Patterns Of Cytokine Responses Research Articles

Patterns of Multidrug Bacterial Clinical Isolates and Cytokine Responses to Antibiotic Misuse in Nnewi, Anambra State, Nigeria

Patterns of Multidrug Bacterial Clinical Isolates and Cytokine Responses to Antibiotic Misuse in Nnewi, Anambra State, Nigeria

P321: PERTURBATIONS IN STIMULUS-INDUCED CYTOKINE RESPONSES IN CHILDREN WITH BCP-ALL

Background: B-cell precursor acute lymphoblastic leukemia (BCP-ALL) is the most common subtype of childhood leukemia with a peak during early childhood when children are exposed to viral infections. About 5% of all newborns carry a preleukemic clone, but only about 0,2% of them develop BCP-ALL later in life. It has been assumed that a dysregulated, altered immune response underlies the outgrowth of a pre-leukemic clone in infection-triggered B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Aims: We investigated whether children with BCP-ALL differ in their cytokine response from healthy, age-matched, non-leukemic children, after immune cells were challenged with viral, fungal or bacterial stimuli. Methods: We set up a functional experimental platform with 73 stimulus-cytokine pairs to comprehensively characterize the immune profile of pediatric patients with BCP-ALL with the two most common genetic subtypes, either carrying the ETV6/RUNX1 (E/R) gene fusion or the high-hyperdiploid (HD) karyotype in comparison with an age-and gender-matched healthy control cohort without BCP-ALL and their healthy parents (in total n = 101 individuals). Children were in stable first remission, at least two years after the end of therapy and had a fully reconstituted immune system. Blood was taken of patients and controls in parallel and peripheral blood mononuclear cells (PBMCs) were isolated using a density gradient centrifugation. Cells had been stimulated with various fungal, bacterial and viral pathogens, such as Candida albicans, Staphylococcus aureus, Influenza virus, Respiratory Syncytial virus and toll-like receptor ligands (TLR), such as lipopolysaccharide (TLR4), R848 (TLR7/8) or CpG (TLR9); subsequently cytokines (IFNa, IL-1b, IL-1Ra, IL-10, IL-12p70, IL-17, IFNg) had been measured in the cell culture supernatant. Results: Cytokine base line level did not differ between the groups, while the challenge with various infectious antigens resulted in an overall elevated cytokine production of the BCP-ALL patients with E/R fusion (n=11) (p < 0.01, Mann-Whitney-Wilcoxon test two-sided with Benjamini-Hochberg correction). By deciphering the characteristics of the immune response, a remarkable difference for the anti-viral immune response (p < 0.01) was identified, while the anti-bacterial and anti-fungal response did not differ: The IFNa-response induced by Influenza virus and R848 and the IL-17-response induced by R848 differed most prominently, although not significantly after correction for multiple testing. Further analysis of the cytokine profile of the E/R parents compared with an age- and gender-matched healthy control group revealed an overall significantly elevated cytokine response (p < 0.05) after stimulation, but the different sub-features, such as the fungal, bacterial and viral immune responses were not different. Most strikingly, we could identify this pattern specifically for the E/R cohort. Comparing the cytokine responses of the HD patients (n=8) with the healthy controls (n=22) did not result in any significant differences. Image:Summary/Conclusion: Our study suggests that an altered pro-inflammatory anti-viral cytokine response pattern in children with E/R-BCP-ALL is specific to those children, who later developed the BCP-ALL. It is still present years after chemotherapy and possibly contributes to the outgrowth of a pre-leukemic clone, consistently generated in utero during fetal hematopoiesis.

