Athanasia Mouzaki | University of Patras (original) (raw)

Papers by Athanasia Mouzaki

Anticancer Research, Jul 1, 2021

Background/Aim: The role of immune cells and PD-L1 in cutaneous squamous carcinogenesis is unclea... more Background/Aim: The role of immune cells and PD-L1 in cutaneous squamous carcinogenesis is unclear. This study examines T-cell populations, Langerhans cells (LCs) and PD-L1 in invasive squamous cell carcinoma (inSCC), adjacent precursors and normal skin (NS) to investigate their participation in tumorigenesis. Materials and Methods: Cases of cutaneous inSCC with adjacent precursors (n=125) were selected. In situ SCC (isSCC) and actinic keratosis (AK) were observed in 53 and 123 cases, respectively, whereas NS was present in 123 lesions. Immunohistochemistry was performed for CD3, CD8, Foxp3, CD1a and PD-L1. Results: T-cells, LCs and PD-L1 gradually increase during the evolution from AK to isSCC and inSCC, with statistical significance between all lesions, except for CD3+ and CD8+ cells between isSCC and inSCC. Epithelial PD-L1 expression correlates with tumor diameter and thickness. Conclusion: The progressive increase of T-cells, LCs and PD-L1 in cutaneous squamous carcinogenesis provides rationale for immunotherapy and identification of predictive biomarkers. Cutaneous invasive squamous cell carcinoma (inSCC), the second most common invasive skin cancer, usually evolves from precursor lesions, namely actinic keratosis (AK) and in situ squamous cell carcinoma (isSCC) (1). Although early stage inSCC is rarely fatal, advanced disease or the treatment can produce significant patient disfigurement (1, 2). AK, a common skin lesion, has an estimated risk of 1-10% to progress to inSCC. Since no progression parameters are defined, each AK should be treated, to prevent development of inSCC and related morbidity (2, 3). During carcinogenesis, tumor-specific neoantigens are recognized by the immune system and an immune response is orchestrated against the tumor (4). Tumor infiltrating lymphocytes (TILs), mostly CD3+ T-cells, CD8+ cytotoxic Tcells (CTLs) and Foxp3+ regulatory T-cells (Tregs), accumulate in the tumor microenvironment (TME) in various tumors, including inSCC (5). Langerhans cells (LCs), the antigen-presenting cells (APCs) of the epidermis, participate in adaptive immunity, which is crucial for tumor elimination (6), but have not been studied in inSCC. T-cell stimulation requires binding of the T-cell receptor (TCR) to the antigen, presented by APCs via molecules of the major histocompatibility complex (MHC). Additional second signals are provided by APC-bound ligands to co-stimulatory receptors on T-cells. These signals, known as immune checkpoints, propagate or inhibit T-cell activation. Programmed Death-1 (PD-1), a member of the CD28/cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) family of co-stimulatory receptors, after binding to its ligands, PD-L1 or PD-L2, inhibits T-cell activation, impedes the immune response in the TME and allows tumor survival (4). PD-L1 is expressed on lymphocytes, macrophages, APCs (7) and tumor cells of various tumors (4, 8, 9). Accumulated knowledge on the role of immune checkpoints in cancer evolution led to the development of immune checkpoint inhibitors (ICIs). Antibodies targeting CTLA-4, PD-1 and PD-L1 have been approved for metastatic 3439

