Bronchial Nodes Research Articles

Phenotypic characterization of a highly pathogenic Italian porcine reproductive and respiratory syndrome virus (PRRSV) type 1 subtype 1 isolate in experimentally infected pigs

Highly pathogenic (HP) isolates of the PRRS virus started emerging in North America and Asia in the late 1990s. More recently, they have emerged in Europe. These isolates are characterized by high viral loads, severe general clinical signs and high mortality, in sows, weaners and growers. Their genome shows a discontinuous aminoacids deletion in the non-structural protein 2 (NSP2). The present study was aimed at characterizing the clinical, pathological and immunological features of a highly pathogenetic, Italian PRRSV-1 subtype 1 isolate (PRRSV1_PR40/2014), following experimental infection in conventional 4-weeks-old pigs. The PRRSV1_PR40/2014 infected group showed severe clinical signs (high fever and dispnoea). Pathological lesions, including severe lymphocytopenia in bronchial lymph-nodes and thymus were also recorded. Higher serum PRRSV genome copies and lower virus neutralizing antibody titer were observed in the PR40 group, when compared to the group infected with a conventional PRRSV strain. The genetic analysis of the strain, and the phenotypic features observed in the field and reproduced in the experimental study, confirmed the high pathogenicity of the Italian PRRSV-1 subtype 1 PR40 isolate.

Ochratoxin A-induced autophagy in vitro and in vivo promotes porcine circovirus type 2 replication

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium. Porcine circovirus type 2 (PCV2) is recognized as the causative agent of porcine circovirus-associated diseases. Recently, we reported that low doses of OTA promoted PCV2 replication in vitro and in vivo, but the underlying mechanism needed further investigation. The present studies further confirmed OTA-induced PCV2 replication promotion as measured by cap protein expression, viral titer, viral DNA copies and the number of infected cells. Our studies also showed that OTA induced autophagy in PK-15 cells, as assessed by the markedly increased expression of microtubule-associated protein 1 light chain 3 (LC3)-II, autophagy-related protein 5 (ATG5), and Beclin-1 and the accumulation of green fluorescent protein (GFP)-LC3 dots. OTA induced complete autophagic flux, which was detected by monitoring p62 degradation and LC3-II turnover using immunoblotting. Inhibition of autophagy by 3-methylademine (3-MA) and chloroquine (CQ) significantly attenuated OTA-induced PCV2 replication promotion. The observed phenomenon was further confirmed by the knock-down of ATG5 or Beclin-1 by specific siRNA. Further studies showed that N-acetyl-L-cysteine (NAC), an ROS scavenger could block autophagy induced by OTA, indicating that ROS may be involved in the regulation of OTA-induced autophagy. Furthermore, we observed significant increases in OTA concentrations in lung, spleen, kidney, liver and inguinal lymph nodes (ILN) and bronchial lymph nodes (BLN) of pigs fed 75 and 150 μg/kg OTA compared with controls in vivo. Administration of 75 μg/kg OTA significantly increased PCV2 replication and autophagy in the lung, spleen, kidney and BLN of pigs. Taken together, it could be concluded that OTA-induced autophagy in vitro and in vivo promotes PCV2 replication.

