• Asian-Australasian Journal of Animal Sciences
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• v.32 no.11
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• pp.1789-1800
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• 2019

Objective: Although alveolar macrophages play a key role in the respiratory immunity of livestock, studies on the mechanism of differentiation and survival of alveolar macrophages are lacking. Therefore, we undertook to investigate changes in the lipid metabolism and survival rate, using 3D4/31 macrophages and Dudleya brittonii which has been used as a traditional asthma treatment. Methods: 3D4/31 macrophages were used as the in vitro porcine alveolar macrophages model. The cells were activated by exposure to phorbol 12-myristate 13-acetate (PMA). Dudleya brittonii extraction was performed with distilled water. For evaluating the cell survival rate, we performed the water-soluble tetrazolium salt cell viability assay and growth curve analysis. To confirm cell death, cell cycle and intracellular reactive oxygen species (ROS) levels were measured using flow cytometric analysis by applying fluorescence dye dichlorofluorescein diacetate and propidium iodide. Furthermore, we also evaluated cellular lipid accumulation with oil red O staining, and fatty acid synthesis related genes expression levels using quantitative polymerase chain reaction (qPCR) with SYBR green dye. Glycolysis, fatty acid oxidation, and tricarboxylic acid (TCA) cycle related gene expression levels were measured using qPCR after exposure to Dudleya brittonii extract (DB) for 12 h. Results: The ROS production and cell death were induced by PMA treatment, and exposure to DB reduced the PMA induced downregulation of cell survival. The PMA and DB treatments upregulated the lipid accumulation, with corresponding increase in the acetyl-CoA carboxylase alpha, fatty acid synthase mRNA expressions. DB-PMA co-treatment reduced the glycolysis genes expression, but increased the expressions of fatty acid oxidation and TCA cycle genes. Conclusion: This study provides new insights and directions for further research relating to the immunity of porcine respiratory system, by employing a model based on alveolar macrophages and natural materials.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.177-183
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• 1997

One cytopathogenic virus was isolated in alveolar macrophages from pig without any apparent respiratory clinical signs. Biophysical properties and electron microscopy of the isolate showed the characteristics of adenovirus. Intracytoplasmic inclusion bodies were seen in virus-inoculated cells. The genetic analysis indicated the presence of DNA with the size of >20Kb. In a serological survey of 40 serum samples collected from two different farms in slaughter house, 9 sera were positive for neutralizing antibody against the isolate. The potential implications of the isolate as the causative agent in respiratory disorder were discussed.

• Korean Journal of Veterinary Pathology
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• v.1 no.2
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• pp.125-134
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• 1997

Porcine Reproductive and Respiratory Syndrome Virus infection (PRRSV) was confirmed by serology histopathology immunohistochemistry and bacteriologic examination in young pigs. Four suckling and six weaned piglets submitted from three different farms showed coughing sneezing labored rapid abdominal respiration lethargy and anorexia. Grossly apical and cardiac lung lobes appeared mottled with pale to dark tan discoloration. Submandibular and bronchial lymph nodes were tan and enlarged. All piglets were seropositive for PRRSV antibodies by the indirect immunofluorescent antibody(IFA) test. Microscopically lung lesions were characterized by hyperplasia and hypertrophy of type 2 pneumocytes infiltration of mononuclear cells in alveolar intersitium accumulation of necrotic debris in alveolar spaces accompanied by proliferation of alveolar multinucleated syncytial cells. Using immunohistochemical technique PRRSV antigens were demonstrated in alveolar macrophages and type 2 pneumocytes in histologic lung tissue sections. Also PRRSV antigens were detected in brain lymph nodes spleen and heart. Additionally piglets showed nonsuppurative meningoencephalitis mandibular necrotic lymphadenopathy splenic atrophy and myocardial necrosis.