Journal of Cutaneous Pathology, Mar 14, 2018

Background: Tumor infiltrating lymphocytes (TILs) represent important regulators of carcinogenesi... more Background: Tumor infiltrating lymphocytes (TILs) represent important regulators of carcinogenesis. Cutaneous invasive squamous cell carcinoma (inSCC) develops through precursor lesions, namely in situ squamous cell carcinoma (isSCC) and actinic keratosis (AK), representing a natural model of carcinogenesis. The study evaluates TIL subpopulations in inSCC and its precursors by comparing two semi-quantitative scoring systems, and assesses the presence of regulatory T-cells (Tregs) in these lesions. Methods: Paraffin sections from 33 cases of AK, 19 isSCCs and 34 inSCCs with adjacent precursor lesions or normal skin were immunostained for CD3, CD4, CD8 and Foxp3. TIL subgroups were evaluated by the semi-quantitative Klintrup-Mäkinen score, and by a more detailed modification of this system. Treg counts were assessed by image analysis quantification. Results: An increase of all TIL subpolulations from precursor lesions towards inSCC was shown by both scoring systems. Treg counts progressively increased from normal skin to AK and isSCC, but decreased in inSCC. Tregs were more numerous in pT2 and around indolent inSCCs compared to T1 and aggressive subtypes. This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as

Immunobiology, Aug 1, 2016

T cells play a pivotal role in controlling the immune response and have been the focus of extensi... more T cells play a pivotal role in controlling the immune response and have been the focus of extensive research. We studied the process of in vitro generation of antigen-specific T effector cells (Teffs) to assess the dynamics of antigen presentation and determine the best conditions for cell therapy. We used a peptidic construct consisting of combined HLA class I and II epitopes of the tumor antigen MAGE-3 as an antigen. Monocytes were isolated from healthy donors and were differentiated to dendritic cells (DCs) in vitro. The peptide was added to the DC culture, the pulsed cells were transferred to a co-culture with lymphocytes from the same donor, either as irradiated feeders or untreated, and were cultured in the presence or absence of IL-2. Several rounds of restimulation followed. The cells were analyzed by Flow Cytometry, and cytokine levels were measured by ELISA and Cytometric Bead Array for Th1/Th2/Th17 profiling. The results showed that the lymphocytes in culture upregulated their activation markers and produced Th1 proinflammatory cytokines in response to the peptide, optimally when it was presented by non-irradiated dendritic cells in the presence of IL-2. In contrast, DC irradiation resulted in low activation potential and a shift toward a suppressive phenotype. After prolonged antigenic stimulation, the culture displayed Th17 polarization. In conclusion, the functional integrity of DCs is necessary for the development of antigen-specific Teffs, and culture conditions can be developed to create Teffs with specific properties for eventual use in cell therapy applications.

European journal of medicinal chemistry, Aug 1, 2015

Properties of myelin altered peptide ligand cyclo(87-99) (Ala91,Ala96)MBP87-99 render it a promis... more Properties of myelin altered peptide ligand cyclo(87-99) (Ala91,Ala96)MBP87-99 render it a promising drug lead for immunotherapy of Multiple Sclerosis,

