Intestinal Lymph Nodes Research Articles

The effect of a novel immunosuppressive drug, a PAK-2 inhibitor, on macrophage differentiation/polarization in a rat small intestinal transplantation model

ObjectivePQA-18 (Prenylated quinolinecarboxylic acid-18) has been reported to be a novel immunosuppressant that attenuates the production of various cytokines, and the differentiation of macrophages by inhibiting PAK2. In this study, we investigated the function of this drug mainly on macrophages using a rat small intestinal transplant model. MethodsMale Dark Agouti (DA) and Lewis rats (LEW), 7–9 weeks of age, were used as donor and recipient, respectively. Approximately 15 cm intestinal grafts were heterotopically transplanted to the recipient rats. The recipient rat was treated with PQA-18 (4 mg/kg/day) by intraperitoneal injection (ip) from postoperative day 1 for 2 weeks. The in vivo effects of this drug were evaluated based on changes in body weight, and the population of each type of blood cell. Mixed lymphocyte reaction (MLR) was also assessed, using the T cells from intestinal mesenteric lymph nodes (MLN) of the grafts on POD6. Total cells from MLN and graft Payer's patch (PP) were next collected on POD6, and the number of infiltrated macrophages was determined. ResultsWhile the survival time was 7.0 ± 0.77 days for the control group (n = 9), that for the PQA-18 group was 10.7 ± 1.26 days (n = 10) (p < .001). Histological examinations showed a relatively clear difference in the grafts for both groups. In addition, the MLR response was significantly lower in recipients treated with PQA-18, suggesting PQA-18 well suppressed the T cells. Moreover, while a significant increase of both MHC class II and CD11b/c positive cells, estimated as differentiated/polarized macrophages, in MLN & PP was observed in the control group, PQA-18-administration significantly suppressed the differentiation of macrophages in the MLN & PP. ConclusionPQA-18 significantly prolonged the survival of the rats with intestinal grafts, and also suppressed the infiltration of lymphocytes, and macrophages to the grafts.

Histopathological Lesions Caused by Experimental Toxocara Canis and Toxascaris Leonina Infections in Farm Mink (Neovison Vison).

IntroductionCanine roundworm T. canis and T. leonina infections were investigated in experimentally infected farm mink (Neovison vison) to describe the pattern of pathological lesions in this paratenic host.Material and MethodsMaterial and MethodsInfections in mink developed following ingestion of embryonated eggs of either parasite or mice tissue infected with both parasite species.ResultsComparative analysis of haematoxylin- and eosin-stained slides showed essential differences among the experimental groups. The lesions observed included eosinophil and mononuclear inflammatory infiltrates of the intestinal wall and local lymph nodes, inflammation and haemorrhages in liver tissues, and interstitial inflammation and mineralisation of the kidneys and lungs. Larvae migrating through the minks’ bodies also caused particularly salient enlargement of lymphoid follicles in the spleen and inflammatory infiltrates of mononuclear cells in skeletal and heart muscles.ConclusionsIt is assumed that histopathological lesions appeared as a local and general host response to invasive L3 T. canis and T. leonina larvae migrating through the tissues of infected farm mink. Interestingly, mink infected with embryonated eggs had more pronounced lesions than animals infected with tissue larvae. Detailed histopathological examinations of parenchymal organs and striated muscles revealed lesions resembling those observed in other paratenic host species due to toxocarosis.

Strain specific maturation of Dendritic cells and production of IL-1β controls CD40-driven colitis.

