shots on day 0 and day 14 (n = 47), group 2, on day 0 and day 28 (n = 50), and group 3, on day 0, 14 and 28 (n = 47). All participants had a booster shot after one year. The seroconversion rate was 40-46% after first shot, 95-100% after the second one; all were anti-HAV positive after the booster injection. All subjects had specific antibodies in a mean concentration of > 400 IU/l two weeks after the second shot. In almost all subjects antibodies were demonstrable up to the 12th month after the first immunization (> 20 IU/l). Mean antibody concentration during the interval between the second and third immunization after basal immunization on day 0 and 14 or 0 and 28, respectively, was comparable. Three initial immunizations 14 days apart did not achieve much higher anti-HAV levels. The vaccine was equally well tolerated in all three groups. Thus basal immunization shortened to two injections two weeks apart presents a good alternative to the standard scheme of two injections four weeks apart. Three injections two weeks apart bring no demonstrable advantage.

Adult↗

Preferential induction of a Th1 immune response and inhibition of specific IgE antibody formation by plasmid DNA immunization.

We compared the antigen-specific antibody isotypes and lymphokine secretion by CD4+ T cells in BALB/c mice immunized intradermally with either Escherichia coli beta-galactosidase (beta-gal) or plasmid DNA (pDNA) encoding beta-gal in a cytomegalovirus-based expression vector (pCMV-LacZ). pCMV-LacZ induced mainly IgG2a, whereas beta-gal in saline or alum induced IgG1 and IgE beta-gal-specific antibodies. In addition, splenic CD4+ T helper (Th) cells isolated from pDNA-immunized mice secreted interferon-gamma but not interleukin (IL)-4 and IL-5, whereas Th cells from beta-gal-injected mice secreted IL-4 and IL-5 but not interferon-gamma after in vitro stimulation with antigen. Together these data demonstrate that pDNA immunization induced a T helper type 1 (Th1) response, whereas protein immunization induced a T helper type 2 (Th2) response to the same antigen. Interestingly, priming of mice with pCMV-LacZ prevented IgE antibody formation to a subsequent i.p. beta-gal in alum injection. This effect was antigen-specific, because priming with pCMV-LacZ did not inhibit IgE anti-ovalbumin antibody formation. Most importantly, intradermal immunization with pCMV-LacZ (but not pCMV-OVA) of beta-gal in alum-primed mice caused a 66-75% reduction of the IgE anti-beta-gal titer in 6 weeks. Also, pCMV-LacZ induced specific IgG2a antibody titers and interferon-gamma secretion by Th cells in the beta-gal in alum-primed mice. The data demonstrate that gene immunization induces a Th1 response that dominates over an ongoing protein-induced Th2 response in an antigen-specific manner. This suggests that immunization with pDNA encoding for allergens may provide a novel type of immunotherapy for allergic diseases.

Animals↗

Effect of dose and immunization schedule on immune response of baboons to recombinant glycoprotein 120 of HIV-1.

To evaluate the immunogenicity of purified recombinant envelope glycoprotein of HIV-1 (rgp120) as a potential vaccine for AIDS, the antibody response of 45 baboons to rgp120 was investigated using an adjuvant (alum) and route of administration (intramuscular) suitable for humans. The primary purpose was to evaluate the effects of different doses and immunization schedules on the antibody response to rgp120 in primates. A secondary objective was to evaluate possible adverse consequences of rgp120 immunization. A liquid-phase radioimmunoprecipitation (RIP) assay for detection of rgp120-reactive antibodies revealed that rgp120 doses of 30-300 micrograms per administration resulted in nearly indistinguishable serum antibody responses. However, significant enhancement of serum antibody titers was observed when the interval between the second and third administrations was increased from 4 to 20 w. Although changing the interval significantly altered the magnitude of resulting peak titers, the kinetics of antibody formation were not changed. Thus, of the three schedules of immunization tested, none resulted in a sustained humoral immune response. The significance of the RIP assay for evaluating immune responses was confirmed by analysis showing that the percentage of immunized baboons that developed in vitro HIV-1 serum neutralizing responses was greatest in groups that also exhibited high anti-rgp120 RIP titers. Immunization with rgp120 had no significant adverse effect on any clinical or laboratory parameter monitored over the course of the study.

