scholarly journals Host-cell apoptosis inTaenia solium-induced brain granulomas in naturally infected pigs

Parasitology ◽  
2008 ◽  
Vol 135 (10) ◽  
pp. 1237-1242 ◽  
Author(s):  
C. S. SIKASUNGE ◽  
I. K. PHIRI ◽  
M. V. JOHANSEN ◽  
A. L. WILLINGHAM ◽  
P. S. LEIFSSON
Keyword(s):  
T Lymphocytes ◽  
Normal Cell ◽  
Taenia Solium ◽  
Cell Types ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Apoptotic Cells ◽  
Host Cell Apoptosis ◽  
Cellular Immune

SUMMARYTo assess whether apoptosis occurs in pig brain granulomas due toTaenia soliumcysticerci, brain tissues from 30 pigs naturally infected withT. soliumcysticercosis were evaluated by terminal deoxynucleotidyl transferase-end labelling (TUNEL) staining. In addition, tissues were stained with CD3 marker to identify T lymphocytes. Examination of TUNEL-stained tissues showed apoptotic cells in early lesions that contained viable cysticerci. Apoptotic cells were primarily found interspersed with normal cell types, and were mostly located in the inflammatory infiltrate. Late or advanced granulomas with disintegrated scolices did not show TUNEL-positive cells. CD3+ cells were found in both early and advanced lesions and apoptosis mainly co-localized with CD3+ T lymphocytes. This suggests that these cells are constantly undergoing apoptosis and thus die as soon as they arrive at the site of infection. Apoptosis indeed may be one way by whichT. soliumcysticerci down-regulate the host's cellular immune response in early cysticercosis. Therefore, further research is needed to establish if other cells besides T-lymphocytes are also a target for destruction by cysticerci in early cysticercosis as well as studies to assess if cysteine protease is expressed by viable cysticerciin situ.

scholarly journals Comparison of Apoptosis Detection Markers Combined with Macrophage Immunostaining to Study Phagocytosis of Apoptotic Cells in Situ

2006 ◽  
Vol 1 ◽  
pp. 117727190600100 ◽  
Author(s):  
Dorien M. Schrijvers ◽  
Guido R.Y. De Meyer ◽  
Mark M. Kockx ◽  
Arnold G. Herman ◽  
Wim Martinet
Keyword(s):  
Lupus Erythematosus ◽  
Caspase 3 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Apoptotic Cells ◽  
Immunological Responses ◽  
Apoptosis Detection ◽  
Apoptosis Markers ◽  
Suitable Marker ◽  
Parp 1

Efficient phagocytosis of cells undergoing apoptosis by macrophages is important to prevent immunological responses and development of chronic inflammatory disorders such as systemic lupus erythematosus, cystic fibrosis and atherosclerosis. To study phagocytosis of apoptotic cells (AC) by macrophages in tissue, we validated different apoptosis markers (DNA fragmentation, caspase-3 activation and cleavage of its substrate poly(ADP-ribose)polymerase-1) in combination with macrophage immunostaining. Human tonsils were used as a model because they show a high apoptosis frequency under physiological conditions as well as efficient phagocytosis of AC by macrophages. On the other hand, advanced human atherosclerotic plaques were examined since plaques show severely impaired phagocytosis of AC. Our results demonstrate that the presence of non-phagocytized terminal deoxynucleotidyl transferase end labelling (TUNEL)-positive AC represents a suitable marker of poor phagocytosis by macrophages in situ. Other markers for apoptosis, such as cleavage of caspase-3 or PARP-1, should not be used to assess phagocytosis efficiency, because activation of the caspase cascade and cleavage of their substrates can occur in AC when they have not yet been phagocytized by macrophages.

scholarly journals Presence of double-strand breaks with single-base 3' overhangs in cells undergoing apoptosis but not necrosis.