Intestinal integrity is maintained by balanced numbers of CD103+ Dendritic cells (DCs), which generate peripherally induced regulatory T cells (iTregs). We have developed a mouse model where DC-specific constitutive CD40 signals caused a strong reduction of CD103+ DCs in the lamina propria (LP) and intestinal lymph nodes (LN). As a consequence, also iTregs were strongly reduced and transgenic mice on the C57Bl/6-background (B6) developed fatal colitis. Here we describe that transgenic mice on a pure Balb/c-background (B/c) do not show any pathologies, while transgenic C57Bl/6 x Balb/c (F1) mice develop weak colon inflammation, without fatal colitis. This graded pathology correlated with the effects of CD40-signalling on DCs in each background, with striking loss of CD103+ DCs in B6, but reduced in F1 and diminished in B/c background. We further show direct correlation of CD103+ DC-numbers with numbers of iTregs, the frequencies of which behave correspondingly. Striking effects on B6-DCs reflected robust loss of surface MHCII, known to be crucial for iTreg induction. Furthermore, elevated levels of IL-23 together with IL-1, found only in B6 mice, support generation of intestinal IFN-γ+IL-17+ Th17 cells and IFN-γ+ Th1 cells, responsible for onset of disease. Together, this demonstrates a novel aspect of colitis-control, depending on genetic background. Moreover, strain-specific environmental sensing might alter the CD103+ DC/iTreg-axis to tip intestinal homeostatic balance to pathology.

Morphological diagnosis of Johne’s disease: a case report in cattle

The present case report deals with the morphological (gross and histopathological) and histochemical characterization of Johne?s disease in a cow. The carcass was presented in a hidebound condition with sunken eyes and with a history of persistent and nonresponsive diarrhea. Necropsy revealed generalized gelatinization of the visceral and peripheral fat. Thorough gross examination unveiled major pathological changes in the digestive tract. The gallbladder was distended, with severely thickened walls and blood mixed with bile. The small intestine was severely congested and the mesenteric lymph nodes were enlarged. The opening of the small intestine showed marked thickening of the intestinal walls, with a corrugated appearance. Diffuse granulomatous lesions were observed in the mesenteric lymph nodes. Histopathological examination revealed severe infiltration of intestinal layers and mesenteric lymph nodes by macrophages, lymphocytes, and epithelioid cells with multiple Langhans giant cells. Ziehl=Neelsen staining demonstrated multiple acid-fast bacilli in the cytoplasm of macrophages from impression smears.

Spread of an Experimental Salmonella Derby Infection in Antibiotic-Treated or Lawsonia intracellularis Vaccinated Piglets.

Simple SummaryToday, pigs repeatedly suffer from diarrhoea which requires treatment with antibiotics. Infections with Lawsonia intracellularis are one of the most common diseases, which, although usually subclinical, have a negative impact on performance. The alternative to antibiotic treatment is vaccinations against Lawsonia intracellularis, not least because antibiotic treatments are suspected of promoting the spread of certain zoonotic pathogens. A study was carried out with piglets from a farm that had a problem with Lawsonia intracellularis infections. In half of the animals, antibiotic treatments with tylosin were carried out in piglet rearing. In the other group, the piglets had been vaccinated against Lawsonia intracellularis as suckling piglets. Individual animals from both groups were subsequently artificially infected with Salmonella as piglets. A total of 72 animals were included in the study, 12 of which were primarily infected. The other animals had the possibility of becoming infected via direct animal contact or the faeces of infected animals. The detection of Salmonella in stool and intestinal lymph nodes was significantly higher in animals previously treated with antibiotics. Treatment with tylosin may significantly increase the spread of the Salmonella infection not observed after early Lawsonia intracellularis vaccination.Lawsonia intracellularis infections are a common reason for antibiotic treatment in pig production. Experimental studies in animals naturally infected with Lawsonia intracellularis comparing the course of an experimental Salmonella infection in piglets previously treated with tylosin or vaccinated against Lawsonia intracellularis are scarce. A total of 72 seven-week-old Salmonella-free pigs were taken from a herd with a Lawsonia intracellularis history in piglet rearing. The pigs were divided into two groups with three replicates each. Animals had either been previously treated with tylosin (10 mg/kg body weight) for seven days (AB+VAC−) or had been vaccinated as suckling pigs by drenching (Enterisol®Ileitis; AB−VAC+). Two animals per replicate were primarily infected with Salmonella Derby (1.04 × 108 colony-forming units per animal). The detection of Salmonella in faeces (p < 0.0001, odds ratio: 3.8364) and in the ileocaecal lymph nodes (p = 0.0295, odds ratio: 3.5043) was significantly more frequent in AB+VAC− animals. Overall, the odds ratio for detecting Salmonella in any substrate or organ was significantly higher in the AB+VAC− group animals (p = 0.0004, odds ratio: 5.9091). Treatment with tylosin can significantly increase the spread of a Salmonella infection, which is not observed after early Lawsonia intracellularis vaccination.