Гістологічні зміни в котів за хламідіозу

The article presents the results of the study of histological changes in organs and tissues of cats for chlamydial infection. The histopathological examination of the postmortem cases of cats (n = 8) of different breeds between the ages of 3 to 6 years old, who lives with the laboratory methods have been diagnosed and identified the pathogen Chlamydia felis. According to historical data from sick animals were recorded various nature and degree of conjunctivitis and pronounced signs of a lesion of the respiratory tract (rhinitis, bronchitis, pneumonia). The made histological sections were stained with hematoxylin and eosin according to routine standard methods. The general histological structure and microstructural changes in histological preparations were studied under a light microscope. The histopathological examination were confirmed and specified the pathoanatomical diagnoses, established after autopsy of dead animals. It is shown that the most pronounced damage and characteristic changes all dead cats fixed contact in lung tissues and regional lymph nodes (mediastinal and bronchial), and in the spleen. The morphological criteria of chlamydial infection in the studied dead animals at the microscopic level were as follows:1) interstitial pneumonia; 2) pulmonary fibrosis; 3) fibrinous-purulent pleuropneumonia; 4) hyperplasia and serous lymphadenitis of the mediastinal and bronchial lymph nodes; 5) hyperplasia of lymphoid nodules of the spleen; 6) passive venous congestion of the liver and kidneys; 7) fatty and granular liver dystrophy. The results of our study, the pathomorphological diagnosis of chlamydial infection in most of the cats that died was based on changes characteristic of interstitial pneumonia followed pneumosclerosis or fibrinous-purulent pleuropneumonia, with deep affection of the bronchial epithelium. Consequently, the features of chlamydial infection are such that the clinical picture does not always correspond to the severity of morphological manifestations and complications of infection. Weak clinical manifestations can be combined with significant destructive, degenerative and necrotic changes in organs and tissues.

Genetic characterization of porcine circovirus type 2 in captive wild boars in southern Brazil.

Porcine circovirus type 2 (PCV2) has been identified in pig population in Brazil since 2000, but scarce studies involving wild boars with PCV2 infection are reported in the country. This study aimed to perform the genetic characterization of PCV2 detected in clinically healthy captive wild boars from farms located in Southern Brazil. Bronchial and mesenteric lymph nodes from 129 clinically healthy captive wild boars were tested by nested PCR for PCV2 detection. Six out of 38 positive samples (29.5%) were submitted to a quantitative real time PCR (qPCR) and genetic sequencing. Viral load up to 1.19×109 viral DNA copies/uL was detected in lymph nodes samples by qPCR. According to the ORF2 gene sequence analysis, all PCV2 samples were classified into PCV2b genotype. Comparisons based on a 702nt region of the ORF2 of all six isolates revealed a high degree of similarity between these isolates. The ORF2 sequences characterized here share 97.1-100% of nucleotide identity and 95.7-100% of amino acid identity with other PCV2b isolated in Brazil from wild boars and feral pigs. This study reports the first detection and genetic characterization of PCV2b in captive wild boars in Brazil and provides important information on PCV2 infection in this domesticated species.

Патоморфологічна характеристика хламідіозу в котів

The results of the study of the morphological features of pathological processes in organs and tissues of dead cats from chlamydial infection. A study of cadavers (n = 8), cats of different breeds aged from 3 to 6 years old, who lives with the laboratory methods have been diagnosed and identified the pathogen Chlamydia felis. According to historical data from sick animals were recorded various nature and degree of conjunctivitis and pronounced signs of a lesion of the respiratory tract (rhinitis, bronchitis, pneumonia). The main research method was mortem examination, during which the fixed and described the macroscopic changes in the affected organs and tissues. The autopsy was carried out partial evisceration, while selected pathological material for further histological studies. It is shown that the most pronounced damage and characteristic macroscopic changes all dead cats fixed contact in lung tissues and regional lymph nodes (mediastinal and bronchial), and in the spleen. Macroscopic picture of lungs affected varied somewhat depending on the degree of damage and stages of morphogenesis. In most cases (n = 6) lesions in the lungs looked like multiple well-defined lesions that are localized in all lobes of the lungs. Morphological manifestations of chlamydial infection in the investigated dead cats on the macroscopic level following features are: 1) productive inflammation in the lungs in a subtotal and total interstitial pneumonia and pulmonary fibrosis; 2) catarrhal bronchitis; 3) hyperplasia and serous lymphadenitis of the mediastinal and bronchial lymph nodes; 4) hyperplasia of lymphoid nodules of the spleen; 5) serous conjunctivitis. Also, a disease accompanied by nonspecific general pathological processes, such as: passive venous congestion of the liver and kidneys; degenerative changes in the liver; dilation of the right ventricle of the heart, which occurs as a complication of the underlying disease. While studies of other organs and tissues of dead animals we have not found any macroscopic changes.

