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Development of mucosal immune function in man: potential for GI disease states.

The human mucosal immune system is structurally mature and has all the necessary cellular components to generate an immune response at birth. However, in the absence of dietary antigens and bacterial flora, there are no secondary follicles in the Peyer's patches and virtually no immunoglobulin A plasma cells in the lamina propria. Reactive follicle centers develop after birth but it takes 2 years for mucosal IgA plasma cell density to reach adult levels. T cells are present in the epithelium and lamina propria at birth, albeit at a lower frequency than later in life and there are major differences in phenotype between T cells in fetal intestine and postnatal intestine. There is no information on the impact of the massive antigenic challenge at birth on the mucosal immune system. Well-documented deficiencies in the ability of the blood T cells of the neonate to produce interleukin-4 and interferon-gamma may also occur in the intestine. It is still an open question whether it is better to try to prevent immunological sensitization of the newborn by avoiding potential allergens (i.e. cow's milk), or whether early exposure (as happens when premature infants are given formula feeds) might tolerize the infant. Hydrolysed cow's milk formulae are probably less antigenic than whole cow's milk and have been widely used in the treatment of cow's milk allergy. Some thought is now being given as to whether the prophylactic use of hydrolysates can reduce cow's milk allergy in 'at-risk' infants.

Gastric Mucosa↗

Mucosal immunity of the adolescent female genital tract.

This study sought to characterize mucosal immunity of the adolescent genital tract during the cycle and determine if adolescents have more suppressed immunoglobulin levels in the follicular phase than adults. Daily from cycle day 9 until ovulation, then every other day until menses, cervical secretions for IgA, IgG, and cytokines were collected via Weck-Cel sponge and serum for luteinizing hormone (LH), estradiol, and progesterone was obtained from three adolescent girls (mean age 16.8 years). Immunoglobulin and cytokine levels varied during the menstrual cycle, reaching their nadir around ovulation. Compared with 13 adults, adolescents had a greater drop in IgG in the follicular phase (mean beta-953 vs. -269 microg/mL/day, p = .045), but a similar rate of rise in IgG in the luteal phase (mean beta +118 vs. +100 microg/mL/day, p = .252). Rates of change in IgA did not differ between adolescents and adults for either phase. Although limited by the small sample size, these findings suggest that adolescents may be more sensitive to unopposed estrogen and warrant further investigation.

Adolescent↗

Mucosal immune response to Human Papilloma Virus (HPV) infection in HIV positive women.

Mucosal immunity plays a central role in the control of genital human papilloma virus (HPV) infection. HIV infection is associated with higher risks of HPV-induced neoplasias. Prevalence, incidence and evolution of genital HPV lesions are correlated with the level of immunodepression. Several changes of the local immune response in the genital tract of HIV infected women have been demonstrated. The influence of HIV on the production of some immunoregulatory cytokines appears of particular interest. The shift from the helper T cells type 1 (Th1) to the helper T cells type 2 (Th2) immune response, which determines the downregulation of the cell-mediated immunity, may explain the frequent loss of immunologic control of HPV and its oncological complications

Down-Regulation↗

Mannosylated niosomes as adjuvant-carrier system for oral mucosal immunization.

The aim of the present study was to develop mannosylated niosomes as oral vaccine delivery carrier and adjuvant for the induction of humoral, cellular, and mucosal immunity. Tetanus toxoid (TT) loaded niosomes composed of sorbiton monostearate (Span 60), cholesterol, and stearylamine were prepared by the reverse-phase evaporation method. They were coated with a modified polysaccharide o-palmitoyl mannan (OPM) to protect them from bile salts caused dissolution and enzymatic degradation in the gastrointestinal tract and to enhance their affinity toward the antigen presenting cells of Peyer's patches. Prepared niosomes were characterized in vitro for their size, shape, entrapment efficiency, ligand binding specificity, and stability in simulated gastric fluid and simulated intestinal fluid. OPM-coated niosomes were found to more stable in simulated gastrointestinal conditions. The immune stimulating activity was studied by measuring serum IgG titer, IgG2a/IgG1 ratio in serum, and sIgA levels in intestinal and salivary secretions following oral administration of niosomal formulations in albino rats. The results were compared with alum-adsorbed TT following oral and intramuscular administration, and it was observed that OPM-coated niosomes produced better IgG levels as compared to plain uncoated niosomes and alum-adsorbed TT upon oral administration. Oral niosomes also elicited a significant mucosal immune response (sIgA levels in mucosal secretions). The developed formulations also elicited a combined serum IgG2a/IgG1 response, suggesting that they were capable of eliciting both humoral and cellular response. The study signifies the potential of OPM-coated niosomes as an oral vaccine delivery carrier and adjuvant. The proposed system is simple, stable, and cost-effective and may be clinically acceptable.