Abstract 4006: Serine protease PRSS8 suppresses colorectal carcinogenesis and metastasis

Abstract PRSS8 is a serine protease and has important physiological and pathophysiological functions, but its roles in cancer initiation and progression are unclear. In this study, a conditional knockout mouse model of Prss8 fl/fl,p-Villin-Cre+ mice were developed and thoroughly characterized at the ages of 3, 6 and 9 months. Intestinal epithelial cell proliferation, migration, and differentiation, were determined. The approaches of gain- and loss of expression of PRSS8 in human colon cancer cells were performed to evaluate anti-cancer and anti-metastasis in vitro and in nude mice by mouse subcutaneous injection, tail vein injection and intra-spleen transplantation. Gene profile and gene set enrichment analysis and mechanistic studies were conducted in vitro and in human colorectal cancer tissues. Results: Genetic deficiency of the Prss8 gene caused spontaneous formation of colitis and inflamed rectum at the early age, but caused intestinal tumors and lymph node metastasis at late age, which was linked to increased intestinal cell proliferation and migration, but decreased cell differentiation. Increased PRSS8 expression showed inhibition of cancer cell growth and metastasis in nude mice, and inhibited migration, invasion, colony formation and tumor sphere formation in vitro, but reduced expression of PRSS8 facilitated malignancies. Mechanistic studies revealed that PRSS8 targets Wnt/β-catenin, epithelial-mesenchymal-transition, and stem cell signaling pathways, which was supported by the results from the Cancer Genome Atlas, and were validated in human colorectal cancers. Conclusion: PRSS8 is a novel tumor suppressor that plays critical roles in the suppression of carcinogenesis and metastasis via targeting Wnt/β-catenin, EMT and stem cell signaling pathway. Citation Format: Yonghua Bao, Yongchen Guo, Yiqiong Yang, Xiaonan Wei, Xiangdong Zhu, Wei Zhang, Wancai Yang. Serine protease PRSS8 suppresses colorectal carcinogenesis and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4006.

Paratuberculose bij melk- en dwerggeiten: een onderschat probleem?

Paratuberculose is een chronische, intestinale aandoening bij geiten en andere herkauwers. Het doel van dit overzichtsartikel is om de huidige kennis omtrent prevalentie, diagnostische mogelijkheden, preventie en mogelijke controlemaatregelen tegen paratuberculose bij melk- en dwerggeiten in Europa samen te vatten. De bedrijfsprevalentie van Mycobacterium avium subspecies paratuberculosis (MAP) is in de ons omringende landen zorgwekkend hoog, namelijk 71%, 63% en 86% in Duitsland, Frankrijk en Nederland, respectievelijk. De prevalentie bij dwerggeiten is onbekend. Antistof ELISA, cultuur en PCR van mest zijn vlot beschikbaar om de diagnose te bevestigen. Een sterk suggestief letsel voor paratuberculose is de aanwezigheid van vergrote intestinale lymfeknopen op abdominale echografie. Om deze aandoening onder controle te krijgen, is een combinatie van “test and cull” en hygiënische maatregelen sterk aangeraden. In tegenstelling tot bij rundvee is vaccineren mogelijk en zou het een bijdrage kunnen leveren tot controleprogramma’s voor paratuberculose bij geiten. Vaccinantistoffen zijn niet meer detecteerbaar één jaar na vaccinatie, waardoor serologische monitoring mogelijk blijft op gevaccineerde bedrijven. De verontrustende prevalentiecijfers in aangrenzende landen tonen de dringende noodzaak van een prevalentiestudie op melkgeitenbedrijven in België aan.

