Influences of Short‐Term Fasting and Carbohydrate Supplementation on Gut Immunity and Mucosal Morphology in Mice

Background: Preoperative carbohydrate (CHO) supplementation has been recommended in enhanced recovery after surgery protocols. However, the effects of CHO supplementation on gut and systemic immunity are not well understood. Methods: Mice (n = 60) were randomized to 1 of the following 5 groups: control (ad lib feeding), 12‐hour fasting without CHO administration (fasting), and 12 hours of fasting with CHO administration at 2, 4, and 8 hours before sacrifice. Then, lymphocytes were isolated from gut‐associated lymphoid tissue, that is, Peyer's patches, the intraepithelial space, and the lamina propria of the small intestine. These lymphocyte numbers and phenotypes were evaluated. IgA levels in respiratory and small‐intestinal washings were determined by ELISA. Morphology, proliferation, and apoptosis of the intestinal epithelium were also evaluated histologically. Results: Although there were no significant differences in IgA levels among the 5 groups, fasting decreased intraepithelial and lamina propria, but not Peyer's patches lymphocyte numbers. CHO at 2 hours prevented lymphocyte loss in intraepithelial, whereas CHO at 4 hours reversed lamina propria lymphocytes numbers. Percentages of lymphocyte phenotypes were similar in each site among the 5 groups. Fasting caused villous atrophy; however, CHO at 2 hours restored villous structure along with maintenance of epithelial cell proliferation rate. Conclusions: Only 12 hours of fasting causes marked gut‐associated lymphoid tissue cell loss along with gut atrophy. However, CHO at 2 hours preserves gut immunity and morphology not completely but moderately.     

Systemic-oral immunization with 56 kDa molecule of Giardia lamblia affords protection in experimental mice.

Prior systemic-oral immunization of inbred mice with Giardia lamblia surface-associated antigen of molecular mass 56 kDa not only significantly blocked colonization but also resulted in elimination of G. lamblia trophozoites by 9-11 days following challenge. The colonization and multiplication of the trophozoites in unprotected animals were accompanied by a pronounced influx of suppressor T cells in intraepithelial or lamina propria of the small intestine and a significant decline in IgA-bearing plasma cells in the lamina propria. An induction of helper/inducer T cells in the intraepithelial and lamina propria and significant enhancement of IgA and IgG-bearing cells in the lamina propria of the small gut resulted in a decline, and eventual elimination, of the trophozoites from the gut. The completion of the immunization of animals with 56 kDa G. lamblia antigen resulted in: significant enhancement of helper/inducer T lymphocytes with no effect on suppressor T cells in the intraepithelial and lamina propria of the small gut; significant enhancement of IgA- and IgG-bearing plasma cells in lamina propria; and significant elevation of antibodies to 56 kDa G. lamblia antigen in the systemic circulation. The stimulation of such effector mechanisms in 56 kDa-immunized animals appears to result in failure of the trophozoites to get established, prevention of multiplication and earlier elimination from the gut. The data suggest that the 56 kDa molecule of G. lamblia immunoregulates the giardial infection.     

Rectal immunization of mice with hepatitis A vaccine induces stronger systemic and local immune responses than parenteral immunization

Systemic (spleen cell (SPLC), serum antibodies) and intestinal mucosal (Peyer's patch cells (PPC), lamina propria lymphocytes (LPLs), coproantibodies) immune responses were compared in mice immunized with varying doses (144, 72, 36, 18 ELISA units [EU]) of HAVRIX, an alum-adsorbed killed hepatitis A virus (HAV) vaccine, delivered either intrarectally (i.r.) or intraperitoneally (i.p.) in three doses at weekly intervals. HAV-specific IgG, IgM, and IgA antibody responses were evaluated by ELISPOT and EIA and HAV-responsive lymphocytes by lymphocyte stimulation assays. Systemic IgG responses were greater in mice immunized intraperitoneally with 144, 72, and 36EU of HAVRIX, while IgM and IgA responses were greater in PPC and LPL cell populations, serum and coproantibodies of rectally immunized mice, particularly at HAVRIX doses of 36 and 18EU. Rectal immunization at lower doses (36, 18EU) also elicited strong cellular responses in all cell populations while parenteral (i.p.) vaccination, did not. Results suggest that rectal immunization may be a highly effective way of inducing both local and systemic immunity to HAV.     

