scholarly journals Ανάπτυξη in vitro συστημάτων καλλιεργειών δενδριτικών κυττάρων για την αντιγονοπαρουσίαση τεχνητών αντιγόνων σε Τ κύτταρα ασθενών με σκλήρυνση κατά πλάκας με απώτερο σκοπό τη δημιουργία θεραπευτικών εμβολίων

2018 ◽  
Author(s):  
Μαρία Ρόδη
Keyword(s):  
Cytometric Bead Array ◽  
Gm Csf ◽  
Tnf Α ◽  
Ifn Γ ◽  
Tgf Β1

Η σκλήρυνση κατά πλάκας (ή πολλαπλή σκλήρυνση, ΠΣ) είναι μια αυτοάνοση νόσος που προσβάλει το κεντρικό νευρικό σύστημα και o παθογενετικός μηχανισμός της ενορχηστρώνεται κυρίως από αυτοαντιδρώντα Τ και Β κύτταρα τα οποία έχουν ξεφύγει από την επιτήρηση του μηχανισμού της ανοσολογικής ανοχής. Η επαγωγή της ανοχής μέσω δενδριτικών κυττάρων (DCs) είναι μία νέα στρατηγική για τη θεραπεία της. Σκοπός της παρούσας ερευνητικής εργασίας ήταν η δημιουργία και η μελέτη ενός συστήματος αντιγονοπαρουσίασης του πεπτιδίου MOG35-55 συζευγμένο με μαννάνη από ανοσοανεκτικά δενδριτικά κύτταρα (tolDCs), τα οποία παρουσιάζουν το πεπτίδιο και επάγουν την ανοχή των Τ κύτταρων. Στο πειραματικό πρωτόκολλο που αναπτύξαμε, απομονώθηκαν περιφερικά μονοπύρηνα κύτταρα του αίματος (PBMCs), από υγιείς δότες και ασθενείς με RR-MS, και τα οποία διαφοροποιήθηκαν σε διάφορους τύπους DCs υπό την επίδραση IL-4 και GM-CSF, παρουσία δεξαμεθαζόνης(DEXA) ή βιταμίνης D3(VitD3) ή και των δύο. Την 6η ημέρα της καλλιέργειας τους τα DCs «φορτώθηκαν» με το πεπτίδιο ή τη μαννάνη και έλαβαν το σήμα ωρίμανσής τους μέσω του LPS. Την 8η ημέρα συνκαλλιεργήθηκαν με αυτόλογα PBMCs παρουσία IL-2 για 3 ή 4 κύκλους αντιγονοπαρουσίασης. Ο φαινότυπος των DCs και των T-κυτταρικών πληθυσμών προσδιορίστηκε με κυτταρομετρία ροής. Τα κυτταροκινικά προφίλ των DC και των Τ-κυττάρων αποτελούμενα από τις κυτταροκίνες: IL-1β/IL-2/IL-4/IL-6/IL-8/IL-10/IL-12p70/IL-17A/TNF-α/IFN-γ, μετρήθηκαν με Cytometric Bead Array και ο TGF-β1 με ELISA.Η μελέτη και η ανάλυση των DCs την 8η ημέρα της καλλιέργειάς τους έδειξε ότι η χαμηλότερη επιφανειακή έκφραση των μορίων CD80 και CD86 και η μεγαλύτερη παραγωγή IL-10 από τα tolDCs σε σύγκριση με τα ώριμα DCs μας επιτρέπει το σαφή φαινοτυπικό και λειτουργικό διαχωρισμό μεταξύ ώριμων και ανοσοανεκτικών DCs.Οι συνκαλλιέργειες των PBMCs με τα διάφορα είδη DCs, μετά από 3 ή 4 κύκλους αντιγονοπαρουσίασης, προήγαγαν τη δημιουργία μνήμης στα CD4 και όχι στα CD8 T κύτταρα. Από τον προσδιορισμό των επίπεδων ενεργοποίησης των CD4 T κυττάρων παρατηρήθηκε καταστολή της ενεργοποίησής τους στις συνκαλλιέργειες με τα tolDCs που αντιγονοπαρουσίασαν το πεπτίδιο. Παράλληλα, παρατηρήσαμε αύξηση των ποσοστών των εξαντλημένων CD4+PD-1+ και των ρυθμιστικών CD4+CD25highFoxP3+ Τ κυττάρων και την έκκριση IL-10 ή και TGF-β1. Τα αποτελέσματα μας υποδεικνύουν ότι τα tolDCs που αναπτύξαμε in vitro επάγουν την ανοχή στα Τ κύτταρα και θα μπορούσαν να χρησιμοποιηθούν σαν θεραπευτικό εμβόλιο για την ΠΣ.

