Terms Of Cytokines Research Articles

Levels of Inflammatory and Bone Metabolic Markers in the Gingival Crevicular Fluid of Individuals Undergoing Fixed Orthodontic Treatment in Comparison to Those Utilizing Invisalign.

Background and Objectives: Evaluation of the levels of cytokine and bone metabolic biomarkers (BMBs) in patients receiving fixed orthodontic therapy (FOT) and Invisalign. Materials and Methods: Sixty participants were enrolled after meeting the predefined inclusion criteria. Patients then underwent either FOT or Invisalign by allocating them randomly to each group (n = 30). The basic periodontal assessment was performed, including the plaque index (PI), gingival index (GI), and bleeding on probing (BoP), at baseline and again after 4 weeks. Gingival crevicular fluid (GCF) samples were taken from each individual at baseline and after 4 weeks. An enzyme-linked immunosorbent assay (ELISA) technique was used to determine the cytokine and BMB levels. An unpaired t-test compared the FOT and Invisalign group's means and SDs. Paired t-tests examined the difference between T0 baseline and T1. Results: Patients treated with either FOT or Invisalign presented no statistically significant difference in terms of periodontal parameters such as PI, GI, and BoP (p > 0.05). The levels of IL-6 were significantly higher in patients treated with FOT as compared to Invisalign at T1 (p < 0.05) The other tested cytokines, IL-10, 13, 17, and GM-CSF, were not significantly different in either the FOT or Invisalign group at baseline and 4 weeks follow-up (p > 0.05). Regarding BMBs, it was detected that NTx and OC levels in both of the investigated groups were not significantly different at baseline and after 4 weeks (p > 0.05). However, NTx levels rose significantly (p < 0.05) and OC levels fell from T0 to T1. Conclusions: FOT and Invisalign displayed comparable outcomes in terms of cytokine and BMB levels. However, only IL-6 and NTx were significantly different at week 4 from baseline.

Impacts of delta and omicron variants on inactivated SARS‐CoV‐2 vaccine‐induced T cell responses in patients with autoimmune diseases and healthy controls

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes coronavirus disease 2019 (COVID-19), which is still devastating economies and communities globally. The increasing infections of variants of concern (VOCs) in vaccinated population have raised concerns about the effectiveness of current vaccines. Patients with autoimmune diseases (PAD) under immunosuppressant treatments are facing higher risk of infection and potentially lower immune responses to SARS-CoV-2 vaccination. Blood samples were collected from PAD or healthy controls (HC) who finished two or three doses of inactivated vaccines. Spike peptides derived from wild-type strain, delta, omicron BA.1 were utilisedto evaluate T cell responses and their cross-recognition of delta and omicron in HC and PAD by flow cytometry and ex vivo IFNγ-ELISpot. We found that inactivated vaccine-induced spike-specific memory T cells were long-lasting in both PAD and HC. These spike-specific T cells were highly conserved and cross-recognized delta and omicron. Moreover, a third inactivated vaccine expanded spike-specific T cells that responded to delta and omicron spike peptides substantially in both PAD and HC. Importantly, the polyfunctionality of spike-specific memory T cells was preserved in terms of cytokine and cytotoxic responses. Although the extent of T cell responses was lower in PAD after two-dose, T cell responses were boosted to a greater magnitude in PAD by the third dose, bringing comparable spike-specific T cell immunity after the third dose. Inactivated vaccine-induced spike-specific T cells remain largely intact against delta and omicron variants. This study expands our understanding of inactivated vaccine-induced T cell responses in PAD and HC, which could have important indications for vaccination strategy.

Effect of dietary sophorolipids on growth performance and gastrointestinal functionality of broiler chickens infected with Eimeria maxima