Absolute counts and distribution of lymphocyte subsets in small intestine of BALB/c mice change during weaning

The gut immune system is an essential part of the barrier function of the gut. At weaning, major changes can be expected in the number and subset composition of lymphocytes in the small intestine since the gut is exposed to a wide variety of food and microbial antigens, especially when human milk is gradually replaced by weaning foods. The purpose of this study was to evaluate the changes in small intestine lymphocyte subsets in mice during weaning. BALB/c male mice at weaning (3 wk old) were fed a nonpurified diet for 18 d and were killed at different times (0, 4, 7, 12 and 18 d). Lymphocyte populations from lamina propria (LPL), Peyer's patches (PPL) and intestinal epithelium (IEL) were isolated. The expression of different antigens (CD3, CD4, CD8alpha, CD8beta, CD22 and CD45R) in those lymphocyte populations was analyzed by flow cytometry. The percentages of cells expressing T-cell antigens, such as CD3, were significantly higher in LPL during weaning compared to d 0. The percentages of cells expressing CD8alpha and CD8beta increased in both IEL and LPL. However, the percentage of CD4+ cells tended (P = 0.07) to decrease in IEL and to increase in LPL. The percentages of cells expressing B-cell antigens, such as CD22 or CD45R in PPL increased. Changes in the specific phenotypes of intestinal lymphocyte populations at weaning are apparently related to the maturation of the intestinal immune system during early life. Thus, B cells increase in PPL and T cell increase in IEL and LPL.     

An investigation into the effect of a probiotic on gut immune function in surgical patients

BACKGROUND: The gut-associated lymphoid tissue (GALT) is an important component of the gut barrier. We have previously demonstrated a significant increase in various parameters of gut immune function in association with bacterial translocation. Animal studies have suggested that the probiotic Lactobacillus plantarum 299v improves the immunological status of the intestinal mucosa. The aim of this study was to determine whether the same is true in humans. METHOD: This was a prospective randomised controlled study, in which immunohistochemical techniques were used to measure the concentrations of plasma cells, IgA positive cells and IgM positive cells in the lamina propria, together with the concentrations of IgA and IgM at the mucosal surface in specimens of normal small bowel obtained from patients undergoing elective abdominal surgery who had consumed an oral preparation containing the probiotic Lactobacillus plantarum 299v (ProViva) during the immediate preoperative period. These were compared with similar specimens obtained from control subjects who did not receive the probiotic. RESULTS: A total of 22 patients were studied (probiotic group n = 11, control group n = 11). The median volume of ProViva consumed was 3250 ml (range 2100-9000 ml), for a median duration of 9 days (range 5-18 days). There were no significant differences between the probiotic and control groups in terms of concentrations of plasma cells, IgA positive cells or IgM positive cells in the lamina propria. There was a significantly higher concentration of IgM at the mucosal surface in the control group (P = 0.02, Fishers Exact test mid P), but no difference in terms of IgA. CONCLUSIONS: The increase in IgA observed in the intestinal mucosa in response to probiotics in animal studies does not occur in humans. The significance of the increase in IgM at the mucosal surface in the controls is unclear.     

Development of hepatitis B oral vaccine using B-cell epitope loaded PLG microparticles

Oral hepatitis B vaccine formulation was prepared by successful encapsulation of immunogenic peptide representing residues 127-145 of the immunodominant B-cell epitope of hepatitis B surface antigen (HBsAg) in poly(D,L-lactide co-glycolide) (PLG) microparticles. The smooth, spherical PLG microparticles with a diameter of around 10 microm was prepared by using W/O/W double emulsion solvent evaporation method. The entrapment efficiency of B-cell epitope peptide (BCEP) into PLG microparticles was 64%. In vitro studies showed B-cell epitope loaded PLG microparticles (BCEM) released the peptide in sustained profile and reached 64.9% efficiency by Day 25. Single oral immunization of mice with BCEM led to the significant induction of specific serum IgG and IgM anti-HB antibodies. After the termination of antibody induction, the orally immunized mice were infected with HBsAg, which resulted in the rapid production of antibodies against HBsAg as a result of secondary immune response. PLG microparticles formulation approach may have potential in increasing the efficacy of microparticulate systems for the oral administration of hepatitis B vaccine.     

