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Anomalous observations on IPV and OPV vaccination.

Anomalies of vaccine performance are events or observations that go counter to the current paradigms on the properties of a vaccine. In the current paradigm, inactivated poliovirus vaccine (IPV) induces serum immunity to protect the individual from poliomyelitis, but not mucosal immunity to prevent subsequent wild poliovirus infection. Therefore, it is believed not to be suitable to interrupt virus transmission in polio-endemic tropical and developing countries, where poliovirus transmission is thought to be predominantly faecal-oral, unlike in countries with excellent sanitation, where it is predominantly respiratory. Oral poliovirus vaccine (OPV), because it is believed to mimic natural (wild) poliovirus infection and to induce serum and mucosal immunity, is considered ideal to interrupt virus transmission in countries with poor sanitation and endemic polio. Moreover, the vaccine viruses are shed in the stools and it is presumed to spread faeco-orally to unvaccinated children in the vicinity, thus increasing its effectiveness in the community. These paradigms on both vaccines may be seen in standard textbooks of virology and paediatrics. In reality, IPV induces very high mucosal immunity in a monkey model, lasting at least 12 months and providing complete protection against oral challenge with wild virus. In field trials, IPV has been found to have very high vaccine efficacy (VE) and to retard significantly wild-virus transmission in communities. OPV, requiring five to seven doses for individual protection, does not seem to induce effective mucosal immunity. Hence, 10-15 doses of OPV and near 100% vaccination coverage are necessary for interruption of virus transmission in countries with poor sanitation. In the monkey model, wild poliovirus infection did not offer any mucosal protection against a second infection, indicating that live virus infection is not the best way to induce mucosal immunity (unless repeated several times). The immunity induced by OPV has two arms, individual protection (by serum immunity) and mucosal protection from later infection by wild virus; these are dichotomous, as evidenced further by protected children participating in wild virus transmission in breakthrough outbreaks in communities well vaccinated with OPV. There is no evidence that polioviruses, wild or vaccine, spread faeco-orally to any meaningful extent. The median age of polio in India, in the pre-vaccine era, and even in recent years was 12-18 months. No other faeco-oral infection has such a low median age. Measles virus transmission is respiratory, and the median age of measles is over two years. Wild virus is shed faecally, but by the age when faecal microbes infect children, most of them are already immune to disease caused by polioviruses. As for vaccine viruses, even when heavy inocula are fed repeatedly, children may not always become infected. That exposure to the small amounts of virus through faecal contamination (unlike microbes that multiply in food) would infect them, is an unrealistic expectation. There are sufficient anomalies demanding our revision of the old paradigms. IPV is a better immunogen than OPV and is completely safe. During the interval between the cessation of wild virus transmission and the global stoppage of polio vaccination, it will be advantageous to use IPV, particularly combined with DPT vaccine, to provide individual protection, and herd effect to prevent spread in case of the introduction of wild polioviruses, whether unintentionally or otherwise, and to bolster the Expanded Programme on Immunisation.

Animals↗

Lymphocyte-mucosal interaction of the middle ear mucosa.

The middle ear mucosa possesses immunologic features similar to those of the peripheral mucosa sites in the common mucosal immune system and after mucosal immunization, antigenspecific IgA-forming cells appear in the inflamed mucosa of the tympanic cavity. Recent investigations suggest that lymphocyte migration to lymphoid tissues is regulated by lymphocyte-high endothelial venules (HEV) interaction. However, the lymphocyte migration mechanism to the middle ear mucosa is still unclear. We investigated whether or not organ-specific determinants which lymphocytes bind with are present on the middle ear mucosa by in vivo and in vitro lymphocyte adherence assays by using fluorescein-labeled lymphocytes from various lymphoid tissues. Many lymphocytes from Peyer's patches and hilar lymphnodes adhered on the inflamed middle ear mucosa with or without mucosal immunization, while these cells were not found on the normal tympanic mucosa. The number of the cells was smaller than that in the gastrointestinal mucosa. Lymphocyte adherence to the middle ear mucosa was not suppressed by anti-T cell antibody. These findings suggest that the middle ear mucosa possesses organ-specific mucosal determinants which B-lymphocytes selectively bind with, and that those determinants which regulate lymphocyte migration to the middle ear mucosa differ from those of other mucosae in the gastrointestinal tract.