Acquired Immunodeficiency Syndrome↗

Cell-mediated immune responses after immunization of healthy seronegative children with varicella vaccine: kinetics and specificity.

Humoral and cell-mediated immune responses were determined in seronegative children immunized with live attenuated Oka strain varicella vaccine. At 2 weeks after immunization, 80% of children had detectable lymphocyte proliferation to varicella-zoster virus (VZV) antigens, while only 40% had antibodies to VZV as detected by ELISA. By 6 weeks after immunization, 97% of children seroconverted, and 95% of these responded to VZV antigens in the proliferation assay. A high proportion of immunized children also responded in the proliferation assay to purified glycoproteins I, II, and III of VZV. These results indicate that most children develop a broad cell-mediated immune response to VZV antigens within weeks after immunization with varicella vaccine.

Antibodies, Viral↗

Costs of immunity: immune responsiveness reduces survival in a vertebrate.

Immune defences are undoubtedly of great benefit to the host, reducing the impact of infectious organisms. However, mounting immune responses also entails costs, which may be measured by inducing immune responses against artificial infections. We injected common eider (Somateria mollissima) females with three different non-pathogenic antigens, sheep red blood cells (SRBC), diphtheria toxoid and tetanus toxoid, early in their incubation period. In the group of females that mounted a humoral immune response against SRBC, the return rate was only 27%, whereas the group of females that did not mount a response against SRBC had a return rate of 72%. Moreover, responding against diphtheria toxoid when also responding against SRBC led to a further reduction in return rate. These results are repeatable, as the same effect occurred independently in two study years. The severely reduced return rate of females producing antibodies against SRBC and diphtheria toxoid implies that these birds experienced considerably impaired long-term survival. This study thus documents severe costs of mounting humoral immune responses in a vertebrate. Such costs may explain why many organisms suppress immunity when under stress or when malnourished, and why infections may sometimes be tolerated without eliciting immune responses.

Animals↗

Oral spray immunization may be an alternative to intranasal vaccine delivery to induce systemic antibodies but not nasal mucosal or cellular immunity.

Sixty-five healthy adult volunteers were immunized four times at 1-week intervals with an inactivated whole-virus influenza vaccine based on the strain A/New Caledonia/20/99 (H1N1) without adjuvant. The vaccine was administered as nasal spray with a newly developed device to secure intranasal delivery (OptiMist, OptiNose AS, Oslo, Norway), as regular nasal spray, nasal drops or as an oral spray. Significant IgA-antibody responses in nasal secretions were induced in volunteers immunized intranasally but not after oral spray immunization. In saliva, IgA antibodies were only marginally amplified even after oral spray immunizations. At least 73% of the volunteers belonging to any group of vaccine delivery reached serum haemagglutination inhibition titres of 40 or higher, considered protective against influenza, after only two vaccine doses. Those who had the vaccine delivered intranasally also showed evidence from in vitro secretion of granzyme B that cytotoxic T cells had been stimulated. Although immunization with the breath-actuated OptiMist device and nasal drops were superior with respect to both mucosal and systemic immune responses, oral spray immunization might still be considered for studies of mucosal adjuvants that are not yet acceptable for intranasal use.

Administration, Intranasal↗

Special feature for the Olympics: effects of exercise on the immune system: exercise effects on mucosal immunity.

The present review examines the effects of exercise on mucosal immunity in recreational and elite athletes and the role of mucosal immunity in respiratory illness. Habitual exercise at an intense level can cause suppression of mucosal immune parameters, while moderate exercise may have positive effects. Saliva is the most commonly used secretion for measurement of secretory antibodies in the assessment of mucosal immune status. Salivary IgA and IgM concentrations decline immediately after a bout of intense exercise, but usually recover within 24 h. Training at an intense level over many years can result in a chronic suppression of salivary immunoglobulin levels. The degree of immune suppression and the recovery rates after exercise are associated with the intensity of exercise and the duration or volume of the training. Low levels of salivary IgM and IgA, particularly the IgA1 subclass, are associated with an increased risk of respiratory illness in athletes. Monitoring mucosal immune parameters during critical periods of training provides an assessment of the upper respiratory tract illness risk status of an individual athlete. The mechanisms underlying the mucosal immune suppression are unknown.

Exercise↗