Potential Role Of Proinflammatory Cytokines Research Articles

Abstract 3442: FXR mediates macrophage intrinsic responses to suppress colon cancer progression

Abstract Bile acids (BAs) affect the intestinal environment by ensuring barrier integrity, regulating epithelium turnover, maintaining microbiota balance, and modulating the immune system. As a master regulator of BAs homeostasis and innate immunity, Farnesoid X Receptor (FXR), is severely compromised in inflammatory bowel disease (IBD) and colitis-associated colorectal carcinoma (CAC) patients. At the front line, gut macrophages react to the microbiota and metabolites that breach the epithelium. We aim to study the role of the BAs-FXR axis in macrophages. To dissect how inflammation drives tumorigenesis, we employed a classic mouse model of CAC, profiled BAs and cytokines, monitored tumorigenesis, and examined the function of macrophages and Th17 cells. Moreover, we investigated the potential of FexD, an FXR agonist, in ameliorating intestinal inflammation, thus inhibiting tumorigenesis. Further, we identified how macrophage intrinsic FXR senses the BAs abnormality and reshapes the gut macrophages in vivo and ex vivo. We also investigated the potential role of pro-inflammatory cytokines in stimulating intestinal stem cells (ISCs)’ stemness using mouse organoids. This study demonstrates that inflammation-induced epithelial abnormalities compromised FXR signaling and altered BAs profile in CAC. Gut macrophage intrinsic FXR senses the aberrant BAs, leading to the secretion of pro-inflammatory cytokines, which promotes ISCs’ proliferation. Notably, FexD reduced intestinal inflammation and tumorigenesis by suppressing pro-inflammatory responses in gut macrophages and Th17 cells. Mechanistically, FXR regulates gut macrophages' recruitment, polarization, maturation, and crosstalk with Th17 cells. In summary, we uncovered a novel role of FXR modulation of gut macrophages, highlighting the potential of FXR as a therapeutic target over biologicals for treating colon cancer. Citation Format: Ting Fu, Xingchen Dong. FXR mediates macrophage intrinsic responses to suppress colon cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3442.

Target Densities in Malignant and Normal Cells Determine CAR T Cell Efficacy and Off-Target Hematotoxicity

Target antigen density has been established as a key factor for CAR activation and there are ongoing investigations into the number of surface molecules required for target cell lysis. A more precise understanding of differential target expression in malignant vs normal cells is needed to select the optimal CAR design, maintaining efficacy while limiting toxicity on normal cells. In AML, the lack of a leukemia-specific surface target antigen requires fine-tuned combinatorial approaches that enable distinctive target elimination based on differential antigen co-expression patterns in AML vs normal cells. Here, we present our target phenotypic profiling of primary AML and normal hematopoietic cells, and link it with CAR efficacy and changes in normal hematopoietic cell populations. We characterized the AML surface target phenotype across a cytogenetically and molecularly heterogeneous AML patient cohort (n>30) with relapsed/refractory disease. Using spectral flow cytometry on bone marrow and peripheral blood from AML patients and healthy control subjects, we quantified differential target expression of several AML targets, including CD33, CD123, CLEC12A and the novel target ADGRE2. We estimated the numbers of surface molecules per cell on AML bulk, LSC, normal HSPC and differentiated lineages. This allowed us to create two-dimensional maps of combinatorial target densities in AML vs normal. In this analysis, the combination of ADGRE2+CLEC12A offered the best therapeutic window. We developed a novel combinatorial CAR+CCR platform, ADCLEC.syn1, consisting of an ADGRE2-targeting 28z1XX-CAR and a CLEC12A-targeting chimeric costimulatory receptor (CCR) providing additional 4-1BB costimulation. This CAR+CCR configuration triggers killing of cells with high CAR target density alone, whereas cells with low CAR target density are only killed if the CCR target is co-expressed (IF-BETTER gate). Both in vitro and in vivo, we used MOLM13 AML cell line variants that we engineered to match a variety of different ADGRE2 and CLEC12A target density combinations found in AML or normal hematopoietic cells. The observed target density thresholds for ADCLEC.syn1-mediated cell lysis demonstrated potential for sparing vital normal hematopoietic cells. In addition, we evaluated ADCLEC.syn1 in several molecularly distinct AML PDX models with clinically representative target phenotypes, achieving long-term AML remissions in all studied PDX models. Next, we assessed the impact of ADCLEC.syn1 on normal hematopoietic cells. In vitro, we found that soluble factors from antigen-activated CAR T cells led to a reduction of CD34+CD38- HSPC, suggesting a potential role of pro-inflammatory cytokines leading to off-target hematotoxicity. To gain further insight, we established a humanized mouse model that allowed to serially monitor normal hematopoiesis by quantification of HSPC and several differentiated populations in the setting of controlled ADCLEC.syn1 or CD19 CAR activation based on an engineered CD19-positive AML cell line. We found that at time of AML remission, both ADCLEC.syn1 and CD19 CAR-treated mice had detectable CD34+ HSPC, indicating preferential killing of AML cells and relative sparing of HSPC. However, both ADCLEC.syn1 and CD19 CAR led to in vivo reduction of CD34+ HSPC as compared to untreated humanized controls, suggesting an indirect effect of inflammation upon CAR T cell activation. The observed HSPC loss was significantly reduced by blocking pro-inflammatory cytokines, further supporting off-target rather than on-target mechanisms of HSPC reduction. Overall, we profiled antigen densities of several CAR target combinations in AML and normal hematopoietic cells, which informed the development of the ADCLEC.syn1 CAR platform. Pre-clinical in vivo efficacy models based on AML with clinically representative target densities demonstrated high anti-leukemic efficacy of ADCLEC.syn1 and established killing thresholds with potential for sparing vital normal hematopoietic cells. Our in vitro and in vivo hematotoxicity models identified target-independent mechanisms of HSPC reduction in the context of both ADCLEC.syn1 and CD19 CAR activation, a finding that may be related to the off-target hematotoxicity observed clinically in some recipients of CD19 and other CAR T cell products.

