| 1. |
- Al-Omari, Mariam, et al.
(författare)
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Beneficial effects of alpha-1 antitrypsin therapy in a mouse model of colitis-associated colon cancer
- 2023
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Ingår i: BMC Cancer. - 1471-2407. ; 23:1
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Tidskriftsartikel (refereegranskat)abstract
- Background: It is widely accepted that chronic inflammatory bowel diseases significantly higher a risk for colorectal cancer development. Among different types of treatments for patients with colon cancer, novel protein-based therapeutic strategies are considered. AIM: To explore the effect of human plasma alpha-1 antitrypsin (AAT) protein in the chemically induced mouse model of colorectal cancer. Methods: BALB/c mice with azoxymethane/dextran sodium sulfate (AOM/DSS)-induced colitis-associated colorectal cancer (CAC), we intraperitoneally treated with commercial preparation of human plasma AAT (4 mg per mouse). Effects of this therapy were evaluated histologically, and by immunohistochemical and gene expression assays. Results: When compared with non-treated controls, AOM/DSS mice receiving AAT therapy exhibited significantly longer colons, and less anal bleeding. Concurrently, AAT-treated mice had significantly fewer polyps, and lower numbers of large colon tumors. Immunohistochemical examinations of colon tissues showed significantly lower neutrophil counts, more granzyme B-positive but fewer MMP9 (gelatinase B)-positive cancer cells and lower numbers of apoptotic cells in mice receiving AAT therapy. The expression levels of IL4 were significantly higher while TNFA was slightly reduced in tumor tissues of AOM/DSS mice treated with AAT than in AOM/DSS mice. Conclusion: Human AAT is an acute phase protein with a broad-protease inhibitory and immunomodulatory activities used as a therapeutic for emphysema patients with inherited AAT deficiency. Our results are consistent with previous findings and support an idea that AAT alone and/or in combination with available anti-cancer therapies may represent a new personalized approach for patients with colitis-induced colon cancer.
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| 2. |
- Esquinas, Cristina, et al.
(författare)
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Gene and miRNA expression profiles in PBMCs from patients with severe and mild emphysema and PiZZ alpha I-antitrypsin deficiency
- 2017
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Ingår i: International Journal of COPD. - 1176-9106. ; 12, s. 3381-3390
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Tidskriftsartikel (refereegranskat)abstract
- Introduction: COPD has complex etiologies involving both genetic and environmental determinants. Among genetic determinants, the most recognized is a severe PiZZ (Glu342Lys) inherited alpha1-antitrypsin deficiency (AATD). Nonetheless, AATD patients present a heterogeneous clinical evolution, which has not been completely explained by sociodemographic or clinical factors. Here we performed the gene expression profiling of blood cells collected from mild and severe COPD patients with PiZZ AATD. Our aim was to identify differences in messenger RNA (mRNA) and microRNA (miRNA) expressions that may be associated with disease severity. Materials and methods: Peripheral blood mononuclear cells from 12 COPD patients with PiZZ AATD (6 with severe disease and 6 with mild disease) were used in this pilot, high-throughput microarray study. We compared the cellular expression levels of RNA and miRNA of the 2 groups, and performed functional and enrichment analyses using the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene-ontology (GO) terms. We also integrated the miRNA and the differentially expressed putative target mRNA. For data analyses, we used the R statistical language R Studio (version 3.2.5). Results: The severe and mild COPD-AATD groups were similar in terms of age, gender, exacerbations, comorbidities, and use of augmentation therapy. In severe COPD-AATD patients, we found 205 differentially expressed genes (DEGs) (114 upregulated and 91 downregulated) and 28 miRNA (20 upregulated and 8 downregulated) compared to patients with mild COPD-AATD disease. Of these, hsa-miR-335-5p was downregulated and 12 target genes were involved in cytokine signaling, MAPK/mk2, JNK signaling cascades, and angiogenesis were much more highly expressed in severe compared with mild patients. Conclusions: Despite the small sample size, we identified downregulated miRNA (hsa-miR-335) and the activation of pathways related to inflammation and angiogenesis on comparing patients with severe vs mild COPD-AATD. Nonetheless, our findings warrant further validation in large studies.