Bone marrow transplantation induces changes in the gut microbiota that chronically increase the cytokine response pattern of splenocytes

Bone marrow transplantation (BMT) involves conditioning regimens which acutely induce side effects, including systemic inflammation, intestinal damage and shifts in the gut microbial composition, some of which may persist chronically. As the gut microbiota affect systemic immune responses, we aimed to investigate whether, post-BMT, the peripheral immune system is modulated as a direct consequence of alterations in the gut microbiota. We show that 24 weeks post-BMT, splenocytes but not peritoneal macrophages display increased cytokine response patterns upon ex-vivo stimulation with various pathogens as compared to untreated controls. The pattern of BMT-induced cytokine responses was transferred to splenocytes, and not to peritoneal macrophages, of healthy controls via co-housing and transferred to germfree mice via transplantation of cecum content. Thus, BMT induces changes in gut microbiota that in their turn increase cytokine responsiveness of splenocytes. Thus, BMT establishes a dominant microbiota that attenuates normalization of the immune-response.

Systems biological assessment of altered cytokine responses to bacteria and fungi reveals impaired immune functionality in schizophrenia.

Evidence suggests that complex interactions between the immune system and brain have important etiological and therapeutic implications in schizophrenia. However, the detailed cellular and molecular basis of immune dysfunction in schizophrenia remains poorly characterized. To better understand the immune changes and molecular pathways, we systemically compared the cytokine responses of peripheral blood mononuclear cells (PBMCs) derived from patients with schizophrenia and controls against bacterial, fungal, and purified microbial ligands, and identified aberrant cytokine response patterns to various pathogens, as well as reduced cytokine production after stimulation with muramyl dipeptide (MDP) in schizophrenia. Subsequently, we performed single-cell RNA sequencing on unstimulated and stimulated PBMCs from patients and controls and revealed widespread suppression of antiviral and inflammatory programs as well as impaired chemokine/cytokine-receptor interaction networks in various immune cell subpopulations of schizophrenic patients after MDP stimulation. Moreover, serum MDP levels were elevated in these patients and correlated with the courseof the disease, suggesting increased bacterial translocation along with disease progression. In vitro assays revealed that MDP pretreatment altered the functional response of normal PBMCs to its re-stimulation, which partially recapitulated the impaired immune function in schizophrenia. In conclusion, we delineated the molecular and cellular landscape of impaired immune function in schizophrenia, and proposed a mutual interplay between innate immune impairment, reduced pathogen clearance, increased MDP translocation along schizophrenia development, and blunted innate immune response. These findings provide new insights into the pathogenic mechanisms that drive systemic immune activation, neuroinflammation, and brain abnormalities in schizophrenia.

Role of Pegylated Interferon in Patients with Chronic Liver Diseases in the Context of SARS-CoV-2 Infection.

The coronavirus 2019 (COVID-19) pandemic has resulted in 168 million cases and about 3.5 million deaths (as of May 26, 2021) during the last 18 months. These 18 months of the COVID-19 pandemic have been characterized by phases or waves of new cases, the emergence of new variants of the deadly virus, and several new complications. After providing emergency approval to several drugs and adherence to several public health measures with frequent full and partial lockdowns, the incidence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) could not be contained till now on a global basis. Although prophylactic vaccines have inspired optimism, the scarcity of vaccines and several vaccine-related regulations indicate that the vaccine's benefit would not be reaching the people of developing countries anytime soon. In the course of our clinical practice, we used pegylated interferon (Peg-IFN) in 35 patients with chronic liver diseases (CLD), and we found that only two of them were infected with SARS-CoV-2 that was mild in nature. These two patients with CLD have a mild course of disease cured without any specific therapy. Patients with CLD are usually immune-compromised. However, three CLD patients remained free of SARS-CoV-2 although they had COVID-19 patients among their family members. Next, we accomplished two studies for assessing the immune-modulatory capacities of Peg-IFN, 1 and 12 injections following administration of Peg-IFN. The data revealed that peripheral blood mononuclear cells (PBMCs) of Peg-IFN-administered CLD patients produced significantly higher levels of some cytokines of innate immunity in comparison with the cytokines produced by PBMC of CLD patients before Peg-IFN intake. The pattern of cytokine responses and absence of infection of SARS-CoV-2 in 33 of 35 CLD patients represent some preliminary observations indicating a possible role of Peg-IFN in patients with CLD. The study may be extended to other chronic infections and cancers in which patients receive Peg-IFN. The role of Peg-IFN for pre- or postexposure prophylaxis in the acquisition of SARS-CoV-2 infection and influencing the natural course of COVID-19 remains to be clarified.How to cite this articleAkbar SMF, Mahtab MA, Aguilar JC, et al. Role of Pegylated Interferon in Patients with Chronic Liver Diseases in the Context of SARS-CoV-2 Infection. Euroasian J Hepato-Gastroenterol 2021;11(1):27–31.