• Korean Journal of Veterinary Service
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• v.37 no.1
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• pp.1-9
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• 2014

Although it has been generally accepted that porcine reproductive and respiratory syndrome virus (PRRSV) induces weak and delayed protective immunity after infection, it is unclear that the same immunological features can be applicable to all PRRS viruses because huge genetic variation exists even among the same genotypes of PRRSV (Type 1 and 2). In the current study, two genetically distinct type 2 PRRSV strains (VR-2332 and JA142) which showed approximately 90% nucleotide homology based on ORF5 sequences were characterized by both in vitro and in vivo assessments to determine the immunological features of the viruses. For in vitro assessment, porcine alveolar macrophages (PAM) were infected with the viruses at $10^{-3}$ multiplicity of infection (MOI) and then supernatants and cells were collected separately at 36 hrs post infection to determine the relative expression levels of IL-$1{\alpha}$, IL-12, TNF-${\alpha}$ and INF-${\alpha}/{\beta}$ by quantitative RT-PCR. In addition, five PRRSV-free pigs were inoculated with either of JA142 or VR2332 for in vivo assessment. Serum samples were collected every week until 6 weeks post challenge. The serum samples were analyzed for the levels of viremia, PRRSV nucleocapsid-specific antibody and virus neutralizing antibody. Based on those assessments, the two viruses showed different patterns of cytokine expression in PAM and immune responses in pigs after infection. These results indicate that genetically distinct PRRSV strains have different immunological features, which might be criteria for virus classification and selection of candidate virus strains for vaccine development in the future.

• BMB Reports
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• v.47 no.1
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• pp.45-50
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• 2014

Aspirin has been demonstrated to be effective in inhibiting COX-2 and $PGE_2$ in Alveolar macrophages (AMs). However, the mechanisms have not been fully understood. In the present study, we found that pretreatment with aspirin inhibited LPS-induced COX-2 and$PGE_2$ upregulation, $I{\kappa}B{\alpha}$ degradation, NF-${\kappa}B$ activation and the increase of PKC activity, but elevated LPS-induced the decrease of PTP activity. The PKC inhibitor calphostin C dramatically reduced the COX-2 mRNA and $PGE_2$ levels, but the PTP inhibitor peroxovanadium (POV) significantly increased the COX-2 mRNA and$PGE_2$ levels. Furthermore, the PTP inhibitor mitigated the inhibitory effect of aspirin on COX-2 and$PGE_2$ upregulation and NF-${\kappa}B$ activation, whereas the PKC inhibitor enhanced the inhibitory effects of aspirin on the production of COX-2 and$PGE_2$. Our data indicate a novel mechanism by which aspirin acts as a potent anti-inflammatory agent in alveolus macrophages and ALI.

• Korean Journal of Veterinary Research
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• v.53 no.4
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• pp.245-251
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• 2013

Porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) have been suspected to have immunosuppressive effects on pigs. To investigate the correlation between these virus infection and the lesions of lymph nodes including sub-mandibular and inguinal lymph node, 44 pigs (PCV2 single, n = 14; PRRSV single, n = 10; PCV2/PRRSV, n = 14; negative control, n = 6) were examined by histopathology and immunohistochemistry. Histopathologically, granulomatous lymphadenitis characterized by lymphoid depletion with histiocytic cells infiltration was observed in PCV-2 single and PCV-2/PRRSV group. Immunohistochemically, there were significant reduction of B and T lymphocytes in lymph nodes of these groups, while the number of macrophages was increased. In only PRRSV infected group, germinal center hypertrophy and lymphoid necrosis were observed. Immunohistochemically, the number of CD3+ T lymphocytes was slightly increased. Severe lymphocytic depletion in PCV-2 infection-related lymph nodes might be associated with producing immunocompromised state in pig. Comparing with PCV-2 infected group, PRRSV produced minor effects on the changes in immune cell population in the lymph nodes of pigs. PRRSV may increase susceptibility of the disease in pigs by disruption of the first defense lines in target organs, such as the alveolar macrophages in lungs.

• Korean Journal of Veterinary Service
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• v.43 no.4
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• pp.251-256
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• 2020