Shock, Jun 2, 2020

ABSTRACT Introduction: Sepsis is a life-threatening syndrome which can progress to multiple organ... more ABSTRACT Introduction: Sepsis is a life-threatening syndrome which can progress to multiple organ dysfunction with high mortality. Intestinal barrier failure exerts a central role in the pathophysiological sequence of events that lead from sepsis to multiple organ dysfunction. The present study investigated the role of hydrocortisone (HC) administration and fecal microbiota transplantation (FMT) in several parameters of the gut barrier integrity, immune activation, and survival, in a model of polymicrobial sepsis in rats. Methods: Forty adults male Wistar rats were randomly divided into four groups: sham (group I), cecal ligation and puncture (CLP) (group II), CLP + HC (2.8 mg/kg, intraperitoneally single dose at 6 h) (group III), and CLP + FMT at 6 h (group IV). At 24 h post-CLP, ileal tissues were harvested for histological and immunohistochemical analyses while endotoxin, IL-6, and IL-10 levels in systemic circulation were determined. In a second experiment the same groups were observed for 7 days for mortality, with daily administration of hydrocortisone (group III) and FMT (group IV) in surviving rats. Results: HC administration and FMT significantly reduced mortality of septic rats by 50%. These interventions totally reversed intestinal mucosal atrophy by increasing villous density and mucosal thickness (μm, mean ± SD: Group I: 620 ± 35, Group II: 411 ± 52, Group III: 622 ± 19, Group IV: 617 ± 44). HC and FMT reduced the apoptotic body count in intestinal crypts whereas these increased the mitotic/apoptotic index. Activated caspase-3 expression in intestinal crypts was significantly reduced by HC or FMT (activated caspase-3 (+) enterocytes/10 crypts, mean ± SD: Group I: 1.6 ± 0.5, Group II: 5.8 ± 2.4, Group III: 3.6 ± 0.9, Group IV: 2.3 ± 0.6). Both treatments increased Paneth cell count and decreased intraepithelial CD3(+) T lymphocytes and inflammatory infiltration of lamina propria to control levels. In the sham group almost the total of intestinal epithelial cells expressed occludin (92 ± 8%) and claudin-1 (98 ± 4%) and CLP reduced this expression to 34 ± 12% for occludin and 35 ± 7% for claudin-1. Administration of HC significantly increased occludin (51 ± 17%) and claudin-1 (77 ± 9%) expression. FMT exerted also a significant restoring effect in tight junction by increasing occludin (56 ± 15%) and claudin-1 (84 ± 7%) expression. The beneficial effects of these treatments on gut barrier function led to significant reduction of systemic endotoxemia (EU/mL, mean ± SD: Group I: 0.93 ± 0.36, Group II: 2.14 ± 1.74, Group III: 1.48 ± 0.53, Group IV: 1.61 ± 0.58), while FMT additionally decreased IL-6 and IL-10 levels. Conclusion: Fecal microbiota transplantation and stress dose hydrocortisone administration in septic rats induce a multifactorial improvement of the gut mechanical and immunological barriers, preventing endotoxemia and leading to improved survival.

Journal of Medicinal Chemistry, Dec 15, 2001

ABSTRACT

Autoimmunity Reviews, Oct 1, 2018

Sjögren's syndrome (SS) is a chronic autoimmune disease that primarily affects the exocrine gland... more Sjögren's syndrome (SS) is a chronic autoimmune disease that primarily affects the exocrine glands, resulting in their functional impairment. In SS, lymphocytic infiltration of salivary and lacrimal glands, and deposition of several types of autoantibodies, mainly anti-SS-A (anti-Ro) and anti-SS-B (anti-La), lead to chronic inflammation, with xerostomia and keratoconjunctivitis sicca. In its primary form (pSS), SS does not involve additional connective tissue diseases, whereas in its secondary and more common form (sSS), SS presents in association with other rheumatic autoimmune diseases, mainly rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and systemic sclerosis (SSc). As in most autoimmune diseases, environmental, hormonal and genetic factors are implicated in SS pathogenesis. In SS T cells predominate in mild lesions, whereas B cells predominate in advanced lesions. Th1, Th2, Th17, follicular helper T (Tfh) cells and regulatory cells (Tregs/Bregs), with their characteristic cytokine profiles, have been implicated in the pathogenesis of SS. It has been suggested that Th1 and Th17 cells initiate SS and, as the disease progresses, Th2 and Tfh cells predominate. It is assumed that, as in all autoimmune and inflammatory conditions, tolerance defects contribute to SS pathogenesis. It is intriguing that in SS it remains unclear which types of regulatory cells are functional and whether they ameliorate or worsen the disease. In this review we present a comprehensive update on SS with emphasis on immune system involvement, and suggest new insights into SS immunopathogenesis.