scholarly journals Γονιδιακή έκφραση και μελέτη παραγόντων που επηρεάζουν την πλαστικότητα των ανθρώπινων Τ βοηθητικών λεμφοκυττάρων σε μεταγγιζόμενους ασθενείς

2016 ◽  
Author(s):  
Παναγιώτα Σπαντιδέα
Keyword(s):  
Cytometric Bead Array ◽  
Tnf Α ◽  
Ifn Γ ◽  
Tgf Β1

Εισαγωγή: Ο πληθυσμός των CD4+ Τ κυττάρων είναι υπεύθυνος για την οργάνωση και ισορροπία του ανοσοποιητικού συστήματος και την αντιμετώπιση ειδικά έναντι σε κάθε αντιγόνο. Αυτό επιτυγχάνεται λόγω της πλαστικότητας των CD4+ κυττάρων μέσα (διαφοροποιούνται τα ρυθμιστικά Τ κύτταρα-Tregs) και κατά την έξοδό τους (παρθενικά CD4+ Τ κύτταρα) από τον θύμο καθώς και τα ερεθίσματα που δέχονται στο μικροπεριβάλλον τους κυρίως από αντιγόνα και κυτταροκίνες. Με τις αλλογενείς μεταγγίσεις αίματος (ABT) τεράστιος αριθμός αντιγόνων εισέρχεται στο σώμα του λήπτη και τον τροποποιεί με μηχανισμό ο οποίος δεν είναι πλήρως κατανοητός μέχρι σήμερα.Σκοπός της εργασίας: Στην παρούσα εργασία, μελετήθηκαν ο τρόπος και οι παράγοντες που επηρεάζουν την πλαστικότητα των CD4+ Τ κυττάρων υπό την επίδραση της μετάγγισης σε ασθενείς που είχαν υποβληθεί σε προγραμματισμένο χειρουργείο αρθροπλαστικής γόνατος ή ισχίου χωρίς άλλες ανοσοτροποποιητικές νόσους. Η διάκριση των ασθενών έγινε με βάση αν έλαβαν μετάγγιση (54 ασθενείς-group1) ή όχι (35 ασθενείς- group2).Ασθενείς, υλικά και μέθοδοι: Ηπαρινισμένα δείγματα ολικού αίματος από 89 ασθενείς συλλέχτηκαν πριν το χειρουργείο (BS) και αμέσως μετά το χειρουργείο μέχρι την πρώτη εβδομάδα (days 0-7) ή μέχρι το εξιτήριο, ένα μήνα μετά το χειρουργείο (1month) και κατά τον επανέλεγχο των ασθενών (>3months). Σε απομονωμένα PBMC’s ασθενών προσδιορίστηκε η μεταβολή των CD4+CD25high/+Foxp3+ (nTreg), CD4+CD25+CD127low/- (iTreg) και άλλων CD3+ πληθυσμών με την μέθοδο FACS και στο πλάσμα των ασθενών προσδιορίστηκαν οι συγκεντρώσεις των κυτταροκινών IL-2, IL-4, IL-5, IL-6, IL-10, TNF-α, IFN-γ με την μέθοδο Cytometric Bead Array (CBA) και των υποδοχέων TNF-RI(p55/p60), TNF-RII(p75/p80), TGF-β1 και TGF-β2 την μέθοδο της ELISA. Επίσης πραγματοποιήθηκαν πειράματα λειτουργικότητας των Tregs σε δείγματα μεταγγισμένων και μη μεταγγισμένων ασθενών, μέσα στην πρώτη μετεγχειρητική εβδομάδα. Σε αυτά καλλιεργήθηκαν απομονωμένα Tregs και Teff από τον ίδιο ασθενή για 72 ώρες σε διάφορους λόγους παρουσία PHA και CFSE. Και τέλος, έγινε συσχέτιση της μετάγγισης με την σοβαρότητα των μετεγχειρητικών επιπλοκών και την παραμονή των ασθενών στο νοσοκομείο. Αποτελέσματα: Από τα παραπάνω πειράματα παρατηρήθηκε σημαντική αύξηση των CD4+CD25high/+Foxp3+ και CD4+CD25+CD127low/- μέχρι την πρώτη μετεγχειρητική εβδομάδα στους μεταγγισμένους αλλά όχι στους μη-μεταγγισμένους ασθενείς. Τα πειράματα λειτουργικότητας απέδειξαν ότι τα Tregs μεταγγισμένων και μη μεταγγισμένων ασθενών είναι λειτουργικά και ικανά να προκαλούν αναστολή του πολλαπλασιασμού των Teff, ωστόσο, τα Tregs των μεταγγισμένων ασθενών μπορούσαν να καταστέλλουν ισχυρότερα και παρουσία μεγαλύτερου αριθμού Teff στην καλλιέργεια. Από τις κυτταροκίνες που ανιχνεύτηκαν στο πλάσμα των ασθενών η IL-6, και οι υποδοχείς TNF-RI(p55/p60) και TNF-RII(p75/p80) εμφάνισαν σημαντική αύξηση ενώ σημαντική μείωση εμφάνισε ο TGF-β1 και ιδιαίτερο ρόλο φαίνεται να παίζουν οι IL-2 και TGF-β2. Παρατεταμένη νοσηλεία καθώς και αυξημένες και πιο σοβαρές μετεγχειρητικές επιπλοκές φαίνεται να εμφανίζουν οι μεταγγισμένοι ασθενείς συγκριτικά με τους μη-μεταγγισμένους.Συμπεράσματα: Οι ασθενείς που δέχονται μετάγγιση κατά την διάρκεια του χειρουργείου είναι ανοσοκατεσταλμένοι για τις πρώτες μέρες μετά την μετάγγιση και αυτό οφείλεται στον αυξημένο αριθμό των ρυθμιστικών τους κυττάρων και την επαγωγή των προφλεγμονωδών αποκρίσεων, έτσι οι ασθενείς αναπτύσσουν Th1 απόκριση και πολλαπλασιασμό των Tregs τα οποία καταστέλλουν τον πολλαπλασιασμό των Teff ισχυρότερα. Έτσι σταδιακά, τα Tregs καταστέλλουν τις προφλεγμονώδης αποκρίσεις μέχρι να επανέλθει η ισορροπία των ληπτών μετά την εγχείρηση. Ωστόσο, η ανοσοκαταστολή οφειλόμενη στην μετάγγιση που παρατηρείται τις πρώτες μέρες, οδηγεί τους ασθενείς σε παρατεταμένη νοσηλεία και αυξημένες μετεγχειρητικές επιπλοκές.

