Sodium butyrate can improve intestinal integrity and immunity in juvenile Chinese mitten crab (Eriocheir sinensis) fed glycinin.

Butyrate is a fermentation byproduct of gut microbiota and is susceptible to chronic oxidative stress. This study investigates the mitigative effects of sodium butyrate (SBT) on growth inhibition and intestinal damage induced by glycinin in juvenile Chinese mitten crab (Eriocheir sinensis). All four experimental diets containing 80 g/kg glycinin were formulated with 0, 10, 20 and 40 g/kg SBT respectively. There was no glycinin or SBT in the control diet. Juvenile crabs (0.33 ± 0.01g) were respectively fed with these five diets for eight weeks. The diets with 10 and 20 g/kg SBT significantly improved the survival and weight gain of the crabs compared with those in the 0 g/kg SBT group, and showed no difference with the control group. The crabs fed diets containing glycinin without SBT had lower glutathione and glutathione peroxidase activities but higher malondialdehyde in the intestine than those in the control group. Moreover, dietary glycinin decreased the lysozyme and phenoloxidase activities and improved the level of histamine in the intestine compared with the control group, while the supplementation of SBT counteracted these negative effects. The addition of SBT could also restore the impaired immunity and morphological structure of the intestine. Dietary SBT could increase the mRNA expression of antimicrobial peptides genes (anti-lipopolysaccharide factor 1 and 2) and decrease the content of pro-inflammatory factor TNF-α. The SBT could restore the intestinal microbial community disorganized by glycinin. The abundance of pathogenic bacteria (Aeromonas, Vibrio and Pseudomonas) decreased significantly and the potential probiotic bacteria (Bacillus, Lactobacillus, Chitinibacter and Dysgonomonas) increased significantly in the 10 g/kg SBT group. This study suggests that sodium butyrate supplementation can mitigate the negative effects induced by glycinin such as growth inhibition, intestinal inflammation and reduction of beneficial flora in the gut.

Identification of the linear immunodominant epitopes in the ß subunit of ß-conglycinin and preparation of epitope antibodies.

ß-conglycinin is one of the major allergens in soybean protein. The purpose of this study was to predict and to identify the major linear epitopes of the ß subunit of ß-conglycinin. Potential linear epitopes were predicted and confirmed by three immunoinformatics tools combined with the Immune Epitope Database (IEDB). Ten potential epitope peptides were synthesized by Fmoc (9-fluorenylmethoxycarbonyl) solid phase peptide synthesis and were validated by the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) using sera from soybean allergic patients. Polyclonal antibodies, which were prepared by immunizing rabbits with synthesized peptides, were used to confirm their binding ability with ß-conglycinin through western blot and dot blot assays. The results showed that 10 peptides were screened as the main epitopes for the ß subunit of ß-conglycinin. All 10 peptides (P1-P10) presented IgG binding activity, and P2 and P6 were also validated as IgE binding peptides. Moreover, the results of dot blot showed that P5 and P8 might be located inside the protein molecule. Western blot indicated that most of polyclonal antibodies were bound effectively to the ß subunit of ß-conglycinin. In addition, few polyclonal antibodies exhibited an immune cross-reaction with the α and α' subunits.

Soybean ß-conglycinin caused intestinal inflammation and oxidative damage in association with NF-κB, TOR and Nrf2 in juvenile grass carp (Ctenopharyngodon idella): varying among different intestinal segments.