Frontiers in Immunology, Jan 13, 2023

Aim: Marathon is a running event in which athletes must cover a distance of 42.195 km. In additio... more Aim: Marathon is a running event in which athletes must cover a distance of 42.195 km. In addition to participating in marathons, marathoners have incorporated extensive running into their lifestyle. In the present study, we investigated the effect of long-term strenuous exercise in the form of marathon running on the immune system. Methods & Results: We collected peripheral blood samples from 37 male marathoners before/after a race and 37 age/sex/body mass index (BMI)matched healthy sedentary controls. Hematological and biochemical tests revealed race-induced leukocytosis attributable to neutrophilia and significant increases in plasma lactate dehydrogenase (LDH), creatine phosphokinase (CPK), and cortisol concentrations. Phenotypic analysis of lymphocytes revealed raceinduced significant decrease in the number of lymphocytes, memory helper T (Th) cells, naive, memory and activated cytotoxic T (Tc) cells, natural killer (NK), NKT, and B1 cells, and a significant increase in the number of activated Th and regulatory Th cells (Tregs). Compared with controls, marathoners maintained significantly lower levels of memory and activated Th cells and higher levels of activated Tc and B1 cells. Measurement of plasma cytokine levels revealed a proinflammatory cytokine polarization that increased after the race. Examination of gene expression of cytokines and Th-cell signature transcription factors in peripheral blood mononuclear cells revealed a significant decrease in tumor necrosis factor a (TNF-a) and interleukin (IL)-17, and a significant increase in IL-6, IL-10 and forkhead box P3 (FoxP3) after the race. Compared with controls, Frontiers in Immunology frontiersin.org 01

Cytokine, Jun 1, 2012

Respiratory distress syndrome (RDS) is a common problem and the leading cause of death in prematu... more Respiratory distress syndrome (RDS) is a common problem and the leading cause of death in premature infants (PI). The introduction of surfactant treatment for RDS management has lowered mortality and morbidity; nevertheless, some neonates do not improve and are at increased risk of pulmonary hemorrhage. Inflammation, not only local but also systemic, seems to play an important role in the pathogenesis of RDS. To determine whether cytokine patterns characterize RDS and its outcome, we measured type-1 (IL-2, TNF-α, IFN-γ, IL-6) and type-2 (IL-4, IL-5, IL-10, TGF-β1) serum cytokines of 47 PI with established RDS and a control group of 30 healthy, appropriate for gestational age, full-term neonates. Cord blood samples were obtained at the time of delivery from PI and controls. Venous blood samples were collected from PI who received surfactant treatment and/or developed pulmonary hemorrhage. Significantly elevated cord blood cytokine levels were observed in PI at time of delivery, compared to controls, except for IL-5 and TNF-α levels that were within control range. The type-1/type-2 cytokine ratio was significantly increased in PI vs controls. Neonates who developed pulmonary hemorrhage between 2 and 3 days of life and/or died, presented the strongest Th1 and type-1 cytokine polarization that was mainly due to increased IFN-γ and TNF-α, and decreased TGF-β1. The majority of these PI were female with very low gestational age. Overall, PI with RDS present a Th1/type-1 cytokine polarization, which persists irrespective of the treatment provided, and is amplified when complications appear. Th1 polarization is associated with poor prognosis.

Oncology, 2004

We report the case of a woman who developed acute thrombocytopenia with hemorrhagic diathesis dur... more We report the case of a woman who developed acute thrombocytopenia with hemorrhagic diathesis during adjuvant treatment of colorectal adenocarcinoma with oxaliplatin, 5-fluorouracil and leucovorin. A 55-year-old woman started adjuvant chemotherapy with oxaliplatin, 5-fluorouracil and leucovorin (mFOLFOX6). Prior to starting the 12th course of chemotherapy, a complete blood cell count showed the following values: neutrophils 1800/mm 3 , platelets 136,000/ m 3 and hemoglobin 11.1 g/dl. A blood count revealed that the platelet levels had dropped to 35,000/mm 3 , with no significant changes in hemoglobin levels following the course. The administration of corticosteroids was begun and the platelet number was recovered. Clinicians should be aware of the possibility of oxaliplatin-induced hematological emergencies during the treatment of colorectal cancer patients in order to optimize supportive treatment and avoid toxic mortality.