Maternal Latent Mycobacterium tuberculosis Does Not Affect the Infant Immune Response Following BCG at Birth: An Observational Longitudinal Study in Uganda.

Background: BCG has low efficacy in tropical countries. We hypothesized that maternal latent Mycobacterium tuberculosis (M.tb) infection (LTBI) results in fetal tolerance to mycobacterial antigens and impaired responses to BCG immunization.Methods: We enrolled 132 LTBI-positive and 150 LTBI-negative mothers and their babies in Entebbe, Uganda. Infants were BCG-immunized at birth. Cord blood and samples at weeks 1, 4, 6, 10, 14, 24, and 52 were analyzed for cytokine/chemokine responses to M.tb antigens by Luminex 17-plex assay in 6-day whole blood cultures and antibody responses by ELISA. Of the 17 Luminex analytes, seven (IL-2, IL-5, IL-10, IL-13, IL-17A, TNF, and IFN-γ) were included in the main analysis as they were considered most likely to represent T cell responses. Immune sensitization was defined as a detectable cord blood cytokine response to PPD for any of the seven cytokines. Patterns of cytokine and antibody responses were compared between infants of mothers with and without LTBI using linear mixed models adjusting for confounders.Results: Most infants (73%) were sensitized in utero to M.tb antigens, with no overall difference seen between infants born to mothers with or without LTBI. Patterns of post-BCG cytokine and antibody responses to mycobacterial antigens were similar between the two infant groups.Conclusions: Our data do not support the hypothesis that maternal LTBI results in an impaired response to BCG immunization, in Ugandan infants. BCG vaccination at or shortly after birth is likely to be beneficial to all infants, irrespective of maternal LTBI status.

A latent class approach for joint modeling of a time-to-event outcome and multiple longitudinal biomarkers subject to limits of detection.

Multiple biomarkers on different biological pathways are often measured over time to investigate the complex mechanism of disease development and progression. Identification of informative subpopulation patterns of longitudinal biomarkers and clinical endpoint may assist in risk stratification and provide insights into new therapeutic targets. Motivated by a multicenter study to assess the inflammatory markers of sepsis in patients with community-acquired pneumonia, we propose a joint latent class analysis of multiple biomarkers and a time-to-event outcome while accounting for censored biomarker measurements due to detection limits. The interrelationship between biomarker trajectories and clinical endpoint is fully captured by a latent class structure, which reveals the subpopulation profiles of biomarkers and clinical outcome. The estimation of joint latent class models becomes more complicated when biomarkers are subject to detection limits. Based on a Metropolis-Hastings method, we develop a Monte Carlo Expectation-Maximization (MCEM) algorithm to estimate model parameters. We demonstrate the satisfactory performance of our MCEM algorithm using simulation studies, and apply our method to the motivating study to examine the heterogeneous patterns of cytokine responses to pneumonia and associated mortality risks.