Cellular Composition of the B- and T-Cell-Dependent Areas in the Small Intestine during the Post-Stress Period (Experimental Study).

A quantitative study of lymphoid cells in the B- and T-cell-dependent areas of intestinal lymphoid nodules and mesenteric lymph nodes in behaviorally passive and active rats was performed at various periods after acute stress on the model of 1-h immobilization with simultaneous electrocutaneous stimulation. Stress exposure is accompanied by a decrease in the number of lymphoid cells in immunogenic structures of the gastrointestinal tract. Post-stress changes in the cytoarchitectonics of B- and T-cell-dependent areas in mesenteric lymph nodes of animals are less pronounced than in lymphoid nodules. Quantitative changes in lymphoid cells of B-cell-dependent areas in the small intestine of rats are greater than in T-cell-dependent areas. Changes in the cellular composition of immunogenic structures in the digestive system are most significant at the early stages of the post-stress period (1st week). Passive rats are characterized by significant changes in the cytoarchitectonics of B- and Tcell-dependent areas in the small intestine after extreme exposure, which illustrates functional exhaustion of the lymphoid tissue in stress-predisposed specimens.

Vaccine potential of antigen cocktails composed of recombinant Toxoplasma gondii TgPI-1, ROP2 and GRA4 proteins against chronic toxoplasmosis in C3H mice

The development of an effective and safe vaccine to prevent Toxoplasma gondii infection is an important aim due to the great clinical and economic impact of this parasitosis. We have previously demonstrated that immunization with the serine protease inhibitor-1 (TgPI-1) confers partial protection to C3H/HeN and C57BL/6 mice. In order to improve the level of protection, in this work, we combined this novel antigen with ROP2 and/or GRA4 recombinant proteins (rTgPI-1+rROP2, rTgPI-1+rGRA4, rTgPI-1+rROP2+rGRA4) to explore the best combination against chronic toxoplasmosis in C3H/HeN mice. All tested vaccine formulations, administered following a homologous prime-boost protocol that combines intradermal and intranasal routes, conferred partial protection as measured by the reduction of brain cyst burden following oral challenge with tissue cysts of Me49 T. gondii strain. The highest level of protection was achieved by the mixture of rTgPI-1 and rROP2 proteins with an average parasite burden reduction of 50% compared to the unvaccinated control group. The vaccine-induced protective effect was related to the elicitation of systemic cellular and humoral immune responses that included antigen-specific spleen cell proliferation, the release of Th1/Th2 cytokines, and the generation of antigen-specific antibodies in serum. Additionally, mucosal immune responses were also induced, characterized by secretion of antigen-specific IgA antibodies in intestinal lavages and specific mesenteric lymph node cell proliferation. Our results demonstrate that rTgPI-1+rROP2 antigens seem a promising mixture to be combined with other immunogenic proteins in a multiantigenic vaccine formulation against toxoplasmosis.

The tumor suppressor phosphatase PP2A-B56α regulates stemness and promotes the initiation of malignancies in a novel murine model.

Protein phosphatase 2A (PP2A) is a ubiquitously expressed Serine-Threonine phosphatase mediating 30–50% of protein phosphatase activity. PP2A functions as a heterotrimeric complex, with the B subunits directing target specificity to regulate the activity of many key pathways that control cellular phenotypes. PP2A-B56α has been shown to play a tumor suppressor role and to negatively control c-MYC stability and activity. Loss of B56α promotes cellular transformation, likely at least in part through its regulation of c-MYC. Here we report generation of a B56α hypomorph mouse with very low B56α expression that we used to study the physiologic activity of the PP2A-B56α phosphatase. The predominant phenotype we observed in mice with B56α deficiency in the whole body was spontaneous skin lesion formation with hyperproliferation of the epidermis, hair follicles and sebaceous glands. Increased levels of c-MYC phosphorylation on Serine62 and c-MYC activity were observed in the skin lesions of the B56αhm/hm mice. B56α deficiency was found to increase the number of skin stem cells, and consistent with this, papilloma initiation was accelerated in a carcinogenesis model. Further analysis of additional tissues revealed increased inflammation in spleen, liver, lung, and intestinal lymph nodes as well as in the skin lesions, resembling elevated extramedullary hematopoiesis phenotypes in the B56αhm/hm mice. We also observed an increase in the clonogenicity of bone marrow stem cells in B56αhm/hm mice. Overall, this model suggests that B56α is important for stem cells to maintain homeostasis and that B56α loss leading to increased activity of important oncogenes, including c-MYC, can result in aberrant cell growth and increased stem cells that can contribute to the initiation of malignancy.