The current study aimed to investigate the effects and mechanisms of dietary soybean ß-conglycinin in immune function and oxidative damage among different intestinal segments of juvenile grass carp (Ctenopharyngodon idella). 240 fish (13.77 ±â€¯0.10 g) were fed control or 8% ß-conglycinin diet for 7 weeks. Dietary ß-conglycinin caused inconsistent suppression effects on the innate immune by decreasing complement component, lysozyme, antimicrobial peptide and acid phosphatase among different intestinal segments. Meanwhile, dietary ß-conglycinin caused inflammation in the mid and distal intestine by raising pro-inflammatory cytokines and declining anti-inflammatory cytokines mRNA levels, while more serious in the distal intestine than in the mid intestine. Furthermore, dietary ß-conglycinin regulating inflammatory cytokines might be associated with transcription factors nuclear factor-κB P65 (NF-κB P65) nucleus translocation and target of rapamycin (TOR) phosphorylation in the distal intestine but only related to TOR phosphorylation in the mid intestine. Interestingly, in the proximal intestine, dietary ß-conglycinin decreased both pro-inflammatory and anti-inflammatory cytokines mRNA level, and did not affect NF-κB P65 nucleus translocation and TOR phosphorylation. For oxidative damage, dietary ß-conglycinin exposure elevated both malondialdehyde (MDA) and protein carbonyl (PC) contents in the distal intestine, which might be attributed to the suppression of the Mn-SOD, catalase (CAT) and glutathione peroxidase (GPx) activities. In the mid intestine, dietary ß-conglycinin only increased PC content in association with the low activities of CAT, GPx and glutathione peroxidase (GR). Unexpectedly, in the proximal intestine, dietary ß-conglycinin did not significantly change MDA and PC contents while decreased antioxidant enzyme activities. Furtherly, dietary ß-conglycinin affect the antioxidant enzyme activity might be regulated by the varying pattern of nuclear factor-erythroid 2-related factor 2 (Nrf2) nucleus translocation among these three intestinal segments. In summary, dietary ß-conglycinin caused intestinal inflammation and oxidative damage in association with NF-κB, TOR and Nrf2 signaling molecules, which were varying among the three intestinal segments of grass carp.

Acidic polypeptides A1a, A3 and A4 of Gly m 6 (glycinin) are allergenic for piglets.

Gly m 6 (glycinin) is one of the major antigenic proteins in soybeans responsible for transient hypersensitivity to soybean meal in weaned piglets. The globulin is a hexamer consisting of subunits containing basic and acidic polypeptides. Multiple acidic polypeptides have long been demonstrated to be allergens for humans and play a key role in the overall allergenicity of Gly m 6. To date, knowledge on the allergenicity of the acidic polypeptides for piglets is very limited. The purpose of this study was to identify the acidic polypeptides that were allergenic for piglets and to characterize these acidic polypeptides by ELISA, western blot, skin prick and basophile histamine release test. The IgG and IgE antibody binding capacities of the acidic polypeptides of Gly m 6 were determined using ELISA and western blot analysis with sera from Gly m 6 sensitized piglets. Skin prick test and basophile histamine release test were conducted to measure the effector cell response to the polypeptides. Specific IgG and IgE antibodies against A1a, A3 and A4 of Gly m 6 were identified in the sera of Gly m 6 sensitized piglets. Meanwhile, positive skin prick test and specific histamine release responses were also induced by the acidic polypeptide A1a, A3 and A4 of Gly m 6 from the basophiles of Gly m 6 sensitized piglets. The results demonstrate that the acidic polypeptide A1a, A3 and A4 of Gly m 6 are allergenic for piglets.

Soybean Glycinin- and ß-Conglycinin-Induced Intestinal Damage in Piglets via the p38/JNK/NF-κB Signaling Pathway.

ß-Conglycinin (7S) and glycinin (11S) are known to induce a variety of hypersensitivity reactions involving the skin, intestinal tract, and respiratory tract. The present study aimed to identify the mechanism underlying the development of allergy to soybean antigen proteins, using piglets as an animal model. Weaned "Duroc × Landrace × Yorkshire" piglets were fed a diet supplemented with 7S or 11S to investigate the signaling pathway involved in intestinal damage in piglets. Results showed that serum nitric oxide (NO), tumor necrosis factor-α (TNF-α), and caspase-3 levels were significantly higher in 7S- and 11S-fed piglets compared to those in suckling or weaned ones. mRNA, protein, and phosphorylation levels of nuclear factor-kappa B (NF-κB), p38, and Jun N-terminal kinase (JNK) were higher in 7S- and 11S-fed piglets than in suckling and weaned ones. Overall, our results indicate that 7S and 11S damaged the intestinal function in piglets through their impact on NF-κB, JNK, and p38 expression.