Adjuvants, Immunologic↗

Assessment of mucosal immune response in genitourinary tract using urine.

A novel method to assess mucosal immune response in the genitourinary mucosa after immunization with a mucosal vaccine has been developed. In this method, secretory IgA antibody is measured by a highly sensitive enzyme immunoassay (immune-complex transfer enzyme immunoassay) using urine as a specimen. The urinary IgA antibody response could be detected by the immune-complex transfer enzyme immunoassay. In contrast, a conventional enzyme immunoassay (enzyme-linked immunosorbent assay (ELISA)) could not detect this response because of its low sensitivity. Because urine samples can be collected easily and nontraumatically, not only from experimental animals but also from humans, both males and females, the present method may be applicable for assessing the protective efficacy of candidates for mucosal vaccines against sexually transmitted microorganisms, such as human immunodeficiency virus. Furthermore, the usefulness of this method for novel mucosal vaccine formulae was shown for a model in which vaccine antigen and Bordetella pertussis adjuvant were adsorbed onto CaCO, and enclosed in enteric coated capsules.

Adjuvants, Immunologic↗

Gastric invasion by Trypanosoma cruzi and induction of protective mucosal immune responses.

Trypanosoma cruzi is an intracellular parasite transmitted from a reduviid insect vector to humans by exposure of mucosal surfaces to infected insect excreta. We have used an oral challenge murine model that mimics vector-borne transmission to study T. cruzi mucosal infection. Although gastric secretions have microbicidal activity against most infectious pathogens, we demonstrate that T. cruzi can invade and replicate in the gastric mucosal epithelium. In addition, gastric mucosal invasion appears to be the unique portal of entry for systemic T. cruzi infection after oral challenge. The mucosal immune responses stimulated by T. cruzi gastric infection are protective against a secondary mucosal parasite challenge. This protective mucosal immunity is associated with increased numbers of lymphocytes that secrete parasite-specific immunoglobulin A. Our results document the first example of systemic microbial invasion through gastric mucosa and suggest the feasibility of a mucosal vaccine designed to prevent infection with this important human pathogen.

Animals↗

The neuropeptide bombesin improves IgA-mediated mucosal immunity with preservation of gut interleukin-4 in total parenteral nutrition-fed mice.

BACKGROUND: The Th2 cytokines, interleukin-4 (IL-4) and interleukin-10 (IL-10), stimulate IgA production. Total parenteral nutrition (TPN) reduces IL-4 and IL-10 messenger RNA in gut lamina propria lymphocytes, total IL-4 and IL-10 in gut homogenates, and IgA-mediated mucosal immunity. Bombesin (BBS) can maintain mucosal immunity in TPN-fed mice, but the effects of BBS on gut IL-4 and IL-10 levels and their mRNA expression in the lamina propria are unknown. METHODS: In experiment 1, mice that were fed chow, TPN, or TPN + BBS (15 microg/kg intravenously-three times a day) for 5 days were killed, and respiratory tract IgA and intestinal IgA, IL-4, and IL-10 levels were measured. In experiment 2, IL-4 and IL-10 mRNA were measured in isolated lamina propria lymphocytes from chow-, TPN-, and TPN+BBS-fed mice by reverse transcriptase-polymerase chain reaction. Intestines were harvested 1 hour after injection of 100 7 microg of lipopolysaccharide intraperitoneally. Samples were standardized to beta-actin. RESULTS: TPN-fed mice had significantly lower respiratory tract IgA levels than chow- or TPN + BBS-fed mice. TPN+BBS did not increase intestinal IL-10 or IL-10 lamina propria mRNA levels but maintained intestinal IL-4 levels and lamina propria IL-4 mRNA expression equal to those of chow-fed mice. CONCLUSIONS: BBS reverses the effects of TPN on intestinal and respiratory tract IgA levels and most effects on gut cytokines. Lamina propria cytokine mRNA levels reflect total gut cytokine concentration.

Animals↗

Mucosal immunization with DNA vaccines.

