STAT6 Expression and IL-13 Production in Association with Goblet Cell Hyperplasia and Worm Expulsion of Gymnophalloides seoi from C57BL/6 Mice

In intestinal helminth infections, Th2 immune respones are generally associated with mucin secretion for worm expulsion from the host intestine. In particular, IL-4 and IL-13 are the important cytokines related with intestinal mucus production via STAT6 signalling in nematode infections. However, this perspective has never been studied in Gymnophalloides seoi infection. The present study aimed to observe the STAT6 signalling and cytokine responses in C57BL/6 mice, a mouse strain resistant to infection with this trematode. The results showed that worm expulsion occurred actively during days 1-2 post-infection (PI), when goblet cells began to proliferate in the small intestine. The STAT6 gene expression in the mouse spleen became remarkable from day 2 PI. Moreover, G. seoi infection induced a significant increase of IL-13 from day 4 PI in the spleen of infected mice. Our results suggested that goblet cell hyperplasia and worm expulsion in G. seoi-infected mice should be induced by STAT6 signalling, in which IL-13 may be involved as a dominant triggering cytokine.

Identificación de las células de paneth en el intestino delgado de crías de alpacas en los primeros 21 días de edad / Identification of paneth's cells in the small intestine of baby alpacas in the first 21 days of age

Las células de Paneth cumplen un rol muy importante en los mecanismos de defensa y protección del tracto gastrointestinal en diferentes especies animales a través de sus secreciones como lisozima, fosfolipasa y A2 secretoria y defensinas. El presente estudio tuvo por objetivo identificar las células de Paneth en el intestino delgado de crías de alpacas; para lo cual se utilizaron 18 animales entre 1 y 21 días de edad. Se tomaron muestras de duodeno, yeyuno e íleon de cada animal, las cuales fueron fijadas en formol al 10 por ciento y procesadas como muestras histológicas. Se prepararon láminas coloreadas con hematoxilina-eosina (H-E) y tricrómico de Masson. Mediante inmunohistoquímica se identificó la presencia de gránulos de lisozima para lo cual se utilizó un anticuerpo policlonal antilisozima. Mediante la coloración con H-E y tricrómico de Masson no se pudo identificar en forma precisa células típicas de Paneth, mientras que mediante inmunohistoquímica se observó células con presencia de gránulos de lisozima en la base de las criptas de Lieberkühn de duodeno, yeyuno e íleon desde 1 día de edad, siendo mayor el número en yeyuno e íleon en alpacas entre los 15 y 21 días de edad. Estas células presentaron áreas comprendidas entre 129.19 y 147.67 µm2, ejes mayores entre 17.96 y 19.92 µm y ejes menores entre 8.68 y 9.79 µm. Por lo tanto, se concluye que las células encontradas desde el primer día de edad en las criptas de Lieberkühn de duodeno, yeyuno e íleon de crías de alpacas son las células de Paneth, siendo mayor su número en yeyuno e ileon entre los 15 y 21 días de edad.

Paneth cells have an important function in the defense and protection mechanisms of gastrointestinal tract in many animal species through its secretions as lysozime, sycretory phospholipase A2 and defensins. The aim of this study was to identify the Paneth cells in the small intestine of baby alpacas; for this, 18 animals between 1 and 21 days of age were used for this purpose. Duodenum, jejunum and ileum samples were removed from each animal. By routinary microscopic light examination and immunohistochemistry, these samples were fixed in 10 per cent phormol and processed for paraffin sections and stained with haematoxylin-eosin (H-E) and Masson trichromic. For immunohistochemistry technique we used a polyclonal antibody anti-lizozime. Typicall Paneth cells were not identified with H-E and Masson trichromic stains, where as by immunohistochemistry technique Iysozime granules conteining cells were identified in the base of the crypts of Lieberkuhn of duodenum, jejunum and ileum since 1 day of age, being the number bigger in jejunum and ileum of alpacas between 15 and 21 days of age. These cells showed areas between 129.19 and 147.67 µm2, major axes between 17.96 and 19.92 µm and minor axes between 8.68 and 9.79 µm. Based in this results, it was concluded that the cells found since the first day of age in the crypts of Lieberkuhn of duodenum, jejunum and ileum of baby alpacas are Paneth cells, being the number bigger in jejunum and ileum between 15 and 21 days of age.