Progress of the application of optical coherence tomography in gastrointestinal tumor surgery

Optical coherence tomography (OCT) is a real-time, cross-sectional optical imaging technology. It is analogous to ultrasonography, except that OCT uses light waves instead of sound waves, and can provide three-dimensional morphological images of living tissues with a micrometer resolution. Through the use of endoscopes, needles, catheters and laparoscopes, OCT has demonstrated tremendous imaging potential in tumor surgery. The current studies suggest that OCT has potential for clinical applications in the following fields of gastrointestinal tumor surgery: (1) Early tumor detection and diagnosis: OCT can distinguish differences between polyp tissue, normal tissue and malignant tissue. It could possibly identify premalignant lesions or conditions potentially predisposing to malignancy, such as gastric and intestinal metaplasia, gastritis associated with Helicobacter pylori, and early gastric cancer involving the mucosa or submucosa. In addition, OCT can differentiate between adenomatous polyps and hyperplastic polyps. (2) Optical biopsy of lymph nodes: As a high-resolution, near-IR imaging modality, OCT is capable of visualizing microscopic features within tissue, distinguishing lymph node tissue from surrounding adipose tissue, revealing nodal structures such as germinal centers and intra-nodal vessels. Consequently, OCT has the ability to show changes in node microarchitecture during metastatic tumor infiltration. (3) Intraoperative guidance for real-time determination of surgical margins: In other tumors such as oral squamous cell carcinoma and breast cancer, it has been demonstrated that OCT can be used to rapidly scan large areas of tissue, to guide at the cellular level the surgical resection of neoplastic disease, and to scan tumor margins for the presence of residual disease, tumor foci, and potentially even metastasizing tumor cells. It implies that colorectal neoplasms surgeons can possibly use the laparoscopic OCT to detect the intestinal tumor margin and lymph nodes during operation in the future, so as to determine the appropriate range of bowel resection and lymph node dissection. At present, there are few reports about the intra-operative application of OCT in the field of gastrointestinal tumor surgery. Thus there is a tremendous opportunity for further research in this field.

Molecular characterization and expression analysis of the NLR family CARD containing five transcripts in the pig.

The NOD-like receptor (NLR) family caspase recruitment domain-containing 5 (NLRC5) is one of the newly discovered and largest NLR family members. The NLRC5 has recently received extensive attention because of its important role in regulating innate and adaptive immune responses. The NLRC5 in many vertebrates, such as humans, mice, cattle, and horses, has already been proven and studied. However, the NLRC5 gene characteristics of pigs remain unclear. Thus, we completely cloned the NLRC5 cDNA sequence of the pig using the rapid amplification of cDNA ends(RACE) technology. A characteristic and tissue expression analysis was also conducted on the pig sequence. The sequence analysis showed that the complete cDNA sequence of the NLRC5 of the pig is 6638 bp, and the open reading frame is 5538 bp which encoded 1846 amino acids. The protein prediction analysis indicates that the overall performance of the NLRC5 protein of the pig is hydrophilic and possesses a typical nucleotide binding and oligomerization domain(NBD) and 20 leucine-rich repeats(LRRs). The homology analysis result indicates that the NLRC5 transcript in pigs is highly homologous to cattle, sheep, macaques, and humans, and accounts for around 80%. The genetic evolutionary tree analysis shows that the NLRC5 transcript in pigs has the closest evolutionary relationship with cattle and sheep. Further tissue expression analysis shows that immune organ systems (e.g., lymph node and spleen) and mucosa organs (e.g., intestinal lymph node, stomach, and lungs) possess high expressions with NLRC5 mRNA. The result of this study indicates that the NLRC5 transcript in pigs is relatively conservative among mammals and may play a vital role in immune reaction, which provides a basis for further studies on the NLRC5 function in the pig immune system and the role in comparative immunity.