Endocrine, metabolic & immune disorders, Jun 12, 2019

ObjectiveIn healthy individuals, leptin is produced from adipose tissue and is secreted into the ... more ObjectiveIn healthy individuals, leptin is produced from adipose tissue and is secreted into the circulation to communicate energy balance status to the brain and control fat metabolism. Corticotropin-Releasing Hormone (CRH) is synthesized in the hypothalamus and regulates stress responses. Among the many adipokines and hormones that control fat metabolism, leptin and CRH both curb appetite and inhibit food intake. Despite numerous reports on leptin and CRH properties and function, little has been actually shown about their association in the adipose tissue environment.MethodsIn this article, we summarized the salient information on leptin and CRH in relation to metabolism. We also investigated the direct effect of recombinant CRH on leptin secretion by primary cultures of human adipocytes isolated from subcutaneous abdominal adipose tissue of 7 healthy children and adolescents, and measured CRH and leptin levels in plasma collected from peripheral blood of 24 healthy children and adolescents to assess whether a correlation exists between CRH and leptin levels in the periphery.Results and ConclusionThe available data indicate that CRH exerts a role in the regulation of leptin in human adipocytes. We show that CRH downregulates leptin production by mature adipocytes and that a strong negative correlation exists between CRH and leptin levels in the periphery, and suggest the possible mechanisms of CRH control of leptin. Delineation of CRH control of leptin production by adipocytes may explain unknown pathogenic mechanisms linking stress and metabolism.

The American Journal of the Medical Sciences, Feb 1, 2020

This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Scientific Reports, Jul 20, 2021

The aim of the present study was to investigate combined effects of cold atmospheric plasma (CAP)... more The aim of the present study was to investigate combined effects of cold atmospheric plasma (CAP) and the chemotherapeutic drug doxorubicin (DOX) on murine and human melanoma cells, and normal cells. In addition to free drug, the combination of CAP with a liposomal drug (DOX-LIP) was also studied for the first time. Thiazolyl blue tetrazolium bromide (MTT) and Trypan Blue exclusion assays were used to evaluate cell viability; the mechanism of cell death was evaluated by flow cytometry. Combined treatment effects on the clonogenic capability of melanoma cells, was also tested with soft agar colony formation assay. Furthermore the effect of CAP on the cellular uptake of DOX or DOX-LIP was examined. Results showed a strong synergistic effect of CAP and DOX or DOX-LIP on selectively decreasing cell viability of melanoma cells. CAP accelerated the apoptotic effect of DOX (or DOX-LIP) and dramatically reduced the aggressiveness of melanoma cells, as the combination treatment significantly decreased their anchorage independent growth. Moreover, CAP did not result in increased cellular uptake of DOX under the present experimental conditions. In conclusion, CAP facilitates DOX cytotoxic effects on melanoma cells, and affects their metastatic potential by reducing their clonogenicity, as shown for the first time. The use of cold atmospheric (-pressure) plasmas (CAPs) is increasingly being considered for various biomedical applications such as wound healing and blood coagulation 1-3 , sterilization and bacterial susceptibility 4-7 , treatment of cancer 6,8 , and various other pathologies 9-11. With regard to cancer treatment, various strategies using different types of CAPs have recently shown promising effects in cellular tumor models of breast cancer 12-14 , cervical cancer 13-15 , liver cancer 16 , lung cancer 17 , skin cancer 18-22 , and other cancers 6,8. Numerous therapeutic advantages of CAPs over treatments with conventional chemotherapeutic agents have been identified as a result of previous studies, the most important being probably the selective effect of cold plasma on normal and carcinoma cells 22-24. Exploration of possible methods to increase the anticancer effect of CAPs, by combinations with nanotechnologies 25-29 and/or chemotherapeutic agents 30-36 has recently been initiated. In several cases, different types of nanotechnologies such as iron oxide or gold nanoparticles have been found to strengthen the therapeutic effects of cold plasmas, and some potential mechanisms of action have been further explored or proposed. Melanoma is a highly resistant and a very aggressive form of skin cancer accounting for only 1% of skin cancers but represents the majority of fatalities related with skin cancer. Early diagnosis and treatment is critical for prognosis/survival; primary melanoma has a 5-year survival rate of 99%, whereas metastatic melanoma only 27% 37,38. Doxorubicin DOX (or Adriamycin), is one of the most potent chemotherapeutic agents with significant therapeutic activity in many cancers. Due to its toxicity (especially its cardiotoxicity) its use is limited. The