B cell subsets are modulated during allergic airway inflammation but are not required for the development of respiratory tolerance in a murine model.

Allergic asthma is a widespread chronic inflammatory disease of the airways. The role of different B cell subsets in developing asthma and respiratory tolerance is not well known. Especially regulatory B (Breg) cells are proposed to be important in asthma regulation. Using wild-type (WT) and B cell-deficient (μMT) mice we investigated how B cells are affected by induction of allergic airway inflammation and respiratory tolerance and whether they are necessary to develop these conditions. WT mice with an asthma-like phenotype, characterized by increased airway hyper reactivity, eosinophilic airway inflammation, mucus hypersecretion and elevated Th2 cytokines, exhibited increased MHCII and CD23 expression on follicular mature B cells in lung, bronchial lymph nodes (bLN) and spleen, which contributed to allergen-specific T cell proliferation in vitro. Germinal center B cell numbers were elevated and associated with increased production of allergen-specific immunoglobulins especially in bLN. In contrast, respiratory tolerance clearly attenuated these B cell alterations and directly enhanced marginal zone precursor B cells, which induced regulatory T cells in vitro. However, μMT mice developed asthma-like and tolerized phenotypes like WT mice. Our data indicate that although B cell subsets are affected by asthma-like and respiratory tolerant phenotypes, B cells are not required for tolerance induction.

SIV Infection Facilitates Mycobacterium tuberculosis Infection of Rhesus Macaques.

Tuberculosis (TB) is a common opportunistic infection and the leading cause of death for human immunodeficiency virus (HIV)-infected patients. Thus, it is necessary to understand the pathogenetic interactions between M.tb and HIV infection. In this study, we examined M.tb and/or simian immunodeficiency virus (SIV) infection of Chinese rhesus macaques. While there was little evidence that M.tb enhanced SIV infection of macaques, SIV could facilitate M.tb infection as demonstrated by X-rays, pathological and microbiological findings. Chest X-rays showed that co-infected animals had disseminated lesions in both left and right lungs, while M.tb mono-infected animals displayed the lesions only in right lungs. Necropsy of co-infected animals revealed a disseminated M.tb infection not only in the lungs but also in the extrapulmonary organs including spleen, pancreas, liver, kidney, and heart. The bacterial counts in the lungs, the bronchial lymph nodes, and the extrapulmonary organs of co-infected animals were significantly higher than those of M.tb mono-infected animals. The mechanistic studies demonstrated that two of three co-infected animals had lower levels of M.tb specific IFN-γ and IL-22 in PBMCs than M.tb mono-infected animals. These findings suggest that Chinese rhesus macaque is a suitable and alternative non-human primate model for SIV/M.tb coinfection studies. The impairment of the specific anti-TB immunity is likely to be a contributor of SIV-mediated enhancement M.tb infection.

Expression of BSE-associated proteins in the CNS and lymphoreticular tissues of cattle and buffalo

Bovine spongiform encephalopathy (BSE), which has been documented in 190,000 cases of BSE-infected cattle, to our knowledge, has not been reported in buffalo. Prion protein (PrP) is critical to susceptibility and development of BSE. Moreover, a new PrP-like protein, Shadoo (Sho), has been shown to have overlapping expression patterns and shared functions with PrP. Therefore, we hypothesize that differences in expression at the transcriptional level and/or the post-transcriptional level of the two genes may be associated with these specific differences between cattle and buffalo. We compared the relative mRNA expression of the prion protein gene (PRNP) and the Shadoo gene (SPRN) in 396 RNA samples using real-time PCR. We also analyzed PrP/Sho protein from 96 samples by Western blot. Our results demonstrated significantly lower PrP expression in the cerebellum, obex, mesenteric lymph node, and bronchial lymph node tissues, but higher relative expression of Sho in the cerebrum and spleen in buffalos compared with cattles. Although these results support our primary assumption, Sho and PrP expressions did not correlate with corresponding mRNA expression, suggesting that the biological modulations of both PrP and Sho proteins are at post-translational levels. Moreover, positive correlations between PRNP and SPRN were found in cattle and buffalo cerebrum as well as in buffalo obex. Conversely, negative correlations between PrP and Sho were detected in buffalo cerebellum and obex tissues. These findings suggest that additional post-transcriptional studies are warranted to elucidate mechanisms behind prion diseases.

