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Aging affects AO rat splenic conventional dendritic cell subset composition, cytokine synthesis and T-helper polarizing capacity
No full textIt is well-established that almost all cellular components of innate and adaptive immunity undergo age-related remodelling. The findings on age-related changes in both human and mouse dendritic cells (DCs) are conflicting, whereas there are no data on the influence of aging on rat DCs. In an attempt to fill this gap, freshly isolated splenic DCs expressing CD103 (alpha(OX-62) integrin), a DC specific marker recognized by MRC OX62 monoclonal antibody, from 3- (young) and 26-month-old (aged) Albino Oxford rats were examined for subset composition, expression of activation/differentiation markers (CD80, CD86 and CD40 and MHC II molecules) and endocytic capacity using flow cytometric analysis (FCA). In addition, splenic OX62+ DCs cultured in the presence or absence of LPS were analysed for the activation marker and TNF-alpha, IL-6, IL-12, IL-23, TGF-beta 1, IL-10 expression using FCA, RT-PCR and ELISA, respectively. Moreover, the allostimulatory capacity of OX62+ DCs and IFN-gamma, IL-4 and IL-17 production by CD4+ T cells in mixed leukocyte reaction was quantified using FCA and ELISA, respectively. It was found that aging: i) shifts the CD4+:CD4- subset ratio in the OX62+ DCs population towards the CD4- subset and ii) influences DCs maturation (judging by activation marker expression and efficiency of endocytosis) by affecting the expression of intrinsic (TNF-alpha and IL-10) and extrinsic maturation regulators. Furthermore, in LPS-matured OX62+ DCs from aged rats expression of TNF-alpha, IL-12, IL-23 and IL-6 was increased, whereas that of IL-10 was diminished compared with the corresponding cells from young rats. Moreover, in MLR, OX62+ DCs from aged rats exhibited enhanced Th1/Th17 driving force and diminished allostimulatory capacity compared with those from young rats
Trichoderma species on Agaricus bisporus farms in Serbia and their biocontrol
No full textTwenty Trichoderma isolates were collected on 13 Serbian Agaricus bisporus farms and one in Bosnia and Herzegovina during 2006–2010. Twelve isolates were classified into five species by standard mycological studies and ITS1/ITS4 sequence analyses, namely Trichoderma atroviride, Trichoderma koningii, Trichoderma virens, Trichoderma aggressivum f. europaeum and Trichoderma harzianum. Eight isolates were not identified to the species level but were shown to be related to T. harzianum. The isolates of T. harzianum exhibited the highest virulence to the harvested A. bisporus pilei and T. virens and T. aggressivum f. europaeum the lowest. Antifungal activity of two biofungicides based on Bacillus subtilis and tea tree oil and the fungicide prochloraz manganese were tested in vitro to all Trichoderma isolates. Prochloraz manganese and B. subtilis were highly toxic to all tested Trichoderma isolates, their ED50 values were below 0.3 and 1.3 mg L−1, respectively. Tea tree oil did not exhibit a significant antifungal activity (ED50 = 11.9–370.8 mg L−1). The effectiveness of biofungicides was evaluated against T. harzianum in a mushroom growing room, and they were applied alone or in combination with the fungicide at a respective proportion of 20:80%. Prochloraz manganese showed higher effectiveness than both tested biofungicides or their respective mixtures. The biofungicide based on B. subtilis demonstrated greater effectiveness in preventing disease symptoms than tea tree oil. B. subtilis combined with the fungicide revealed less antagonism in effectiveness against pathogen than tea tree oil
Age-associated shift in rat dendritic cell T-helper polarizing capacity
Get PDFAlmost all cellular components of innate and adaptive immunity undergo age-related remodeling. The findings on age-related
changes in human and mouse dendritic cells (DCs) are conflicting, whereas there is no data on the influence of aging on rat DCs. In
attempt to fill this gap, freshly isolated splenic conventional OX62+ DCs from 3- (young) and 26-month-old (aged) Albino Oxford rats
were examined for subset composition, cell surface expression of activation markers (CD80, CD86 and CD40 and MHC II molecules)
and endocytic capacity using flow cytometric analysis (FCA). In addition, splenic OX62+ DCs isolated from rats of both ages were
cultured in the presence or in the absence of LPS. These cells were examined for the activation marker and TNF-α, IL-6, IL-12, IL-23,