Identification of the 7S and 11S globulins as percutaneously sensitizing soybean allergens as demonstrated through epidermal application of crude soybean extract.

Cutaneous exposure to food allergens can predispose individuals to food allergies. Soybean, a major allergenic food, is an ingredient in various cosmetic products. However, the types of soybean proteins that are percutaneously sensitizing in humans or animal models remain unknown. In this study, BALB/c mice were dorsally shaved and epicutaneously exposed to a crude soybean extract including sodium dodecyl sulfate or distilled water alone. Specific IgEs secreted in response to 7S globulin (Gly m 5), 11S globulin (Gly m 6), Gly m 3, and Gly m 4 were measured using enzyme-linked immunosorbent assays or immunoblots. Exposure to soybean extract elicited the secretion of soybean-specific IgEs. Of the soybean proteins, 7S and 11S globulins acted as percutaneous sensitizers in 6/9 mice (67%). Additionally, IgE bound specifically and preferentially to the 7S globulin ß subunit. In conclusion, this is the first report to identify percutaneously sensitizing soybean allergens in a mouse model.

Selection of lactic acid bacteria strains for the hydrolysis of allergenic proteins of wheat flour.

BACKGROUND: Wheat flour is one of the most common causative agents of food allergy. The study presents the selection and characterization of lactic acid bacteria (LAB) strains capable of hydrolyzing/modifying allergenic proteins of wheat flour. Hydrolysis of wheat proteins was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with sera from patients with food allergy to gluten. RESULTS: The analysis of electrophoretic profiles of protein extracted from sourdough shows the capability of selected LAB strains for proteolytic degradation of wheat proteins that belong to two factions: albumin/globulin (hydrolysis of 13 polypeptides with a molecular weight between 103 and 22 kDa); and gliadin (seven polypeptides with a molecular weight between 39 and 24 kDa). All analyzed strains were capable of hydrolyzing some IgE-binding epitopes of wheat allergens. The lack of such changes in control samples indicates that they were induced rather by the proteolytic activity of bacterial strains than endogenous enzymes of wheat flour. The gluten proteins were susceptible to hydrolysis by sequential digestion with pepsin and trypsin. CONCLUSION: The selected strains exhibit proteolytic activity, which leads to a reduction in allergenicity of wheat sourdoughs. These strains may be applied as specific starter cultures to prepare bakery products of special nutritional use. © 2015 Society of Chemical Industry.

Glomerulonephritis Induced by Heterologous Anti-GBM Globulin as a Planted Foreign Antigen.

The glomerulonephritides are diseases characterized by immune-mediated glomerular inflammation. Most severe and rapidly progressive forms of glomerulonephritis feature the participation of injurious leukocytes that localize to glomeruli. This unit describes classical models of rapidly progressive glomerulonephritis in mice, induced by injecting heterologous globulin (raised in sheep) that binds to the glomerular basement membrane. These models have been particularly useful in defining the participation of effector leukocytes in severe glomerular disease. In these models, injury typically occurs in two phases. In the initial, heterologous phase, injury is mediated by the globulin bound within the glomerulus acting as an antibody. The later, autologous phase of injury is mediated by the host's adaptive immunity to the heterologous globulin now functioning as a planted foreign antigen within glomeruli. As autologous phase injury is driven by immunity to sheep globulin, assessment of antigen-specific systemic immunity to sheep globulin is critical when using this model.

Soybean ß-conglycinin induces inflammation and oxidation and causes dysfunction of intestinal digestion and absorption in fish.