Bovine whey protein concentrate supplementation modulates maturation of immune system in suckling rats

During neonatal life, challenges from breast milk and microbial flora promote immune system maturation. Immunonutrition in these stages may become an important way to increase natural defence systems. The aim of this study was to determine the effect of a daily bovine milk whey protein concentrate (WPC) supplement on the intestinal and systemic immune systems in suckling rats. The composition of intraepithelial and lamina propria lymphocytes (IEL and LPL) was analysed by flow cytometry. Systemic and intestinal humoral immune responses were determined by sera Ig levels and Ig-secreting cell quantification by ELISA and ELISPOT, respectively. From birth, suckling Wistar rats were supplemented with WPC or standard infant formula (SIF). The WPC group showed the same proportion of most of the main mucosal cell subsets as the reference animals. However, in the first days of life WPC enhanced the innate immunity by increasing the NK cell proportion in both epithelial and lamina propria (LP) compartments. A rise in intestinal CD8alphaalpha+ IEL was also induced by WPC supplementation. A time-course of sera Ig levels and spontaneous IgA, IgM and IgG production by LPL and mononuclear cells from blood and spleen, in the WPC group, exhibited a similar pattern to those pups fed only by dam's milk. In summary, the present results show the effects of WPC on enhancing mucosal innate immunity during early life.     

Different kinetic of antibody responses following infection of newly weaned pigs with an F4 enterotoxigenic Escherichia coli strain or an F18 verotoxigenic Escherichia coli strain

To develop a vaccine against Escherichia coli-induced post-weaning diarrhea and edema disease, insights in the induction of the protective immune response following infection with these pathogenic E. coli is needed. Therefore, the fimbriae-specific antibody response of newly weaned pigs following infection with the Shiga-like toxin type II variant (SLT-IIv) producing F18(+) verotoxigenic E. coli (VTEC) (strain 107/86) was compared with the response following an infection with LT producing F4(+) enterotoxigenic E. coli (ETEC) (strain GIS 26). F4(+) ETEC were able to colonize the gut very soon after infection, as peak excretion of F4(+) E. coli bacteria was seen 2 days post-infection (dpi), but had already disappeared 7dpi. On the other hand, F18(+) VTEC infection resulted in a slower colonization of the gut as the peak excretion of F18(+) E. coli was observed between 3 and 5dpi, but this colonization remained longer as F18(+) E. coli were detected till 9dpi in feces. Furthermore, this fast colonization pattern of F4(+) ETEC is accompanied with the presence of F4-specific antibodies in mucosal tissues and serum from 4dpi onward, with maximal amounts of F4-specific IgA in the jejunal lamina propria and serum 7dpi. In contrast, F18-specific IgA was only readily detected in the jejunal lamina propria 15dpi and showed a maximum serum titer 21dpi. Besides this faster induction and higher antibody response, the switch from IgM to IgA and IgG was also earlier following the F4(+) ETEC infection.     

Consumption of rice bran increases mucosal immunoglobulin A concentrations and numbers of intestinal Lactobacillus spp

Gut-associated lymphoid tissue maintains mucosal homeostasis by combating pathogens and inducing a state of hyporesponsiveness to food antigens and commensal bacteria. Dietary modulation of the intestinal immune environment represents a novel approach for enhancing protective responses against pathogens and inflammatory diseases. Dietary rice bran consists of bioactive components with disease-fighting properties. Therefore, we conducted a study to determine the effects of whole dietary rice bran intake on mucosal immune responses and beneficial gut microbes. Mice were fed a 10% rice bran diet for 28 days. Serum and fecal samples were collected throughout the study to assess total immunoglobulin A (IgA) concentrations. Tissue samples were collected for cellular immune phenotype analysis, and concentrations of native gut Lactobacillus spp. were enumerated in the fecal samples. We found that dietary rice bran induced an increase in total IgA locally and systemically. In addition, B lymphocytes in the Peyer's patches of mice fed rice bran displayed increased surface IgA expression compared with lymphocytes from control mice. Antigen-presenting cells were also influenced by rice bran, with a significant increase in myeloid dendritic cells residing in the lamina propria and mesenteric lymph nodes. Increased colonization of native Lactobacillus was observed in rice bran-fed mice compared with control mice. These findings suggest that rice bran-induced microbial changes may contribute to enhanced mucosal IgA responses, and we conclude that increased rice bran consumption represents a promising dietary intervention to modulate mucosal immunity for protection against enteric infections and induction of beneficial gut bacteria.     