Mediastinoscope and laparoscope-assisted esophagectomy.

Mediastinoscope-assisted transhiatal esophagectomy (MATHE) is a minimally invasive option for thoracic esophageal cancer with the potential benefit of decreasing pulmonary complications by avoiding one-lung ventilation or a transthoracic procedure. However, the conventional MATHE procedure is less radical than transthoracic esophagectomy due to operative view limitations and insufficient mediastinal lymphadenectomy. In upper mediastinal dissection, the conventional MATHE procedure only provides esophageal mobilization with or without lymph node sampling. We developed a novel MATHE procedure with en bloc mediastinal lymphadenectomy by introducing a single-port laparoscopic technique. The patient was placed in a supine position with bilateral lung ventilation. The upper mediastinal dissection, using a left cervical approach, was performed with a single-port mediastinoscopic technique. A laparoscope was used as a 'mediastinoscope'. The lymph nodes along the right recurrent laryngeal nerve (RLN) were dissected under direct vision using a right cervical approach. Bilateral cervical approaches were followed by hand-assisted laparoscopic transhiatal esophagectomy. A single-port technique provides a favorable expansion of the mediastinal space by carbon dioxide insufflation, and improves the visibility and handling in the deep mediastinum around the aortic arch, allowing for en bloc lymphadenectomy in the upper mediastinum including the subaortic arch lymph nodes. In addition, a hand-assisted laparoscopic transhiatal procedure allows for en bloc lymphadenectomy in the middle and lower mediastinum including the subcarinal and bilateral main bronchial lymph nodes. Cervical and transhiatal procedures were performed safely and carefully under video-assisted magnified vision according to the standardized procedure with an appropriate operative field expansion using retractors. Single-port MATHE is feasible as a novel minimally invasive surgery for esophageal squamous cell carcinoma (ESCC) or thoracic esophageal cancer.

RNA expression of TLR10 in normal equine tissues.

BackgroundToll like receptors are one of the major innate immune system pathogen recognition systems. There is little data on the expression of the TLR10 member of this family in the horse.ResultsThis paper describes the genetic structure of the Equine TLR10 gene and its RNA expression in a range of horse tissues. It describes the phylogenetic analysis of the Equine TLR1,6,10,2 annotations in the horse genome, firmly identifying them in their corresponding gene clades compared to other species and firmly placing the horse gene with other TLR10 genes from odd-toed ungulates. Additional 3’ transcript extensions to that annotated for TLR10 in the horse genome have been identified by analysis of RNAseq data. RNA expression of the equine TLR10 gene was highest in peripheral blood mononucleocytes and lymphoid tissue (lymph nodes and spleen), however some expression was detected in all tissues tested (jejunum, caudal mesenteric lymph nodes, bronchial lymph node, spleen, lung, colon, kidney and liver). Additional data on RNAseq expression of all equine TLR genes (1–4 and 6–10) demonstrate higher expression of TLR4 than other equine TLRs in all tissues.ConclusionThe equine TLR10 gene displays significant homology to other mammalian TLR10 genes and could be reasonably assumed to have similar fuctions. Its RNA level expression is higher in resting state PBMCs in horses than in other tissues.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-2161-9) contains supplementary material, which is available to authorized users.