Two experiments were conducted to evaluate the effects of dietary sophorolipids (SLs) supplementation as antibiotic alternatives on growth performance and gut health of chickens infected with Eimeria maxima. In experiment 1, 336 (zero-day-old) male broilers were used. The chickens were weighed and randomly allocated to the following 6 treatments groups with 7 chickens/cage and 8 cages/treatment: control group that received a basal diet (NC), positive control group that received a basal diet and was challenged with E. maxima (PC), PC+C18:1 lactonic diacetyled SL (SL1), PC+C18:1 deacetyled SL (SL2), PC+C18:1 monoacetyled SL (SL3), and PC+C18:1 diacetyled SL (SL4). Each SL (200 mg/kg feed) was added to the corresponding treatment group. In experiment 2, 588 (zero-day-old) male broilers were used. The chickens were randomly allocated to the following experimental groups with 10 or 11 chickens/cage and 8 cages/treatment: NC, PC, PC+ monensin at 90 mg/kg feed (MO), PC+SL1 at 200 mg/kg feed (SL1 200), PC+SL1 at 500 mg/kg feed (SL1 500), PC+SL4 at 200 mg/kg feed (SL4 200), and PC+SL4 at 500 mg/kg of feed (SL4 500). The chickens and feed were weighed at 0, 7, 14, 20, and 22 d to determine growth performance. In both experiments, all chickens except the NC group were orally infected with E. maxima (10,000 oocysts/chicken) at d 14. One chicken per cage was euthanized at d 20 to sample jejunal tissue to measure lesion scores, cytokines, and tight junction (TJ) proteins. Excreta samples were collected daily between d 20 and 22 to measure oocyst numbers. Data were analyzed using Mixed Model (PROC MIXED) in SAS. In experiment 1, SLs did not affect the growth of broiler chickens, but SL4 decreased (P < 0.05) the lesion score and oocyst number compared to PC chickens. In terms of cytokines and TJ protein gene expression, SLs increased (P < 0.05) IL-1β, IL-6, IL-17F, IL-4, IL-13, occludin, and ZO1 levels compared to PC chickens. In experiment 2, monensin increased (P < 0.05) body weight, and decreased (P < 0.05) the lesion score and oocyst number compared to the PC group. SL4 500 increased (P < 0.05) average daily gain and feed conversion ratio but decreased (P < 0.05) lesion score and fecal oocyst number. SL4 decreased (P < 0.05) IL-6, IL-17F, TNFSF-15, IL-2, and IL-10 levels but increased (P < 0.05) occludin and ZO-1 levels. Overall, dietary SL supplementation, especially SL4, improved growth and gastrointestinal functionality of young broiler chickens, demonstrating significant potential as an antibiotic alternative.

Immune-Mediated Mechanisms in Patients Testing Positive for SARS-CoV-2: Protocol for a Multianalysis Study.

BackgroundThe novel coronavirus has a high mortality rate (over 1% for patients older than 50 years). This can only be partially ascribed to other comorbidities. A possible explanation is a factor that assures a prompt response to SARS-CoV-2 in younger people, independent from the novelty of the virus itself. A factor is believed to stimulate the immune system and provide immunity against more antigens. The only external stimulation received by healthy people is vaccination (eg, the diphtheria, tetanus, and pertussis [DTP] vaccine). One hypothesis is that vaccination helps develop specific immunity but generates sprouting immunity against antigens in transit. The underlying immunological phenomena are the “bystander effect” and “trained immunity.” The developed immunity gives protection for years until it naturally fades out. After the fifth decade of life, the immune system is almost incompetent when a viral infection occurs, and thus, at this stage, the novel coronavirus can enter the body and cause acute respiratory distress syndrome.ObjectiveThe initial aim is to demonstrate that blood monocytes and natural killer cells show overpowering hyperactivity, while CD4+ and CD8+ T cells experience impediments to their defensive functions in patients with severe SARS-CoV-2 infection. The secondary objectives are to correlate clinical data and vaccination history with laboratory immune patterns in order to identify protective factors. Subsequently, we are also interested in characterizing the phenotypes and state of the degree of activation of peripheral blood mononuclear cells, including monocytes, natural killer cells, and CD4+ and CD8+ T cells, in healthy subjects vaccinated with the Pfizer vaccine.MethodsData will be collected using the following 3 approaches: (1) an experimental analysis to study the innate immune response and to identify genetic profiles; (2) an epidemiological analysis to identify the patients’ vaccination history; and (3) a clinical analysis to detect the immunological profile.ResultsThe protocol was approved by the Ethics Committee on April 16, 2020, and the study started on April 27, 2020. As of February 2021, enrollment has been completed. Immunological analysis is ongoing, and we expect to complete this analysis by December 2022.ConclusionsWe will recognize different populations of patients, each one with a specific immunological pattern in terms of cytokines, soluble factor serum levels, and immune cell activity. Anamnestic data, such as preceding vaccinations and comorbidities, biochemical findings like lymphocyte immunophenotyping, and pre-existing persistent cytomegalovirus infection, allow depicting the risk profile of severe COVID-19. Proof of the roles of these immunological phenomena in the development of COVID-19 can be the basis for the implementation of therapeutic immunomodulatory treatments.Trial RegistrationClinicalTrials.gov NCT04375176; https://clinicaltrials.gov/ct2/show/NCT04375176International Registered Report Identifier (IRRID)DERR1-10.2196/29892