ß-Conglycinin has been identified as one of the major feed allergens. However, studies of ß-conglycinin on fish are scarce. This study investigated the effects of ß-conglycinin on the growth, digestive and absorptive ability, inflammatory response, oxidative status and gene expression of juvenile Jian carp (Cyprinus carpio var. Jian) in vivo and their enterocytes in vitro. The results indicated that the specific growth rate (SGR), feed intake, and feed efficiency were reduced by ß-conglycinin. In addition, activities of trypsin, chymotrypsin, lipase, creatine kinase, Na(+),K(+)-ATPase and alkaline phosphatase in the intestine showed similar tendencies. The protein content of the hepatopancreas and intestines, and the weight and length of the intestines were all reduced by ß-conglycinin. ß-Conglycinin increased lipid and protein oxidation in the detected tissues and cells. However, ß-conglycinin decreased superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPx) and glutathione reductase (GR) activities and glutathione (GSH) content in the intestine and enterocytes. Similar antioxidant activity in the hepatopancreas was observed, except for GST. The expression of target of rapamycin (TOR) gene was reduced by ß-conglycinin. Furthermore, mRNA levels of interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and transforming growth factor-ß (TGF-ß) genes were increased by ß-conglycinin. However, ß-conglycinin increased CuZnSOD, MnSOD, CAT, and GPx1b gene expression. In conclusion, this study indicates that ß-conglycinin induces inflammation and oxidation, and causes dysfunction of intestinal digestion and absorption in fish, and finally reduces fish growth. The results of this study provide some information to the mechanism of ß-conglycinin-induced negative effects.

Epitopes from two soybean glycinin subunits are antigenic in pigs.

BACKGROUND: Glycinin is a seed storage protein in soybean (Glycine max) that is allergenic in pigs. Glycinin is a hexamer composed of subunits consisting of basic and acidic portions joined by disulfide bridges. There are five glycinin subunit isoforms designated Gy1-Gy5. The purpose of this study is to identify epitopes from selected glycinin subunits that are antigenic in pigs. RESULTS: Twenty-seven out of 30 pigs had antibodies against glycinin in their sera. Ten of these sera had immunoglobulin G (IgG) against the Gy4 (A5A4B3) or Gy1 (A1aBx) subunit. Three sera recognised overlapping regions between the two subunits tested, though no serum stained both A5A4B3 and A1aBx. Two sera stained a highly conserved region between A5A4B3 and A1aBx, though again neither serum stained both peptides. The basic part of the A1aBx subunit was not recognised by any of the sera tested even though immunoblot data indicated that the basic and acidic subunits of glycinin are nearly equally antigenic. CONCLUSION: Two antigenic regions of A5A4B3 and A1aBx were identified that bound antibodies in half of the sera that reacted with these two proteins. Half of the sera reacted with unique regions of A5A4B3 and A1aBx. The failure of the basic portion of A1aBx to bind pig antibodies may indicate that it is less antigenic than the basic portion of A5A4B3 and other glycinin subunits.

Enhanced antiviral activity of soybean ß-conglycinin-derived peptides by acylation with saturated fatty acids.

Peptide mixtures prepared from soybean ß-conglycinin (7S-peptides) were acylated with saturated fatty acids of different chain length (6C-18C) in order to improve their antiviral activity against Feline calicivirus (FCV) strain F9 which is a typical norovirus surrogate. Among the fatty acids varieties, it was revealed that 7S-peptides acylated with myristic and palmitic acids potently inhibited FCV replication. Myristorylation and palmitoylation of 7S-peptides kept host cells viability at 91.51% and 98.90%, respectively. The infectivity of FCV on Crandell-Reese feline kidney cells was further determined after exposure of initial titer of 10(6.47) TCID50/mL. Myristoylated and palmitoylated 7S-peptides significantly (P < 0.006) reduced FCV infectivity as compared to native 7S-peptides. Native 7S-peptides showed 25% FCV inhibitory activity while myristoylated and palmitoylated 7S-peptides exhibited 98.59% and 99.98% reduction in FCV infectivity, respectively. Myristoylated and palmitoylated 7S-peptides demonstrated higher anti-FCV activity in a wide range of concentration with complete reduction at 25 µg/mL. Surface hydrophobicity was significantly (P < 0.05) increased after attachment of long hydrocarbon fatty acids to 7S-peptides as supported by changes in fluorescence intensity. Enzymatic hydrolysis together with acylation will give an insight into surface and physiological functional lipopeptides derived from soy ß-conglycinin.