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| 3. |
- Janciauskiene, Sabina, et al.
(författare)
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Allergen-specific immunotherapy increases plasma gelsolin levels
- 2014
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Ingår i: American Journal of Rhinology & Allergy. - : SAGE Publications. - 1945-8924 .- 1945-8932. ; 28:3, s. 136-140
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Tidskriftsartikel (refereegranskat)abstract
- Background: It has been observed that patients with allergic asthma/rhinitis have increased apoptosis of peripheral blood cells. This study was designed to explore the idea that the markers of apoptosis may help predict the response of allergen immunotherapy. Methods: The Allergy Department of University Hospital, Malmo, Sweden, recruited a total of 58 young adults (<35 years) with a history of birch pollen/grass pollen-induced allergic rhinitis. Their diagnoses were verified by positive skin-prick tests and the presence of serum-specific immunoglobulin E antibodies toward birch and/or grass pollen. Plasma samples were obtained from 34 patients before the start of immunotherapy and 24 patients after treatment. The control group consisted of 38 nonallergic individuals. The levels of plasma gelsolin, soluble forms of Fas (sFas) and Fas ligand (Fas-L), the chemokine CCL17 (thymus- and activation-regulated chemokine), and tissue inhibitor of metalloprotease (TIMP) 1, were measured by enzyme-linked immunosorbent assay. Results: In patients receiving immunotherapy plasma gelsolin levels were higher relative to those without immunotherapy (the median level was 23.97 mu g/mL [range, 18-35.8 mu g/mL] versus 21.2 mu g/mL [range, 13.9-29.8 mu g/mL]; p = 0.012) and were similar to those of healthy controls (24.7 mu g/mL [range, 17.4-35.3 mu g/mL]). Plasma levels of sFas, Fas-L, CCL17, and TIMP-1 did not differ between study groups. Only in controls did the plasma gelsolin levels inversely correlate to the levels of soluble Fas. Conclusion: Allergen-specific immunotherapy increases plasma levels of gelsolin, an antioxidant and antiapoptotic protein.
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| 4. |
- Janciauskiene, Sabina, et al.
(författare)
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Indirect effect of alpha-1-antitrypsin on endotoxin-induced IL-1β secretion from human PBMCs
- 2022
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Ingår i: Frontiers in Pharmacology. - : Frontiers Media SA. - 1663-9812. ; 13
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Tidskriftsartikel (refereegranskat)abstract
- Human alpha-1-antitrypsin (AAT) encoded by the SERPINA1 gene, is an acute phase glycoprotein that regulates inflammatory responses via both protease inhibitory and non-inhibitory activities. We previously reported that AAT controls ATP-induced IL-1β release from human mononuclear cells by stimulating the release of small bioactive molecules. In the current study, we aimed to elucidate the identity of these putative effectors released from human PBMCs in response to AAT, which may inhibit the LPS-induced release of IL-1β. We pre-incubated human PBMCs alone or with different preparations of AAT (4 mg/ml) for 30 min at 37°C, 5% CO2, and collected cell supernatants filtered through centrifugal filters (cutoff 3 kDa) to eliminate AAT and other high molecular weight substances. Supernatants passed through the filters were used to culture PBMCs isolated from the autologous or a heterologous donors with or without adding LPS (1 μg/ml) for 6 h. Unexpectedly, supernatants from PBMCs pre-incubated with AAT (Zemaira®), but not with other AAT preparations tested or with oxidized AAT (Zemaira®), lowered the LPS-induced release of IL-1β by about 25%–60% without affecting IL1B mRNA. The reversed-phase liquid chromatography coupled with mass spectrometry did not confirm the hypothesis that small pro-resolving lipid mediators released from PBMCs after exposure to AAT (Zemaira®) are responsible for lowering the LPS-induced IL-1β release. Distinctively from other AAT preparations, AAT (Zemaira®) and supernatants from PBMCs pre-treated with this protein contained high levels of total thiols. In line, mass spectrometry analysis revealed that AAT (Zemaira®) protein contains freer Cys232 than AAT (Prolastin®). Our data show that a free Cys232 in AAT is required for controlling LPS-induced IL-1β release from human PBMCs. Further studies characterizing AAT preparations used to treat patients with inherited AAT deficiency remains of clinical importance.