The Effect of Obesity on Insulin Resistance in Terms of Cytokines and Hormones

With the increasing incidence and mortality of obesity, obesity-related health problems have become a worldwide priority. Clinical observation shows that obesity related to adipocyte differentiation is an important pathogenic factor of insulin resistance, and weight loss can reduce insulin resistance, indicating that obesity is related to insulin resistance. As the understanding of mechanism between obesity, cytokines, hormones and insulin resistance becomes clear, it is possible that these cytokines or hormones could be used in the use of biomarkers and the design of targeted therapies for insulin resistance. This review provides an overview of how obesity effect adipokines ， hepatokines and inflammatory cytokines whose changes result in or exacerbate insulin resistance.

Small Extracellular Vesicles from adipose derived stromal cells significantly attenuate in vitro the NF-\u03baB dependent inflammatory/catabolic environment of osteoarthritis

The therapeutic ability of Mesenchymal Stem/Stromal Cells to address osteoarthritis (OA) is mainly related to the secretion of biologically active factors, which can be found within their secreted Extracellular Vesicles including small Extracellular Vesicles (sEV). Aim of this study was to investigate the effects of sEV from adipose derived stromal cells (ADSC) on both chondrocytes and synoviocytes, in order to gain insights into the mechanisms modulating the inflammatory/catabolic OA environment. sEV, obtained by a combined precipitation and size exclusion chromatography method, were quantified and characterized, and administered to chondrocytes and synoviocytes stimulated with IL-1β. Cellular uptake of sEV was evaluated from 1 to 12 h. Gene expression and protein release of cytokines/chemokines, catabolic and inflammatory molecules were analyzed at 4 and 15 h, when p65 nuclear translocation was investigated to study NF-κB pathway. This study underlined the potential of ADSC derived sEV to affect gene expression and protein release of both chondrocytes and synoviocytes, counteracting IL-1β induced inflammatory effects, and provided insights into their mechanisms of action. sEV uptake was faster in synoviocytes, where it also elicited stronger effects, especially in terms of cytokine and chemokine modulation. The inflammatory/catabolic environment mediated by NF-κB pathway was significantly attenuated by sEV, which hold promise as new therapeutic strategy to address OA.

The effect of laparoscopic and open surgery on oxidative DNA damage and IL-37 in children with acute appendicitis.

Although laparoscopic appendectomy (LA) is considered to be minimally invasive compared with open appendectomy (OA), there are few studies showing this in terms of cytokines and oxidative stress markers. LA and OA techniques have been compared in terms of different conditions such as operation time and complication rate. Here, we compared how oxidative DNA damage and some cytokines levels change in relation to LA and OA. This study was performed in children diagnosed with acute appendicitis in our hospital. The study was conducted on 30 children with acute appendicitis in each group. Oxidative DNA damage and malondialdehyde levels were determined by high-performance liquid chromatography, and interleukin-1β, interleukin-37, ischaemia-modified albumin and total thiol levels were determined by enzyme-linked immunosorbent assays. There was a dramatic decrease in ischaemia-modified albumin (IMA) levels after LA. However, there were no statistically significant differences in pre- and postoperative IMA levels in the OA group. Oxidative DNA damage and malondialdehyde levels were low significantly according to preoperative levels after the LA and OA. Total thiol levels were high in children who underwent LA while they were low after OA. In addition, postoperative interleukin-1β levels were low in both groups. Furthermore, IL-37 levels postoperatively were low in the LA group, while there was no significant change in the OA group. LA had a more positive effect on oxidative DNA damage, IL-1β and IL-37 than the OA. In addition, surgical stress was reduced with LA.

Abstract C118: Tumor immune response in colon cancer African American patients and its role in cancer disparities