The Potential Role of Proinflammatory Cytokines and Complement Components in the Development of Drug-Induced Neuropathy in Patients with Multiple Myeloma.

The launch of novel chemotherapeutic agents—in particular, proteasome inhibitors and immunomodulatory drugs—dramatically changed multiple myeloma (MM) therapy, improving the response rate and prolonging progression-free survival. However, none of the anti-MM drugs are deprived of side effects. Peripheral neuropathy (PN) seems to be one of the most pressing problems. Despite extensive research in this area, the pathogenesis of drug-induced peripheral neuropathy (DiPN) has not yet been fully elucidated. In the present study, we aimed to assess the potential relationship between proinflammatory factors and the development of PN in MM patients with particular emphasis on the application of VTD (bortezomib, thalidomide, dexamethasone) regimen. Our analysis identified increased concentrations of CCL2, IL-1β, and IFN-γ in plasma of MM patients during treatment, both with and without symptoms of PN, compared with untreated neuropathy-free MM patients. At the same time, the plasma concentration of IL-1β in patients with neuropathy was significantly increased compared with patients without PN before and during treatment. Moreover, the results were enhanced at the transcript level by performing global mRNA expression analysis using microarray technology. The most significant changes were observed in the expression of genes responsible for regulating immunological and apoptotic processes. An in-depth understanding of the mechanisms responsible for the development of DiPN might in the future reduce the incidence of PN and accelerate diagnosis, allowing the choice of neuropathy-free treatment strategies for MM.

Pro-inflammatory cytokines and microRNAs in male infertility

BackgroundMale infertility is a problem that affects 10–15% of men of reproductive age. In particular, gametogenesis is a complex process in which inflammation may play a central role through the secretion of cytokines and the expression of microRNAs. We assessed the potential role of proinflammatory cytokines (TNF-α, IL-6 and IL-1α) and microRNAs (miR-146a-5p, miR-34a-5p and miR-23a-3p) in the seminal plasma of infertile men compared to controls, evaluating their correlation with seminal and biochemical parameters.Methods and resultsExpression of cytokines and microRNAs was analyzed by ELISA and q-PCR. Our data shows that IL-1α was significantly increased in the azoospermic group compared to controls, TNF-α mRNA was more expressed in the oligozoospermic group than controls. There were no significant differences in miRNAs expression among the three groups. The correlations between sperm parameters and inflammatory markers were evaluated, however no significance was highlighted.ConclusionsThe determination of each inflammatory marker separately in the seminal plasma of subfertile men, despite some significant differences, does not have a diagnostic value in male infertility even if an assay of selective pro-inflammatory cytokines and microRNAs in the semen may improve the diagnosis of male infertility.