Animals↗

Gluten-sensitive disease with mild enteropathy.

BACKGROUND & AIMS: Celiac disease is a permanent gluten intolerance strongly associated with HLA class II antigens and possibly showing milder changes of mucosal architecture. Ten patients with symptoms suggesting celiac disease and serum antiendomysium antibodies with normal mucosal architecture were studied. METHODS: Immunohistochemical detection of mucosal immune activation and HLA typings were performed. RESULTS: Mucosal immune activation, with normal mucosal architecture and normal gamma/delta+ intraepithelial lymphocytes counts, was found on a gluten-containing diet. In 3 of 6 patients, multiple biopsy specimens showed one sample with severe villous atrophy. Clinical and immunomorphologic features were strictly gluten dependent. The mucosal immune activation was elicited in vitro by gliadin. Only 4 patients had the typical HLA typing of celiac disease. CONCLUSIONS: Gluten-sensitive celiac-like symptoms may occur in patients with serum antiendomysium antibodies, apparently normal intestinal mucosa, and HLA typing not commonly associated with celiac disease. These patients should undergo multiple biopsies, and signs of immunologic activation should be sought accurately; in the presence of mucosal immune activation, a trial with a gluten-free diet should be encouraged to detect gluten dependency. In vitro immunologic response of small intestinal mucosa to gliadin may support the diagnosis of gluten-sensitive enteropathy.

Adult↗

Protective mucosal Th2 immune response against Toxoplasma gondii by murine mesenteric lymph node dendritic cells.

Toxoplasma gondii, an obligate intracellular parasite pathogen which initially invades the intestinal epithelium before disseminating throughout the body, may cause severe sequelae in fetuses and life-threatening neuropathy in immunocompromised patients. Immune protection is usually thought to be performed through a systemic Th1 response; considering the route of parasite entry it is important to study and characterize the local mucosal immune response to T. gondii. Despite considerable effort, Toxoplasma-targeted vaccines have proven to be elusive using conventional strategies. We report the use of mesenteric lymph node dendritic cells (MLNDCs) pulsed ex vivo with T. gondii antigens (TAg) as a novel investigation approach to vaccination against T. gondii-driven pathogenic processes. Using a murine model, we demonstrate in two genetically distinct mouse strains (C57BL/6 and CBA/J) that adoptively transferred TAg-pulsed MLNDCs elicit a mucosal Toxoplasma-specific Th2-biased immune response in vivo and confer strong protection against infection. We also observe that MLNDCs mostly traffic to the intestine where they enhance resistance by reduction in the mortality and in the number of brain cysts. Thus, ex vivo TAg-pulsed MLNDCs represent a powerful tool for the study of protective immunity to T. gondii, delivered through its natural route of entry. These findings might impact the design of vaccine strategies against other invasive microorganisms known to be delivered through digestive tract.

Adoptive Transfer↗

Inflammatory bowel disease and irritable bowel syndrome: separate or unified?

Both irritable bowel syndrome and inflammatory bowel diseases share symptoms of altered bowel habits associated with abdominal pain or discomfort. Irritable bowel syndrome has been referred to as a functional bowel disorder, which is diagnosed by a characteristic cluster of symptoms in the absence of detectable structural abnormalities. Inflammatory bowel disease is a heterogeneous group of disorders characterized by various forms of chronic mucosal and/or transmural inflammation of the intestine. In this review, the authors discuss recent evidence suggesting several potential mechanisms that might play a pathophysiologic role in both syndromes. Possible shared pathophysiologic mechanisms include altered mucosal permeability, an altered interaction of luminal flora with the mucosal immune system, persistent mucosal immune activation, alterations in gut motility, and a role of severe, sustained life stressors in symptom modulation. It is proposed that similarities and differences between the two syndromes can best be addressed within the framework of interactions between the central nervous system and the gut immune system. Based on recent reports of low-grade mucosal inflammation in subpopulations of patients meeting current diagnostic criteria for irritable bowel syndrome, therapeutic approaches shown to be effective in inflammatory bowel disease, such as probiotics, antibiotics, and antiinflammatory agents, have been suggested as possible therapies for certain patients with irritable bowel syndrome.