Breakdown of mucosal immunity in gut by 2,3,7,8-tetraclorodibenzo-p-dioxin (TCDD)

Objectives Mucosal immunity plays a pivotal role for body defense against infection and allergy. The aim of this study was to clarify the effects of 2,3,7,8-tetraclorodibenzo-p-dioxin (TCDD) on mucosal immunity in the gut. Methods Fecal IgA level and oral tolerance induction were examined in TCDD-treated mice. Flow cytometric and histological analyses were also performed. Results Single oral administration of low dose 2,3,7,8-TCDD resulted in a marked decrease in IgA secretion in the gut without any effects on the cellular components of gut-associated lymphoid tissues (GALT) including Peyer’s patches (PPs) and mesenteric lymph nodes (LNs). Decressed IgA secretion by TCDD was not observed in aryl hydrocarbon receptor (AhR)-deficient mice. Flow cytometric analysis revealed that IgA B cells in PPs and the mesenteric LNs remained unchanged in the TCDD-treated mice. An immunofluorescence study also demonstrated that a significant number of cytoplasmic IgA cells were present in the lamina propria of the gut in the TCDD-treated mice. Furthermore, oral tolerance induction by ovalbumin (OVA) was impaired in the TCDD-treated mice and OVA-specific T cell proliferation occurred in the peripheral lymphoid tissues including the spleen and LNs. Conclusions These results suggest that a relatively low dose of TCDD impairs mucosal immunity in the gut and induces systemic sensitization by oral antigens.     

5-Fluorouracil infusion reduces gut-associated lymphoid tissue cell number and mucosal immunoglobulin A levels

BACKGROUND: Anticancer drugs have been demonstrated to affect gut mucosal morphology and cause gastrointestinal symptoms. We hypothesized that even small doses of 5-fluorouracil (5-FU) would reduce gut-associated lymphoid tissue (GALT) mass and function. METHODS: Mice underwent IV cannulation and received continuous infusion of normal saline or 10 mg/kg of 5-FU for 5 days. GALT cell numbers, phenotypes, and mucosal immunoglobulin A (IgA) levels were measured. RESULTS: During the infusion, there were no significant differences in food intake or body weight change between the 2 groups. Cell yields from the intraepithelial space and lamina propria of the small intestine were lower in the 5-FU than the control group. The lamina propria CD4/CD8 ratio was reduced in the 5-FU compared with the control group. Intestinal and respiratory tract IgA levels were lower in the 5-FU than in the control group. CONCLUSIONS: A small dose of 5-FU reduces GALT cell number and mucosal IgA levels, regardless of food intake.     

Intestinal immune function is unaffected by parenteral nutrition in man.

Animal studies have demonstrated intestinal immunoglobulin production is decreased when luminal nutrition is withheld and nutrition is provided solely on the basis of total parenteral nutrition (TPN).

Eight normal volunteers were hospitalized in the Clinical Research Center for three weeks. The subjects received TPN as an exclusive means of nutritional support for 14 days followed by 5 days of enteral feeding with either standard or a glutamine- and arginine-supplemented formula in which the protein source was primarily free amino acids and peptides. Endoscopic jejunal biopsies obtained before and after TPN and following enteral refeeding were evaluated by immunofluorescence for the number of IgA, IgM and IgG-producing cells; T and B cells as well as intraepithelial and lamina propria lymphocytes were also counted. Serum immunoglobulins and the molecular forms of serum IgA were determined at the same intervals.