Blockade of STAT3 in T Cells Inhibits Germinal Center Reactions against Intranasal Allergens.

Understanding the developmental mechanisms of humoral immunity against intranasal antigens is essential for the development of therapeutic approaches against air-borne pathogens as well as allergen-induced pulmonary inflammation. Follicular helper T (Tfh) cells expressing CXCR5 are required for humoral immunity by providing IL-21 and ICOS costimulation to activated B cells. However, the regulation of Tfh cell responses against intranasal antigens remains unclear. Here, we found that the generation of Tfh cells and germinal center B cells in the bronchial lymph node against intranasal proteinase antigens was independent of TGF-β. In contrast, administration of STAT3 inhibitor STA-21 suppressed the generation of Tfh cells and germinal center B cells. Compared with wild-type OT-II T cells, STAT3-deficient OT-II T cells transferred into recipients lacking T cells not only showed significantly reduced frequency Tfh cells, but also induced diminished IgG as well as IgE specific for the intranasal antigens. Co-transfer study of wild-type OT-II and STAT3-deficient OT-II T cells revealed that the latter failed to differentiate into Tfh cells. These findings demonstrate that T cell-intrinsic STAT3 is required for the generation of Tfh cells to intranasal antigens and that targeting STAT3 might be an effective approach to ameliorate antibody-mediated pathology in the lung.

Bilobectomy in non-small cell lung carcinoma: an analysis of indications and outcome with a review of the literature

Background: The gold standard of treatment for non-small cell lung cancer (NSCLC) is anatomic pulmonary resection. Wider resection methods may be preferred due to the size or anatomic location of the tumor. One method of choice is bilobectomy due of the anatomy of the right lung. Materials and Methods: This study retrospectively analyzed 93 patients who were diagnosed with NSCLC and had bilobectomy, complete resection, and mediastinal lymph node dissection at our center between January 2005 and April 2013. Results: Forty-seven patients underwent superior bilobectomy (sBL), and 46 patients underwent inferior bilobectomy (iBL). Bilobectomy was performed due to fissure invasion in 51 (58.1%) patients, internal or external bronchial tumor invasion in 31 (33.3%) patients, external bronchial lymph node invasion in six (6.4%) patients, and vascular invasion in two (2.2%) patients. The bronchial invasion-based indications were significantly higher in the iBL group compared to the fissure invasion-based indications in the sBL group (P < 0.001). Conclusions: Bilobectomy leads to a substantial amount of parenchymal loss in the right lung, but it is a procedure that should be performed under the necessary conditions. It is obvious that performing bilobectomy under proper indications would result in good outcomes for lung cancer patients.

Lung deposition and clearance of microparticle and nanoparticle C60 fullerene aggregates in B6C3F1 mice and Wistar Han rats following nose-only inhalation for 13 weeks.

C60 fullerenes (C60) are spherical structures consisting of 60 carbon atoms that are generated via combustion from both natural and anthropogenic sources. C60 are also synthesized intentionally for industrial applications. Individual C60 structures have an approximate diameter of 1nm; however, C60 readily forms aggregates and typically exist as larger particles that range from nanometers to micrometers in diameter. In this report, lung and extrapulmonary tissue deposition and lung clearance of C60 nanoparticles (nano-C60, 50nm) and microparticles (micro-C60, 1μm) were examined in Wistar Han rats and B6C3F1/N mice after nose-only inhalation for 90 days. Exposure concentrations were 0.5 and 2mg/m(3) (nano-C60) and 2, 15, and 30mg/m(3) (micro-C60). For both C60 particle sizes, the C60 lung burden increased proportionally to exposure concentration. The C60 lung burden was greater in both species at all time points following exposure to nano-C60 particle exposure compared to micro-C60 exposure at the common exposure concentration 2mg/m(3). The calculated C60 particle lung retention half-times were similar for both nano-C60 and micro-C60 exposure at 2mg/m(3) in male mice (15-16 days). In contrast, in male rats, the half-time of C60 particles following nano-C60 exposure (61 days) was roughly twice as long as the half-time following micro-C60 exposure (27 days) at the same exposure concentration (2mg/m(3)) and was similar to the clearance following micro-C60 exposure at higher exposure concentrations (15 and 30mg/m(3)). C60 was detected in bronchial lymph nodes but the burden was not quantified due to the high variability in the data. C60 concentrations were below the experimental limit of quantitation (ELOQ) in liver, spleen, blood, brain and kidney tissues. These tissue burden data provide information for comparison between nanometer and micrometer sized C60 particle exposure and will aid in the interpretation of toxicity data.