Journal Article↗

Cutting edge: dichotomy of homing receptor dependence by mucosal effector B cells: alpha(E) versus L-selectin.

The common mucosal immune system may be compartmentalized because lymphocyte homing to the upper respiratory tract appears to be mediated by L-selectin interactions rather than alpha(4)beta(7) interactions, as is the case for gut-associated lymphoreticular tissue. To assess the role of L-selectin in effector B cell immunity, L-selectin-deficient mice were intranasally immunized with cholera toxin (CT), and mucosal immune responses were compared with C57BL/6 mice. The absence of L-selectin correlated with a reduction in CT-specific secretory-IgA responses in nasal passages and reproductive tract, but not intestinal lamina propria. Cell sorting experiments showed that an L-selectin-dependent subset was responsible for CT-specific responses in nasal passages and reproductive tract, whereas an alpha(E)beta(7)(+) B cell subset was responsible for L-selectin-independent intestinal immunity. This study provides evidence for compartmentalization of the common mucosal immune system into "intestinal" vs "nonintestinal" effector sites.

Animals↗

Design and tests of an HIV vaccine.

It is likely that a successful vaccine against HIV will need to stimulate the innate immune system, generate high levels of neutralising antibody, strong cellular immune responses, and mucosal immunity. Early efforts to develop HIV vaccines attempted to use the virus glycoprotein, gp120, to induce neutralising antibody, but did not take into account the trimeric structure of the native glycoprotein or the complex nature of the CD4 and chemokine receptor binding sites. Recently, attention has been focused on cellular immune responses, particularly T-cell cytotoxicity, based on evidence from the SIV model and from exposed and uninfected humans. Recent experiments in macaques and man suggest that a prime boost regimen using DNA and recombinant pox virus is highly effective at stimulating cellular immunity. However, in addition to the problems of generating neutralising antibodies and mucosal immunity, the difficulty of inducing broad cellular responses able to protect against all clades of HIV, remains an important issue.

AIDS Vaccines↗

Effects of intranasal immunization on protective immunity against otitis media.

It has been reported that intranasal immunization can induce mucosal immune responses. However, the efficacy of intranasal immunization on otitis media caused by non-typeable Haemophilus influenzae (NTHi) is not yet elucidated. Mice were intranasally, orally, intratracheally or intraperitoneally immunized with outer membrane protein (OMP) isolated from NTHi, and antigen-specific immune responses were determined by enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immuno-spot assay (ELISPOT). Cytokine production from splenic CD4+ T cells was examined by ELISA. Following the immunization, the clearance of NTHi from the nasal and nasopharyngeal cavity was examined. OMP-specific IgA antibody titers in nasal washes and the numbers of specific IgA-producing cells in nasal passages were significantly increased in intranasally immunized mice. Cytokine analysis showed that interferon-gamma (IFN-gamma) and interleukins IL-6 and IL-10 were predominantly produced from CD4+ T cells. The clearance of NTHi was significantly enhanced in the intranasal immunization group. Intranasal immunization is an effective vaccination regimen for the induction of OMP-specific mucosal immune responses.

Animals↗

Enhancement of pulmonary clearance of Moraxella (Branhamella) catarrhalis following immunization with outer membrane protein CD in a mouse model.

Moraxella (Branhamella) catarrhalis is an important human respiratory tract pathogen. Outer membrane protein (OMP) CD is highly conserved among strains and has characteristics that indicate it may be an effective vaccine antigen. This study investigated the effect of immunization with OMP CD on pulmonary clearance following intratracheal challenge of mice with M. catarrhalis. Two routes of immunization were studied: mucosal immunization (intra-Peyer's patch followed by intratracheal boost) and intramuscular immunization with OMP CD. Both resulted in enhanced pulmonary clearance of M. catarrhalis compared with sham-immunized controls. Immunization with OMP CD induced specific antibodies in serum and bronchoalveolar lavage fluid and induced a specific lymphocyte proliferative response in T cells from mesenteric lymph nodes from mice mucosally immunized with OMP CD. On the basis of these results, OMP CD should undergo continued testing to determine whether it will induce a protective immune response in humans.