The number of intestinal IgA-, IgM- and IgG-producing cells was unaffected by TPN (676 +/? 58 vs. 643 +/? 38, 101 +/? 14 vs. 98 +/? 18, 10 +/? 1 vs. 11 +/? 2 per low power field). The total number of intestinal lymphocytes, and CD3+ lymphocytes in the intraepithelial area was unaffected by TPN (10.4 +/? 0.4 vs. 10.2 +/? 1.3, 7.3 +/? 0.8 vs. 8.6 +/? 1.6 per 100 epithelial cells). Similarly, the total number of lymphocytes and CD3+ lymphocytes in the intestinal lamina propria was unaffected by TPN (4.4 +/? 0.2 vs. 6.2 +/? 0.8, 3.3 +/? 0.7 vs. 4.5 +/? 0.8). A small, but statistically significant increase in serum IgA and IgM was seen with TPN 314 +/? 11 vs. 342 +/? 16 mg/dL and 154 +/? 25 vs. 226 +/? 47 mg/dL, although IgG remained unchanged (1262 +/? 69 vs. 1207 +/? 57 mg/dL). The proportion of polymeric and monomeric serum IgA remained unchanged after TPN (19.2 vs. 22.1% polymeric).

The use of TPN is not associated with intestinal immune dysfunction in man. A small, but statistically significant increase in serum IgM, and a borderline statistically significant increase in serum IgM were associated with TPN. The etiology and clinical significance of these observations is unclear.     

Oral vaccination with liposome-encapsulated recombinant fusion peptide of urease B epitope and cholera toxin B subunit affords prophylactic and therapeutic effects against H. pylori infection in BALB/c mice

A new fusion peptide CtUBE of cholera toxin B subunit and Helicobacter pylori urease B subunit epitope was expressed in Escherichia coli. With this fusion peptide, an oral liposome vaccine against H. pylori infection was prepared and evaluated in BALB/c mice. Based on the results of urease tests, quantitation of culturable bacteria colonies in mice stomachs and histological identification of gastritis, the mice were protected significantly after intragastric vaccination with this CtUBE liposome vaccine, which increased the content levels of specific anti-urease serum IgG and mucosal IgA for both prophylactic and therapeutic vaccination protocols. These results showed that the fusion peptide CtUBE retained immunogenicity and could be used as antigen in the development of an oral vaccine against H. pylori infection.     

Mucosal immunization with purified flagellin from Salmonella induces systemic and mucosal immune responses in C3H/HeJ mice

This study investigated the immune response elicited in C3H/HeJ mice after oral, parenteral and nasal immunization with purified flagellin from Salmonella enterica serovar Enteritidis alone or conjugated to starch microparticles as adjuvant or together with the uptake-enhancer recombinant cholera toxin B-subunit (rCTB). Systemic (IgM-IgG, IgA, IgG2a, IgG2b, IgG1) and local (s-IgA) humoral immune responses in the mice were analyzed using enzyme-linked immunosorbent assays (ELISA). Primed splenocytes were also stimulated in vitro with flagellin and the supernatants analyzed for cytokine production. Finally, immunized mice were challenged orally with live Salmonella. A high flagellin-specific IgM-IgG response was seen in all groups, especially in mice immunized nasally with flagellin plus rCTB or subcutaneously, but a strong systemic antibody response was also induced when free antigen was given orally. Intranasal or subcutaneous immunization of mice with flagellin plus rCTB or oral immunization with flagellin plus microparticles resulted in a significantly greater mucosal response (higher s-IgA titers in feces) than seen in the control group (P <0.05). The mucosal IgA responses were significantly correlated with the serum IgA titers. The subclass profile in serum revealed a mixed Th1/Th2-type response, with a predominance of Th1-type, as indicated by the subclass ratio (IgG1/IgG2a + IgG2b). The splenocytes stimulated in vitro produced interferon (IFN)-gamma, at levels, which increased with time. The group immunized with flagellin plus rCTB subcutaneously had a relatively higher IFN-gamma response than the other groups. Interleukin (IL)-2 was also produced, especially in mice immunized nasally or subcutaneously with flagellin conjugated to microparticles. However, neither IL-4 nor IL-5 was produced in any of the groups. After oral challenge with live serovar Enteritidis, the groups immunized orally or nasally with free flagellin had significantly lower degree of infection than the control group (P <0.05).     

