Apoptosis in normal lymphoid organs from healthy normal, conventional pigs at different ages detected by TUNEL and cleaved caspase-3 immunohistochemistry in paraffin-embedded tissues

Resendes, A. R.; Majó, Natàlia; Segalés, Joaquím; Espadamala, Josep; Mateu, Enric; Chianini, Francesca; Nofrarías, Miquel; Domingo, Mariano

doi:10.1016/j.vetimm.2004.02.001

Cited by 62 publications

(58 citation statements)

References 18 publications

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“…Therefore, results obtained on field cases need to be viewed with caution. Studies conducted by several research groups have identified apoptosis by TUNEL assay (Shibahara et al, 2000;Krakowka et al, 2004;Mandrioli et al, 2004;Resendes et al, 2004a;Seeliger et al, 2007) and CCasp3 IHC (Resendes et al, 2004a,b;Seeliger et al, 2007) in conventional, clinically-affected pigs (Rosell et al, 1999(Rosell et al, , 2000b. A recent investigation on conventional pigs using the TUNEL assay, CCasp3 IHC and PCV2 in situ hybridization (ISH) provided evidence of virus-induced apoptosis in endothelial cells in pigs diagnosed with PCV2-associated cerebellar vasculitis (Seeliger et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

“…A recent study compared the differences between the TUNEL and the CCasp3 IHC assays on lymphoid tissues of healthy conventional pigs of different ages (Resendes et al, 2004a); however, to the authors' knowledge no experimental studies have been performed to compare these assays in hepatic tissues. Our study concluded that both methods detected significantly (P < 0.05) higher numbers of apoptotic cells in clinically-affected pigs compared to clinically-unaffected pigs and sham-inoculated control pigs.…”

Section: Discussionmentioning

confidence: 99%

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Singular PCV2a or PCV2b infection results in apoptosis of hepatocytes in clinically affected gnotobiotic pigs

Sinha

Schalk

Lager

et al. 2012

Research in Veterinary Science

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Porcine circovirus type 2 (PCV2) is clinically associated with respiratory disease, failure-to-thrive, hepatitis, and diarrhea; however, the precise pathogenesis of PCV2-associated disease and in particular its involvement in apoptosis is still controversial. The objectives of this study were (1) to determine whether PCV2 is associated with apoptosis by examining and comparing hepatic tissues from clinically affected or unaffected gnotobiotic pigs that were experimentally infected with PCV2, (2) to determine if there are differences between PCV2a and PCV2b in inducing hepatocyte apoptosis, and (3) to determine if there are differences between apoptosis detection systems. Forty-eight gnotobiotic pigs were separated into five groups based on inoculation status and development of clinical disease: (1) sham-inoculated, clinically-unaffected (n = 4), (2) inoculated with PCV2a, clinically-unaffected (n = 10), (3) inoculated with PCV2a, clinically-affected (n = 6), (4) inoculated with PCV2b, clinically-unaffected, (n = 13) and (5) inoculated with PCV2b, clinically-affected (n = 15). Formalin-fixed, paraffin-embedded sections of liver from all pigs were analyzed for signs of apoptosis [presence of single strand DNA breaks in the nucleus by the terminal transferase dUTP nick end labeling (TUNEL) assay or presence of intra-nuclear cleaved caspase 3 (CCasp3) demonstrated by CCasp3 immunohistochemistry (IHC)]. In addition, the liver tissues were also tested for presence of cytoplasmic and intra-nuclear PCV2 antigen by an IHC assay. Specific CCasp3 and TUNEL labeling was detected in the nucleus of hepatocytes in PCV2a and PCV2b infected pigs with significantly (P < 0.05) higher levels of apoptotic cells in clinically-affected pigs. Regardless of PCV2 subtype (PCV2a; PCV2b), there were higher levels of PCV2 antigen in clinically-affected pigs compared to clinically-unaffected pigs. There was no significant difference in detection rate of apoptotic cells between the TUNEL assay and CCasp3 IHC. When high amounts of PCV2 antigen were present, the incidence of CCasp3 and TUNEL staining also increased regardless of the PCV2 genotype. This suggests that PCV2-induced apoptosis of hepatocytes is important in the pathogenesis of PCV2-associated lesions and disease. RightsWorks produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted. b s t r a c tPorcine circovirus type 2 (PCV2) is clinically associated with respiratory disease, failure-to-thrive, hepatitis, and diarrhea; however, the precise pathogenesis of PCV2-associated disease and in particular its involvement in apoptosis is still controversial. The objectives of this study were (1) to determine whether PCV2 is associated with apoptosis by examining and comparing hepatic tissues from clinically affected or unaffected gnotobiotic pigs that were experimentally infected with PCV2, (2) to determine if there are differences between PCV2a and PCV2b in inducing hepatocyte ...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Singular PCV2a or PCV2b infection results in apoptosis of hepatocytes in clinically affected gnotobiotic pigs