Abstract Introduction: Colorectal cancer (CRC) is the third most common cancer among African Americans (AA) and when compared to Caucasian Americans (CA), they present more advanced CRC disease and lower survival rates. Our previous findings suggest that this may be related to the differential expression in genes linked to cell recruitment and immune response. Therefore, we aimed to investigate the cellular antitumor activity and mutational profile of colon tumors from AAs. We also examined the secretion of cytokines characteristic of immune responses by different effector T helper cells (Th) subsets in AA and CA patients, as well as cell lines, to see if these differences play a role in the health disparities observed between these populations. Lastly, we observed the expression of the Program Death Ligand 1 (PD-L1) in response to the cytokines IL-17A and TNF-α in a microsatellite-unstable (MSI) AA and a microsatellite-stable (MSS) CA colon cancer cell line. Methods: Using IHC, we evaluated the cell recruitment and activation of T and natural killer cells in AA tumors. For mutational analysis, we utilized the TruSight Tumor 170 RUO kit (Illumina). ELISA assays (RayBiotech) were used to examine the secretion of cytokines linked to Th subsets (Th1, Th2, Th17) and inflammation in plasma from the AA and CA CRC patients, as well as in supernatants from the AA and CA colon cancer cell lines. Western blots were used to observe the expression of PD-L1 in the in vitro models. Results: ELISAs of plasma of CA and AA patients revealed a differential Th cytokines production patterns between early-stages (I, II) and late-stage (III) disease. The MSI AA cell line showed an increase on PD-L1 protein expression in response to IL-17A and TNF-α with an additive effect when combined in equal concentrations. Lastly, the mutational sequencing allowed us to further investigate the potential alterations that are responsible for the differences that we observed in gene expression between AAs and CAs in our RNA and cytokine expression profiling. Conclusions: Our results indicate that the immune profiles of AA patients differ from CA in terms of cytokines' production; AAs expressed elevated IL-17A, whereas CA expressed elevated IFN-γ, the latter indicative of Th1 immunity that has a more favorable prognosis. As such, these differences could be used as biomarkers and to guide therapeutic strategy for these populations. The mutational sequencing will help us to elucidate the impaired tumor immune response in AAs with colon cancer when compared to CAs that we observed in terms of cell recruitment and cytokine secretion in our previous findings. Importantly, our data indicate that IL-17A and TNF-α promote the protein production of PD-L1 in an MSI AA cell line, which may result in the impairment of T cells' antitumor activity. Taken together, the differences in the immunologic profiles in AA when compared to CA suggest a deficiency of the appropriate immune defense mechanisms in this population that may contribute to the cancer health disparities among CRC patients. Citation Format: Jenny E. Paredes, Ping Ji, Jone Garai, Marzia Spagnardi, Maria Munoz-Sagastibelza, Sayed Imtiaz, Gayle Mendez, Mubarak Akadri, Raavi Gupta, Mohamed Alshal, Maksim Agaronov, Henry Talus, Ellen Li, Jovanny Zabaleta, Laura Martello-Rooney, Jennie Williams. Tumor immune response in colon cancer African American patients and its role in cancer disparities [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C118.

Cathelicidin bovine myeloid antimicrobial peptide (BMAP) 28 is involved in the inflammatory response against alpha-herpesviruses in the bovine nervous system

The role of the local innate immune response in the neuropathogenesis of bovine herpesvirus (BoHV) type 1 and 5 remains largely unknown. This study determined the gene transcriptional expression of relevant bovine cathelicidins, TNFα and IFNβ in the nervous system of experimentally-infected cattle during the different stages of BoHV-1 and BoHV-5 infectious cycle. We studied the modulation of bovine myeloid antimicrobial peptide (BMAP) 27 and 28 by alpha-herpesviruses during acute infection of the central nervous system (CNS). However, BMAP28 was the main cathelicidin modulated. BoHV-5 supressed BMAP28 expression mainly in frontal cortex and cervical medulla whereas BoHV-1 slightly induced the expression of cathelicidins in the olfactory and posterior cortex. The differences in the regulation of the innate response are likely related to distinct replication rates of both alpha-herpesviruses in the CNS. During latency and reactivation, BoHV-1 and -5 decreased BMAP28 and BMAP27 expression, accompanied by high levels of TNFα and IFNβ transcripts in the posterior brain region and medulla during BoHV reactivation. In terms of cytokines, a remarkably overexpression of IFNβ was induced by BoHV-5 (133.8-fold). In trigeminal ganglion (TG) both alpha-herpesviruses induced cathelidicins gene expression at all stages of the infection cycle, while only acute BoHV-5 infection increased TNFα (129-fold) mRNA levels. This study suggests that the pronounced downregulation of BMAP28 in BoHV-5-acutely-infected CNS is due to a decreased immune stimulation during viral infection, favouring its establishment in the CNS with a low replication rate until latency. Thus, cathelicidins, together with IFNβ and TNFα, are differentially regulated by BoHV-5 and BoHV-1 infections and this regulation is dependent on the stage of virus infection in the bovine nervous system.