TGF-β1, IL-10 expression using FCA, and RT-PCR and ELISA, respectively. Moreover, the allostimulatory capacity of OX62+ DCs and
allogeneic CD4+ T cell cytokine (IFN-γ, IL-4 and IL-17) production in MLR was quantified using FCA and ELISA, respectively. It was
found that aging: i) in OX62+ DCs population leads to a shift in CD4+:CD4- cell ratio towards CD4- cells and ii) influences OX62+
DCs maturation capacity (judging by activation marker expression and efficiency of endocytosis) by affecting action of intrinsic (TNF-
α and IL-10) and extrinsic regulatory factor expression. Furthermore, in LPS-matured OX62+ DCs from aged rats TNF-α, IL-12, IL-23
and IL-6 expression was increased, while IL-10 expression was diminished. Moreover, in MLR, OX62+ DCs from aged rats exhibited
enhanced Th1/Th17 driving force and diminished allostimulatory capacity
Morphological changes in lymph nodes and spleen upon EAE induction in C57BL/6 mic
Get PDFMyelin oligodendrocyte glycoprotein (MOG) is a protein widely used in the induction of experimental autoimmune encephalomyelitis (EAE) for studying human multiple sclerosis (MS). In C57BL/6 female mice aged eight weeks, we administered subcutaneously MOG35-55 peptide in CFA (complete Freund's adjuvant) along with pertussis vaccine injected intraperitoneally. We observed the sign of flaccid tail as early as thirteen days post-immunization in five of twelve animals. Hematoxylin and eosin staining of paraffin-embedded sections of lymph nodes and spleen revealed the presence of germinal centers in the immunized animals. In the control group of animals, lymphoid follicles without germinal centers were observed. Immunohistochemical staining of spleen sections revealed an expression of MHC II molecules in the EAE-induced group. We would like to point out that even though the clinical signs are mild, the morphological changes are apparent in the lymph nodes and spleen of MOG35-55-immunized mice
Effects of astaxanthin supplementation on DNA stability and paraoxonase 1 activity in young soccer players
No full textPeritoneal mast cell degranulation differently affected thioglycollate-induced macrophage phenotype and activity in Dark Agouti and Albino Oxford rats
No full textAims: Macrophages are heterogeneous population of inflammatory cells and, in response to the microenvironment, become differentially activated. The objective of the study was to explore macrophage effector functions during different inflammatory conditions in two rat strains. Main methods: We have investigated the effects of in vivo treatment with mast cell-degranulating compound 48/80 and/or thioglycollate on peritoneal macrophage phagocytosis and capacity to secrete hydrogen peroxide (H2O2), tumor necrosis factor-alpha (INF-alpha) and nitric oxide (NO) in Dark Agouti (DA) and Albino Oxford (AO) rat strains. Besides, fresh peritoneal cells were examined for the expression of ED1, ED2 and CD86 molecules. Key findings: In thioglycollate-elicited macrophages, increased proportion of ED1 + cells was accompanied with elevated phagocytosis of zymosan (DA strain), whereas increased expression level of CD86 molecule on ED2 + macrophages matched elevated secretory capacity for H2O2, TNF-alpha and NO (AO rats). Although mast cell degranulation induced by compound 48/80 increased the percentages of ED2 + macrophages in both rat strains, the proportion of ED2 + cells expressing CD86 molecule was decreased and increased in DA and AO rats, respectively. Furthermore, in DA strain compound 48/80 diminished macrophage secretion of NO, but stimulated all macrophage functions tested in AO strain. If applied concomitantly, the compound 48/80 additively increased macrophage activity induced by thioglycollate in AO rats. Significance: Macrophages from DA and AO rat strains show different susceptibility to mediators released from mast cells, suggesting that strain-dependant predisposition(s) toward particular activation pattern is decisive for the macrophage efficacy in response to inflammatory agents. (c) 2013 Elsevier Inc. All rights reserved
The intriguing mission of neuropeptide Y in the immune system