ГІСТОЛОГІЧНІ ЗМІНИ В ТЕЛЯТ ЗА ГОСТРОГО ПЕРЕБІГУ САЛЬМОНЕЛЬОЗУ

The results of study of histological changes in calves by age of 20 – 40 days at acute salmonellosis infection are presented. At a histological research the most expressed changes were registered in the liver, thin intestine and lymphoid organs that contingently reproduction here of the causative agent. At acute form of salmonellosis in calves of this age-grade characteristic histological changes include: submilliar, intralobular, coagulativ, areactive foci of necrosis in a liver; cell granuloma in a liver; the acute diffusive serous-catarrhal enteritis with a hemorrhagic accent; serous lymphadenitis of intestinal lymph nodes; the acute diffusive catarrhal abomasitis. Also nonspecific, but constant morphological signs of salmonellosis are hyperplasia Peyer’s patches of a small intestine, intestinal lymph nodes, spleen and dystrophic changes of parenchymal organs.An analysis and theoretical ground of pathogenetic mechanisms of the educed microstructural changes are conducted. It is shown that in pathogenesis of salmonellosis an important role is played bacteriaemia and toxinaemia, which result to the defeat of endothelia of vessels and blood cells with subsequent violation of mechanisms of haemostasis гand development of the infectiously-toxic shock.

Hepatic tuberculosis with the development of fibrosis and concomitant steatohepatitis

Tuberculous lesions of the liver frequently occur in autopsy material, even though most of them are clinically unapparent. Pathogenetically, hepatic tuberculosis is a manifestation of general military tuberculosis with the infection brought in through the hematogenous route (through the hepatic artery or the portal system) as a result of intestinal or mesenteric lymph node tuberculosis. We report a case of combination of hepatic tuberculosis and diabetes mellitus in a young woman with reference to the clinical course of the disease, its diagnostics and treatment.

Characterization of tuberculous granulomas in different stages of progression and associated tertiary lymphoid tissue in goats experimentally infected with Mycobacterium avium subsp. hominissuis.

Oral infection of goats with Mycobacterium avium subsp. hominissuis (MAH) resulted in a large variety of granulomas in organized gut-associated lymphatic tissues and intestinal lymph nodes. To characterize the cellular composition of granulomas, CD4(+), CD8(+), γδ, B lymphocytes and plasma, CD25(+), CD68(+), MHC-II(+), Ki67(+) and endothelial cells were labeled in consecutive frozen sections by immunohistochemistry and acid fast bacilli (AFB) by Kinyoun stain. Granulomas with extensive necrosis, little mineralization and variable numbers of AFB surrounded by many CD4(+) T cells, but only few epitheloid macrophages were observed in severely sick goats at 2-3mpi. They were interpreted as exuberant immune reaction. Organized granulomas with very few AFB were seen in clinically healthy goats at 13mpi. The necrotic cores were surrounded by a zone of granulomatous infiltrate with many epitheloid macrophages and few lymphocytes. This zone was initially wide and highly vascularized and became progressively smaller. It was enclosed by an increasing layer of connective tissue. All organized granulomas were surrounded by compartimentalized tertiary lymphoid tissue. The granulomas in experimental infection of goats with MAH reflect the heterogeneity of lesions seen in mycobacterial infections of humans and ruminants and are therefore valuable for comparative research.

Listeria monocytogenes in Different Specimens from Healthy Red Deer and Wild Boars.