1996 ◽  
Vol 135 (5) ◽  
pp. 1369-1376 ◽  
Author(s):  
V V Didenko ◽  
P J Hornsby
Keyword(s):  
Dna Damage ◽  
Double Strand Breaks ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Apoptotic Cells ◽  
Dna Fragments ◽  
Strand Breaks ◽  
Single Base ◽  
Double Stranded Dna

Apoptotic cells in rat thymus were labeled in situ in paraffin-embedded and frozen tissue sections by ligation of double-stranded DNA fragments containing digoxigenin or Texas red. Two forms of double-stranded DNA fragments were prepared using the polymerase chain reaction: one was synthesized using Taq polymerase, which yields products with single-base 3' overhangs, and one using Pfu polymerase, which produces blunt-ended products. Both types of fragment could be ligated to apoptotic nuclei in thymus, indicating the presence in such nuclei of DNA double-strand breaks with single-base 3' overhangs as well as blunt ends. However, in nuclei with DNA damage resulting from a variety of nonapoptotic processes (necrosis, in vitro autolysis, peroxide damage, and heating) single-base 3' overhangs were either nondetectable or present at much lower concentrations than in apoptotic cells. Blunt DNA ends were present in such tissues, but at lower concentrations than in apoptotic cells. In contrast, in all of these forms of DNA damage, nuclei contained abundant 3'-hydroxyls accessible to labeling with terminal deoxynucleotidyl transferase. Thus, although single-base 3' overhangs and blunt ends are present in apoptotic nuclei, the specificity of the in situ ligation of 3'-overhang fragments to apoptotic nuclei indicates that apoptotic cells labeled in this way can readily be distinguished from cells with nonapoptotic DNA damage. These data are consistent with the involvement of an endonuclease similar to DNase I in apoptosis, which is predicted to leave short 3' overhangs as well as blunt ends in digestion of chromatin.

Protective effects of vitamin E on aluminium sulphate-induced testicular damage

2020 ◽  
Vol 36 (4) ◽  
pp. 215-227 ◽  
Author(s):  
Ozal Ulfanov ◽  
Nazli Cil ◽  
Esat Adiguzel
Keyword(s):  
Vitamin E ◽  
Male Infertility ◽  
Germinal Epithelium ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Apoptotic Cells ◽  
Protective Effects ◽  
Testicular Damage ◽  
Cell Series ◽  
Control Sham

Male infertility can be caused by environmental factors, genetic defects, physiological and endocrine deficiencies and testicular pathologies. Aluminium (Al) can cause male infertility through a number of mechanisms. The aim of our study was thus to determine whether vitamin E (VitE) has protective effects on Al-induced testicular damage, which was determined according to sperm counts and morphology and using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. Thirty-four male Wistar rats (250–300 g) were randomly assigned to control (no procedures performed; n = 6) or 0.2 mL intraperitoneal injection group ( n = 7 each; three times per week for 4 weeks): sham (distilled water), 10 mg/kg Al, 500 mg/kg VitE and 10 mg/kg Al plus 500 mg/kg VitE (Al + VitE). Sperm samples were evaluated for andrological parameters. The testes were examined by haematoxylin/eosin. The epithelial thickness and areas were calculated and Johnsen scores were determined for the germinal epithelium; the apoptotic indices were determined from TUNEL staining. For Al, the bonds between the germinal epithelial cells were broken in some tubules, and there were unidentified cells in the lumen of some tubules. For control, sham and VitE, normal morphology of the germinal epithelium was generally preserved. With Al + VitE, the full germinal epithelium cell series was maintained, with only mature sperm in the lumen. TUNEL-positive cells were significantly higher with Al compared to control and sham ( p < 0.05). For Al + VitE, the number of apoptotic cells was reduced compared to Al alone and was therefore similar to control, sham and VitE ( p > 0.05). Our findings show that Al caused testicular damage. VitE reduced the number of apoptotic cells during the damage caused by Al.

scholarly journals Expression of Fas ligand by human gastric adenocarcinomas: a potential mechanism of immune escape in stomach cancer

Gut ◽  
1999 ◽  
Vol 44 (2) ◽  
pp. 156-162 ◽  
Author(s):  
M W Bennett ◽  
J O’Connell ◽  
G C O’Sullivan ◽  
D Roche ◽  
C Brady ◽  
...  
Keyword(s):  
Cell Death ◽  
Stomach Cancer ◽  
Fas Ligand ◽  
Immune Escape ◽  
Immune Privilege ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Gastric Adenocarcinomas