Pro-inflammatory cytokines as potential predictors for intradialytic hypotension

Background Intradialytic hypotension (IDH) is a common complication in maintaining hemodialysis (MHD) patients. Immune activation might be part of the mechanisms. However, the association between pro-inflammatory cytokines and blood pressure (BP) has not been deeply explored. So we aim to evaluate the potential role of pro-inflammatory cytokines in IDH. Methods MHD patients starting hemodialysis before January 2016 were enrolled in our retrospective study. Patients' characteristics, laboratory results, and intradialytic BP were collected. IDH was defined as nadir systolic BP ≤ 90 mmHg during hemodialysis. The definition of IDH group was that those who suffered from more than one hypotensive event during one month after the enrollment (10% of dialysis treatments). Spearman correlation analysis and logistic regression were employed to explore the relationship between pro-inflammatory cytokines and IDH. Results Among 390 patients, 72 were identified with IDH (18.5%). High levels of serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were observed in the IDH group (p < 0.001). Both TNF-α and IL-1β positively correlated with predialysis BP (p < 0.01). Receiver operating characteristic curve (ROC) analysis was used to evaluate the diagnostic accuracy of serum IL-1β and TNF-α for IDH. The area under the curve of IL-1β was 0.772 (95% CI: 0.708-0.836, p < 0.01), and that of TNF-α was 0.701 (95% CI: 0.620-0.781, p < 0.01). After adjusting for patients' characteristics, biochemical parameters, comorbid conditions, predialysis BP, and medications, elevated TNF-α and IL-1β were still risk factors for IDH. Conclusion Pro-inflammatory cytokines (TNF-α and IL-1β) could be potential predictors for IDH.

The role of the blood-brain barrier in hypertension.

What is the topic of this review? This review highlights the importance of the blood-brain barrier in the context of diseases involving autonomic dysfunction, such as hypertension and heart failure. What advances does it highlight? It highlights the potential role of pro-inflammatory cytokines, leucocytes and angiotensin II in disrupting the blood-brain barrier in cardiovascular diseases. Advances are highlighted in our understanding of neurovascular unit cells, astrocytes and microglia, with a specific emphasis on their pathogenic roles within the brain. The blood-brain barrier (BBB) is a crucial barrier that provides both metabolic and physical protection to an immune-privileged CNS. The BBB has been shown to be disrupted in hypertension. This review addresses the importance of the BBB in maintaining homeostasis in the context of diseases related to autonomic dysfunction, such as hypertension. We highlight the potentially important roles of the immune system and neurovascular unit in the maintenance of the BBB, whereby dysregulation may lead to autonomic dysfunction in diseases such as heart failure and hypertension. Circulating leucocytes and factors such as angiotensinII and pro-inflammatory cytokines are thought ultimately to downregulate endothelial tight junction proteins that are a crucial component of the BBB. The specific mechanisms underlying BBB disruption and their role in contributing to autonomic dysfunction are not yet fully understood but are a growing area of interest. A greater understanding of these systems and advances in our knowledge of the molecular mechanisms causing BBB disruption will allow for the development of future therapeutic interventions in the treatment of autonomic imbalance associated with diseases such as heart failure and hypertension.

Peripheral proinflammatory cytokines in Chinese patients with generalised anxiety disorder

BackgroundInflammatory responses and inflammatory cytokines have been implicated in the pathogenesis of affective disorders, particularly major depression. Given the limited evidence relating to the potential role of proinflammatory cytokines in generalised anxiety disorder (GAD), we aimed to examine peripheral proinflammatory cytokines in Chinese patients with GAD. MethodsA case-controlled cross-sectional study design, with recruitment of 48 patients with first episode GAD and 48 matched healthy controls. All participants completed measures of anxiety using well-established questionnaires, and serum levels of pro-inflammatory cytokines were measured using multiplex technology. ResultsSerum levels of CRP, IL-1α, IL-2, IL-6, IL-8, IL-12, IFN-γ, and GM-CSF were significantly higher in the GAD group in comparison to the control group (p < 0.05). Pearson correlation revealed significant positive correlations between anxiety measures and serum levels of CRP, IL-1α, IL-6, IL-8, IFN-γ, and GM-CSF (p < 0.05). LimitationsThe cross-sectional study design does not permit definite conclusions on causal directions between inflammation and GAD. The study was limited to a panel of 8 cytokines and does not exclude the possibility of other important cytokines being involved. ConclusionsThese findings indicate an elevated peripheral proinflammatory response, and provide further support for low grade inflammation in GAD. Further research may identify an ‘inflammatory signature’ for diagnosis and treatment response, and guide the search for novel pharmacological interventions.