In the past, Listeria monocytogenes has been isolated from game feces and meat. However, less information is available on the occurrence of L. monocytogenes in other specimens originating from game animals. Hence, the aim of this study was to get an overview of the occurrence and distribution of L. monocytogenes in game animals by characterization of isolates from different matrices. For that purpose, samples were collected from red deer (Cervus elaphus), wild boars (Sus scrofa), and feed during the hunting season 2011-2012 in three different regions of Germany and Austria. Six samples from each animal were examined: tonsils, content of the rumen or the stomach, liver, intestinal lymph nodes, cecum content, and feces. Nineteen of 45 red deer and 12 of 49 wild boars were found to be positive for L. monocytogenes as well as 4 of 22 pooled feed samples. L. monocytogenes was isolated most frequently from the rumen of red deer (14 of 19) and the tonsils of wild boars (7 of 12). Serotypes 1/2a, 1/2b, 4a, and 4b were detected in samples of game animals and feed, and serotypes 1/2a and 4b were the most prevalent serotypes. The presence of L. monocytogenes serotype 4a had not yet been described in red deer. This might be due to the fact that it was only isolated from the content of rumen and that no other study has yet examined ruminal content. Pulsed-field gel electrophoresis showed a wide variety of strains. Some strains occurred in both species and feed samples, but one strain was dominant in one region. The results show that red deer and wild boars can be carriers of L. monocytogenes in different matrices, although the feces samples can be negative.

Transcript analysis of a goat mesenteric lymph node by deep next-generation sequencing.

Deep RNA sequencing (RNA-seq) provides a practical and inexpensive alternative for exploring genomic data in non-model organisms. The functional annotation of non-model mammalian genomes, such as that of goats, is still poor compared to that of humans and mice. In the current study, we performed a whole transcriptome analysis of an intestinal mucous membrane lymph node to comprehensively characterize the transcript catalogue of this tissue in a goat. Using an Illumina HiSeq 4000 sequencing platform, 9.692 GB of raw reads were acquired. A total of 57,526 lymph transcripts were obtained, and the majority of these were mapped to known transcriptional units (42.67%). A comparison of the mRNA expression of the mesenteric lymph nodes during the juvenile and post-adolescent stages revealed 8949 transcripts that were differentially expressed, including 6174 known genes. In addition, we functionally classified these transcripts using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) terms. A total of 6174 known genes were assigned to 64 GO terms, and 3782 genes were assigned to 303 KEGG pathways, including some related to immunity. Our results reveal the complex transcriptome profile of the lymph node and suggest that the immune system is immature in the mesenteric lymph nodes of juvenile goats.

Gastrointestinal granuloma due to Candida albicans in an immunocompetent cat

A 3.5 year-old cat was admitted to the University of Melbourne Veterinary Teaching Hospital for chronic vomiting. Abdominal ultrasonography revealed a focal, circumferential thickening of the wall of the duodenum extending from the pylorus aborally for 3cm, and an enlarged gastric lymph node. Cytology of fine-needle aspirates of the intestinal mass and lymph node revealed an eosinophilic inflammatory infiltrate and numerous extracellular septate acute angle branching fungal-type hyphae. Occasional hyphae had globose terminal ends, as well as round to oval blastospores and germ tubes. Candida albicans was cultured from a surgical biopsy of the duodenal mass. No underlying host immunodeficiencies were identified. Passage of an abrasive intestinal foreign body was suspected to have caused intestinal mucosal damage resulting in focal intestinal candidiasis. The cat was treated with a short course of oral itraconazole and all clinical signs resolved.

SOLiD SAGE sequencing shows differential gene expression in jejunal lymph node samples of resistant and susceptible red deer (Cervus elaphus) challenged with Mycobacterium avium subsp. paratuberculosis