BackgroundDespite being immunogenic, gastric cancers overcome antitumour immune responses by mechanisms that have yet to be fully elucidated. Fas ligand (FasL) is a molecule that induces Fas receptor mediated apoptosis of activated immunocytes, thereby mediating normal immune downregulatory roles including immune response termination, tolerance acquisition, and immune privilege. Colon cancer cell lines have previously been shown to express FasL and kill lymphoid cells by Fas mediated apoptosis in vitro. Many diverse tumours have since been found to express FasL suggesting that a “Fas counterattack” against antitumour immune effector cells may contribute to tumour immune escape.AimTo ascertain if human gastric tumours express FasL in vivo, as a potential mediator of immune escape in stomach cancer.SpecimensThirty paraffin wax embedded human gastric adenocarcinomas.MethodsFasL protein was detected in gastric tumours using immunohistochemistry; FasL mRNA was detected in the tumours using in situ hybridisation. Cell death was detected in situ in tumour infiltrating lymphocytes using terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL).ResultsPrevalent expression of FasL was detected in all 30 resected gastric adenocarcinomas examined. In the tumours, FasL protein and mRNA were co-localised to neoplastic gastric epithelial cells, confirming expression by the tumour cells. FasL expression was independent of tumour stage, suggesting that it may be expressed throughout gastric cancer progression. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumour.ConclusionsHuman gastric adenocarcinomas express the immune downregulatory molecule, FasL. The results suggest that FasL is a prevalent mediator of immune privilege in stomach cancer.

scholarly journals Bovine Herpesvirus 1 Can Infect CD4+ T Lymphocytes and Induce Programmed Cell Death during Acute Infection of Cattle

1999 ◽  
Vol 73 (10) ◽  
pp. 8657-8668 ◽  
Author(s):  
M. T. C. Winkler ◽  
A. Doster ◽  
C. Jones
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Acute Infection ◽  
Bovine Herpesvirus ◽  
Cell Mediated Immunity ◽  
Apoptotic Cells ◽  
Bovine Herpesvirus 1 ◽  
Herpesvirus 1 ◽  
Pharyngeal Tonsil

ABSTRACT Acute infection of cattle with bovine herpesvirus 1 (BHV-1) represses cell-mediated immunity, which can lead to secondary bacterial infections. Since BHV-1 can induce apoptosis of cultured lymphocytes, we hypothesized that these virus-host interactions occur in cattle. To test this hypothesis, we analyzed lymph nodes and peripheral blood mononuclear cells (PBMC) after calves were infected with BHV-1. In situ terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) staining of lymphoid tissues (pharyngeal tonsil, cervical, retropharyngeal, and inguinal) was used to detect apoptotic cells. Calves infected with BHV-1 for 7 days revealed increased apoptotic cells near the corticomedullary junction in lymphoid follicles and in the subcapsular region. Increased frequency of apoptotic cells was also observed in the mucosa-associated lymphoid tissue lining the trachea and turbinate. Immunohistochemistry of consecutive sections from pharyngeal tonsil revealed that CD2+ T lymphocytes were positive for the BHV-1 envelope glycoprotein gD. The location of these CD2+ T lymphocytes in the germinal center suggested that they were CD4+ T cells. Electron microscopy and TUNEL also revealed apoptotic and herpesvirus-infected lymphocytes from this area. Fluorescence-activated cell sorting analyses demonstrated that CD4+ and CD8+ T cells decreased in lymph nodes and PBMC after infection. The decrease in CD4+ T cells correlated with an increase in apoptosis. CD4+ but not CD8+ lymphocytes were infected by BHV-1 as judged by in situ hybridization and PCR, respectively. Immediate-early (bovine ICP0) and early (ribonucleotide reductase) transcripts were detected in PBMC and CD4+ lymphocytes prepared from infected calves. In contrast, a late transcript (glycoprotein C) was not consistently detected suggesting productive infection was not efficient. Taken together, these results indicate that BHV-1 can infect CD4+T cells in cattle, leading to apoptosis and suppression of cell-mediated immunity.

scholarly journals The anti-apoptotic, antioxidant and anti-inflammatory effects of curcumin on acrylamide-induced neurotoxicity in rats