Comparison of ELISA and flow cytometry for measurement of interleukin-1 beta, interleukin-6 and tumor necrosis factor-α

Abstract Background Although majority of the previous studies have shown a good correlation between enzyme linked immuno sorbent assay (ELISA) and flow cytometry in terms of cytokines, two laboratory methods usually were compared with the regression analysis and correlation in the literature. This study aimed at comparing the ELISA and flow cytometry assay for measuring cytokines by using two different statistical methods, regression analysis and Bland-Altman plot. Materials and methods Fifty patients, diagnosed with hypercholesterolemia and expecting high level serum cytokines, and 30 healthy volunteers, expecting normal level serum cytokines, were enrolled in the study. The interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using ELISA, and compared with obtained levels using flow cytometric method. Results Although regression analysis showed that the two methods are compatible with measurements of IL-1β, IL-6 and TNF-α, they tended to show dissimilarity with measurements of IL-1β and TNF-α based on Bland-Altman graphs. Conclusion According to Bland-Altman plot, our results providing evidence of ELISA and flow cytometry assays were compatible with each other for IL-1β and IL-6 measurements compared to TNF-α measurement. However, our study has a small number of participants, hence this study need to be confirmed by investigations involving more participants.

Amorphous nanosilica particles block induction of oral tolerance in mice

The mucosal immune system is exposed to non-self antigens in food and the gut microbiota. Therefore, the recognition of orally ingested non-self antigens is suppressed in healthy individuals to avoid excessive immune responses in a process called “oral tolerance”. The breakdown of oral tolerance has been cited as a possible cause of food allergy, and amorphous silica nanoparticles (nSP) have been implicated in this breakdown. As nSP are widely used in foodstuffs and other products, exposure to them is increasing; thus, investigations of any effects of nSP on oral tolerance are urgent. This study evaluated the effects of nSP30 (particle diameter = 39 nm) on immunological unresponsiveness induced in mice with oral ovalbumin (OVA). Specifically, production of OVA-specific antibodies, splenocyte proliferation in response to OVA, and effects on T-helper (TH)-1, TH2, and TH17 responses (in terms of cytokine and IgG/IgE subclass expression) were evaluated. nSP30 increased the levels of OVA-specific IgG in OVA-tolerized mice and induced the proliferation of OVA-immunized splenocytes in response to OVA in a dose-related manner. nSP30 also increased the expression of OVA-specific IgG1, IgE, and IgG2a, indicating stimulation of the TH1 and TH2 responses. The expression of interferon (IFN)-γ (TH1), interleukin (IL)-4 and IL-5 (TH2), and IL-17 (TH17) was also stimulated in a dose-related manner by nSP30 in splenocytes stimulated ex vivo with OVA. The induction of tolerance by OVA, the production of anti-OVA IgG antibodies, and proliferation of splenocytes in response to OVA was inhibited by nSP30 in conjunction with OVA and was dose-related. The nSP30 enhanced TH1 and TH2 responses that might prevent the induction of oral tolerance. Overall, this study showed that the abrogation of OVA-induced oral tolerance in mice by exposure to nSP30 was dose-related and that nSP30 stimulated TH1, TH2, and TH17 responses.

Sepsis induces incomplete M2 phenotype polarization in peritoneal exudate cells in mice.

BackgroundMacrophages can differentiate into pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes upon exposure to a pathogen or a cytokine microenvironment. However, M1/M2 macrophage polarization in polymicrobial sepsis has not been fully characterized.MethodsThe polarity of peritoneal exudate (PE) cells from mice that had undergone cecal ligation and puncture (CLP) and the response of those cells to lipopolysaccharide (LPS) in terms of cytokine and chemokine expression were examined.ResultsPE cells from CLP mice demonstrated a shift toward the M2 phenotype in terms of marker enzyme expression. In addition, the CLP-derived PE cells showed apparent unresponsiveness to LPS stimulation with regard to expression of pro-inflammatory cytokines such as TNF-α, while the expression of anti-inflammatory cytokines such as IL-10 was induced. Nevertheless, the CLP-PE cells failed to express M2 chemokines including chemokine (C-C motif) ligand 17 (CCL17), CCL22, and CCL24, all of which are important for T cell recruitment.ConclusionsThe results suggested that a shift of naïve monocytes/macrophages to the M2 phenotype, along with the lack of M2 chemokine expression in septic monocytes/macrophages, might be responsible for immunosuppression after sepsis.Electronic supplementary materialThe online version of this article (doi:10.1186/s40560-015-0124-1) contains supplementary material, which is available to authorized users.