DNA vaccines can induce potent humoral and cellular immune responses in numerous animal models. Most DNA vaccines have been administered parenterally; however, more effective protection against mucosal pathogens could be achieved with mucosal immunization. This review concentrates on the use of DNA vaccines for the induction of mucosal immunity.

Animals↗

The ocular surface as part of the mucosal immune system: conjunctival mucosa-specific lymphocytes in ocular surface pathology.

The mucosal immune system includes mucus membranes of the gut, respiratory and urogenital tracts. Mucosa-specific, intraepithelial lymphocytes (IELs), that correspond to the suppressor/cytotoxic subset and also express the human mucosal lymphocyte antigen (HML-1), are a unique component of this system. We have recently demonstrated these cells in the human conjunctiva, establishing the ocular surface as an integral part of the mucosal immune system. In this study we examined the distribution of lymphocyte subsets, with particular attention to mucosa-specific lymphocytes, in two ocular surface disorders, namely conjunctival intraepithelial neoplasia (CIN) and ocular cicatricial pemphigoid (OCP). Cryosections of biopsy specimen were immunostained using a panel of monoclonal antibodies against different lymphocyte subsets. In CIN, the CD8/HML ratio was decreased (1 +/- 0) and CD8/CD4 ratio was reversed (0.54 +/- 0.21). HML-1+ cells were distributed throughout the epithelial layers of dysplastic tissue. Biopsy specimens of OCP showed normal ratios of CD8/HML (1.4 +/- 0.16) but the CD8/CD4 was low (1.29 +/- 0.88). Association of HML-1+ cells with the basal layer of normal epithelium and with all layers of dysplastic epithelium suggests that expression of HML-1 antigen may be induced by actively dividing cells. HML-1+ cells may have a role in immune mechanisms associated with ocular surface disorders.

Adult↗

Genetically engineered Bifidobacterium animalis expressing the Salmonella flagellin gene for the mucosal immunization in a mouse model.

BACKGROUND: A critical component of the host defense against enteric infections is the immunological response of the mucosal membrane, a major starting point of infectious disease, such as typhoid fever. The mucosal immune system consists of an integrated network of lymphoid tissues, mucous membrane-associated cells, and effector molecules. In the present study, we developed a recombinant Bifidobacterium animalis (B. animalis) genetically modified with the Salmonella flagellin gene for mucosal immunization as an oral typhoid vaccine. METHODS: We constructed an oral vaccine against Salmonella typhimurium, consisting of recombinant B. animalis containing the flagellin gene of Salmonella. The recombinant B. animalis was administered orally to mice every other day for 6 weeks. Anti-flagellin antibodies in the serum and stools were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: We detected significantly higher levels of flagellin-specific IgA in the serum and stools of the mice treated with the recombinant B. animalis containing the flagellin gene than was seen in those treated with parental B. animalis. CONCLUSIONS: Our findings suggest that an oral vaccination using recombinant B. animalis genetically modified with the flagellin gene of Salmonella may be effective against Salmonella infections.

Administration, Oral↗

Impact of aging on gastrointestinal mucosal immunity.

There is considerable evidence that the mucosal or secretory immune response in the gastrointestinal tract is compromised by aging. The generation of a mucosal immune response is an extremely complex process that involves antigenic stimulation of a specific subpopulation of immunologically competent cells in the Peyer's patches, differentiation and migration of these cells to the small intestinal lamina propria, initiation and regulation of local antibody production in the intestinal wall, and mucosal epithelial cell receptor-mediated transport of antibodies to the intestinal lumen. Available data suggest that gastrointestinal mucosal immunosenescence reflects deficits in: (1) the differentiation and/or migration (homing) of immunoglobulin A immunoblasts to the intestinal lamina propria, and (2) the initiation and/or regulation of local antibody production. The significant age-related increases in the incidence and severity of gastrointestinal infectious diseases, coupled with the potential for immunopharmacologic manipulation of the mucosal immune compartment, substantiate the merit of studies designed to resolve the etiology of mucosal immunodeficiency in the elderly.

Aging↗

Mucosal immunization with DNA-liposome complexes.