Abstract 1555: Monoclonal antibodies targeting the TAM family of receptor tyrosine kinases

Abstract The TAM receptors (Tyro3/Axl/MerTK) family of receptor tyrosine kinases (RTKs) are important negative regulators of innate immunity. TAM receptor activation in myeloid cells by its ligands Gas6 or Protein S (PROS) promotes phosphatidylserine-dependent efferocytosis of apoptotic cells, inducing a tolerogenic state and mediating resolution of inflammation. TAM-deficient mice exhibit phenotypes consistent with systemic inflammation and autoimmunity. Importantly, individual ablation of TAM receptors can confer tumor immunity, increased pro-inflammatory cytokines and tumor lymphocyte infiltration, leading to the proposal that TAM receptors act as checkpoints of innate immunity. We hypothesize that pharmacological targeting of this family of receptors with monoclonal antibodies (mAbs) may lead to a similar pro-inflammatory response and recapitulate the antitumor effects observed in TAM-deficient mice. From a panel of human anti-MerTK, Axl or Tyro3 mAbs derived from phage-display libraries or human IgG expressing mice we identified unique mAbs that markedly enhanced cytokine and chemokine release from primary human immune cells, alone or in the presence of inflammatory stimuli. Interestingly, the qualitative and quantitative pattern of cytokine response from dendritic cells was very similar using antibodies targeting the individual TAM receptors, and also similar to activation of dendritic cells using a CD40 agonist mAb, suggesting that TAM-targeting mAbs can promote immune activation. We identified surrogate mAbs targeting mouse TAM receptors that elicited similar responses in vivo, and demonstrated antitumor activity when dosed alone, or in combination with PD-1/L1 blockade in syngeneic tumor models. In addition, human MerTK transgenic, and TAM knockout mice have been generated and characterized in order to establish in vivo proof-of-concept with human TAM mAbs. Overall, pharmacological modulation of TAM receptors with mAbs enhances cytokine production in human and murine model systems consistent with the published role of TAMs as negative regulators of innate immunity. Mechanistic and proof-of-concept studies support further development of these mAbs as novel approaches to overcome these checkpoints of the innate immune response. Citation Format: Diego Alvarado, Laura Vitale, Mike Murphy, Thomas O'Neill, Andrew Proffitt, Jay Lillquist, Gwenda Ligon, Komal Patel, Anna Wasiuk, Jeff Weidlick, Jenifer Widger, Laura Mills-Chen, Andrea Crocker, Colleen Patterson, Russell A. Hammond, Li-Zhen He, Joel Goldstein, Lawrence J. Thomas, Henry C. Marsh, Tibor Keler, Richard Gedrich. Monoclonal antibodies targeting the TAM family of receptor tyrosine kinases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1555.

Ontogeny of cytokine responses to PHA from birth to adulthood.

Altered production of cytokines is believed to contribute to early childhood susceptibility to infection. The aim of this study was to get further insight into the developmental patterns of cytokine responses from birth to adulthood. The expression levels of 13 cytokines were compared in the supernatants of phytohemaggluttinin (PHA)-stimulated whole blood from healthy neonates (cord blood, n = 8), infants ( < 1-year-old, n = 20), and school-aged children (3-15 y; n = 20). Five adults were used as reference. While Th1, Th2, and Th17 cytokine levels increased progressively from birth to childhood (Mann-Whitney, p < 0.003), high IL-10 secretion at birth dropped to low adult levels in infants (p < 0.004) such that a negative correlation between IL-10 and Th1, Th2, and Th17 cytokine levels at birth (Spearman's correlation, r < -0.70, p < 0.01) converted to a positive correlation in infants (r > 0.60, p < 0.001). Finally, high IL-2, IL-7, and Granulocyte-Colony Stimulating factor (G-CSF) cytokine levels at birth decreased steadily over the first year of life (Mann-Whitney, p ≤ 0.001). The most noticeable result of the study is the rapid shift from enhanced IL-10 secretion capacity at birth toward balanced IL-10/Th1/Th2/Th17 cytokine levels early in life. This change appears an essential precondition to fight pathogens and at the same time to avoid overwhelming inflammatory reactions.

Cytokine Responses to Rhinovirus and Development of Asthma, Allergic Sensitization, and Respiratory Infections during Childhood.