Expression of type 1 (interferon gamma) and type 2 (interleukin-13, interleukin-5) cytokines at distinct stages of natural killer cell differentiation from progenitor cells

Blood ◽  
2002 ◽  
Vol 99 (4) ◽  
pp. 1273-1281 ◽  
Author(s):  
Matthew J. Loza ◽  
Loris Zamai ◽  
Livio Azzoni ◽  
Emanuela Rosati ◽  
Bice Perussia
Keyword(s):  
Cell Differentiation ◽  
Nk Cells ◽  
Interferon Gamma ◽  
Gm Csf ◽  
Type 2 Cytokines ◽  
Tnf Α ◽  
Ifn Γ

To determine whether production of type 1 and type 2 cytokines defines discrete stages of natural killer (NK) cell differentiation, cytokine expression was analyzed in human NK cells generated in vitro in the presence of interleukin-15 (IL-15) and/or IL-2 from umbilical cord blood hematopoietic progenitors. Like peripheral NK cells, the CD161+/CD56+ NK cells from these cultures contained a tumor necrosis factor alpha (TNF-α)+/granulocyte macrophage–colony-stimulating factor (GM-CSF)+ subset, an interferon gamma (IFN-γ)+ subset, mostly included within the former, and very few IFN-γ−/IL-13+ cells. Instead, most immature CD161+/CD56− NK cells, detectable only in the cultures with IL-2, produced IL-13, TNF-α, and GM-CSF, but not IFN-γ, and contained an IL-5+ subset. In short-term cultures with IL-12 and feeder cells, a proportion of the immature cells acquired the ability to produce IFN-γ. Part of these produced both IFN-γ and IL-13, irrespective of induced CD56 expression. These in vitro data indicate that ability to produce the type 2 cytokines IL-13 and IL-5 defines CD161+ NK cells at intermediate stages of differentiation, and is lost upon terminal functional differentiation, concomitant with acquired ability to produce IFN-γ.