Antiviral Therapy, Aug 1, 2011

World journal of transplantation, 2016

Kidney transplantation is recognised as the most effective treatment for patients with end-stage ... more Kidney transplantation is recognised as the most effective treatment for patients with end-stage renal disease (ESRD). Kidney transplantation continues to face several challenges including long-term graft and patient survival, and the side effects of immunosuppressive therapy. The tendency in kidney transplantation is to avoid the side effects of immunosuppresants and induce immune tolerance. Regulatory T-cells (Tregs) contribute to self-tolerance, tolerance to alloantigen and transplant tolerance, mainly by suppressing the activation and function of reactive effector T-cells. Additionally, Tregs are implicated in the pathogenesis of diabetes, which is the leading cause of ESRD, suggesting that these cells play a role both in the pathogenesis of chronic kidney disease and the induction of transplant tolerance. Several strategies to achieve immunological tolerance to grafts have been tested experimentally, and include combinations of co-stimulatory blockade pathways, T-cell depletion, in vivo Treg-induction and/or infusion of exvivo expanded Tregs. However, a successful regimen that induces transplant tolerance is not yet available for clinical application. This review brings together certain key studies on the role of Tregs in ESRD, diabetes and kidney transplantation, only to emphasize that many more studies are needed to elucidate the clinical significance and the therapeutic applications of Tregs.

Infectious diseases, Jun 29, 2021

Abstract Background N-acetyl-cysteine (NAC) has been previously shown to exert beneficial effects... more Abstract Background N-acetyl-cysteine (NAC) has been previously shown to exert beneficial effects in diverse respiratory diseases, through antioxidant and anti-inflammatory actions. Our aim was to evaluate NAC potential impact in hospitalised patients with COVID-19 pneumonia, in terms of progression to severe respiratory failure (SRF) and mortality. Patients and Methods This retrospective, two-centre cohort study included consecutive patients hospitalised with moderate or severe COVID-19 pneumonia. Patients who received standard of care were compared with patients who additionally received NAC 600 mg bid orally for 14 days. Patients’ clinical course was recorded regarding (i) the development of SRF (PO2/FiO2 <150) requiring mechanical ventilation support and (ii) mortality at 14 and 28 days. Results A total of 82 patients were included, 42 in the NAC group and 40 in the control group. Treatment with oral NAC led to significantly lower rates of progression to SRF as compared to the control group (p < .01). Patients in the NAC group presented significantly lower 14- and 28-day mortality as compared to controls (p < .001 and p < .01 respectively). NAC treatment significantly reduced 14- and 28-day mortality in patients with severe disease (p < .001, respectively). NAC improved over time the PO2/FiO2 ratio and decreased the white blood cell, CRP, D-dimers and LDH levels. In the multivariable logistic regression analysis, non-severe illness and NAC administration were independent predictors of 28-days survival. Conclusion Oral NAC administration (1200 mg/d) in patients with COVID-19 pneumonia reduces the risk for mechanical ventilation and mortality. Our findings need to be confirmed by properly designed prospective clinical trials.