No full textFor many years, the central nervous system and the immune system were considered two autonomous entities. However, extensive research in the field of neuroimmunomodulation during the past decades has demonstrated the presence of different neuropeptides and their respective receptors in the immune cells. More importantly, it has provided evidence for the direct effects of neuropeptides on the immune cell functions. Neuropeptide Y (NPY) is generally considered the most abundant peptide in the central and peripheral nervous system. However, it is also distinguished by exhibiting pleiotropic functions in many other physiological systems, including the immune system. NPY affects the functions of the cells of the adaptive and innate immunity. In this respect, NPY is known to modulate immune cell trafficking, T helper cell differentiation, cytokine secretion, natural killer cell activity, phagocytosis and the production of reactive oxygen species. The specific Y receptors have been found in immune cells, and their expression is amplified upon immune stimulation. Different Y receptor subtypes may mediate an opposite effect of NPY on the particular function, thus underlining its regulatory role. Since the immune cells are capable of producing NPY upon appropriate stimulation, this peptide can regulate immune cell functions in an autocrine/paracrine manner. NPY also has important implications in several immune-mediated disorders, which affirms the clear need for further investigation of its role in either the mechanisms of the disease development or its possible therapeutic capacity. This review summarises the key points of NPY's mission throughout the immune system
Hormoni ovarijuma imaju različit uticaj na modelovanje apoptoze i proliferacije unutar različitih subpopulacija timocita tokom involucije timusa
Get PDFThe study was aimed to define the putative role of ovarian hormones in shaping thymocyte apoptosis and proliferation during thymic involution. Thymocytes from young adult and middle-aged rats ovariectomized (Ox) before puberty were examined for apoptosis and proliferation. Apoptosis and proliferation were measured in fresh thymocyte suspensions and in their 18-hour cultures, and fresh thymocyte suspensions, respectively. The thymocyte population and the major thymocyte subsets were analyzed following triple staining using anti-CD4 and anti-CD8 monoclonal antibodies and 7-AAD to label apoptotic or proliferating cells. The frequency of apoptotic cells was lower in thymocyte suspensions and cultures from Ox rats of both ages. This reflected in a diminished frequency of apoptotic cells amongst CD4+CD8+ double positive (DP) and CD4+CD8- single positive (SP), and DP cells in young and middle-aged Ox rats, respectively. Additionally, in thymocyte cultures from Ox rats the frequency of apoptotic cells amongst CD4+CD8- and CD4-CD8+ SP cells decreased with age, but increased within DP and CD4-CD8- double negative (DN) subsets, reaching in the former subset from middle-aged Ox rats higher values than in age-matched controls. The frequency of proliferating cells was also lower in Ox rats than in controls. This reflected the lower frequency of cycling cells amongst CD4+CD8- SP and CD4-CD8+ SP thymocytes in young rats, and DP and CD4-CD8+ SP thymocytes in middle-aged rats. Besides, in both SP and DP thymocyte subsets from Ox rats the frequency of proliferating cells declined with age. In conclusion, thymocyte apoptosis and proliferation exhibit ovarian hormone-dependent thymocyte subset specific alterations during thymic involution.Cilj istraživanja je bio da se definiše značaj hormona ovarijuma za razvoj promena u apoptozi i proliferaciji timocita tokom involucije timusa. U tom cilju apoptoza i proliferacija timocita ispitivana je kod prepubertetno ovariektomisanih (Ox) mladih (uzrasta 2 meseca) i sredovečnih pacova (uzrasta 11 meseci). Apoptoza je određivana u suspenziji sveže izolovanih timocita i nakon njihove 18- časovne kultivacije, a proliferacija u suspenziji sveže izolovanih timocita. Procenat apoptotičnih i proliferišućih ćelija je određivan u celokupnoj populaciji timocita, i unutar glavnih subpopulacija ovih ćelija, koje su razdvojene na osnovu ekspresije CD4/CD8 molekula, metodom protočne fluorocitometrije, korišćenjem 7-aminoaktinomicina D (7-AAD). Procenat ćelija u apoptozi je bio značajno manji u suspenzijama svežih timocita koji su izolovani iz Ox životinja i u njihovim kulturama nego u onim izolovanim iz kontrolnih životinja. Ovaj nalaz je odražavao smanjenu učestalost ćelija u apoptozi u CD4+CD8+ dvostruko pozitivnoj (DP) i CD4+CD8- jednostruko pozitivnoj (JP) subpopulaciji timocita kod mladih i u DP subpopulaciji kod sredovečnih Ox pacova. U kulturama timocita koji su izolovani iz sredovečnih Ox pacova uočeno je smanjenje učestalosti ćelija u apoptozi unutar subpopulacija CD4+CD8- i CD4-CD8+ JP timocita, a povećanje unutar DP i CD4-CD8- dvostruko negativne (DN) subpopulacije ovih ćelija. Učestalost proliferišućih ćelija je takođe bila niža u suspenzijama timocita izolovanih iz Ox pacova nego u onim izolovanim iz kontrolnih životinja. Ovo je odražavalo smanjenu proliferaciju CD4+CD8- i CD4-CD8+ JP timocita kod mladih, a DP i CD4-CD8+ JP timocita kod sredovečnih pacova. Procentualna zastupljenost proliferišućih ćelija u subpopulacijama JP i DP timocita je bila veća kod mladih nego kod sredovečnih Ox pacova. U zaključku, tokom involucije timusa dolazi do promena u apoptozi i proliferaciji timocita koje su specifične za pojedine subpopulacije timocita i zavisne od prisustva hormona ovarijuma
Role of ovarian hormones in T-cell homeostasis: From the thymus to the periphery
No full textThe study explored the putative role of ovarian hormones in the peripubertal remodelling of peripheral T-cell compartment. Ovariectomy at age of 1 month enhanced the peripubertal rise in CD4+ and CD8+ cell numbers in peripheral blood (PB) and spleen from 2-month-old rats. This reflected maintenance of thymopoietic efficiency at the prepubertal level (judging by numbers of the most mature CD4+ and CD8+ thymocytes and recent thymic emigrants) and alterations in T-cell survival/proliferation in the periphery. Compared with age-matched controls, the frequency of apoptotic cells among CD8+ peripheral blood lymphocytes (PBLs) and CD4+ and CD8+ splenocytes was diminished in ovariectomized (Ox) rats, at least partly, due to lower CD95 surface density. The diminished frequency of the apoptotic T splenocytes could also be associated with the rise in the amount of splenic IL-7 mRNA. Additionally, the latter finding was consistent with the augmented proliferation of CD4+ and CD8+ splenocytes. However, the enhanced proliferation of these cells could also be linked to the rise in IL-2 receptor surface density. This increase was related to the enhanced splenic TNF-alpha mRNA expression. Additionally, ovariectomy led to the phenotypic alterations in the major PBL and splenic T-cell subsets by diminishing/preventing the peripubertal changes in the frequency of cells at distinct stages of post-thymic differentiation/maturation (recent thymic emigrants, mature naive and memory cells), and by decreasing the frequency of NKT cells within peripheral CD8+ subsets. In addition to numerical and phenotypic changes in T-cell compartment (due to the lack of ovarian hormone action at both the thymic and peripheral level), Ox rats exhibited a much larger delayed-type hypersensitivity (DTH) response compared with age-matched controls. This suggested the augmented T-cell-mediated immune response in Ox rats compared with aged-matched controls. (C) 2012 Elsevier GmbH. All rights reserved
Structure and antioxidant activity of beta-lactoglobulin-glycoconjugates obtained by high-intensity-ultrasound-induced Maillard reaction in aqueous model systems under neutral conditions
Get PDFSonication is a new processing technology in the dairy industry. The aim of this study was to test glycation of beta-lactoglobulin (BLG) in Maillard reaction (MR) induced by high-intensity ultrasound in aqueous solution under neutral conditions at 10-15 degrees C, which is not favourable for the MR. BLG was sonicated in the presence of glucose, galactose, lactose, fructose, ribose and arabinose. Formation of Maillard reaction products (MRPs) was monitored by mass spectrometry, spectrophotometry and fluorimetry. Ultrasound treatment resulted in formation of MRPs with all tested carbohydrates. Ribose induced the highest degree of modification resulting in 76% of BLG modified and an average of three anhydroribose units attached. Circular dichroism spectra analyses indicated only minor alterations in secondary and tertiary structures. MRP obtained by ultrasound exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and possessed increased iron-chelating activity and reducing power. High-intensity ultrasound efficiently promotes BLG-glycoconjugates formation by MR in aqueous solutions under non-denaturing conditions.This is the peer-reviewed version of the following article: Stanic-Vucinic, D.; Prodic, I.; Apostolovic, D.; Nikolic, M.; Cirkovic Velickovic, T. Structure and Antioxidant Activity of β-Lactoglobulin-Glycoconjugates Obtained by High-Intensity-Ultrasound-Induced Maillard Reaction in Aqueous Model Systems under Neutral Conditions. Food Chemistry 2013, 138 (1), 590–599. [https://doi.org/10.1016/j.foodchem.2012.10.087]