2020 ◽  
Author(s):  
Jie Guo ◽  
Xiaolu Cao ◽  
Xianmin Hu ◽  
Shulan Li ◽  
Jun Wang
Keyword(s):  
Oxidative Stress ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Apoptotic Cells ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Factor Α ◽  
Necrosis Factor

Abstract Background: Acrylamide (ACR) formed during heating of tobacco and carbohydrate-rich food as well as widely applied in industries has been known as a well-established neurotoxic pollutant. Although the precise mechanism is unclear, enhanced apoptosis, oxidative stress and inflammation have been demonstrated to contribute to the ACR-induced neurotoxicity. In this study, we assessed the possible anti-apoptotic, antioxidant and anti-inflammatory effects of curcumin, the most active component in a popular spice known as turmeric, on the neurotoxicity caused by ACR in rats.Methods: Curcumin at the dose of 50 and 100 mg/kg was orally given to ACR- intoxicated Sprague-Dawley rats exposed by ACR at 40mg/kg for 4 weeks. All rats were subjected to behavioral analysis. The HE staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) staining were used to detect histopathological changes and apoptotic cells, respectively. The mRNA and protein expressions of apoptosis-related molecule telomerase reverse transcriptase (TERT) were detected using real-time PCR and immunohistochemistry, respectively. The contents of malondialdehyde (MDA) and glutathione (GSH) as well as the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured as the indicators for evaluating the level of oxidative stress in brain. The levels of pro-inflammatory cytokinestumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the cerebral homogenates were detected using ELISA assay.Results: ACR-induced weigh loss, deficits in motor function as well as pathological alterations in brains were significantly improved in rats administrated with 50 and 100 mg/kg curcumin. TUNEL-positive apoptotic cells in curcumin-treated ACR intoxicated brains were less than those in the ACR model group. Curcumin administration especially at the dose of 100 mg/kg upregulated the TERT mRNA expression and enhanced the number of TERT-positive cells in ACR-intoxicated cortex tissues. Moreover, curcumin treatment reduced the concentrations of TNF-α, IL-1β and MDA, while increased the GSH contents as well as the SOD and GSH-Px activities in the cerebral homogenates, in comparison to ACR control group.Conclusions: These data suggested the anti-apoptotic, antioxidant and anti-inflammatory effects of curcumin on ACR-induced neurotoxicity in rats. Maintaining TERT-related anti-apoptotic function might be one mechanism underlying the protective effect of curcumin on ACR-intoxicated brains.

scholarly journals In situ cellular immune response in non-ulcerated skin lesions due to Leishmania (L.) infantum chagasi infection

2021 ◽  
Author(s):  
Carmen Sandoval ◽  
Gabriela Araujo ◽  
Wilfredo Sosa ◽  
Sara Avalos ◽  
Fernando Silveira ◽  
...  
Keyword(s):  
Immune Response ◽  
T Lymphocytes ◽  
Cutaneous Leishmaniasis ◽  
Cellular Immune Response ◽  
Defense Mechanisms ◽  
Skin Lesions ◽  
Cd8 T Lymphocytes ◽  
Ifn Γ ◽  
Cellular Immune

Background Skin lesions of patients affected by non-ulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi are characterized by lymphohistiocytic inflammatory infiltrate associated with epithelioid granuloma and scarce parasitism. However, the in situ cellular immune response of these patients is unclear. Therefore, the aim of the present study was to characterize the cellular immune response in the skin lesions of patients affected by NUCL. Methods Twenty biopsies were processed by immunohistochemistry using primary antibodies to T lymphocytes (CD4, CD8), NK cells, B lymphocytes, macrophages, nitric oxide synthase and interferon-gamma. Results Immunohistochemistry revealed higher expression of all cellular types and molecules (IFN-γ, iNOS) in the dermis of diseased skin compared to the skin of healthy individuals (p < 0.05). Morphometric analysis performed in the skin lesions sections showed the predominance of CD8+ T lymphocytes in the mononuclear infiltrate, followed by macrophages, mostly iNOS+, a response that could be mediated by IFN-γ. Conclusion Our study improves knowledge of the cellular immune response in non-ulcerated or atypical cutaneous leishmaniasis caused by L. (L.) infantum chagasi in Central America and pointed to the pivotal participation of CD8+ T lymphocytes in the host defense mechanisms against the parasite in patients with NUCL.