Enhanced osteogenic gene expression in microcarrier-bound scalably expanded fetal bone marrow MSCS

Mesenchymal stem cell (MSC) therapy is a promising strategy to address the unmet clinical need for improved bone regeneration. However, challenges to the scalable stem cell production hinder the clinical translation of MSC therapy. Furthermore, the effects of scalable microcarrier-based culture and enzymatic cell harvest on the phenotype and molecular identity of MSCs upon directed osteogenic differentiation are not well understood. In order to address these issues, we expanded fetal bone marrow MSCs on either commercially-available microcarriers in spinner flask culture or on control tissue culture plastic (TCP) monolayer surfaces. At the end of this expansion, we characterized the phenotype of microcarrier- or TCP monolayer-expanded MSCs in terms of cytokine, gene and surface marker expression. We then differentiated microcarrier-bound, microcarrier-harvested or plastic-harvested MSCs which were seeded onto collagen-coated plates or polycaprolactone-tricalcium phosphate scaffolds. In both cases, microcarrier-bound MSCs displayed increased expression of osteogenic genes such as RUNX2 and ALPL compared to microcarrier-harvested and plastic-harvested cells upon osteogenic induction. These results indicate that microcarrier-bound MSCs undergo enhanced osteogenic differentiation. Our findings also suggest that compared to conventional methods of delivering MSC cell suspensions that were harvested either from microcarriers cultured in agitated bioreactors or from TCP surfaces, in vivo delivery of MSC-bound microcarriers may present a promising method for bone regeneration.

Staphylococcus aureus biofilms induce apoptosis and expression of interferon-γ, interleukin-10, and interleukin-17A on human sinonasal explants.

Staphylococcus aureus is one of the most common bacteria associated with chronic rhinosinusitis (CRS). Although S. aureus biofilms have been correlated with disease severity in CRS, little is known about the initial immune response that biofilms induce in the sinonasal mucosa. The aim of this study was to evaluate the innate immune response (in terms of cytokines) of nondiseased human sinonasal tissue to S. aureus biofilms. Full-thickness sinonasal explant cultures (n = 7 donors) were challenged with established S. aureus biofilms for 24 hours. The expression profiles of 17 cytokines were measured using multiplex analysis, real-time quantitative reverse transcription polymerase chain reaction, and immunohistochemistry. Differences in expression were evaluated using Student's t-test. Interleukin (IL)-1β, IL-10, TNF, IL-17A, and interferon (IFN)-γ were up-regulated at the RNA and protein levels in biofilm-treated tissues compared with controls. Elevation of caspase-3 in biofilm-treated samples indicates S. aureus biofilms induce apoptosis on the sinonasal mucosa. S. aureus biofilms induced apoptosis and a predominant proinflammatory immune response on normal sinonasal mucosal explants. This immune response appeared to be triggered by intrinsic bacterial elements but also by components of the biofilm matrix. Live biofilms were present on the mucosa at the end of the challenge, suggesting an inability of the induced immune response to eliminate the S. aureus biofilms.

In vitro T-cell profile induced by BCG Moreau in healthy Brazilian volunteers

Tuberculosis (TB) remains the world's leading cause of morbidity and mortality. Although Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only vaccine currently in use, its efficacy is highly variable. It has been suggested that early antigenic presentation is a pivotal event leading to a better immune response in TB vaccine models. To investigate this further, we compared in vitro cell-mediated immune responses in the context of early sensitization with TB (i.e. healthy adults vaccinated with BCG when they were young, HD; n = 25) to those in its absence (i.e., newborns with naïve immunity to TB, UV; n = 10) by challenging mononuclear cells with BCG Moreau. After 48 hours, CD4+ and CD8+ T cells were harvested from both groups and stained for PD-1/CD25/ FOXP3. In addition, supernatants were assayed for a broad range of cytokines using an array system. The HD group showed robust reactivity to Protein Purified Derivative and BCG while the naïve, UV group did not. Similarly, in terms of PD-1 expression and Treg cells (CD4+/CD25high+/FOXP3+), only the HD group showed higher levels in CD4 lymphocytes. Otherwise, only the UV group showed expression of CD25dim+ as an activation marker dependent on BCG infection. In terms of cytokines, the HD group showed higher levels of Th1 (IL-2/TNF-α/IFN-γ) and regulatory (IL-10) profiles, with monocytes, but not Tr1 cells, acting as the main source of IL-10. Taken together, our results highlight critical roles of early sensitization with TB in mounting cell-mediated immune responses.