Inbred Chinese Wuzhishan (WZS) minipig model for soybean glycinin and beta-conglycinin allergy.

Validated murine models have been built to assess the potential allergenicity of novel proteins. Large animals, such as pigs, share more similarities to humans in physiology and immunology than murine. Among Chinese minipigs, Wuzhishan (WZS) minipigs have the highest inbreeding coefficient, more stable heredity, and less variability, which were gastrically sensitized and excitated with diets containing 4% glycinin or 4% beta-conglycinin or neither to induce anaphylactic reactions in the present study. In glycinin- and beta-conglycinin-sensitized animals, diarrhea symptoms and skin wheal and flare responses were observed. In comparison to the control, after oral excitation with glycinin or beta-conglycinin, the serum IgE was increased by 34.4 or 38.4% and the serum histamine was increased by 42.1 or 46.9%, respectively. In addition, the serum IFN-gamma were reduced by 12.4 or 30.0%, respectively. The jejunum histamine level of beta-conglycinin-sensitized animals was increased by 196.6%, while the number of mast cells in the submucosa of jejunum and ileum of the glycinin-sensitized animals declined by 48.1 and 45.0%. In conclusion, the WZS minipig allergy models induced by soybean glycinin and beta-conglycinin represent type-I hypersensitivity reactions mediated by IgE, which could potentially be useful in determining the potential allergenicity of novel proteins.

[Anaphylactic reactions in WZS minipig orally induced by glycinin].

OBJECTIVE: To explore the suitable dosage and observation period of anaphylactic reactions in WZS minipigs induced by glycinin (11S). METHODS: Twelve 45-day-old WZS minipigs were randomly divided into three groups (control group, 4% 11S group and 8% 11S group, n = 4), which were respectively orally sensitized (0 - 10d) and orally challenged (16 - 18d, 31d) with diets containing 0%, 4% 11S or 8% 11S soybean proteins to induce anaphylactic reactions. Clinical symptoms, skin prick reactions, serum IgG, serum IgE, histamine, mast cell in the small intestine were observed or measured. RESULTS: Diarrhea and positive skin prick reactions at different degrees were observed in 11S groups. Compared with the control group, serum IgG, IgE and histamine levels of 4% 11S group at 19d and 32d were significantly higher (P < 0.05), and reached the peak when 11S groups were orally challenged at 19d. The jejunum histamine level was negatively correlated with the mast cell numbers in the jejunum mucosa and sub-mucosa (P < 0.05). Compared with the control, the mast cell number in the jejunum sub-mucosa of 4% 11S group was decreased (P < 0.05). CONCLUSION: The WZS minipig allergic reaction orally induced by glycinin could be type I hypersensitivity mediated by IgE, reaction induced by 4% dosage glycinin could be more strong than that of 8% dosage, and stimulating phase could be a suitable observation period for anaphylactic reactions which could be related with the activation of mast cells in the small intestine.

Treatment with thymoglobulin as the cause of acuted emyelinating polyneuropathy in arenal transplant patient

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Soybean beta-conglycinin as the main allergen in a patient with food-dependent exercise-induced anaphylaxis by tofu: food processing alters pepsin resistance.