A predictive model for the level of sIgA based on IgG levels following the oral administration of antigens expressed in Sacchromyces cerevisiae

Oral vaccination may be the most efficient way of inducing an immune response at the remote mucosal site through the common mucosal immune network. Antigenspecific secretory IgA (sIgA) is the major immunoglobulin type generally detected in the secretions of experimental animals following an effective oral immunization. Actinobacillus pleuropneumoniae causing disease in the lung of pig initially interacts, colonizes, and infects the host tissues at the mucosal surface of the respiratory tract. Also, importantly for A. pleuropneumoniae protection, the quantity of sIgA in the lung had merits associated with the mucosal immunity. However, there is no simple method to monitor the level of sIgA as an indicator for the induction of local immune responses by an oral vaccination in the target tissue. Therefore, the relationship between sIgA and IgG was analyzed to evaluate the induction of local immune responses by an oral immunization with Saccharomyces cerevisiae expressing the apxIA and apxIIA genes of A. pleuropneumoniae in this study. The correlation coefficient of determination (r2 x 100) for paired samples in both vaccinated and control groups showed a significant positive-relationship between IgG in sera and sIgA in the lung or intestine. These results indicated that IgG antibody titers in sera could be useful to indirectly predict local immune response, and sIgA, in the lung or intestine to evaluate the efficacy of an oral vaccination.

Estudo das alterações das citocinas inflamatórias na rejeição aguda do transplante intestinal em ratos / Cytokine participation in the acute rejection of intestinal transplantation in rats

RACIONAL: O transplante de intestino delgado é procedimento cirúrgico em estudo visando sua aplicação no tratamento dos pacientes portadores da síndrome do intestino curto, com vistas à reabilitação oral. Porém a grande barreira se deve à "rejeição" pela grande quantidade de tecido linfóide presente no intestino delgado. OBJETIVO: Avaliar a atuação das citocinas, interleucina-6 e interferon-gama em alotransplante heterotópico intestinal. MATERIAL E MÉTODOS: Realizaram-se 24 alotransplantes intestinais em ratos da raça Brown-Norway (doador) para Lewis (receptor), sendo subdivididos em três subgrupos de oito animais, sacrificados respectivamente no terceiro dia de pós-operatório (Tx(3)), no quinto dia de pós-operatório (Tx(5)) e no sétimo dia de pós-operatório (Tx(7)) para coleta das biopsias dos enxertos intestinais. Enquanto que no grupo isotransplante (C) envolveu oito animais da raça Lewis (doador) para Lewis (receptor), porém neste grupo realizaram-se biopsias seriadas no mesmo animal, sendo subdivididos em três momentos: biopsia no terceiro dia de pós-operatório (C(3)), no quinto dia de pós-operatório (C(5)) e sacrificados no sétimo dia de pós-operatório (C(7)) para coleta da biopsia. Realizou-se inicialmente análise intragrupo entre os momentos C(3), C(5) e C(7) para todos os parâmetros de rejeição citados anteriormente, como também para os três subgrupos Tx(3), Tx(5) e Tx(7). Posteriormente, realizou-se a análise intergrupo de forma transversal e pareada comparando-se o grupo isotransplante com o grupo alotransplante (C(3) com Tx(3); C(5) com Tx(5) e C(7) com Tx(7)). RESULTADOS: No grupo isotransplante não houve diferença estatisticamente significante quanto à imunoexpressão das citocinas estudadas, todavia no grupo alotransplante observou-se que alterações da interleucina-6 e de interferon-gama ocorreram a partir do quinto dia de pós-operatório.

Protracted Diarrhea: Results of the Five-year Survey in a Tertiary Hospital in Korea

The syndrome of protracted diarrhea (PD) includes several diseases with diverse etiologies. This study was conducted to characterize the spectrum of causes, clinical manifestations, and the outcomes of PD. A retrospective analysis of the clinical and pathological findings was performed on 25 patients with diarrhea starting within the first 2 yr of life and a requirement of parenteral nutrition (PN). According to the intestinal histopathology, patients were classified into four groups: immune enteropathy (12 cases), lymphangiectasia (6 cases), epithelial dysplasia (5 cases), and unclassified (2 cases). All patients with epithelial dysplasia had earlier onset of diarrhea and longer duration of PN than those in the other groups. Three patients (12%) had an evidence of a familial condition. Five patients (three with microvillous inclusion disease and two with immune enteropathy) died. Sixteen patients recovered, and three (two with primary lymphangiectasia and one with microvillous inclusion disease) still had diarrhea. One patient underwent intestinal transplantation for tufting enteropathy. In conclusion, infants with PD should be referred to specialized centers where advanced diagnostic and therapeutic facilities are available, because histological analysis is critical for the diagnosis of PD, and PN or intestinal transplantation is the only therapeutic option in a subset of cases.