This study compared in vivo lymph node gene expression levels between six young red deer that were either relatively resistant (R) or susceptible (S) to paratuberculosis following experimental challenge with Mycobacterium avium subsp. paratuberculosis. Intestinal lymph nodes were biopsied at 4, 12 and 50 weeks post challenge (pc) and parallel changes in histopathology, immunology and bacterial load monitored.SOLiD SAGE (serial analysis of gene expression) next generation sequencing of biopsied lymph node samples generated a total of 373 million transcript tags 26–28bp in length after filtering. A total of 36,632 unique transcripts were identified and 14,325 of these were able to be annotated. The copy number of each transcript was counted, averaged and compared for R and S animals (R–S). P values and False Discovery Rates (FDR) were calculated for each transcript. Genes differentially upregulated ≥2 fold (FDR<0.5) totalled 9, 40 and 32 in R animals (+ values) and 23, 164 and 47 in S animals (− values) at weeks 4, 12, and 50pc, respectively.Transcripts displaying greatest differential expression between R and S animals at each time point were IFIT2 (189 fold) and S100A8 (−32.7 fold) at week 4, LRR1 (52.7 fold), SERPINF2 (−214.6 fold) at week 12 and CEACAM8 (84.6 fold), and STK31 (−129.5 fold) at week 50, respectively.All 9 genes significantly upregulated at week 4 in R animals relate specifically to host defence and all involve Type I interferon stimulated genes. By contrast genes upregulated in S animals at week 4, relate predominantly to inflammation, but also involve adaptive immune responses, mitochondrial function and apoptosis regulation.At week 12, the genes differentially upregulated in R animals are linked predominantly to regulation of adaptive immunity and mucosal immunity, while many of the genes in S animals are associated with pro-inflammatory interleukins involved with innate and adaptive immunity. These correlated with greater lesion severity and higher MAP numbers in lymph nodes of S animals.By week 50 the number of upregulated genes declined in both groups. A number of genes upregulated in R animals appear to be associated with host resistance and regulation of adaptive immunity, especially CEACAM8. Genes upregulated in S animals involve antigen presentation (ENDOD1) and gut associated immune pathology (HSH2D).In conclusion, gene expression in jejunal lymph nodes of resistant and susceptible deer infer that the resistant phenotype is associated with pathways of adaptive immunity, while susceptibility is linked with upregulated non-protective pro-inflammatory responses, following experimental MAP infection.

Chronic alcohol increases CD8+ T-cell immunosenescence in simian immunodeficiency virus-infected rhesus macaques

Activated CD8+ T-cells correlate with viral load and may foretell antiretroviral therapy (ART) failure. HIV infection has been suggested to accelerate immunosenescence through chronic persistent inflammation. Alcohol-use disorders (AUD) are prevalent in persons living with HIV/AIDS (PLWHA). We tested the hypothesis that hazardous alcohol consumption accelerates immune activation and immunosenescence. Immune activation and immunosenescence were examined in CD8+ T lymphocytes (CD3+CD4−CD8+) isolated from intestinal biopsies, axillary lymph nodes, and peripheral blood mononuclear cells (PBMCs) of chronic binge alcohol (CBA)-consuming simian immunodeficiency virus (SIV)-infected male rhesus macaques with and without antiretroviral therapy (ART; CBA/ART+, CBA/ART−) and in PBMCs isolated from a cohort of PLWHA. Polychromatic flow cytometry was used to phenotype cells isolated from intestinal biopsies, lymph nodes, and peripheral blood from rhesus macaques and PLWHA. The Alcohol Use Disorders Identification Test (AUDIT) identified hazardous alcohol drinking in PLWHA. Viral load was determined by RT-qPCR and telomere length was measured using qPCR. PBMC CD8+ T-cell activation (CD38+HLA-DR+) and immunosenescence (CD28−) were increased over baseline levels (857% ± 334, p < 0.05; 398% ± 80, p < 0.05, respectively) only in CBA animals not receiving ART. Viral load correlated with CD8+ T-cell immunosenescence in macaque PBMCs (rs = 0.49, p = 0.02). Activated immunosenescent T-cell (CD8+CD38+CD28−) frequencies in PBMCs from PLWHA significantly correlated with AUDIT scores (rs = 0.75, p = 0.001), while no correlation was observed with CD4+ T-cell and AUDIT scores (rs = −0.24, p = 0.38). Activated immunosenescent T-cells had shorter telomeres than CD8+ T-cells (CD8+CD28+) from PLWHA. Our results suggest that CBA and AUD augment immune activation and immunosenescence in SIV-infected macaques and PLWHA.