scholarly journals 684 The in vitro effects of 5-Azacitidine on the immunophenotype of monocyte-derived dendritic cells from patients with higher-risk myelodysplastic syndromes

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A712-A712
Author(s):  
Randy Tsai ◽  
Hannah Fields ◽  
Xinlian Zhang ◽  
Valentina Ferrari ◽  
Soo Park ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Trypan Blue ◽  
Cultured Cells ◽  
Poly I:C ◽  
Gm Csf ◽  
Tnf Α ◽  
Before And After ◽  
Ifn Γ ◽  
Marker Expression

BackgroundMyelodysplastic syndromes (MDS) are the most common acquired cause of bone marrow failure. Though DNA hypomethylating agents (HMAs) such as 5-Azacitidine (5-Aza) may increase survival of patients with higher-risk MDS, their mechanistic effects on hematopoiesis and immune cell function remain unclear. Using whole exome sequencing analysis, we previously identified MDS-related mutations within monocyte-derived dendritic cells (moDCs) from patients with higher-risk MDS. Here we examine the effect of 5-Aza on the phenotype of moDCs from the same cohort of patients with higher-risk MDS.MethodsPurified CD14+ cells were magnetically isolated from peripheral blood mononuclear cells from 6 patients with IPSS-R Intermediate/High/Very High-risk MDS (herein collectively referred to as higher-risk MDS). Cells were cultured in complete medium with IL-4 (800 U/mL) and GM-CSF (1200 U/mL) for 5 days. Freshly prepared 5-Aza or dimethylsulfoxide (DMSO) vehicle was added to cultures every 24 hours for a total of three 1 μM doses starting on Day 1. Immature moDCs were then stimulated with poly(I:C) (20 ng/mL), IL-1β (25 ng/mL), IFN-α (3000 U/mL), IFN-γ (1000 U/mL), and TNF-α (50 ng/mL) for 48 hours to generate moDCs. Flow cytometry analyses were performed with Guava easyCyte 8HT before and after addition of maturation cocktail.ResultsBased on trypan blue staining, in vitro addition of 5-Aza to CD14+ cells from 6 patients with higher-risk MDS did not result in a significant reduction in the percentage of cell survival on Day 5 and Day 7 in culture (figure 1a, p=0.8765 and p=0.7109, respectively). Treatment with 5-Aza significantly reduced the percentage of CD14-CD209+ moDCs on Day 7 following the addition of maturation cocktail (figure 1b, p<0.0001). Flow cytometry assessment showed comparable expression of common maturation and co-stimulatory markers such as CD80, CD83, CD86, HLA-DR, CD209, CD141, CD40, and CCR7 between 5-Aza and DMSO-treated immature moDCs on Day 5 (figure 1c). Similarly, 5-Aza treatment had no significant effect on marker expression on mature moDCs generated with maturation cocktail on Day 7.ConclusionsThere was no significant difference in maturation and co-stimulatory marker expression of immature and mature moDCs from patients with higher-risk MDS following in vitro treatment with 5-Aza. Though recent studies have identified important immunoregulatory effects of 5-Aza, functional changes that may occur within the dendritic cell population are not fully understood. Further studies are planned, including cytokine analyses and transcriptome sequencing of mature moDCs, and may help elucidate the immunological mechanisms underlying the therapeutic effects of 5-Aza in patients with higher-risk MDS.Ethics ApprovalThe study is being conducted as per the Declaration of Helsinki and was approved by the University of California San Diego Institutional Review Board (#161345) and registered with ClinicalTrials.gov (NCT02667093). All patients were provided written informed consent.Abstract 684 Figure 15-Aza and DMSO vehicle-treated moDCs from patients with higher-risk MDS were evaluated for phenotypic markers before and after stimulation with maturation cocktail. Purified CD14+ cells were magnetically isolated from PBMC from 6 higher-risk MDS patients and cultured with IL-4 and GM-CSF for 5 days followed by addition of poly(I:C), IL-1β, IFN-α, IFN-γ, and TNF-α for 48 hours at 37°C in a 5% CO2 incubator. Freshly prepared 5-Aza or DMSO vehicle was added to cultures every 24 hours for a total of three 1 μM doses starting on Day 1. (A) Cultured cells were stained with trypan blue to determine the percentage of cell survival on Day 5 and Day 7 in culture. (B) Treatment with 5-Aza significantly reduced the percentage of CD14-CD209+ moDCs on Day 7 following addition of maturation cocktail (p<0.0001). (C) The percentage of CD14-CD83+ cells is comparable between 5-Aza and vehicle-treated immature moDCs on Day 5 and mature moDCs on Day 7 (p=0.2434 and p=0.5846, respectively). (D) Cultured cells were stained with fluorochrome-conjugated antibodies to determine the expression of common maturation and co-stimulatory markers using flow cytometry. Cells were gated on CD14-CD11c+ to distinguish moDCs, and scatterplots represent the geometric mean fluorescence intensity (gMFI) of marker expression pre- and post-maturation. Individual dots represent one of three experimental replicates performed for the 6 higher-risk MDS patient samples. Each dot is labeled by MDS patient sample. Statistical analysis was performed by Welch's t-test using GraphPad Prism.