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| 5. |
- Menzel, Mandy, et al.
(författare)
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Oxidative Stress Attenuates TLR3 Responsiveness and Impairs Anti-viral Mechanisms in Bronchial Epithelial Cells From COPD and Asthma Patients
- 2019
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Ingår i: Frontiers in Immunology. - : Frontiers Media SA. - 1664-3224. ; 10
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Tidskriftsartikel (refereegranskat)abstract
- COPD and asthma exacerbations are commonly triggered by rhinovirus infection. Potentially promoting exacerbations, impaired anti-viral signaling and attenuated viral clearance have been observed in diseased bronchial epithelium. Oxidative stress is a feature of inflammation in asthma and COPD and is prominent during exacerbations. It is not known whether oxidative stress affects the anti-viral signaling capacity. Bronchial epithelial cells from asthmatic and COPD donors were infected with rhinovirus or treated with the oxidative stressor H2O2 followed by exposure to the synthetic viral replication intermediate poly(I:C). Poly(I:C) was used to ascertain a constant infection-like burden. Gene and protein levels of antioxidants as well as anti-viral responses were measured 3 and 24 h post poly(I:C) exposure. Rhinovirus infection and poly(I:C) stimulation induced protein levels of the antioxidants SOD1 and SOD2. In asthmatic bronchial epithelial cells pre-treatment with H2O2 dose-dependently decreased the antioxidant response to poly(I:C), suggesting exaggerated oxidative stress. Further, poly(I:C)-induced IFN beta gene expression was reduced after pre-treatment with H2O2. This epithelial effect was associated with a reduced expression of the pattern recognition receptors RIG-I, MDA5 and TLR3 both on gene and protein level. Pre-treatment with H2O2 did not alter antioxidant responses in COPD bronchial epithelial cells and, more modestly than in asthma, reduced poly(I:C)-induced IFN beta gene expression. Knockdown of TLR3 but not RIG-I/MDA5 abrogated impairment of poly(I:C)-induced IFN beta gene expression by H2O2. We developed a method by which we could demonstrate that oxidative stress impairs anti-viral signaling in bronchial epithelial cells from asthmatic and COPD patients, most pronounced in asthma. The impairment apparently reflects reduced responsiveness of TLR3. These present findings shed light on molecular mechanisms potentially causing reduced interferon responses to rhinovirus infection at exacerbations in asthma and COPD. Together, our findings suggest a possible self-perpetuating vicious cycle underlying recurrent exacerbations, leading to an impaired anti-viral response, which in turn leads to viral-induced exacerbations, causing more airway inflammation.
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| 6. |
- Neuzil, J., et al.
(författare)
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Induction of cancer cell apoptosis by a-tocopheryl succinate : Molecular pathways and structural requirements
- 2001
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Ingår i: The FASEB Journal. - : Wiley. - 0892-6638 .- 1530-6860. ; 15:2, s. 403-415
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Tidskriftsartikel (refereegranskat)abstract
- The vitamin E analog a-tocopheryl succinate (a-TOS) can induce apoptosis. We show that the proapoptotic activity of a-TOS in hematopoietic and cancer cell lines involves inhibition of protein kinase C (PKC), since phorbol myristyl acetate prevented a-TOS-triggered apoptosis. More selective effectors indicated that a-TOS reduced PKCa isotype activity by increasing protein phosphatase 2A (PP2A) activity. The role of PKCa inhibition in a-TOS-induced apoptosis was confirmed using antisense oligonucleotides or PKCa overexpression. Gain- or loss-of-function bcl-2 mutants implied modulation of bcl-2 activity by PKC/PP2A as a mitochondrial target of a-TOS-induced proapoptotic signals. Structural analogs revealed that a-tocopheryl and succinyl moieties are both required for maximizing these effects. In mice with colon cancer xenografts, a-TOS suppressed tumor growth by 80%. This epitomizes cancer cell killing by a pharmacologically relevant compound without known side effects.