Animals↗

Dissociation between systemic and mucosal humoral immune responses in coeliac disease.

We examined humoral immunity in coeliac disease as expressed in serum (systemic immunity), and in saliva, jejunal aspirate, and whole gut lavage fluid (mucosal immunity). The aims were to define features of the secretory immune response (IgA and IgM concentrations and antibody values to gliadin and other food proteins measured by enzyme linked immunosorbent assay (ELISA)) in active disease and remission, and to establish whether secretions obtained by relatively non-invasive techniques (saliva and gut lavage fluid) can be used for indirect measurements of events in the jejunum. Serum, saliva, and jejunal aspirate from 26 adults with untreated coeliac disease, 22 treated patients, and 28 immunologically normal control subjects were studied, together with intestinal secretions obtained by gut lavage from 15 untreated and 19 treated patients with coeliac disease and 25 control subjects. Jejunal aspirate IgA and IgM and gut lavage fluid IgM concentrations were significantly raised in patients with untreated coeliac disease; the lavage fluid IgM concentration remained higher in patients with treated coeliac disease than in controls. Serum and salivary immunoglobulin concentrations were similar in the three groups. Patients with untreated coeliac disease had higher values of antibodies to gliadin compared with treated patients and control subjects in all body fluids tested; these were predominantly of IgA and IgG classes in serum, and of IgA and IgM classes in jejunal aspirate and gut lavage fluid. Values of salivary IgA antibodies to gliadin were significantly higher in untreated coeliacs, though antibody values were generally low, with a large overlap between coeliac disease patients and control subjects. In treated patients, with proved histological recovery on gluten free diet, serum IgA antigliadin antibody values fell to control values, though serum IgG antigliadin antibody values remained moderately raised. In contrast, there was persistence of secretory antigliadin antibodies in treated patients (particularly IgM antibody) in both jejunal aspirate and gut lavage fluid. Antibody responses to betalactoglobulin and ovalbumin were similar to those for gliadin, including persistence of high intestinal antibody values in patients with treated coeliac disease. There was a positive correlation between antibody values in jejunal aspirate and gut lavage fluid, but not between saliva and jejunal aspirate; thus salivary antibodies do not reflect intestinal humoral immunity.

Adolescent↗

How the gut links innate and adaptive immunity.

Mucosal surfaces represent the main sites in which environmental microorganisms and antigens interact with the host. Sentinel cells, including epithelial cells, lumenal macrophages, and intraepithelial dendritic cells, continuously sense the environment and coordinate defenses for the protection of mucosal tissues. The mucosal epithelial cells are crucial actors in coordinating defenses. They sense the outside world and respond to environmental signals by releasing chemokines and cytokines that recruit inflammatory and immune cells to control potential infectious agents and to attract cells able to trigger immune responses. Among immune cells, dendritic cells (DC) play a key role in controlling adaptive immune responses, due to their capacity to internalize foreign materials and to present antigens to naive T and B lymphocytes, locally or in draining organized lymphoid tissues. Immune cells recruited in epithelial tissues can, in turn, act upon the epithelial cells and change their phenotype in a process referred to as epithelial metaplasia.

Adaptation, Physiological↗

Mucosal immunoregulation: transcription factors as possible therapeutic targets.

Much progress has been recently made with regard to our understanding of the mucosal immune system in health and disease. In particular, it has been shown that uncontrolled mucosal immune responses driven by lymphocytes or non-lymphoid cells may lead to immunological diseases such as allergy, hypersensitivity and inflammation. Thus, a more detailed understanding of mucosal immune regulation and decision making at mucosal surfaces is essential for a better understanding of mucosal immune responses in health and disease. Antigen presenting cells and T lymphocytes play a key role in controlling mucosal immune responses. To deal with this key task, T helper cells differentiate into functionally distinct subsets: TH1 (CD4+ T Helper cells), TH2, TH3, Tr1, and CD4+CD25+ T (Treg) cells. This review summarizes the role of antigen presenting cells, eosinophils, mast cells and T-cell subsets in the pathogenesis of allergic inflammation and intestinal inflammation. Furthermore, we discuss novel immunological treatment modalities for allergic inflammation (e.g. allergic asthma) and chronic intestinal inflammation (e.g. inflammatory bowel diseases (IBD)) such as the control of the expression of transcription factors to redirect pathological immune responses.