Interleukin-7 dose-dependently restores parenteral nutrition-induced gut-associated lymphoid tissue cell loss but does not improve intestinal immunoglobulin a levels

BACKGROUND: Without enteral nutrition, the mass and function of gut-associated lymphoid tissue (GALT), a center of systemic mucosal immunity, are reduced. Therefore, new therapeutic methods, designed to preserve mucosal immunity during parenteral nutrition (PN), are needed. Our recent study revealed that exogenous interleukin-7 (IL-7; 1 microg/kg twice a day) restores the GALT cell mass lost during intravenous (IV) PN but does not improve secretory immunoglobulin A (IgA) levels. Herein, we studied the IL-7 dose response to determine the optimal IL-7 dose for recovery of GALT mass and function during IV PN. We hypothesized that a high dose of IL-7 would increase intestinal IgA levels, as well as GALT cell numbers. METHODS: Male mice (n = 42) were randomized to chow, IL-7-0, IL-7-0.1, IL-7-0.33, IL-7-1 and IL-7-3.3 groups and underwent jugular vein catheter insertion. The IL-7 groups were fed a standard PN solution and received IV injections of normal saline (IL-7-0), 0.1, 0.33, 1, or 3.3 microg/kg of IL-7 twice a day. The chow group was fed chow ad libitum. After 5 days of treatment, the entire small intestine was harvested and lymphocytes were isolated from Peyer's patches (PPs), intraepithelial (IE) spaces, and the lamina propria (LP). The lymphocytes were counted and phenotypes determined by flow cytometry (alphabetaTCR, gammadeltaTCR, CD4, CD8, B cell). IgA levels of small intestinal washings were also examined using ELISA (enzyme-linked immunoabsorbent assay). RESULTS: IL-7 dose-dependently increased total lymphocyte numbers in PPs and the LP. The number of lymphocytes harvested from IE spaces reached a plateau at 1 microg/kg of IL-7. There were no significant differences in any phenotype percentages at any GALT sites among the groups. IgA levels of intestinal washings were significantly higher in the chow group than in any of the IL-7 groups, with similar levels in all IL-7 groups. CONCLUSIONS: Exogenous IL-7 dose-dependently reverses PN-induced GALT cell loss, with no major changes in small intestinal IgA levels. IL-7 treatment during PN appears to have beneficial effects on gut immunity, but other therapeutic methods are needed to restore secretory IgA levels.     

The jejunal Peyer's patches are the major inductive sites of the F4-specific immune response following intestinal immunisation of pigs with F4 (K88) fimbriae

A recently developed oral immunisation model in pigs in which F4 (K88) fimbriae of enterotoxigenic Escherichia coli are administered to induce a protective intestinal immunity, was used to determine the optimal inductive sites of the F4-specific intestinal immune response. Hereto, pigs were immunised with F4 orally, in the lumen of the mid-jejunum, ileum or mid-colon. Throughout the small intestine, the highest number of ASC was found following jejunal immunisation, followed by ileal, oral and colonic immunisation. To determine the signifance of Peyer's patches in the induced immune response, F4 was injected into the jejunal Peyer's patches (JPP), lamina propria (LP) and ileal Peyer's patches (IPP). Immunisation in the JPP induced the highest number ASC in the small intestine, whereas immunisation in the LP and IPP resulted in lower intestinal antibody responses. In conclusion, we have shown that the JPP are the major inductive sites of the F4-specific intestinal antibody response. This knowledge could be important when using the pig as an animal model for vaccination studies.     

Induction of immune responses in pigs following oral administration of purified F4 fimbriae

An effective way of stimulating the mucosal immune system was examined in piglets, using F4 fimbriae of enterotoxigenic Escherichia coli (ETEC). It was demonstrated that purified F4 fimbriae, as opposed to ovalbumin (OVA), are powerful oral immunogens. Indeed, oral administration of purified F4 induced antigen-specific antibody-secreting cells (ASC) in the Peyer's patches, mesenteric lymph nodes (LN), blood and lamina propria 4, 7, 9 and 11 days postimmunization, respectively, indicating a stimulation of the mucosal immune system, whereas upon oral administration of OVA, no immune response was observed. Moreover, the induced F4-specific IgA and IgG antibody responses were comparable with those obtained upon oral infection with viable E. coli and intramuscular (i.m.) F4 injection, respectively. Furthermore, a priming of the mucosal immune system is better obtained by oral infection (ASC localized in mesenteric LN) than by i.m. F4 injection (ASC localized in spleen and retropharyngeal LN) since an oral boost with purified F4 induced a secondary response in the orally infected animal (mainly IgA and IgG ASC, rapid increase of IgA antibodies) while in the i.m. primed animal a secondary (more circulating antigen-specific ASC than in the unprimed animal) as well as a primary IgM and IgA response (mainly IgM ASC, slow increase of IgA antibodies), suggesting a primary mucosal response, were seen. An oral challenge of the naive control displayed a primary response (mainly IgM ASC, slow increase of IgA and IgG antibodies). The capacity of purified F4 to activate the mucosal immune system on oral administration, is of importance for the development of oral vaccines against ETEC infections.     