Journal of Clinical Oncology, May 20, 2011

e18059 Background: Lung cancer is rarely cured by the current therapeutic approaches. Although nu... more e18059 Background: Lung cancer is rarely cured by the current therapeutic approaches. Although numerous studies have implicated Foxp3 positive T regulatory cells in cancer pathogenesis, the role of Foxp3 in lung cancer pathogenesis remains unknown. METHODS We determined Foxp3 expression, using immunohistochemistry in 44 NSCLC tissue specimens, 20 samples from adjacent non neoplastic lung parenchyma and 5 normal lung tissue specimens. Immunohistochemical reactivity was graded on a scale of 0-3 according to intensity of staining and percentage of immunopositive cells. Foxp3 expression in cancer cells was categorized in three groups (high vs median vs low) using as a cut-off point the 33rd and 66th percentile. The total score for each slide was the sum of the intensity and distribution (between 0 and 6). Statistical analysis was performed with SPSS 17.0. RESULTS Nuclear Foxp3 immunostaining was detected in al tissues assessed. Foxp3 expression levels were higher in neoplastic cells than in adjacent non neoplastic tissue and normal lung parenchyma. Moreover, Foxp3 expression in tumor cells correlated with lymphocytic Foxp3-immunopositivity and the presence of lymph node metastasis. Furthermore, Foxp3 lymphocytic expression was negatively associated with the age of the patients. CONCLUSIONS Foxp3 is overexpressed in NSCLC cells and tumor infiltrating lymphocytes. Moreover, tumor Foxp3 expression correlates with lymph node metastasis while lymphocytic Foxp3 levels may be age-related.

Blood, Nov 15, 2013

Purpose/Objective Clinical and experimental studies have established that allogeneic blood transf... more Purpose/Objective Clinical and experimental studies have established that allogeneic blood transfusion (ABT) can cause immunosuppression. To identify immune parameters that contribute to this effect, we determined the effect of ABT on peripheral blood (PB) cytokine profiles and Treg numbers and function in a cohort of patients with no underlying pathologies. Materials and Methods Heparinized PB samples were collected from 46 patients (7M/39F, 28-88 yo) that underwent joint replacement surgery. The samples were collected immediately before surgery (BS), and after surgery (AS) on days 0, 7, 1 month, and 3 months to 1 year. Thirty six patients received ABT and 10 did not. PBMC were isolated, and the numbers and % of CD4+CD25+Foxp3+ Tregs and CD4+CD25high/+CD127low/- Tregs were determined by FACS. Tregs and T effectors (Teff) were isolated from patients on days 0-7 and Treg functional assays were performed by culturing Tregs with PHA-stimulated Teff at different ratios for 72h with CFSE, and analyzed by FACS for proliferation. Cytokine levels were determined in plasma by a cytometric bead array assay for IL-2, IL-4, IL-5, IL-6, IL-10, TNF-á, IFN-ã, and ELISA for TNF-á, TNF-RI(p55/p60) and II(p75/p80), TGF-â1 and â2. Results Both, CD4+CD25+Foxp3+ and CD4+CD25high/+CD127low/- Treg populations increased significantly on d0 AS and decreased on d7 AS to below BS levels, to achieve homeostasis &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;3 months in transfused patients. In contrast, Treg levels remained the same between BS and AS in non-transfused patients. Functional assays showed that Tregs were functional post-ABT and could suppress Teff proliferation efficiently. In culture, isolated Tregs secreted TGF-â1. All cytokines and TNF-RI and II plasma levels increased on d0 AS (IL-6, TNF-RI and II significantly), in transfused patients immediately AS, and decreased by d7 AS. In contrast, TGF-â1 levels decreased on d0 AS and increased by d7. No differences in cytokine or receptor levels were observed in non-transfused patients AS. Conclusion ABT induces the immediate production of plasma cytokines, especially pro-inflammatory IL-6 and TNF-RI and II that decreases by d7. In parallel, ABT induces the proliferation of Tregs. These Tregs are functional, and secrete TGF-â1 that reaches BS plasma levels by d7. The Tregs seem to be Th3 inducible Tregs. Homeostasis is reached by 3 months post-ABT. Disclosures: No relevant conflicts of interest to declare.

Critical Care, 2008

Introduction In order to find out the frequency rates of domestic and wild animal bites as well a... more Introduction In order to find out the frequency rates of domestic and wild animal bites as well as the evaluation of the prevalence rates of rabies disease in the human population in the Province of Kerman, a retrospective study was designed to analyze statistically the collected recorded data related to this project. Methods This study was conducted within the framework of MPVM student research projects by means of collaboration between