Animals↗

Genetic modification of a vaginal strain of Lactobacillus fermentum and its maintenance within the reproductive tract after intravaginal administration.

Many micro-organisms cause important diseases of the female genital tract. Because systematic vaccination does not usually provide a good immune response at mucosal sites, commensal lactobacilli from the female genital tract were developed as vehicles to deliver continued doses of foreign antigen directly to the genital mucosal surface with the aim of stimulating strong local mucosal immune responses. Lactobacilli were shown to be common inhabitants of the genital tract of the animal model studied, the guinea-pig. One species, Lactobacillus fermentum, was found in all guinea-pigs studied and was chosen for genetic manipulation. Improved methods of electroporation were developed to enable the routine transformation of L. fermentum BR11 strain with the broad host range plasmid pNZ17. This recombinantly modified Lactobacillus strain was shown to possess good segregational stability over 120 generations in the absence of antibiotic selection. When this recombinant L. fermentum strain was administered to the vaginal tract of three guinea-pigs it persisted for only 5 days. Despite the relatively short period of persistence in these initial experiments, this novel vaccine approach could provide an effective means of stimulating mucosal immunity in the female genital tract.

Animals↗

Effect of Bacillus firmus on antibody formation after mucosal and parenteral immunization in mice.

Immunostimulatory properties of B. firmus, a nontoxic, nonpathogenic G + bacterium of external environment, were described previously. Antiinfectious and antitumor activity, macrophage activation and strong polyclonal stimulation of B lymphocytes were proved in human, mice and rats. The adjuvant effect of B. firmus on specific antibody response to ovalbumin in BALB/c mice is the topic of the present study. Against our expectation, B. firmus exerts more suppressive than stimulatory effect on specific antibody response. Formolized B. firmus decreased anti-ovalbumin response after subcutaneous immunization and only slightly increased serum antibodies after intraperitoneal immunization. After mucosal immunization, both oral and rectal, ovalbumin itself did not cause a significant systemic response but induced IgA anti-ovalbumin response in the intestine. B. firmus applied together with ovalbumin increased systemic serum response but absolutely eliminated intestinal response. The rectal route of antigen administration has been found less convenient because of less precise dosing of antigen in this mode of immunization.

Adjuvants, Immunologic↗

The effect of sodium cromoglycate on the induction of experimental IgA nephropathy.

Mesangial IgA nephropathy was experimentally induced in ddY mice by oral and parenteral administration of the poliomyelitis vaccine (POLIO), and we then tried to investigate if IgA deposition could be prevented by the concurrent use of sodium cromoglycate (SCG), which is known to inhibit the local mucosal immune reaction. Mucosal and systemic immunity could be induced by the administration of POLIO; proteinuria, increased serum IgA levels, mesangial cell proliferation, mesangial matrix widening, mesangial deposits of IgA, and large electron dense deposits in the mesangium were observed. Concurrent administration of SCG and POLIO resulted in a significant decrease in the serum IgA level and mesangial IgA deposits. The later addition or abstinence of SCG after the 70th day did not influence the glomerular mesangial IgA deposition. But the serum IgA level was still decreased by the continuous treatment of SCG even after the 70th day. Thus, mesangial IgA nephropathy simulating IgA nephropathy in humans could be induced in ddY mice using POLIO and its induction could largely be prevented by the concurrent use of SCG. However mesangial IgA deposits already present could not be cleared by the late administration of SCG.

Animals↗

Timing, localization, and persistence of colonization by segmented filamentous bacteria in the neonatal mouse gut depend on immune status of mothers and pups.