The mucosal surfaces represent the primary site for transmission of several viruses including HIV. To prevent mucosal transmission and dissemination to the regional lymph nodes, an effective HIV vaccine may need to stimulate immune responses at the genital and rectal mucosa. Optimal induction of mucosal immunity in general requires targeting antigens to the specialized antigen presenting cells of mucosal associated lymphoid tissues. The nasal mucosa may provide a simple, non-invasive route to deliver DNA encoding the introduced gene to stimulate mucosal immunity. As a first step to evaluate the feasibility of this approach, we have investigated as a model system, systemic and mucosal immune responses elicited to firefly luciferase generated by DNA immunization. Incorporating DNA into liposomes with cationic lipids enhanced luciferase expression in nasal tissue, and was associated with induction of a humoral response in serum and vaginal fluids and also a proliferative and cytotoxic T lymphocyte response in the spleen and iliac lymph nodes draining the genital and rectal mucosa.

AIDS Vaccines↗

Immunophysiology of the gut: a research frontier for integrative studies of the common mucosal immune system.

This review highlights work that, within the past decade, transformed mucosal immunophysiology from a hypothetical concept to a fully recognized interdiscipline. The regulation of epithelial and smooth muscle functions by the mucosal immune system represents an exquisitely sensitive adaptation to local antigenic challenge. Furthermore, immunologic cells communicate with nerves via paracrine secretions to rapidly transduce antigenic signals into panmucosal changes in function. These local immunocyte-nerve interactions are modulated by the autonomic and central nervous systems. Because of the common mucosal immune system, antigen-induced changes similar to those occurring in the intestine and colon are predicted to occur in mucosa of all hollow organs. The drawing together of fields as diverse as medicine and agriculture underscores the scope of areas encompassed by immunophysiology. Newly acquired knowledge has positioned the field to advance rapidly in both basic and applied directions. Forces that will remodel the field in the next decade will be derived from public concerns about human health maintenance and the explosive and novel use of new research tools stemming from molecular biology. These forces will draw on and advance our knowledge in areas as diverse as vaccine development and prevention of allergic reactions to foods, bioengineered foods in particular.

Animals↗

Functional attributes of mucosal immunity in cervical intraepithelial neoplasia and effects of HIV infection.

The role of mucosal immunity in human papillomavirus (HPV)-related cervical diseases is poorly understood. To characterize the local immune microenvironment in cervical intraepithelial neoplasia (CIN) 2/3 and determine the effects of HIV infection, we compared samples from three groups: normal cervix, CIN 2/3 from immunocompetent women (HIV- CIN 2/3), and CIN 2/3 from HIV seropositive women (HIV+ CIN 2/3). CIN 2/3 lesions contained increased numbers of immune cells from both the acquired and innate arms of the immune response in stroma [CD4+ and CD8+ T cells, macrophages, mast cells, B cells, neutrophils, and natural killer (NK) cells] and dysplastic epithelium (CD4+ T cells, macrophages, and NK cells). Immune cells in CIN 2/3 expressed activation markers, as measured by interleukin-2 receptor (IL-2R) and transcription factor T bet. Interferon-gamma production was significantly up-regulated in CIN lesions and was expressed by CD4+ and CD8+ T cells and NK cells, indicating the activation of immune cells. Abundant presence of transforming growth factor-beta+ CD25+ cells in the infiltrates associated with CIN lesions, and of immature CD1a+ dendritic cells expressing IL-10 and transforming growth factor-beta, indicate that CIN is associated with an influx of immune cells that produce a mixture of proinflammatory and regulatory cytokines. In HIV+ CIN, immune cell densities (CD4+ T cells, macrophages, neutrophils, and NK cells) and expression of interferon-gamma were significantly decreased compared with HIV- CIN. Regulatory cytokines were also down-regulated in this group. Therefore, both pro- and anti-inflammatory responses present in CIN 2/3 lesions are suppressed in HIV-seropositive women.

Adult↗

Induction of systemic and mucosal immune responses by intranasal administration of alginate microspheres encapsulated with tetanus toxoid and CpG-ODN.