BACKGROUND: Food-dependent exercise-induced anaphylaxis (FDEIA) due to soybeans is a rare disorder. The allergen responsible for FDEIA due to soybeans has not yet been determined. OBJECTIVE: We characterized the clinical features of a patient with FDEIA due to tofu, who was well tolerant to drinking soy milk. We then sought to identify the responsible soybean allergen(s) in that patient. We further studied whether different stabilities of the allergen(s) to pepsin digestion between two soybean products are related to their clinical allergenicity. METHODS: Skin prick tests and provocation tests using soybean products were performed to detect the responsible food and other factors that induced the allergic symptoms. Specific IgE to various soybean allergens were examined by ImmunoCAP, ELISA and protein microarray assays. Immunoblotting for soybeans and soybean products using the patient's serum was also performed. Soybean products were serially digested by pepsin to disclose the stability of the allergens. RESULTS: Provocation with ingestion of tofu and exercise induced the allergic symptoms, while ingestion of soy milk and exercise did not. Immunoblot analysis, ELISA and protein microarray assay revealed that beta-conglycinin mainly reacts with IgE antibodies in the patient's serum. By immunoblot analysis, beta-conglycinin in soy milk completely disappeared after pepsin digestion within 20 min, whereas beta-conglycinin in tofu was almost intact after more than 120 min of pepsin digestion. CONCLUSION: We identified beta-conglycinin as the causative allergen in a patient with FDEIA induced by tofu. The difference in resistance to pepsin digestion between tofu and soy milk suggests that the presence of undigested allergens in the digestive tract is a prerequisite for the development of FDEIA.

[A WZS miniature swine food hypersensitivity model orally induced by soybean beta-conglycinin].

OBJECTIVE: To establish WZS miniature swine model of beta-conglycinin (7 S) allergy for evaluating the potential allergenicity of genetically modified food. METHODS: Twelve 45-day-old WZS miniature swines from three litters were randomly divided into three groups (control group; 4% 7 S group and 8% 7 S group, n = 4), which were respectively gastric sensitized (day 0 - 10) and oral challenged (day 6 - 18, 31) to induce anaphylactic reactions. Clinical symptoms, skin prick reactions were recorded. At day 10, 19 and 32, serum IgG, IgE, histamine and cytokines levels were measured by ELISA. RESULTS: Diarrhea at different degrees were observed in 4% and 8% 7 S groups. The skin erythema reactions in grade "-", "+/-", "+", "++" of control group respectively were 2/4, 2/4, 0/4, 0/4, of 4% 7 S group were 0/4, 0/4, 2/4, 2/4 and of 8% 7 S group were 0/4, 0/4, 1/4, 3/4. The serum IgE and histamine levels of day 11, 19 and 32 were all significantly and positively correlated (Pearson coefficients = 1, P = 0.000). The serum IgG, IgE and histamine levels all reached the peak after 7 S groups were oral challenged at day 19.Compared with the control group, serum IgG (lg IgG: 2.95 +/- 0.31 vs 2.29 +/- 0.25, t = 3.19, P = 0.011), IgE (lg IgE: 2.45 +/- 0.30 vs 1.77 +/- 0.23, t = 3.31, P = 0.009) and histamine levels(lg histamine:2.13 +/- 0.30 vs 1.45 +/- 0.23, t = 3.34, P = 0.009) of 4% 7 S group at day 19 were all significantly higher, while the serum IFN-gamma content [(35.78 +/- 6.42) pg/ml vs (51.10 +/- 9.67) pg/ml, t = -2.33, P = 0.045] of 4%7 S group was significantly lower. CONCLUSION: The WZS miniature swine model orally induced by soybean beta-conglycinin is type I hypersensitivity mediated by IgE, which can be used to predict the potential allergenicity of genetically modified food.

Allergic and anaphylactic response to sesame seeds in mice: identification of Ses i 3 and basic subunit of 11s globulins as allergens.