Role of Human NK Cells in Dentric-Cell Based Vaccine.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1550-1550
Author(s):  
Gerard M.J. Bos ◽  
Janine CHMJ Van Elssen ◽  
Joris Vanderlocht ◽  
Brigitte LMG Senden-Gijsbers ◽  
Wilfred LMG Germeraad
Keyword(s):  
Nk Cells ◽  
Human Study ◽  
Dependent Manner ◽  
Gm Csf ◽  
Tnf Α ◽  
Toll Like Receptor 2 ◽  
Ifn Γ ◽  
Th1 Polarization ◽  
Dc Maturation

Abstract Figure Figure Besides their prominent role in the destruction of altered self-cells, natural killer (NK) cells have been shown to potentiate T cell responses by interacting with dendritic cells (DC). In mouse models as well as in a recent human study it has been demonstrated that DC might activate NK cells. In the context of dendritic cell-based vaccines – i.e. optimising the optimal maturation cocktail - it remains to be determined if and how NK-DC interactions depend on differential DC maturation and what factors influence the NK activation.. By comparing differential DC differentiation (IL-4/GM-CSF and IL-13/GMCSF) and maturation cocktails (IFN-γ/FMKp and PGE2/TNF-α), we show that the ability of human DCs to attract NK cells is imprinted during DC maturation. Only FMKP/IFN-γ (stimulation Toll like receptor 2 and 4) maturated DCs have the capacity to actively recruit NK cells in vitro and our data indicate that CCR5 is the dominant chemokine receptor in this recruitment (Figure 1). Furthermore, in contrast to PGE2/TNF-α matured DC, FMKP/IFN-γ maturated DCs activate NK cells to produce IFN-γ in a IL-12/IL18 dependent manner, of which we show it contributes to strong TH1 polarization. In addition upon contact with these DCs NK cells upregulate their lymph node homing receptors, possibly inducing secondary migration to the lymph nodes. In conclusion, besides the identification of a superior DC maturation cocktail which contributes to NK-DC interactions, we identified a novel recruitment mechanism for peripheral human NK cells which may contribute to secondary, central DC-NK interactions and strong TH1 polarization.

Tumour microenvironment markers in spontaneous and induced incubation of breast cancer biopsies

2021 ◽  
Vol 12 (1) ◽  
pp. 50-59
Author(s):  
Yu. S. Gergenreter ◽  
N. B. Zakharova ◽  
O. L. Morozova
Keyword(s):  
Breast Cancer ◽  
Tumour Microenvironment ◽  
Exact Test ◽  
Gm Csf ◽  
Spontaneous Production ◽  
Tnf Α ◽  
Ifn Γ ◽  
Group 2 ◽  
Tgf Β1 ◽  
Group 1

