The farming environment protects mice from allergen-induced skin contact hypersensitivity.

BACKGROUND: Being born and raised in a farm provides a long-lasting protection for allergies. The microbial environment provided by farm animals is crucial to induce this protective effect, although underlying immune mechanisms remain elusive. OBJECTIVE: To establish a mouse model of global exposure to the farming environment and to study immunologic changes linked to protection of allergy. METHODS: Mice colonies were bred in parallel in a farm cowshed and the university animal facility (AF). Mice from both locations were subjected to a skin contact allergy model. Peripheral blood cells and cell cytokine production were assessed in both populations. In addition, the gut microbiome at various ages was characterized. RESULTS: Mice born in the farm were less prone to develop allergy than mice bred in the AF. Mice transfers between the AF and the farm showed a better protection when mice were moved to the farm early in life. As compared to AF-bred mice, farm mice displayed early immune activation with higher CD4+ T cell population, in particular CD4+ CD25+ FoxP3- (activated cells). The cytokine profile of mice from the farm was skewed towards an IL-17 and IL-22 secreting cell profile accompanied by increased IL-10 secretion. These differences were mostly seen within a specific age window between birth and 8 weeks of age. Microbiome analysis showed differences between 4 and 20 weeks old mice and between farm and AF mice with an increased number of Murine mastadenovirus B in young farm mice exclusively. CONCLUSION: The farming environment provides a strong, allergy protective IL-22 stimulus and generates activated CD4+ T cells. Exposure to the farm environment early in their life may also provide a better protection for contact skin allergy. Whether a viral trigger might decisively influence protection for allergies remains to be determined.

Gut T cell receptor-γδ(+) intraepithelial lymphocytes are activated selectively by cholera toxin to break oral tolerance in mice.

The gut immune system is usually tolerant to harmless foreign antigens such as food proteins. However, tolerance breakdown may occur and lead to food allergy. To study mechanisms underlying food allergy, animal models have been developed in mice by using cholera toxin (CT) to break tolerance. In this study, we identify T cell receptor (TCR)-γδ(+) intraepithelial lymphocytes (IELs) as major targets of CT to break tolerance to food allergens. TCR-γδ(+) IEL-enriched cell populations isolated from mice fed with CT and transferred to naive mice hamper tolerization to the food allergen ß-lactoglobulin (BLG) in recipient mice which produce anti-BLG immunoglobulin (Ig)G1 antibodies. Furthermore, adoptive transfer of TCR-γδ(+) cells from CT-fed mice triggers the production of anti-CT IgG1 antibodies in recipient mice that were never exposed to CT, suggesting antigen-presenting cell (APC)-like functions of TCR-γδ(+) IELs. In contrast to TCR-αß(+) cells, TCR-γδ(+) IELs bind and internalize CT both in vitro and in vivo. CT-activated TCR-γδ(+) IELs express major histocompatibility complex (MHC) class II molecules, CD80 and CD86 demonstrating an APC phenotype. CT-activated TCR-γδ(+) IELs migrate to the lamina propria, where they produce interleukin (IL)-10 and IL-17. These results provide in-vivo evidence for a major role of TCR-γδ(+) IELs in the modulation of oral tolerance in the pathogenesis of food allergy.

Intestinal lamina propria TcRgammadelta+ lymphocytes selectively express IL-10 and IL-17.

BACKGROUND: The characteristics and roles of gut lymphocytes have been only partly elucidated, in particular with regard to activation patterns. OBJECTIVES: To characterize lymphocytes from various parts of the gut and examine their activation pattern as a network. METHODS: Lymphocytes were isolated from the epithelium, the lamina propria, Peyer's patches, mesenteric lymph nodes, the spleen, and peripheral blood of naive mice. They were then characterized for T cell phenotype, T cell receptors (TcRs), activation markers, and cytokine production. RESULTS: The results showed a gradient of cells with an increasing proportion of TcRgammadelta+, CD8alphaalpha+ cells towards the gut lumen, with the highest number found in intraepithelial lymphocytes. These cells, together with lamina propria lymphocytes (LPLs) were also characterized by a memory-like phenotype (CD25- CD45RB(low) and CD44(high)) and CD69 expression. CD8+ TcRgammadelta+ LPLs produced IL-10 and IL-17, while TcRalphabeta+ LPLs were FoxP3 positive. CONCLUSIONS: Gut lymphocytes express various receptors and cytokines according to their location. These specific features suggest a differential function for gut lymphocytes depending on their location.