Immunophenotypes of antiviral responses, and their relationship with asthma, allergy, and lower respiratory tract infections, are poorly understood. We characterized multiple cytokine responses of peripheral blood mononuclear cells to rhinovirus stimulation, and their relationship with clinical outcomes. In a population-based birth cohort, we measured 28 cytokines after stimulation with rhinovirus-16 in 307 children aged 11 years. We used machine learning to identify patterns of cytokine responses, and related these patterns to clinical outcomes, using longitudinal models. We also ascertained phytohemagglutinin-induced T-helper cell type 2 (Th2)-cytokine responses (PHA-Th2). We identified six clusters of children based on their rhinovirus-16 responses, which were differentiated by the expression of four cytokine/chemokine groups: interferon-related (IFN), proinflammatory (Inflam), Th2-chemokine (Th2-chem), and regulatory (Reg). Clusters differed in their clinical characteristics. Children with an IFNmodInflamhighestTh2-chemhighestReghighest rhinovirus-16-induced pattern had a PHA-Th2low response, and a very low asthma risk (odds ratio [OR], 0.08; 95% confidence interval [CI], 0.01-0.81; P = 0.03). Two clusters had a high risk of asthma and allergic sensitization, but with different trajectories from infancy to adolescence. The IFNlowestInflamhighTh2-chemlowRegmod cluster exhibited a PHA-Th2lowest response and was associated with early-onset asthma and sensitization, and the highest risk of asthma exacerbations (OR, 1.37; 95% CI, 1.07-1.76; P = 0.014) and lower respiratory tract infection hospitalizations (OR, 2.40; 95% CI, 1.26-4.58; P = 0.008) throughout childhood. In contrast, the IFNhighestInflammodTh2-chemmodReghigh cluster with a rhinovirus-16-cytokine pattern was characterized by a PHA-Th2highest response, and a low prevalence of asthma/sensitization in infancy that increased sharply to become the highest among all clusters by adolescence (but with a low risk of asthma exacerbations). Early-onset troublesome asthma with early-life sensitization, later-onset milder allergic asthma, and disease protection are each associated with different patterns of rhinovirus-induced immune responses.

Systemic Responses of Inflammation-Related Factors Following Eccentric Exercise in Humans

Background: Exercise-induced muscle damage is followed by muscle adaptation which has been associated with an inflammatory response and is influenced by a crucial balance between pro- and anti-inflammatory cytokines. This study investigated the pattern of systemic cytokine responses for several hours after muscle-damaging exercise. Methods: Nine healthy, young men volunteers performed 50 maximal eccentric muscle actions with each leg using the knee extensors. Serum levels of interleukin (IL)-1α, IL-2, IL-6 and IL-10, transforming growth factor (TGF)-β1 and tumor necrosis factor (TNF)-α were measured by ELISA before and at 6, 48 and 120 hours post-exercise. Results: Volunteers reported significant muscle soreness and their serum creatine kinase (CK) activity increased gradually up to 120 hrs post-exercise (p<0.05). Circulating levels of IL-1a remained unaltered and TGF-β1 increased slightly over time, while IL-2 showed a moderate increase 48 hrs following eccentric exercise (p>0.05). Levels of TNF-α and IL-10 exhibited a similar pattern of response over time, showing a nearly 50% and 100% increase, respectively, 6 hrs post-exercise, while IL-6 increase significantly 6 and 48 hrs post-exercise (p<0.05). Conclusion: These findings suggest that eccentric exercise might trigger a systemic, predominantly anti-inflammatory, acute cytokine response as part of the adaptation process to muscle damage, where IL-6 may be especially involved.

Tumor Necrosis Factor-Alpha and Interleukin-1 Alpha as Predictors of Survival in Peritonitis: A Pilot Study.