Оцінка стану імунної системи у хворих на хіміорезистентний туберкульоз легень залежно від клінічної форми

Метою роботи була оцінка стану імунної системи у хворих на хіміорезистентний туберкульоз легень залежно від клінічної форми за показниками цитокінів, лейкоцитарної формули та гематологічними індексами. Оцінку стану імунітету проводили на початку лікування у 60 хворих на хіміорезистентний туберкульоз легень. Дослідження рівнів інтерлейкінів-2, -6, -4, -10 та туморнекротичного фактора альфа (TNF-α) у сироватці крові проведено методом твердофазного імуноферментного аналізу. У всіх хворих визначалось прогресування активності специфічного процесу зі значним переважанням гуморального імунітету та максимальним пригніченням клітинного, переважанням продукції раннього прозапального цитокіну — TNF-α.

Innate Immune Receptors in Human Airway Smooth Muscle Cells: Activation by TLR1/2, TLR3, TLR4, TLR7 and NOD1 Agonists

BackgroundPattern-recognition receptors (PRRs), including Toll-like receptors (TLRs), NOD-like receptors (NLRs) and RIG-I-like receptors (RLRs), recognize microbial components and trigger a host defense response. Respiratory tract infections are common causes of asthma exacerbations, suggesting a role for PRRs in this process. The present study aimed to examine the expression and function of PRRs on human airway smooth muscle cells (HASMCs).MethodsExpression of TLR, NLR and RLR mRNA and proteins was determined using real-time RT-PCR, flow cytometry and immunocytochemistry. The functional responses to ligand stimulation were investigated in terms of cytokine and chemokine release, cell surface marker expression, proliferation and proteins regulating the contractile state.ResultsHASMCs expressed functional TLR2, TLR3, TLR4, TLR7 and NOD1. Stimulation with the corresponding agonists Pam3CSK4, poly(I:C), LPS, R-837 and iE-DAP, respectively, induced IL-6, IL-8 and GM-CSF release and up-regulation of ICAM-1 and HLA-DR, while poly(I:C) also affected the release of eotaxin and RANTES. The proliferative response was slightly increased by LPS. Stimulation, most prominently with poly(I:C), down-regulated myosin light chain kinase and cysteinyl leukotriene 1 receptor expression and up-regulated β2-adrenoceptor expression. No effects were seen for agonist to TLR2/6, TLR5, TLR8, TLR9, NOD2 or RIG-I/MDA-5.ConclusionActivation of TLR2, TLR3, TLR4, TLR7 and NOD1 favors a synthetic phenotype, characterized by an increased ability to release inflammatory mediators, acquire immunomodulatory properties by recruiting and interacting with other cells, and reduce the contractile state. The PRRs might therefore be of therapeutic use in the management of asthma and infection-induced disease exacerbations.

Assessment of immune function in Down syndrome patients

In Down syndrome (DS), trisomy 21 leads to overexpression of gene coding for specific enzymes. This overexpression translates directly into biochemical aberrations that affect multiple interacting metabolic pathways which culminates in cellular dysfunction and contributes to the unique pathogenesis of DS. The aim of this study is to evaluate parameters of immune response in terms of cytokines [tumor necrosis factor-α (TNF-α) and interlukin-2 (IL-2)] together with the quantitative expression of cystathionine beta synthase (CBS), whose transsulfuration pathway generates cysteine and hydrogen sulfide (H2S). H2S is known to boost host defense at physiological concentrations and to display cytotoxic activity at higher concentrations. Calcineurin activity (CAN) was also measured as its dysregulation has been shown to cause immune suppression. Subjects were 60 DS patients vs. 30 age and socioeconomic matching healthy controls. In their blood, the cytokines: TNF-α and IL-2, together with CBS and its by product H2S as well as CAN activity, were measured. Results showed that CBSmRNA relative expression (0.56±.06 vs. 0.32±.02), plasma H2S (72±8.5 vs. 50.8±4.1) and TNF-α (8.11±.01 vs. 3.6±0.9) were significantly higher among DS patients compared to controls, while CAN (0.27±0.1 vs. 0.45±0.2 units), was significantly decreased in blood of DS patients compared to controls. IL-2 (36.4±2.6 vs. 37.4±0.9) showed no significant difference between DS patients and controls. Accordingly it can be concluded that excessive synthesis of multiple gene products derived from overexpression of the genes present on chromosome 21 may be the cause for decreased immunity in DS patients compared to controls.