Avirulant Salmonella typhimurium strains prevent food allergy in mice.

Oral tolerance to foods can be regulated by microorganisms in the gut lumen. We hypothesized that pretreatment with avirulent Salmonella typhimurium strains could prevent food allergy in mice. Mice were administered S. typhimurium PhoPc (STPhoPc) or S. typhimurium AroA prior to oral sensitization to beta-lactoglobulin in the presence of cholera toxin. An oral antigen challenge after sensitization assessed antigen-induced anaphylaxis. Antigen-specific antibody titres were measured by enzyme-linked immunosorbent assay in the serum and enzyme-linked immunospot (ELISPOT) in the spleen, and cytokine-secreting cells were measured by ELISPOT in the Peyer's patches, lamina propria and epithelium cells. We showed first that S. typhimurium could up-regulate interleukin (IL)-12 and IL-10 secretion by gut T cells. Mice pretreated with STPhoPc had decreased anaphylaxis upon challenge, along with decreased immumoglobulin G1 (IgG1) and IgE antibody titres. Mice having received S. typhimurium AroA had partly decreased anaphylaxis as well as decreased serum IgG1 antibody titres in the serum, and increased serum IgA antibody titres. Antibody titres could be correlated with increased numbers of spleen and Peyer's patches antibody-producing cells. STPhoPc-treated mice showed significantly decreased anaphylaxis when compared with the control mice, while S. typhimurium AroA-pretreated mice had a similar immune response together with increased secretory IgA titres. Our experiments have proved a potential immunomodulatory protective effect by two avirulent S. typhimurium strains.

Oral carrageenan induces antigen-dependent oral tolerance: prevention of anaphylaxis and induction of lymphocyte anergy in a murine model of food allergy.

Immunosuppressive effects of carrageenan, a high-molecular-weight polysaccharide, on antibody and T cell responses have been previously demonstrated. However, its effect on anaphylaxis is unknown. Our objectives were to test carrageenan-mediated oral tolerance induction in young mice subsequently sensitized to a common cow's milk antigen. C3H/HeJ mice were fed or not lambda-carrageenan (0.5 g/L) and/or 0.01 mg/mL beta-lactoglobulin (BLG) for 5 d before oral sensitization with BLG and cholera toxin. Subsequently, the mice were challenged with BLG and symptom scores of anaphylaxis were recorded. Mesenteric lymph node cells, spleen cells, Peyer's patches cells, intraepithelial lymphocytes, and lamina propria lymphocytes were isolated and stimulated in vitro with BLG, IL-2, or left unstimulated. BLG-specific IgG, IgG(1), and IgG(2a) antibodies were measured. Pretreatment with carrageenan and BLG, but not pretreatment with either carrageenan or BLG alone or omission of pretreatment, diminished significantly the number of anaphylactic mice after BLG challenge (6.3 % versus 53 % in mice without pretreatment, p = 0.006). Mesenteric lymph nodes and spleen cells from pretreated mice proliferated less in presence of BLG or IL-2 than cells from sensitized control mice. Antigen-specific antibody production and passive cutaneous anaphylaxis was not suppressed by carrageenan and BLG pretreatment. In conclusion, carrageenan administered to young mice in conjunction with low doses of allergen before sensitization efficiently prevents anaphylaxis.

CD8(+) T cells secreting type 2 lymphokines are defective in protection against viral infection.

Effector T cells secreting type 1 and/or type 2 lymphokines (Tc1, Tc0, Tc2) were generated in vitro from CD8(+) T cells of mice with a transgenic TCR recognizing lymphocytic choriomeningitis virus (LCMV) glycoprotein to compare their effector function in vitro and in vivo. Tc1, Tc2, and Tc0 showed similar Fas- and perforin-mediated cytotoxicity in vitro. Upon adoptive transfer, Tc2 and Tc0 effectors were less efficient than Tc1 at controlling LCMV or recombinant vaccinia virus expressing the LCMV glycoprotein in vivo. Tc2 and Tc0 had decreased surface VLA-4 density and deficient activation-induced LFA-1/ICAM-1-dependent homotypic adhesion in vitro. Therefore, the reduced antiviral activity in vivo of Tc2 and Tc0 compared with Tc1 is not due to reduced cytotoxic activity or IFN-gamma secretion but may be explained by defective homing to the target organ due to decreased expression and/or lower activity of adhesion molecules.