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| 7. |
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| 8. |
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| 9. |
- Olejnicka, Beata Teresa
(författare)
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Insulin-Producing Cells, Iron, Oxidative Stress, and Lysosomal Pathology
- 1999
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Accumulating evidence suggests that injnries caused by oxygen-derived radicals contribute to the destruction of pancreatic islet ß-cells in autoinnnune diabetes mellitus (diabetes type I, or IDDM). Oxidative stress may be caused by an enhanced production of oxygen-derived radicals, or by a decreased scavenging of such molecules. It was recently suggested that iron-mediated intralysosomal oxidative reactions result in the destabilization of lysosomal membranes, leakage of lysosomal contents to the cytosol, cellular destruction and, moreover, that such mechanisms may operate in IDDM.In the present study, we have investigated the mechanisms by which hydrogen peroxide induces cell damage, and its possible relationship to intralysosomal iron. The work was done on three insulin-producing insulinoma cell lines: HIT-TI5, NIT-1, and RINmF cells, on mouse pancreatic islets ß-cells, and the macrophage-like J-774 cells. In particular, we studied the influence of induced autophagocytosis (by glucose- and amino acid starvation) on the sensitivity to oxidative stress; the influence of high-glucose growth media on hydrogen peroxide cytotoxicity; the protective effects by starvation-stimulated intracellular fertitin synthesis against oxidative stress; the possible relationship between oxidative stress, lysosomal destabilization and apoptotic/necrotic cell death; and the impact of iron chelation on lysosomal stability, and insulinoma- and ß-cell survival.A higb susceptibility to oxidative stress was demonstrated for all the insulin-producing cells. Starvation-induced autophagocytosis increased the concentration of desfertioxarnine-available lowmolecular- weight iron in HIT-T15 cells, as assayed by HPLC. Tbe iron was mainly found in secondary lysosomes, as shown by the autometallography technique when applied at electron nticroscopical level. Starvation enhanced oxidative stress-induced damage of the IDT-T15, RINm5F and J-774 cells, as assayed by the trypan blue dye exclusion test and tests for lysosomal stability (the actidine orange relocation/uptake tests). In contrast, the pronounced starvationinduced autophagocytosis that was shown by the most vulnerable insulinoma cell line (NIT -1) was paralleled by enhanced resistance to oxidative stress, and by increased lysosomal stability as well. A rapid NIT -1 fertitin synthesis was demonstrated by inununocytochentistry under conditions of starvation. It is believed that autophagocytotic lysosomal uptake of non-iron-saturated fertitin will allow such fertitin to act as an iron chelator and stabilize lysosomes against oxidative stress. NIT -1 and B-cells which were subjected to a low level of oxidative stress (30 J.LM H20 2 for 15 min) were still largely intact at the light nticroscopical level ,but 10-20% of the cells exhibited nuclear chromatin condensation as an early sign of apoptosis when examined by the Ho334/PI staining technique, or by TEM, 0.5-1 h after the insult. At the same point of time, a decrease in the number of intact lysosomes was also observed. The rate of oxidative stress-induced lysosomal destabilization progressed with time, and a widespread apoptotic/necrotic-type degeneration/fragmentation ensued, as demonstrated by SEM, TEM, and the TUNEL-reaction. The ntitochondria revealed a mixture of lamelliform and swollen cristae, indicating altered properties of the mitochondrial membranes. Pre-treatment with the iron chelator desfenioxamine attenuated the lysosomal destabilization, and increased cell viability, following exposure to oxidative stress.At high-glucose conditions, the ~02-sensitivity of HIT-T15, NIT-I, and B-cells was reduced which, was consistent with a moderately enhanced stability of their lysosomes, as measured by the acridine orange-relocation test, and with reduced amounts of desfenioxamine-available iron.We conclude that the decisive role of free lysosomal iron in oxidative stress is strongly supported by the following lines of evidence, provided by the present study (a) glucose- and amino acid-starvation promotes autophagic/crinophagic activity of the cells, resulting in enrichment of intracellular (intralysosomal) desfenioxamine-available iron; (b) high-glucose conditions depress autophagic/crinophagic activity and, consequently, the occurrence of intralysosomal iron; (c) starvation-stimulated fenitin synthesis enhances lysosomal stability during oxidative stress by limiting lysosomal redox-active iron; (d) lysosomal destabilization and related apoptotic cell death are associated with the amounts of intralysosomal iron in redox -active form,
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| 10. |
- Tumpara, Srinu, et al.