In the induction of systemic and mucosal immunity, particulate antigens are more effective than soluble antigens; possibly because they are more efficiently endocytosed by mucosal-associated lymphoid tissue (MALT) M cells. In this study, we determined the systemic and mucosal immune responses in rabbits following intranasal immunization with encapsulated tetanus toxoid (TT) and CpG-ODN in alginate microspheres. The microspheres were less than 4 microm in diameter. Encapsulation efficiency of TT and CpG-ODN was determined as 47.7+/-6.6 and 34.2+/-7.4, respectively. Release of TT and CpG-ODN in a simulated model with nasal cavity was 14.2+/-3.06 and 36.7+/-2.4% after 4 h. Encapsulated TT preserved its intact structure, but its immunoreactivity was decreased to about 91+/-5%. The highest serum IgG and antitoxin, and nasal lavage IgA titers were observed in groups immunized with microsphere formulations. CpG-ODN as an adjuvant could increase the serum IgG and antitoxin titers when co-administered with TT solution, but its co-encapsulation with TT in alginate microspheres failed to potentiate the systemic immune response while induced high IgA titers in nasal lavages. No hemolysis was occurred on incubation of alginate microspheres and human RBCs. Also after nasal administration of plain microspheres to human volunteers, no local irritation was observed. Intranasal administration of microspheres encapsulated with vaccines showed to be an effective way for inducing a variety of immune responses and that a strong systemic IgG and mucosal IgA responses can be induced in rabbits with intranasal administration of alginate microspheres encapsulated with TT.

Adjuvants, Immunologic↗

Cytokines as adjuvants for the induction of mucosal immunity.

Safe nasal vaccines capable of promoting both mucosal and systemic immunity are needed for effective protection against bacterial and viral pathogens. While parenteral cytokine treatment could lead to unwanted toxicity, the nasal delivery route results in low but biologically active serum cytokine levels. Interleukin (IL)-6, IL-1 and IL-12, which promote either Th2- or Th1-type responses, respectively, also enhance systemic immunity to co-administered antigens. The chemoattractants lymphotactin (Lptn), RANTES and defensins also exerted adjuvant activity for systemic immunity when nasally administered with antigens. However, each cytokine or innate factor promoted a distinct pattern of T helper cell responses and corresponding IgG subclass response. Interleukin-12, IL-1, and the chemokines Lptn and RANTES promote mucosal immunity. In contrast, nasal IL-6 and defensins failed to induce mucosal S-IgA Ab responses, suggesting that mechanisms more complex than T cell activation and chemotaxis are required for the development of mucosal immunity after nasal delivery of cytokines.

Adjuvants, Immunologic↗

Cellular dissemination of priming for a mucosal immune response to cholera toxin in rats.

Using CT as the test antigen, we sought 1) to learn whether primary immunization at 1 mucosal site caused priming of distant nonstimulated mucosae, 2) to study the role of migrating memory cells in the dissemination of mucosal priming, and 3) to compare disseminated priming with priming that occurs at the site of initial immunization. CT given i.c. or i.d. caused priming in tracheal and nonexposed enteric mucosae; i.t. immunization, however, did not cause detectable enteric priming. Adoptive transfer of immune TDLs showed that priming was conveyed by migrating memory cells. These appeared to be of 2 types: those that recirculated briefly before settling in MALT, and those that continued to recirculate until recruited by antigen to the site of mucosal challenge. Both types were required for secondary responses at mucosae distant from the site of priming. The time-course of disseminated mucosal priming resembled that of priming at the site of initial CT exposure, both lasting at least 16 wk. Disseminated priming persisted better in jejunal than tracheal mucosa, suggesting that the subgroup of memory cells that did not continue to recirculate settled preferentially in jejunal MALT. Disseminated priming supported smaller challenge responses than priming at the site of initial CT exposure did, suggesting that sessile memory cells also contributed to the latter process. These observations extend the concept of a "common mucosal immune system" to include cellular dissemination of mucosal priming, but also show quantitative differences between local and disseminated priming that probably reflect the patterns of distribution of migrating and sessile memory cells.

Animals↗

Mucosal immune system: implications in otitis media with effusion.

The ontogeny of the mucosal immune system as it relates to the development of lymphoid tissue in the respiratory tract and the gastrointestinal tract has been studied quite extensively over the past few years. It is apparent now that the bronchus-associated lymphoid tissue and gut-associated lymphoid tissue are the major sources of immunocompetent precursor B lymphocytes. After the induction of antigens in the respiratory tract or the gastrointestinal tract, precursor lymphoid cells in these sites are preferentially activated to undergo significant proliferation. Such antigen-sensitized cells eventually migrate to other mucosa sites, such as mammary glands, genital tract, conjuctiva, etc. Recent evidence has suggested that the immunocompetent tissue observed in the middle ear cleft during otitis media with effusion may function as an extension of the mucosal immune system in the upper respiratory tract. The implications of these observations relative to middle ear disease are discussed.

Antibody Formation↗