Two suckling piglets, 4 days and 10 days of age, showed lethargy and dyspnea after birth and mortality had been increased after incoming gilts from breeding farm. At necropsy, the lungs showed diffuse fail to collapse with rubbery consistency, edematous dilatation of interlobular septa, and lobular consolidation with purple red color. Heart was diffuse pale in color and had several irregular linear-shaped macules or patches. Histopathologically, diffuse interstitial pneumonia with the proliferation of type II pneumocytes was present in the lungs of 2 piglets. Alveolar lumens contained necrotic cellular debris derived from neutrophils and macrophages. Multifocal hemorrhage and necrotizing pneumonia with protozoan tachyzoites were observed in the lungs. Severe multifocal to confluent necrotic myocarditis, necrotic encephalitis, and necrotic adrenalitis with intralesional protozoan tachyzoites were observed in piglets. According to immunohistochemical analysis (IHC), Toxoplasma (T.) gondii tachyzoites antigens were confirmed in lung, heart, brain, and adrenal gland. And porcine reproductive and respiratory syndrome virus (PRRSV) antigens were also detected in the cytoplasm of macrophages in lungs using IHC. Based on the gross, histopathologic and immunohistochemical features, two suckling piglets were diagnosed as co-infection of T. gondii and PRRSV.

• Korean Journal of Veterinary Research
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• v.58 no.1
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• pp.9-16
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• 2018

A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, $CD3^+CD4^+$, $CD3^+CD8^+$, $CD3^+CD4^+CD8^+$ T-lymphocytes, and PRRSV-specific interferon $(IFN)-{\gamma}$ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages ($CD172{\alpha}^+PRRSV-N^+\;PAM$) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of $CD172{\alpha}^+PRRSV-N^+\;PAM$ (p < 0.05) as well as with macroscopic lung lesion (p < 0.01). Plasma and tissue viral loads had significant (p < 0.05) negative correlations with $CD3^+CD4^+CD8^+$ T-lymphocyte percentage in PBMC. Frequencies of $CD3^+CD8^+$ and $CD3^+CD4^+$ T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads. $IFN-{\gamma}$-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002). However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.63-69
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• 2022

Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in post-weaning pigs. In this study, we investigated the presence of swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) and Aujeszky's disease virus (ADV) in PNP lesions in Jeju pigs. Based on the histopathologic criteria for PNP, a total of 50 cases were selected in Jeju pigs between 2008 and 2010. Coupled with histopathological examinations, the presence of ADV and SIV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) and PRRSV and PCV2 by immunohistochemical (IHC) methods were investigated. Based on the PCR and RT-PCR methods, ADV and SIV nucleic acids were not detected in all cases. According to IHC, PRRSV was detected in 38 of the 50 cases examined (76%) and PCV2 in 25 cases (50%). PRRSV or PCV2 were detected in 19 (38%) or 6 (12%) cases, respectively. Both PRRSV and PCV2 were identified in other 19 cases (38%). Antigens of PRRSV and PCV2 were commonly observed in the cytoplasm of macrophages and clusters of necrotic cells in alveolar cavities. The results of the present study demonstrate that PRRSV is predominantly associated with PNP in Jeju pigs. Co-infection with PRRSV and PCV2 may enhance the severity of PNP lesions in affected pigs.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.793-807
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• 1997

We tried to develop detection system of porcine reproductive and respiratory syndrome virus(PRRSV) by in situ hybridization(ISH) in the piglets experimentally infected with KPRRS-2, the Korean isolate(12 piglets) or Mn-1b, the American isolate(4 piglets), and in the natural infection suspected 6 piglets. Twelve 30-days-old piglets(two pigs per each inoculated group) were inoculated by nasal instillation of KPRRS-2 virus(total dose $10^{4.5}TCID_{50}$), Six piglets(one pig per each group) were induced contact infection with inoculated piglets, during the experiment, and two piglets were used as control. Inoculated or contacted piglets were euthanized at 1, 3, 5, 7, 14 and 21 days postinoculation(DPI). The respiratory signs such as coughing and nasal discharge were observed on day 3 DPI, and ear cyanosis were on day 5 DPI, including contacted piglets. Through the necropsy, purple discolorization of dorsal part of lung, and hypertrophy of local lymph nodes were observed. The histopathological lesions of lung were interstitial pneumonia characterized by type 2 pneumocyte hyperplasia. We prepared the probe for ISH by RNA isolation from KPRRS-2, RT-PCR, and biotin labeling. We performed the ISH within only 1~2 hours using $Microprobe^{TM}$ capillary action system. As the results, the strong red specific positive signals, means PRRSV distribution, was mainly observed in the cytoplasm of alveolar macrophages. And also signals were detected in some type 2 pneumocytes and bronchiolar epithelium of lung, myocardium, liver, kidney, tonsil, spleen, gastrointestinal mucosa, testis and lymph nodes.