Context:Peritonitis induces an inflammatory response characterized by the elevation of various cytokine levels. Included in this cascade of cytokines are tumor necrosis factor-alpha (TNF-α) and interleukin-1 alpha (IL-1α). The outcome of patient care may be associated with the pattern of elaboration of these cytokines.Aim:The aim of this study was to describe the pattern of cytokine response (TNF-α and IL-1α) in the course of peritonitis and evaluate them as predictors of mortality in peritonitis.Setting and Design:This was a prospective study conducted in the Division of Gastrointestinal Surgery, University College Hospital, Ibadan.Methods:Consenting patients with clinical diagnosis of generalized peritonitis over a 6-month period (July to December 2015) were recruited. The serum samples of these patients were obtained at presentation, immediately after surgery, and 24 h and 48 h after surgery with a follow-up period of 30 days.Results:Twenty-six samples out of thirty could be analyzed. Serum TNF-α and IL-1α levels were both elevated at presentation in all patients. However, the patterns of change after intervention varied between the survivors and nonsurvivors.Conclusion:Peritonitis triggers a simultaneous increase in serum levels of TNFα and IL-1α. Lower serum level of TNF-α is associated with survival, while on the contrary, higher level of IL-1α is associated with survival.

Postnatal Innate Immune Development: From Birth to Adulthood.

It is well established that adaptive immune responses are deficient in early life, contributing to increased mortality and morbidity. The developmental trajectories of different components of innate immunity are only recently being explored. Individual molecules, cells, or pathways of innate recognition and signaling, within different compartments/anatomical sites, demonstrate variable maturation patterns. Despite some discrepancies among published data, valuable information is emerging, showing that the developmental pattern of cytokine responses during early life is age and toll-like receptor specific, and may be modified by genetic and environmental factors. Interestingly, specific environmental exposures have been linked both to innate function modifications and the occurrence of chronic inflammatory disorders, such as respiratory allergies. As these conditions are on the rise, our knowledge on innate immune development and its modulating factors needs to be expanded. Improved understanding of the sequence of events associated with disease onset and persistence will lead toward meaningful interventions. This review describes the state-of-the-art on normal postnatal innate immune ontogeny and highlights research areas that are currently explored or should be further addressed.

EARLY PULMONARY LESIONS IN INFLUENZA

Comparative analysis of the results of clinical-laboratory and instrumental data from 379 patients hospitalized during 2009—2016, with influenza complicated with pneumonia, showed that the most severe lung lesions were statistically significantly more likely to develop in the first three days caused by shift changed influenza agent in patients with comorbid diseases — chronic diseases of heart and vessels (in isolation or in association with other factors). The differences of the main characteristics of pneumonia and the patterns of cytokine responses between survivors and deceased patients with a severe form of influenza.

Cytokine response patterns to complex biofilms by mononuclear cells discriminate patient disease status and biofilm dysbiosis

ABSTRACTLocalized aggressive periodontitis (LAP) is a rare form of periodontal disease with site-specific rapid tissue destruction. A lipopolysaccharide (LPS) hyper-inflammatory response was shown in LAP using peripheral whole blood, although responses to other bacterial surface components or complex oral biofilms have not been evaluated. Peripheral blood mononuclear cells (PBMCs) from 14 LAP patients, 15 healthy siblings (HS), and 13 unrelated healthy controls (HC) were stimulated with: LPS, lipoteichoic acid, or peptidoglycan; intact or sonically dispersed in vitro–grown biofilms from a LAP disease site, a LAP healthy site, or a healthy control site. Cell culture supernatants were assayed for 14 cyto/chemokines. Discriminant function analysis determined cyto/chemokines that discriminate disease status by response patterns to different stimuli. Qualitative differences in the cytokine response pattern among patient groups were observed to intact and dispersed biofilms, yet responses to healthy and diseased biofilms could not be discriminated. Despite an equivalent magnitude of response, LAP-derived PBMCs demonstrated a qualitatively different pattern of response to LPS and dispersed biofilms. PMBCs from each group responded distinctly to stimulation withsubgingival biofilms. Multiple underlying mechanisms related to bacterial-induced inflammatory responses can culminate in LAP disease initiation and/or progression, and biofilm homeostasis could play an important role.