As a member of the indigenous gut mucosal microbiota, segmented filamentous bacteria (SFB) colonize the guts of a variety of vertebrates and invertebrates. They are potent microbial stimuli of the gut mucosal immune system. In the small intestines of mice and rats, it has been observed that SFB are absent during the suckling period and appear in high numbers shortly after weaning, then quickly retreat to the cecum and large intestine. In this study, we explored whether this microecological phenomenon resulted from the interaction between SFB and the passively acquired maternal mucosal immunity and/or the actively acquired mucosal immunity. We set up a mouse model by reciprocal crossings and backcrossings of SFB-monoassociated, formerly germ-free, immunocompetent (+/+) BALB/c mice and immunodeficient (scid/scid) mice to produce pups which are either immunocompetent (scid/+) or immunodeficient (scid/scid) and are born either to immunocompetent (scid/+) mothers or to immunodeficient (scid/scid) mothers. We monitored the number of SFB on the mucosa of the small intestine in the four different groups of mice after birth, as well as the level of passively acquired antibodies, the active gut mucosal immune responses, and immunoglobulin A (IgA) coating of SFB in the gut. The results showed that, irrespective of whether the pups were scid/scid or scid/+, SFB could be found earlier on the mucosa of the small intestine in pups born to scid/scid mothers, appearing from day 13 and rapidly reaching a climax around weaning time on day 28, compared to the significantly delayed colonization in the pups of scid/+ mothers, starting from day 16 and peaking around days 28 to 32. After the climax, SFB quickly declined to very low levels in the small intestines of scid/+ pups of either scid/scid mothers or scid/+ mothers, whereas they remained at high levels in scid/scid pups at least until day 70, the last observation time in this study. The dynamic changes in SFB colonization of the small intestines of the different groups of pups may be related to the dynamic changes in the levels of SFB coated with secretory IgA (sIgA), which resulted from the significantly different levels of sIgA obtained from the mothers' milk during the suckling period and, later, of self-produced sIgA in the small intestine. Nevertheless, it is evident that the timing, localization, and persistence of colonization of the neonatal gut by SFB depends on the immune status of both mothers and pups.

Animals↗

A comparison of natural and recombinant cholera toxin B subunit as stimulatory factors in intranasal immunization.

Cholera toxin B (CTB) is often envisaged and used as an immune stimulating agent in protocols for mucosal immunization. However, the nature of the CTB used (natural vs recombinant) is frequently not taken in consideration. This is important since the usage of natural CTB in mucosal immunization regimen and the mucosal response resulting from such an immunization can be effected by the presence of the CTA subunit in commercial CTB preparations. To clarify this, we have compared natural vs recombinant CTB in an intranasal (i.n.) mucosal immunization procedure using ovalbumin (OVA) as antigen. The results show that recombinant CTB induces similar immune responses like natural CTB. Furthermore, our experiments show that covalent coupling of OVA to CTB is not required for the induction of OVA specific mucosal and systemic immune responses upon i.n. immunization.

Adjuvants, Immunologic↗

Bilayer films for mucosal (genetic) immunization via the buccal route in rabbits.

PURPOSE: The oral buccal mucosa may be an ideal site for mucosal immunization, allowing for the needle-free administration of cost-effective vaccines. A novel mucoadhesive bilayer film was developed to test the feasibility of this route of immunization in rabbits. METHODS: Bilayer films were developed using different ratios of Noveon and Eudragit S-100 as the mucoadhesive layer and a pharmaceutical wax as the impermeable backing layer. Optimal 3/8-inch films were post-loaded with 100 microg of plasmid DNA (CMV-beta-gal) or beta-galactosidase protein. The in vitro release rates and stability of the postloaded antigens were determined. The films were applied to the buccal pouch of rabbits on days 0. 7, and 14, and the humoral and splenocyte proliferative immune responses to beta-gal were determined through day 28 and compared to those responses after conventional subcutaneous injection of adjuvanted protein. RESULTS: The weight ratio of Noveon and Eudragit S-100 had a significant effect on adhesion time of bilayer films. Postloaded plasmid DNA and beta-gal remained stable after being released from bilayer films (release of -60-80% in 2 h for both). Buccal immunization using novel bilayer films (109 +/- 6-microm thickness) containing plasmid DNA led to comparable antigen-specific IgG titer to that of subcutaneous protein injection. All rabbits immunized with plasmid DNA via the buccal route but none by the subcutaneous route with protein antigen demonstrated splenocyte proliferative immune responses. CONCLUSION: The feasibility of buccal (genetic) immunization with these novel bilayer films was demonstrated.

Adhesives↗