BACKGROUND: Allergy to sesame seeds is an emerging food allergy of a serious nature due to a high risk of systemic anaphylaxis. Although a mouse model to study sesame anaphylaxis is desirable, currently it is not available. Here, using a transdermal exposure model system, we tested the hypothesis that sesame seed elicits IL-4-associated IgE antibody response with consequent clinical sensitization in mice. METHODS: Groups of BALB/c mice were exposed to sesame seed extract or saline or a control food (vanilla bean extract) by transdermal applications. Systemic IgE, IgG1 and IgG2a antibody responses were examined using preoptimized ELISA. Type 2 and type 1 cytokine responses were evaluated by ex vivo antigen-mediated activation of spleen cells. Clinical response to oral sesame challenge was studied. Western blot and N-terminal amino acid sequence analyses were performed to identify the sesame allergens. RESULTS: Transdermal exposure to sesame elicited robust IgE and IgG1 but very little IgG2a antibody responses. IgE response to transdermal exposure in two high-IgE responder mice strains with disparate MHC confirmed the intrinsic allergenicity of sesame seed. Transdermal sensitization was associated with activation of IL-4 but not IFN-gamma. Furthermore, oral exposure to sesame resulted in clinical signs of systemic anaphylaxis. Western blot and sequence analysis identified four allergens including Ses i 3 and the basic subunit of 11s globulins. CONCLUSION: These data argue that transdermal exposure to sesame seed can result in IL-4 activation, IgE response and clinical sensitization for systemic anaphylaxis.

Structural, biological, and evolutionary relationships of plant food allergens sensitizing via the gastrointestinal tract.

The recently completed genome sequence of the model plant species Arabidopsis has been estimated to encode over 25,000 proteins, which, on the basis of their function, can be classified into structural and metabolic (the vast majority of plant proteins), protective proteins, which defend a plant against invasion by pathogens or feeding by pests, and storage proteins, which proved a nutrient store to support germination in seeds. It is now clear that almost all plant food allergens are either protective or storage proteins. It is also becoming evident that those proteins that trigger the development of an allergic response through the gastrointestinal tract belong primarily to two large protein superfamilies: (1) The cereal prolamin superfamily, comprising three major groups of plant food allergens, the 2S albumins, lipid transfer proteins, and cereal alpha-amylase/trypsin inhibitors, which have related structures, and are stable to thermal processing and proteolysis. They include major allergens from Brazil nut, peanuts, fruits, such as peaches, and cereals, such as rice and wheat; (2) The cupin superfamily, comprising the major globulin storage proteins from a number of plant species. The globulins have been found to be allergens in plant foods, such as peanuts, soya bean, and walnut; (3) The cyteine protease C1 family, comprising the papain-like proteases from microbes, plants, and animals. This family contains two notable allergens that sensitize via the GI tract, namely actinidin from kiwi fruit and the soybean allergen, Gly m Bd 30k/P34. This study describes the properties, structures, and evolutionary relationships of these protein families, the allergens that belong to them, and discusses them in relation to the role protein structure may play in determining protein allergenicity.

Effects of CTLA4-Fc on glomerular injury in humorally-mediated glomerulonephritis in BALB/c mice.

The effect of cytotoxic T-lymphocyte-associated molecule 4-immunoglobulin fusion protein (CTLA4-Fc) on humorally-mediated glomerulonephritis was studied in accelerated anti-glomerular basement membrane (anti-GBM) glomerulonephritis induced in BALB/c mice. This strain of mice develops antibody and complement dependent glomerulonephritis under this protocol. Sensitized BALB/c mice developed high levels of circulating autologous antibody titres, intense glomerular deposition of mouse immunoglobulin and complement, significant proteinuria, renal impairment, significant glomerular necrosis and a minor component of crescent formation 10 days after challenge with a nephritogenic antigen (sheep anti-GBM globulin). Early treatment during the primary immune response, or continuous treatment throughout the disease with CTLA4-Fc, significantly suppressed mouse anti-sheep globulin antibody titres in serum, and immunoglobulin and complement deposition in glomeruli. The degree of glomerular necrosis was improved and proteinuria was reduced, particularly in the earlier stages of disease. Late treatment by CTLA4-Fc starting one day after challenge with sheep anti-mouse GBM did not affect antibody production and did not attenuate glomerulonephritis. The low level of crescent formation found in BALB/c mice developing glomerulonephritis was not prevented by the administration of CTLA4-Fc. These results demonstrate that CTLA4-Fc is of benefit in this model of glomerulonephritis by its capacity to attenuate antibody production, without affecting the minor degree of cell-mediated glomerular injury.