Cellular and population plasticity of helper CD4+ T cell responses

Vertebrates are constantly exposed to pathogens, and the adaptive immunity has most likely evolved to control and clear such infectious agents. CD4+ T cells are the major players in the adaptive immune response to pathogens. Following recognition of pathogen-derived antigens naïve CD4+ T cells differentiate into effectors which then control pathogen replication either directly by killing pathogen-infected cells or by assisting with generation of cytotoxic T lymphocytes (CTLs) or pathogen-specific antibodies. Pathogen-specific effector CD4+ T cells are highly heterogeneous in terms of cytokines they produce. Three major subtypes of effector CD4+ T cells have been identified: T-helper 1 (Th1) cells producing IFN-γ and TNF-α, Th2 cells producing IL-4 and IL-10, and Th17 cells producing IL-17. How this heterogeneity is maintained and what regulates changes in effector T cell composition during chronic infections remains poorly understood. In this review we discuss recent advances in our understanding of CD4+ T cell differentiation in response to microbial infections. We propose that a change in the phenotype of pathogen-specific effector CD4+ T cells during chronic infections, for example, from Th1 to Th2 response as observed in Mycobactrium avium ssp. paratuberculosis (MAP) infection of ruminants, can be achieved by conversion of T cells from one effector subset to another (cellular plasticity) or due to differences in kinetics (differentiation, proliferation, death) of different effector T cell subsets (population plasticity). We also shortly review mathematical models aimed at describing CD4+ T cell differentiation and outline areas for future experimental and theoretical research.

PP54 PROBIOTIC VSL#3 MAY BE EFFECTIVE TO CHANGE THE PROFILE OF CYTOCHINES AND IMMUNOGLOBULINS IN BREAST MILK?

The first month of life represents a critical period for the maturation of the infant’s immune system and, thus, this is a window of opportunity to adopt measures to reduce the risk of diseases. Aim of Study: To examine the specific effects of maternal supplementation with the oral probiotic VSL#3 on breast milk health promoting agents (cytokines and immunoglobulins). Patients andMethods: The study population consisted of 18 healthy pregnant women recruited from the Policlinico Hospital-Department of Obstetrics, Gynecology and Neonatology, University of Bari (Italy). Exclusion criteria were the use of antibiotics or other probiotics during the study period, diabetes and other chronic illnesses. Half of the mothers received daily VSL#3 probiotic supplementation starting from 1 month before delivery until 1 month after delivery while the other 9 pregnant women were used as control. VSL#3 (VSL Pharmaceuticals, Inc, Towson, MD) consisted of sachets containing 900 billion live lyophilized bacteria of 4 strains of Lactobacillus (L. paracasei, L. plantarum, L. acidophilus, and L. delbrueckii subsp. bulgaricus), 3 strains of bifidobacteria (B. longum, B. breve, and B. infantis), and 1 strain of Streptococcus thermophilus. Milk and stool samples were collected 3 days (T0) and one month (T30) after delivery in order to assess cytokines values (Il-6, IL-1β, Il-10 and TGFβ-1) and immunoglobulin values (IgA, IgG, IgM) in breast milk of healthy mothers and immunoglobulins values (IgA, IgG, IgM) in stool samples of their newborns,.The concentrations of TGF-β1, IL-6, IL-10 and IL-1β were analyzed using commercial kits (R & D System Inc., Minneapolis, MN, USA; Research Diagnostics Inc., Flandern, NJ, USA). Statistical analysis: Analysis was done by SPSS. A non parametrical test for independent samples (U-Mann-Whitney) was performed for each variable examined. Results: TGF-β values (pg/ml) were higher at T0 in VSL#3 group (mean: 637±520 vs 381±143.31 (p<0.006). and increased significantly at T30 in the VSL#3 group (1754±1151.16) but not in control group (273±53.81) (p<0.001). IL10: in both groups, there was significant decrease between T0 and T30. No group difference was observed at T0 but at T30, IL10 levels were significantly higher in the VSL#3 group (85±9.81 vs 72±17.93) (p=0.01). IL-1β: significantly higher at T0 in VSL#3 group (284±448.63 vs 46±68.01) (p=0.02). Immunoglobulins. IgA and IgM levels were significantly higher (p<0.05) at T30 and IgG levels were significantly higher at both T0 and T30 in the milk samples of the VSL#3 group (p<0.05). Regarding the stool samples, IgM levels were significantly higher at T0 and T30 and IgA and IgG were significantly higher at T30 in the VSL#3 group (p<0.05) Conclusion: These data indicate that VSL#3 may be a powerful immunomodulator in terms of cytokines and immunoglobulin production, mostly at mucosal surface. Preliminary experiments also indicate that VSL#3 can represent a good supplement in the diet of pregnant women.