Enterocolite necrosante em neonatos a termo, lactentes e pré-escolares / Necrotizing enterocolítis in term neonates, older children and preschools

Os autores analisam 300 autopsias em que foram identificados 71 casos de enterocolite necrosante em recém-nascidos a termo e pré-escolares. Diarréia e desnutriçäo foram fatores predisponentes comumente associados. Dois tipos histopatológicos distintos da doença foram demonstrados: enterocolite necrosante difusa, cujo padräo é semelhante ao descrito em recé-nascidos pré-termo, e enterocolite necrosante pápulo-necrótica, padräo menos freqüente que o difuso, e que se apresenta principalmente como lesöes crônicas isoladas em crianças maiores. Peritonite purulenta ou fibrinosa ocorreu mais comumente por contigüidade à lesäo intestinal, sendo identificados apenas quatro casos de perfuraçäo. Constatou-se que, em analogia ao recém-nascido pré-termo com asfixia perinatal, cuja suscetivilidade a infecçöes é maior devido à imaturidade do seu sistema imunológico, diarréia crônica e desnutriçäo em recém-nascidos a termo e crianças maiores causam deficiência imunológica e alteraçäo na barreira intestinal, que associadas a hipovolemia (por desidrataçäo) e sépsis levam a isquemia e invasäo da mucosa intestinal pela flora micobacteriana local produzindo, freqüentemente, um padräo histológico distinto daquele descrito para enterocolite necrosante em recém-nascidos pré-termo de risco. Os autores chamem atençäo para a desnutriçäo como fator predisponente para o desenvolvimento da doença

Estudo das imunoglobinas intestinais na estrongiloidíase / Intestinal immunoglobulin study in clinical strongyloidiasis

Foram selecionados 32 pacientes infectados pelo Strongyloides stercoralis, sem imunodeficiência prévia ou outra doença associada. Foram divididos em três grupos: assintomáticos, sintomáticos leves e sintomáticos graves, com o objetivo de estudar a resposta imunológica do hospedeiro ao parasita. Os pacientes do grupo sintomático grave apresentavam esteatorréia, emagrecimento acentuado, albuminemia menor que 3 por cento e alteraçöes radiológicas. Doze voluntários sadios, sem verminose, formaram o grupo controle. A biópsia peroral do intestino delgado foi o método utilizado para obtençäo de fragmento da mucosa intestinal, ao nível do ângulo de Treitz. O estudo da concentraçäo das imunoglobulinas locais (IgA, IgC e IgM), pela técnica da imunofluorescência direta, demonstrou diminuiçäo significativa da concentraçäo de IgA nos pacientes sintomáticos graves e diminuiçäo da concentraçäo de IgM nos três grupos, quando comparados ao grupo controle. Näo houve alteraçöes da concentraçäo da concentraçäo de IgC

Gut associated immune responses in clinical and experimental giardiasis: an overview.

Giardia lamblia, the protozoan parasite, first described by Von Leewenhoek in 1681, has come into prominence in last quarter century because of mounting awareness that it may cause significant morbidity and loss of man power. Earlier thought to be a commensal organism, it has been recognised as a true intestinal pathogen in the past three decades. Nevertheless, the mechanism of the disease caused by this protozoan parasite (in the human host's small intestine) continues to remain unexplained. The infection with G. lamblia is worldwide with an average prevalence of 12.5 per cent and is especially common in children and may cause failure of child to thrive. The G. lamblia infection has been implicated in a number of water borne epidemics and is important cause of traveller's diarrhoea all over the world. Infection with G. lamblia may be entirely asymptomatic, may produce a mild, self limiting illness or chronic diarrhoea with or without malabsorption. The reasons for such variations in severity are not clearly understood. However, interplay of virulence of parasite, nutritional status and type of the host immune responses and its effect on intestinal mucosa appear to modulate the infection.

Functional characterization of intestinal intraepithelium & lamina propria lymphocytes from Giardia lamblia infected mice.

Quantitation of T cell subsets in intraepithelium and lamina propria during the course of experimental G. lamblia infection in the inbred mice revealed increased influx of Thy 1.2+ (T cells) and Lyt 2.2+ (suppressor/cytotoxic T cells) during the establishment (3-5 day post-inoculation) and acute (9-11 day post-inoculation) phases of infection. The influx of these cells reduced as the parasite load declined. In contrast, no significant changes were noticed in lamina propria and intraepithelium Lyt 1.1+ (helper T cell) cells during the establishment or acute phase but such cells increased significantly in the decline (17-21 day post-inoculation) phase of infection. Further, both intraepithelium and lamina propria lymphocytes isolated from uninfected or infected animals failed to kill G. lamblia trophozoites in vitro in the absence or presence of antigiardial antibodies. Our data suggest that the clearance of G. lamblia trophozoites was not mediated by cytotoxic T cells. However, the induction of helper T cells during the declining phase of infection might be an important mechanism for the induction of parasite specific antibody response leading to the immune elimination of G. lamblia trophozoites from the gut.