Aim. To study the spontaneous and stimulated production of cytokines in biopsies of breast cancer (BC) depending on the cancer stage.Materials and methods. An experimental study was carried out with cell cultures of breast cancer biopsies of stages I–II (group 1, n = 15) and III–IV stages (group 2, n = 15). The control consisted of 6 healthy women who underwent mastopexy. We used enzyme immunoassay method to access spontaneous and induced by a complex of polyclonal activators (PA: phytohemagglutinin 4 μg / ml, concanavalin A 4 μg / ml, lipopolysaccharide 2 μg / ml) concentration of TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF, MCP-1, TGF-β1. The index of the effect of polyclonal activators (IVPA) on cytokine production (induced production / spontaneous production) was calculated. To compare groups, the Mann-Whitney test and the median test, the chi-square test and the Fisher’s exact test were used.Results. Groups 1 and 2 did not differ in age, histological variant and immunohistochemical type of tumour, predominantly invasive cancer without signs of specificity prevailed. In group 2, a pronounced vascularization was more often observed: in 6 (40%) patients versus 1 (7%) in group 1 (p < 0.05). In both groups, compared with the control, there was a statistically sig-nificant (p < 0.05) increase in spontaneous production of TNF-α by 4.2 and 4.8 times, MCP-1 by 6.7 and 6.3 times, TGF-β1 – 2.2 and 2.5 times, VEGF 11.9 and 14.6 times; GM-CSF 15.6 and 13.4 times, G-CSF 96.8 and 79.5 times, respectively. The concentration of MCP-1 and IFN-γ was higher in group 1 (p < 0.05), VEGF and TGF-β1 – in group 2 (p < 0.05). IVPA in group 2 exceeded similar values   in group 1 for G-CSF, VEGF, TGF-β1 (p < 0.05).Conclusion. The production of cytokines (TNF-α, MCP-1, GM-CSF, G-CSF, VEGF, TGF-β1) in breast cancer biopsies is significantly higher than in biopsies of the unchanged mammary gland and depends on the stage of the tumour process.

scholarly journals Decitabine Promotes Modulation in Phenotype and Function of Monocytes and Macrophages That Drive Immune Response Regulation

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 868
Author(s):  
Fabiana Albani Zambuzi ◽  
Priscilla Mariane Cardoso-Silva ◽  
Ricardo Cardoso Castro ◽  
Caroline Fontanari ◽  
Flavio da Silva Emery ◽  
...  
Keyword(s):  
Immune Response ◽  
Hematological Malignancies ◽  
M2 Macrophage ◽  
M2 Polarization ◽  
Tnf Α ◽  
Monocytes And Macrophages ◽  
Ifn Γ ◽  
And Function ◽  
The Impact

Decitabine is an approved hypomethylating agent used for treating hematological malignancies. Although decitabine targets altered cells, epidrugs can trigger immunomodulatory effects, reinforcing the hypothesis of immunoregulation in treated patients. We therefore aimed to evaluate the impact of decitabine treatment on the phenotype and functions of monocytes and macrophages, which are pivotal cells of the innate immunity system. In vitro decitabine administration increased bacterial phagocytosis and IL-8 release, but impaired microbicidal activity of monocytes. In addition, during monocyte-to-macrophage differentiation, treatment promoted the M2-like profile, with increased expression of CD206 and ALOX15. Macrophages also demonstrated reduced infection control when exposed to Mycobacterium tuberculosis in vitro. However, cytokine production remained unchanged, indicating an atypical M2 macrophage. Furthermore, when macrophages were cocultured with lymphocytes, decitabine induced a reduction in the release of inflammatory cytokines such as IL-1β, TNF-α, and IFN-γ, maintaining IL-10 production, suggesting that decitabine could potentialize M2 polarization and might be considered as a therapeutic against the exacerbated immune response.

scholarly journals TNF-α -308 G>A and IL10 -1082A>G polymorphisms as potential risk factors for lymphoproliferative disorders in autoimmune rheumatic diseases

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Manal Y. Tayel ◽  
Aida Nazir ◽  
Ibtessam M. Abdelhamid ◽  
Myriam A. S. Helmy ◽  
Nadia E. Zaki ◽  
...  
Keyword(s):  
Risk Factors ◽  
Rheumatic Diseases ◽  
Lymphoproliferative Disorders ◽  
Group Iii ◽  
Autoimmune Rheumatic Diseases ◽  
Group I ◽  
Tnf Α ◽  
Ifn Γ ◽  
Tgf Β1 ◽  
Distribution Frequency