CD8+ T Cells Mediate Female-Dominant IL-4 Production and Airway Inflammation in Allergic Asthma.

The prevalence and severity of bronchial asthma are higher in females than in males after puberty. Although antigen-specific CD8+ T cells play an important role in the development of asthma through their suppressive effect on cytokine production, the contribution of CD8+ T cells to sex differences in asthmatic responses remains unclear. In the present study, we investigated the sex-specific effect of CD8+ T cells in the suppression of asthma using an ovalbumin mouse model of asthma. The number of inflammatory cells in bronchoalveolar lavage (BAL) fluid, lung type 2 T-helper cytokine levels, and interleukin-4 (IL-4) production by bronchial lymph node cells were significantly higher in female wild-type (WT) mice compared with male mice, whereas no such sex differences were observed between male and female cd8α-disrupted mice. The adaptive transfer of male, but not female, CD8+ T cells reduced the number of inflammatory cells in the recovered BAL fluid of male recipient mice, while no such sex difference in the suppressive activity of CD8+ T cells was observed in female recipient mice. Male CD8+ T cells produced higher levels of IFN-γ than female CD8+ T cells did, and this trend was associated with reduced IL-4 production by male, but not female, CD4+ T cells. Interestingly, IFN-γ receptor expression on CD4+ T cells was significantly lower in female mice than in male mice. These results suggest that female-dominant asthmatic responses are orchestrated by the reduced production of IFN-γ by CD8+ T cells and the lower expression of IFN-γ receptor on CD4+ T cells in females compared with males.

Host gene expression response to specific BRD pathogens

Bovine respiratory disease complex (BRDC) is an important cause of economic loss to the cattle industry. Despite the development of numerous vaccines and much scientific investigation, the impact of BRDC on cattle producers remains high. Increased pressure to decrease the use of antibiotics in livestock medicine contributes to the urgency to find a better way to decrease the incidence of BRDC. A consortium of investigators funded by the USDA National Institute of Food and Agriculture was assembled to address this issue using the most recent genetic technology. The work presented herein summarizes a series of experiments in which steers were infected with single pathogens of BRDC (bovine respiratory syncytial virus, infectious bovine rhinotracheitis, bovine viral diarrhea virus, Mannheimia haemolytica, Pasteurella multocida, and Mycoplasma bovis) and their bronchial lymph node RNA transcriptomes were then analyzed to determine gene usage in infected vs control animals. Results showed activation of multiple pathways related to innate and acquired immune responses.

Clinico-Pathological Studies in Some Respiratory Diseases in Newly Born Buffalo Calves