Sinha

Schalk

Lager

et al. 2012

Research in Veterinary Science

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“…Caspase-3 is synthesized as inactive proenzyme that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by other upstream proteases (e.g., caspases 8, 9, and 10). The detection of active caspase-3 is widely used in many cell lines undergoing apoptosis (Jakob et al 2008;Resendes et al 2004). As shown in Fig.…”

Section: Cholesterol Loading Induces Apoptosis Of Min6mentioning

confidence: 99%

Cholesterol induces pancreatic β cell apoptosis through oxidative stress pathway

Liu

Hou

et al. 2011

Cell Stress and Chaperones

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Type 2 diabetes is often associated with high blood cholesterol. Here, we investigated the effect of cholesterol loading on MIN6 cells derived from pancreatic β cells. Exposure of MIN6 cells to cholesterol-induced apoptosis in time-and dose-dependent manner. Treatment with methyl-β-cyclodextrin that removes cholesterol from plasma membrane prevented the cells from cholesterolinduced apoptosis. Western blot analysis revealed that the levels of phosphorylated-p38 mitogen-activated protein kinase (P-p38 MAPK) and c-Jun N-terminal kinases (P-JNK) were significantly increased after the cholesterol loading, suggesting that the stress-activated protein kinase signaling was stimulated. A specific p38 inhibitor rescued MIN6 cells from cholesterol-induced apoptosis, while JNK inhibitor failed, suggesting the importance of activation of p38 MAPK signaling in response to cholesterol. The expression of Bip and CHOP, the endoplasmic reticulum (ER) stress markers, remained unaffected, indicating that the ER stress may not be involved in the cytotoxicity of cholesterol on the MΙΝ6 cells. The intracellular concentration of reactive oxygen species measured by use of 2′,7′-dichlorofluorescin diacetate was significantly increased after cholesterol loading, demonstrating the induced apoptosis was mediated through oxidative stress. Addition of reduced form of glutathione in the medium rescued MIN6 cells from apoptosis induced by cholesterol loading. Taken together, these results demonstrate that the free cholesterol loading can induce apoptosis of MIN6 cells mediated by oxidative stress and the activation of p38 MAPK signaling.

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“…This situation provides great ease and benefits in various ways while examining the steps of immune system development in newly born piglets (10). Nevertheless, studies on lymphocyte biology in pigs are mostly conducted with lymphoid organs obtained from pigs that have reached the age of slaughter (11). Pigs at this stage has a lower chance of providing reliable information for researchers due to the diseases they have experienced, antigen accumulations, and the stress they experience prior to slaughtering.…”

Section: Introductionmentioning

confidence: 99%

Ultrastructure of apoptotic T lymphocytes and thymicepithelial cells in early postnatal pig thymus

Ceylan¹,

Alabay²

2017

Turk J Vet Anim Sci

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The thymus is a primary lymphoid organ that provides the microenvironment required for the development of T lymphocytes. Thymic epithelial cells (TECs) are among the most critical components in the thymic microenvironment supporting thymocyte selection and maturation. Apoptosis is primarily a physiologic process that plays a major role during the development of T lymphocytes. The first purpose of this study was to examine morphological changes in apoptotic thymocytes by transmission electron microscope in piglets. The second purpose was to reveal the morphological typology of TECs in piglets. Tissue samples of the thymus were taken from healthy Yorkshire race pigs of two different age groups (7 days and 1 month). Ultrastructurally, apoptotic lymphocytes showed similar morphologies, but they also had differences in distribution. In both groups a high density of apoptotic and proliferating thymocytes was found in the cortex. The rate of apoptotic thymocytes in the cortex and medulla increased with age, respectively. It was observed that the TECs had various morphological subtypes in both the cortex and the medulla. A similar cellular distribution pattern of TECs was seen in both groups. It was concluded that the morphological diversity in the TECs may be related to the form-function relationship and the metabolic process.

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Apoptosis in normal lymphoid organs from healthy normal, conventional pigs at different ages detected by TUNEL and cleaved caspase-3 immunohistochemistry in paraffin-embedded tissues

Cited by 62 publications

References 18 publications

Singular PCV2a or PCV2b infection results in apoptosis of hepatocytes in clinically affected gnotobiotic pigs

Singular PCV2a or PCV2b infection results in apoptosis of hepatocytes in clinically affected gnotobiotic pigs

Cholesterol induces pancreatic β cell apoptosis through oxidative stress pathway

Ultrastructure of apoptotic T lymphocytes and thymicepithelial cells in early postnatal pig thymus

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