Abstract Background Chronic inflammation with sustained unregulated immune stimulation in autoimmune rheumatic diseases (ARD) may be a risk factor for developing lymphoproliferative disorders (LPD). Markers of ARD activity as high erythrocyte sedimentation rate or erosive joint diseases and the development of B-symptoms were accounted as risk factors for LPD development. We investigated the association of five inflammatory cytokine genes single nucleotide polymorphisms (SNPs): TNF-α -308G>A; TGF-β1 gene codon 10 T>C and 25 G>C; IL-10 promoter SNPs -1082 A>G, -819T>C, and -592A>C; IL-6 -174G>C; and IFN-γ 874 T>A with the risk of LPD development in ARD patients. The study was conducted on 70 patients divided into group I, 25 ARD patients diagnosed as RA (n = 15) and SLE (n = 10) and with no history of malignancy; group II, 25 patients diagnosed with LPD and had no ARD; and group III, 20 patients diagnosed with both diseases: ARD and LPD. Cytokine genotyping was analyzed by PCR-sequence-specific primer (PCR-SSP). Results ARD+LPD patients had significantly higher frequency of TNF-α -308A allele and AA+AG genotype (high TNF-α producers) and IL-10 -1082A allele and AA genotype (low IL-10 producers) than ARD patients (p = 0.003, p = 0.024, p = 0.003, p = 0.03, respectively) with a significantly increased risk of LPD development in ARD patients expressing the corresponding alleles and genotypes. No significant differences were detected in the distribution frequency of either TGF-β1, IL-6, or IFN-γ SNPs between groups I and III or any of the studied SNPs between groups II and III. The distribution frequency of IL-10 ATA haplotype was significantly increased in group III as compared to group I (p = 0.037). Conclusion The significantly increased frequency of the high-TNF-α- and low-IL-10-producing alleles and genotypes in ARD patients may participate in the provision of a proinflammatory milieu that eventually increases the risk of LPD development.

scholarly journals Water Extract of Rubus coreanus Prevents Inflammatory Skin Diseases In Vitro Models

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1230
Author(s):  
Sumin Pyeon ◽  
Ok-Kyung Kim ◽  
Ho-Geun Yoon ◽  
Shintae Kim ◽  
Kyung-Chul Choi ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Skin Diseases ◽  
Water Extract ◽  
Total Phenolic ◽  
Hacat Cells ◽  
Rubus Coreanus ◽  
Tnf Α ◽  
Ifn Γ ◽  
Inflammatory Skin

Atopic dermatitis (AD) is a chronic inflammatory skin disease caused by immune hypersensitivity reaction. The cause of AD is unclear, but its symptoms have a negative effect on quality of life; various treatment methods to alleviate these symptoms are underway. In the present study, we aimed to evaluate in vitro antioxidant and anti-inflammatory effects of Rubus coreanus water extract (RCW) on AD. Total phenolic compounds and flavonoid content of RCW were 4242.40 ± 54.84 mg GAE/g RCE and 1010.99 ± 14.75 mg CE/g RCW, respectively. RCW reduced intracellular reactive oxygen species level and increased the action of antioxidant enzymes, such as catalase, superoxide dismutase, and glutathione peroxidase in tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ)-stimulated HaCaT cells. Moreover, mRNA expression of the pro-inflammatory cytokines, including TNF-α, interleukin-1β, and interleukin-6, was downregulated by RCW in the TNF-α/IFN-γ-stimulated cells. The levels of inflammatory chemokines (thymus- and activation-regulated chemokine; eotaxin; macrophage-derived chemokine; regulated on activation, normal T-cell expressed and secreted; and granulocyte-macrophage colony-stimulating factor) and intercellular adhesion molecule-1 were decreased in the TNF-α/IFN-γ-stimulated HaCaT cells after RCW treatment. Additionally, the mRNA expression levels of filaggrin and involucrin, proteins that form the skin, were increased by RCW. Furthermore, RCW inhibited the nuclear factor kappa-light-chain-enhancer of the activated B cells pathway in the TNF-α/IFN-γ-stimulated HaCaT cells. Collectively, the present investigation indicates that RCW is a potent substance that inhibits AD.