Maternal BCG scar is associated with increased infant proinflammatory immune responses

IntroductionPrenatal exposures such as infections and immunisation may influence infant responses. We had an opportunity to undertake an analysis of innate responses in infants within the context of a study investigating the effects of maternal mycobacterial exposures and infection on BCG vaccine-induced responses in Ugandan infants. Material and methodsMaternal and cord blood samples from 29 mother-infant pairs were stimulated with innate stimuli for 24h and cytokines and chemokines in supernatants were measured using the Luminex® assay. The associations between maternal latent Mycobacterium tuberculosis infection (LTBI), maternal BCG scar (adjusted for each other’s effect) and infant responses were examined using linear regression. Principal Component Analysis (PCA) was used to assess patterns of cytokine and chemokine responses. Gene expression profiles for pathways associated with maternal LTBI and with maternal BCG scar were examined using samples collected at one (n=42) and six (n=51) weeks after BCG immunisation using microarray. ResultsMaternal LTBI was positively associated with infant IP-10 responses with an adjusted geometric mean ratio (aGMR) [95% confidence interval (CI)] of 5.10 [1.21, 21.48]. Maternal BCG scar showed strong and consistent associations with IFN-γ (aGMR 2.69 [1.15, 6.17]), IL-12p70 (1.95 [1.10, 3.55]), IL-10 (1.82 [1.07, 3.09]), VEGF (3.55 [1.07, 11.48]) and IP-10 (6.76 [1.17, 38.02]). Further assessment of the associations using PCA showed no differences for maternal LTBI, but maternal BCG scar was associated with higher scores for principal component (PC) 1 (median level of scores: 1.44 in scar-positive versus −0.94 in scar-negative, p=0.020) in the infants. PC1 represented a controlled proinflammatory response. Interferon and inflammation response pathways were up-regulated in infants of mothers with LTBI at six weeks, and in infants of mothers with a BCG scar at one and six weeks after BCG immunisation. ConclusionsMaternal BCG scar had a stronger association with infant responses than maternal LTBI, with an increased proinflammatory immune profile.

Cytokine Profile in Patients Infected with Anisakis simplex in Endemic Areas: Dietary Intervention with Polyphenols: A Working Hypothesis.

Anisakiasis is a zoonosis due to infection with the nematode Anisakis (A.) simplex. Humans represent an accidental host after ingesting raw fish infected with A. simplex. A. simplex larvae lead to a robust host immune response mostly at gastro-intestinal level and, usually, T helper (h)2 lymphocytes play a protective against this nematode. However, larvae can elude immunity triggering activation of T regulatory (Treg) cells, which, in turn, suppress Th2 responses via production of interleukin (IL)-10. According to our own experience in the area of Bari (south Italy) where anisakiasis is endemic, chronic urticaria (CU) is very often associated to A. simplex infection. The cytokine profile in CU patients with Anisakis infection is characterized by an elevated release of interferon-γ and IL-17, while IL-10 production is very low. This pattern of pro-inflammatory cytokine response was consistently detectable in patients who underwent a raw fish free diet regimen over a period of six months. In the light of the above considerations, we hypothesize that treating these patients with polyphenols it may be beneficial in terms of activation of Treg cell function. In fact, polyphenols have the capacity to expand Treg cell response in vitro. In this regard, activation of human Treg cells, by analogy to other helminthiasis, may lead to a reduction of allergic/inflammatory reaction in the course of human anisakiasis.

A dynamic relationship between mucosal T helper type 17 and regulatory T-cell populations in nasopharynx evolves with age and associates with the clearance of pneumococcal carriage in humans