A Total of 40 buffalo calves aged from 2 weeks to 3 months were used in this study. The animals were divided into two main groups the first group (group I) represented as a control group. The second group (group II) showed the clinical signs of pneumonia (natural diseased non- treated). Group (II) was subdivided into two subgroups: the first subgroup (group A) naturally diseased and treated with florofenicol (Nuflor), and the second subgroup (group B) naturally diseased and treated with tulathromycin (Draxxin). Concerning the hematological findings of the diseased non treated group (II) revealed anemia in addition to neutrophilic leukocytosis with increase in both TNF- α, IL-6 and decrease in both IgG and IgM levels was observed compared to control healthy group. While subgroups (A & B) revealed increase in RBCs count, Hb concentration, PCV, MCHC, IgG and IgM and decrease in MCV, MCH, TNF- α and IL-6 compared with diseased non treated group (II). Diseased treated subgroups (A&B) revealed decrease in T.L.C, neutrophil and monocytes with increase of lymphocytes, basophils, eosinophils and platelets count compared with diseased non treated group (II). The diseased treated subgroup (B) with Draxxin exhibited improvement in hematological and immunological parameters analysis compared with the diseased treated subgroup (A) with Nuflor. The lungs of dead calves with respiratory diseases were examined microscopically. The obtained results were lung edema, hemorrhage, collapse and different types of pneumonia such as catarrhal, suppurative, fibrinous and interstitial pneumonia. Associated bronchial lymph nodes showing congestion of blood vessels and hyperplasia of lymphocyte cells.

Immunological Response to Single Pathogen Challenge with Agents of the Bovine Respiratory Disease Complex: An RNA-Sequence Analysis of the Bronchial Lymph Node Transcriptome

Susceptibility to bovine respiratory disease (BRD) is multi-factorial and is influenced by stress in conjunction with infection by both bacterial and viral pathogens. While vaccination is broadly used in an effort to prevent BRD, it is far from being fully protective and cases diagnosed from a combination of observed clinical signs without any attempt at identifying the causal pathogens are usually treated with antibiotics. Dairy and beef cattle losses from BRD are profound worldwide and genetic studies have now been initiated to elucidate host loci which underlie susceptibility with the objective of enabling molecular breeding to reduce disease prevalence. In this study, we employed RNA sequencing to examine the bronchial lymph node transcriptomes of controls and beef cattle which had individually been experimentally challenged with bovine respiratory syncytial virus, infectious bovine rhinotracheitis, bovine viral diarrhea virus, Pasteurella multocida, Mannheimia haemolytica or Mycoplasma bovis to identify the genes that are involved in the bovine immune response to infection. We found that 142 differentially expressed genes were located in previously described quantitative trait locus regions associated with risk of BRD. Mutations affecting the expression or amino acid composition of these genes may affect disease susceptibility and could be incorporated into molecular breeding programs. Genes involved in innate immunity were generally found to be differentially expressed between the control and pathogen-challenged animals suggesting that variation in these genes may lead to a heritability of susceptibility that is pathogen independent. However, we also found pathogen-specific expression profiles which suggest that host genetic variation for BRD susceptibility is pathogen dependent.

PTH-159 The relevance of lymph node metastases in the proximal field of the chest in patients with adenocarcinoma of the gastro-oesophageal junction treated with neoadjuvant therapy followed by transthoracic oesophagectomy

<h3>Introduction</h3> The prognosis of gastroesophageal junction adenocarcinoma is unquestionably related to the extent of nodal involvement. Not only the amount of positive lymph nodes but also the location of postive nodes negatively influence survival. Earlier small studies revealed a poor survival for patients with positive nodes in the proximal field of the chest when treated with surgery alone. It is thus far unknown what the relevance of these nodes is after neoadjuvant therapy. The aim of the present study was to identify the incidence and prognostic significance of lymph node metastases in the proximal part of the chest in patients who underwent a transthoracic oesophagectomy after neoadjuvant therapy. <h3>Method</h3> From a prospectively collected database, a consecutive series of patients in two high volume centres in Europe was analysed. All patients with potentially curable adeno carcinoma of the gastro-oesophageal junction were treated with neoadjuvant chemo (radiation)therapy therapy followed by transthoracic oesophagectomy and two-field lymphadenectomy. <h3>Results</h3> Between January 2000 and September 2013 a consecutive series of 208 patients underwent an oesophagectomy after neoadjuvant therapy for adenocarcinoma of the gastro-oesophageal junction. 80 (38.5%) patients had no evidence of lymph node metastases (N0). There were 128 patients (61.5%) with positive nodes (N1=41 (19.7%), N2=44 (21.2%), N3=43 (20.7%)). Of the node positive patients, 25 (19.5%) had positive nodes in the proximal field of the chest (paratracheal, subcarinal, bronchial and aorto-pulmonary window lymph nodes). More advanced N-stage was significantly associated with positive nodes in the proximal field of the chest (p &lt; 0.001). Median survival was significantly (p = 0.003) worse for patients with positive nodes in the proximal part of the chest (12.4 months (95% CI: 11.4–13.32)) compared with patients with positive nodes on other locations (28.3 months (95% CI: 22.7–33.9)). <h3>Conclusion</h3> Lymph node metastases in the proximal part of the chest in patients is a common phenomonemon and a sign of advanced disease. Radical surgical resection can only be achieved with an extended transthoracic resection. Even after neoadjuvant therapy followed by transthoracic resection long-term survival is poor. <h3>Disclosure of interest</h3> None Declared.