scholarly journals Cellular Immune Suppressor Activity Resides in Lymphocyte Cell Clusters Adjacent to Granulomata in Human Coccidioidomycosis

2005 ◽  
Vol 73 (7) ◽  
pp. 3923-3928 ◽  
Author(s):  
Lijin Li ◽  
Sharon M. Dial ◽  
Monika Schmelz ◽  
Margaret A. Rennels ◽  
Neil M. Ampel
Keyword(s):  
T Lymphocytes ◽  
B Lymphocytes ◽  
Lymphocyte Subsets ◽  
Interleukin 10 ◽  
Suppressor Activity ◽  
Ifn Γ ◽  
Cellular Immune ◽  
Lymphocyte Cell ◽  
Immune Suppressor

ABSTRACT The in situ immunologic response in human coccidioidomycosis remains undefined. To explore this further, pulmonary necrotizing coccidioidal granulomata were examined using immunohistochemical staining for lymphocyte subsets and for the cytokines interleukin-10 (IL-10) and gamma interferon (IFN-γ). Discrete perigranulomatous lymphocytic clusters were seen in eight of nine tissues examined. In these tissues, T lymphocytes (CD3+) significantly outnumbered B lymphocytes (CD20+) in the mantle area of the granulomata (P = 0.028), whereas the clusters were composed of roughly equal numbers of T and B lymphocytes. While the number of cells in the mantle expressing IL-10 was similar to those in the perigranulomatous clusters, there were significantly more cells expressing IFN-γ in the mantle than in the clusters (P = 0.037). Confocal microscopy revealed that CD4+ T lymphocytes and B lymphocytes are associated with IL-10 production. CD4+CD25+ T lymphocytes were identified in the perigranulomatous clusters but were not associated with IL-10 production. This is the first report noting perigranulomatous lymphocyte clusters and IL-10 in association with human coccidioidal granulomata and suggests that down-regulation of the cellular immune response is occurring within coccidioidal granulomata.

Both B and T lymphocytes may be clonally involved in myelofibrosis with myeloid metaplasia

Blood ◽  
2003 ◽  
Vol 101 (5) ◽  
pp. 1981-1983 ◽  
Author(s):  
Terra L. Reeder ◽  
Richard J. Bailey ◽  
Gordon W. Dewald ◽  
Ayalew Tefferi
Keyword(s):  
T Lymphocytes ◽  
Cell Types ◽  
Interphase Cytogenetics ◽  
Myeloid Metaplasia ◽  
Karyotypic Abnormality ◽  
Normal Limits ◽  
Myelofibrosis With Myeloid Metaplasia ◽  
Lymphocyte Populations ◽  
B And T Lymphocytes

A combination of magnetic cell sorting (MACS) and fluorescent in situ hybridization (FISH) techniques was used to detect clonal cytogenetic markers in different myeloid and lymphoid cell types of the peripheral blood from 4 patients with myelofibrosis with myeloid metaplasia (MMM) that was associated with either a 13q− or a 20q− karyotypic abnormality. Interphase cytogenetics studies demonstrated abnormal clonal FISH signal patterns in neutrophil, myeloid, erythroid, megakaryocyte, and B- and T-cell preparations in 3 of the 4 patients. In one patient, FISH results were within normal limits in T cells and slightly abnormal in B cells. In general, the percentage of abnormal nuclei was variable in both lymphocyte populations but always higher in B lymphocytes compared with T lymphocytes. The current study provides direct evidence for the clonal involvement of both B and T lymphocytes in MMM. A larger study is needed to clarify the relevance of the observed interpatient heterogeneity in clonal constitution.

Author response for "Antigen‐driven PD‐1 + TOX + BHLHE40 + and PD‐1 + TOX + EOMES + T lymphocytes regulate juvenile idiopathic arthritis in situ"

2020 ◽  
Author(s):  
Patrick Maschmeyer ◽  
Gitta Anne Heinz ◽  
Christopher Mark Skopnik ◽  
Lisanne Lutter ◽  
Alessio Mazzoni ◽  
...  
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
T Lymphocytes ◽  
Author Response
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