Pneumococcal carriage is common in young children, which may account for the high incidence of disease in this age group. Host factors determining the clearance of carriage in humans remain unclear. We aimed to study the relationships between T helper type 17 (Th17) and Foxp3+ regulatory T (Treg) cells in nasopharynx-associated lymphoid tissue (NALT) and carriage in children and adults. Frequencies of Th17 and Treg cells in NALT were analysed by flow cytometry in association with age and pneumococcal carriage status. Cytokine responses following pneumococcal stimulation were analysed by cytometric beads array. The frequencies of Th17 and Treg cells in NALT were inversely correlated (R –0.60). Whereas Treg cell frequency decreased with age (R –0.63), both Th17 and the Th17: Treg ratio increased with age (R 0.62 and R 0.64, respectively). Also, the Th17: Treg ratio was higher in carriage-negative than in carriage-positive children (p <0.01). Pneumococcal stimulation of tonsillar cells increased both Th17 and Treg cell numbers, but the Th17: Treg ratio and pattern of cytokine responses differed between carriage-negative and carriage-positive children. The former showed markedly higher Th17: Treg and interleukin-17A: interleukin-10 ratios than in the latter (p <0.01). Pneumococcal stimulation also induces Th17, although the capacity of this Th17 differentiation from naive T cells of young children was low, but increased with age. We demonstrated a dynamic relationship between Th17 and Treg cells in human nasopharynx that evolves with age. The balance between Th17 and Treg cells in NALT appears to be a major host factor closely associated with the clearance of Streptococcus pneumoniae from the nasopharynx.

Prognostic value of TH1/TH2 cytokines in infants with wheezing in a three year follow-up study.

The immune system is known to play a key role in the development of viral-induced wheeze, but the pattern of cytokine response is not clear. The aim of the study was to determine whether plasma cytokine levels during the acute wheezing illness in infants are associated with the subsequent development of persistent recurrent wheezing (PRW). 43 infants admitted to Iashvili Central Children Hospital, Tbilisi, Georgia, were selected. The concentrations of IFN-g, TNF-a and IL-6 were determined by enzyme-linked immunosorbent assay (ELISA). Total serum IgE level was also determined. IgM and IgG antibodies to RSV, Chlamydophila pneumonia and Mycoplasma pneumonia were tested by ELISA. All children were followed-up during 3 years period. There was no significant association with PRW and atopy in the infant or parental atopy, low maternal age, low birth weight, serum level of IL-6, serum level of IFN-g, serum level of total IgE and the particular pathogen. There was a tendency for the association with male sex, bottle feeding and household cigarette smoking, although the relation was not significant. There was just TNF-a which was strongly associated with the risk of PRW after three years of follow up. We have identified TNF-a, as a biomarker, which can be detected in children under 3 years of age and may be useful in the prediction of development of persistence of wheezing later. To further evaluate the usefulness of this and other markers and establish cutoff levels for prediction and use in clinical practice, a larger prospective follow-up study is needed.

COPD disease severity and innate immune response to pathogen-associated molecular patterns.

The airways of COPD patients are often colonized with bacteria leading to increased airway inflammation. This study sought to determine whether systemic cytokine responses to microbial pathogen-associated molecular patterns (PAMPs) are increased among subjects with severe COPD. In an observational cross-sectional study of COPD subjects, PAMP-induced cytokine responses were measured in whole blood ex vivo. We used PAMPs derived from microbial products recognized by toll-like receptors 1, 2, 4, 5, 6, 7, and 8. Patterns of cytokine response to PAMPs were assessed using hierarchical clustering. One-sided Student’s t-tests were used to compare PAMP-induced cytokine levels in blood from patients with and without severe COPD, and for subjects with and without chronic bronchitis. Of 28 male patients, 12 had moderate COPD (FEV1 50%–80%) and 16 severe COPD (FEV1 <50%); 27 participants provided data on self-reported chronic bronchitis, of which 15 endorsed chronic bronchitis symptoms and 12 did not. Cytokine responses to PAMPs in severe COPD were generally lower than in subjects with milder COPD. This finding was particularly strong for PAMP-induced interleukin (IL)-10, granulocyte colony stimulating factor, and IL-1β. Subjects with chronic bronchitis showed higher PAMP-induced IL-1RA responses to most of the PAMPs evaluated. COPD patients with more severe disease demonstrated a diminished cytokine response to PAMPs, suggesting that chronic colonization with bacteria may dampen the systemic innate immune response.