Honeybee (Apis mellifera) Venom Reinforces Viral Clearance during the Early Stage of Infection with Porcine Reproductive and Respiratory Syndrome Virus through the Up-Regulation of Th1-Specific Immune Responses.

Porcine reproductive and respiratory syndrome (PRRS) is a chronic and immunosuppressive viral disease that is responsible for substantial economic losses for the swine industry. Honeybee venom (HBV) is known to possess several beneficial biological properties, particularly, immunomodulatory effects. Therefore, this study aimed at evaluating the effects of HBV on the immune response and viral clearance during the early stage of infection with porcine reproductive and respiratory syndrome virus (PRRSV) in pigs. HBV was administered via three routes of nasal, neck, and rectal and then the pigs were inoculated with PRRSV intranasally. The CD4+/CD8+ cell ratio and levels of interferon (IFN)-γ and interleukin (IL)-12 were significantly increased in the HBV-administered healthy pigs via nasal and rectal administration. In experimentally PRRSV-challenged pigs with virus, the viral genome load in the serum, lung, bronchial lymph nodes and tonsil was significantly decreased, as was the severity of interstitial pneumonia, in the nasal and rectal administration group. Furthermore, the levels of Th1 cytokines (IFN-γ and IL-12) were significantly increased, along with up-regulation of pro-inflammatory cytokines (TNF-α and IL-1β) with HBV administration. Thus, HBV administration—especially via the nasal or rectal route—could be a suitable strategy for immune enhancement and prevention of PRRSV infection in pigs.

Dynamic control of Th2 cell responses by STAT3 during allergic lung inflammation in mice

Signal transducer and activator of transcription (STAT) family molecules play essential roles during the differentiation of helper T cells from naïve precursors. Although the role of STAT3 in driving Th17 cell polarization has been well established, its role on Th2 responses to allergens remains incompletely understood. By employing T cell-specific STAT3 deficient mice, we demonstrate that STAT3 in T cells plays diverse role on Th2 cells depending on their locations in an animal model of allergic asthma. In the bronchial lymph nodes, STAT3-deficient T cells produced significantly reduced levels of Th2 cytokines. The frequencies of Th2 cells among CD4+ T cells in the lung were comparable between STAT3-sufficient and STAT3-deficient T cells. By contrast, STAT3-deficient T cells in the airway exhibited significantly enhanced production of Th2 cell cytokines compared to STAT3-sufficient T cells. Interestingly, a major population of IL-4/5 producers among STAT3-deficient T cells in the airway co-produced IFNγ. The frequency of Th17 cells was significantly diminished whereas that of Th1 cells was increased in all the lung-associated tissues. Our results demonstrate the dynamic and opposing roles of STAT3 during the development of Th2 cells from bronchial lymph nodes to the airway and propose the need of careful consideration on STAT3-targeting approaches for the treatment of lung diseases.