scholarly journals Identification of a microRNA signature in renal fibrosis: role of miR-21

2011 ◽  
Vol 301 (4) ◽  
pp. F793-F801 ◽  
Author(s):  
Abolfazl Zarjou ◽  
Shanzhong Yang ◽  
Edward Abraham ◽  
Anupam Agarwal ◽  
Gang Liu
Keyword(s):  
Epithelial Cells ◽  
Renal Fibrosis ◽  
Transforming Growth Factor ◽  
Response To Treatment ◽  
Tubular Epithelial Cells ◽  
Altered Expression ◽  
Tnf Α ◽  
Tgf Β1

Renal fibrosis is a final stage of many forms of kidney disease and leads to impairment of kidney function. The molecular pathogenesis of renal fibrosis is currently not well-understood. microRNAs (miRNAs) are important players in initiation and progression of many pathologic processes including diabetes, cancer, and cardiovascular disease. However, the role of miRNAs in kidney injury and repair is not well-characterized. In the present study, we found a unique miRNA signature associated with unilateral ureteral obstruction (UUO)-induced renal fibrosis. We found altered expression in UUO kidneys of miRNAs that have been shown to be responsive to stimulation by transforming growth factor (TGF)-β1 or TNF-α. Among these miRNAs, miR-21 demonstrated the greatest increase in UUO kidneys. The enhanced expression of miR-21 was located mainly in distal tubular epithelial cells. miR-21 expression was upregulated in response to treatment with TGF-β1 or TNF-α in human renal tubular epithelial cells in vitro. Furthermore, we found that blocking miR-21 in vivo attenuated UUO-induced renal fibrosis, presumably through diminishing the expression of profibrotic proteins and reducing infiltration of inflammatory macrophages in UUO kidneys. Our data suggest that targeting specific miRNAs could be a novel therapeutic approach to treat renal fibrosis.

scholarly journals Granulocyte-Macrophage Colony-Stimulating Factor-Deficient Mice Have Impaired Resistance to Blood-Stage Malaria

2001 ◽  
Vol 69 (1) ◽  
pp. 129-136 ◽  
Author(s):  
Julie Riopel ◽  
MiFong Tam ◽  
Karkada Mohan ◽  
Michael W. Marino ◽  
Mary M. Stevenson
Keyword(s):  
Blood Stage ◽  
Colony Stimulating Factor ◽  
Necrosis Factor Alpha ◽  
Gm Csf ◽  
Tnf Α ◽  
Macrophage Colony Stimulating Factor ◽  
Ifn Γ ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Blood Stage Malaria

ABSTRACT The contribution of granulocyte-macrophage colony-stimulating factor (GM-CSF), a hematopoietic and immunoregulatory cytokine, to resistance to blood-stage malaria was investigated by infecting GM-CSF-deficient (knockout [KO]) mice with Plasmodium chabaudi AS. KO mice were more susceptible to infection than wild-type (WT) mice, as evidenced by higher peak parasitemia, recurrent recrudescent parasitemia, and high mortality. P. chabaudiAS-infected KO mice had impaired splenomegaly and lower leukocytosis but equivalent levels of anemia compared to infected WT mice. Both bone marrow and splenic erythropoiesis were normal in infected KO mice. However, granulocyte-macrophage colony formation was significantly decreased in these tissues of uninfected and infected KO mice, and the numbers of macrophages in the spleen and peritoneal cavity were significantly lower than in infected WT mice. Serum levels of gamma interferon (IFN-γ) were found to be significantly higher in uninfected KO mice, and the level of this cytokine was not increased during infection. In contrast, IFN-γ levels were significantly above normal levels in infected WT mice. During infection, tumor necrosis factor alpha (TNF-α) levels were significantly increased in KO mice and were significantly higher than TNF-α levels in infected WT mice. Our results indicate that GM-CSF contributes to resistance to P. chabaudi AS infection and that it is involved in the development of splenomegaly, leukocytosis, and granulocyte-macrophage hematopoiesis. GM-CSF may also regulate IFN-γ and TNF-α production and activity in response to infection. The abnormal responses seen in infected KO mice may be due to the lack of GM-CSF during development, to the lack of GM-CSF in the infected mature mice, or to both.

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