Phycocyanin enhances secretary IgA antibody response and suppresses allergic IgE antibody response in mice immunized with antigen-entrapped biodegradable microparticles

In the present study, we have investigated the effects of phycocyanin, a biliprotein of Spirulina platensis, on mucosal and systemic immune responses and allergic inflammation in C3H/HeN and BALB/cA mice. To induce the antigen-specific antibodies in the peripheral lymphoid tissues such as Peyer's patches and mesenteric lymph nodes, biodegradable ovalbumin-entrapped poly (DL-lactide-co-glycolide) particles were used as an antigen. Two weeks after the onset of phycocyanin ingestion, mice were immunized with an aqueous ovalbumin (OVA) solution. Starting at one week after the primary immunization, the mice were subjected to oral immunization with the biodegradable OVA microparticles twice a week. IgA, IgE and IgG1 antibodies were determined by ELISA. The OVA microparticles of 4-microm diameter successfully induced antigen-specific antibodies. In the mice that received phycocyanin treatment for 6 wk, a marked increase in the antigen-specific, as well as the total, IgA antibody level was observed in the Peyer's patches, mesenteric lymph nodes and intestinal mucosa as well as in the spleen cells. Both antigen-specific IgG1 and IgE antibody levels in the serum were suppressed by ingestion of phycocyanin for 8 wk. However, inflammation of the small intestine, monitored as vascular permeability by the Evans blue-leaking method was reduced by phycocyanin at 6 wk, which preceded the suppression of antigen-specific IgG1 and IgE antibody production by 2 wk. These results suggest that phycocyanin enhances biological defense activity against infectious diseases through sustaining functions of the mucosal immune system and reduces allergic inflammation by the suppression of antigen-specific IgE antibody.     

Targeting lymphocyte Peyer's patch adhesion molecule-1: a relay approach to gut immunization

Targeting vaccines to dendritic cells (DCs) can enhance responses to weak vaccine antigens. Although there are molecules that are relatively specific for the various DC subsets, there are none that are both region-specific and DC-specific. This has provided some limitation to targeting regional DC populations. We proposed that these limits could be overcome by targeting antigens not to the DC subsets directly but to cells that persistently seek out and closely interact with DCs, namely lymphocytes. To investigate this hypothesis, we targeted antigens to a unique population of gut-homing lymphocytes and then looked at the induction of immune responses at this site. Using an anti-LPAM-1 (Lymphocyte Peyer's patch adhesion molecule-1; alpha(4)beta(7) integrin) monoclonal antibody (mAb) as a model antigen, we found that targeting gut-homing lymphocytes could significantly elevate the gut mucosal IgA response. Moreover, such a strategy greatly elevated the systemic IgG as well as IgA response. We found that LPAM-1-targeting enhanced the localization of antigen to both the systemic and mucosal lymphoid compartments where both IgA and IgG responses were induced. We also found that any parenteral route of delivery sufficed. Overall, targeting unique populations of lymphocytes may provide a strategy for ferrying antigen to sites that such lymphocytes home to.     

Serum-derived IgG1-mediated immune exclusion as a mechanism of protection against H. pylori infection

The induction of protective immunity against Helicobacter challenge in a murine model was found to correlate with the magnitude of IgG (serum and gastric lavage) responsiveness to intra-nasal (i.n.) immunisation. IgG1-secreting hybridoma backpacks in Helicobacter pylori (H. pylori)-infected mice revealed serum transudation into the stomach. A Lpp20-specific monoclonal antibody was associated with significantly reduced H. pylori colonisation. Histology revealed aggregates of the remaining H. pylori in these mice, suggesting a role for IgG1-mediated immune exclusion of the bacteria. In vitro immunogold electron microscopy supported this hypothesis, but also suggested that a threshold of H. pylori-specific antibody needs to be maintained if immune exclusion by the host is to overcome immune evasion by the bacteria.