(författare)
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A novel mouse monoclonal antibody c42 against c-terminal peptide of alpha-1-antitrypsin
- 2021
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Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 22:4
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Tidskriftsartikel (refereegranskat)abstract
- The C-terminal-fragments of alpha1-antitrypsin (AAT) have been identified and their diverse biological roles have been reported in vitro and in vivo. These findings prompted us to develop a monoclonal antibody that specifically recognizes C-36 peptide (corresponding to residues 359–394) resulting from the protease-associated cleavage of AAT. The C-36-targeting mouse mono-clonal Immunoglobulin M (IgM) antibody (containing κ light chains, clone C42) was generated and enzyme-linked immunosorbent assay (ELISA)-tested by Davids Biotechnologie GmbH, Germany. Here, we addressed the effectiveness of the novel C42 antibody in different immunoassay formats, such as dot-and Western blotting, confocal laser microscopy, and flow cytometry. According to the dot-blot results, our novel C42 antibody detects the C-36 peptide at a range of 0.1–0.05 µg and shows no cross-reactivity with native, polymerized, or oxidized forms of full-length AAT, the AAT-elastase complex mixture, as well as with shorter C-terminal fragments of AAT. However, the C42 antibody does not detect denatured peptide in SDS-PAGE/Western blotting assays. On the other hand, our C42 antibody, unconjugated as well as conjugated to DyLight488 fluorophore, when applied for immunofluorescence microscopy and flow cytometry assays, specifically detected the C-36 peptide in human blood cells. Altogether, we demonstrate that our novel C42 antibody successfully recognizes the C-36 peptide of AAT in a number of immunoassays and has potential to become an important tool in AAT-related studies.
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| 11. |
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| 12. |
- Zendehrokh, Nooreldin, et al.
(författare)
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Liquid-based cytology using cytorich Red/Tripath is diagnostically equivalent to conventional smears for bronchial washings and brushings and reduces the cost.
- 2013
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Ingår i: Diagnostic Cytopathology. - : Wiley. - 8755-1039. ; 41:10, s. 876-884
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Tidskriftsartikel (refereegranskat)abstract
- A split sample study was performed on 109 bronchial brushings and washings and evaluated from conventional slides (CS) and CytoRich Red/Tripath preparations (CRR/Tripath). Unassessable bronchial washings were significantly more frequent in CS (5 vs. 0), but as all brushings were assessable with both methods, no overall diagnostic advantage was found. CS and CRR/Tripath gave discordant diagnoses in one case with a final benign diagnosis and in six cases with final malignant diagnoses. In the benign case, atypia was assessed in CS. In the malignant cases, suspected malignancy was found in one CRR/Tripath and one CS, atypia vs. benign assessment was also balanced, with three atypias in CRR/Tripath and two in CS. The better preserved cells in CRR/Tripath facilitated correct diagnosis in some cases, but might also lead to false positive diagnoses. In small cell carcinomas diagnostic hints such as smearing and moulding were less pronounced in CRR/Tripath but this did not affect the diagnostic accuracy. Overall, the diagnostic performance with CRR/Tripath was at least as good as with conventional slides, although statistically no difference could be seen. The number of slides and screening time, and thereby cost was significantly reduced with CRR/Tripath, thus the liquid-based method is preferred for bronchial washings and brushings. Diagn. Cytopathol. 2013. © 2013 Wiley Periodicals, Inc.
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