Potential Role of Proinflammatory Cytokines in the Pathogenetic Mechanisms of Vascular Lesions in Goats Naturally Infected with Bluetongue Virus Serotype 1

In vitro studies have demonstrated that bluetongue virus (BTV)-induced vasoactive mediators could contribute to the endothelial cells dysfunction and increased vascular permeability responsible of lesions characteristic of bluetongue (BT) like oedema, haemorrhages and ischaemic necrosis in different tissues. However, few in vivo studies have been carried out to clarify the causes of these lesions. The aim of this study was to elucidate in vivo the pathogenetic mechanisms involved in the appearance of vascular lesions in different organs during BT. For this purpose, tissue samples from goats naturally infected with bluetongue virus serotype 1 (BTV-1) were taken for histopathological and immunohistochemical studies to determine the potential role of proinflammatory cytokines (tumour necrosis factor alpha, TNFα and interleukin one alpha, IL-1α) in the increased vascular permeability and their relationship with the presence of virus. Gross and histopathological examination revealed the presence of vascular damage leading to generalized oedema and haemorrhages. Immunohistochemical studies displayed that endothelial injury may have been due to the direct pathogenic effect of BTV infection on endothelial cells or may be a response to inflammatory mediators released by virus-infected endothelial cells and, possibly, other cell types such as monocytes/macrophages. These preliminary results of what appears to be the first in vivo study of tissue damage in small BT-infected ruminants suggest a direct link between the appearance of vascular changes and the presence of BTV-induced vasoactive cytokines.

Injurious effects of acute ethanol exposure during late gestation on developing white matter in fetal sheep

Fetal exposure to maternal alcohol intake can be harmful to the developing brain but the effects of acute exposures are less well documented. Our objective was to determine the effects of acute alcohol exposure on developing white matter and to investigate the potential role of pro-inflammatory cytokines. Fifteen pregnant ewes underwent surgery at 110.0±1.0 days of the 147 day gestation for fetal catheterization. Ethanol (1g/kg maternal weight) was administered intravenously to 8 ewes for 1h on 3 consecutive days at 116.0±1.0 days of gestation (0.8 of full term); 7 pregnant control ewes received saline. Fetal brains were collected at necropsy 5 days after the initial ethanol exposure and processed for structural analysis. Maternal and fetal blood ethanol concentrations reached maximal values (0.11±0.01g/dL) 1h after infusions commenced, declining to zero thereafter. Ethanol exposure did not cause fetal hypoxemia, acidemia, hypercapnia, hypoglycemia or hypotension. Subcortical white matter injury, defined as microglia/macrophage infiltration, axonal disruption, increased apoptosis, astrogliosis and altered glial cell morphology, was observed in 4 of the 8 ethanol-exposed fetuses. The injury occupied 6.6–18.3% of the cross-sectional area of cerebral white matter examined and was substantial in 2/8 and modest in 2/8 ethanol-exposed fetuses. Three remaining fetuses exhibited astrogliosis and elevated levels of apoptosis in cerebral white matter. There was a positive correlation between maternal and fetal blood ethanol concentrations and the extent of brain damage. There was no significant elevation in concentrations of the pro-inflammatory cytokines tumor necrosis factor-α, interleukin-1β and interleukin-6 in fetal plasma. Developing white matter in the late gestation fetus is vulnerable to acute alcohol exposure, but mechanisms remain unclear.

Coinfection of pigs with porcine respiratory coronavirus and Bordetella bronchiseptica

Coinfection with two or more pathogens is a common occurrence in respiratory diseases of most species. The manner in which multiple pathogens interact is not always straightforward, however. Bordetella bronchiseptica and porcine respiratory coronavirus (PRCV) are respiratory pathogens of pigs whose relatives, B. pertussis and the SARS virus, cause respiratory disease in humans. In an initial experiment, the effect of coinfection of PRCV and B. bronchiseptica was examined in thirty, 4-week-old pigs (10 pigs/group) that were infected with either PRCV or B. bronchiseptica, or both PRCV and B. bronchiseptica. An additional 10 pigs served as sham infected controls. Five pigs from each group were euthanized at 4 and 10 days post-infection. Gross and histopathological lung lesions were more severe in the coinfected group as compared to the groups infected with B. bronchiseptica or PRCV alone. In order to investigate the potential role of proinflammatory cytokines in disease severity after coinfection, a second experiment was performed to examine cytokine transcription in alveolar macrophages from single and dually infected pigs. A total of 48 pigs were divided equally into groups as above, but 4 pigs from each group were euthanized at 1, 4 and 10 days post-infection. Coinfected pigs showed a greater and more sustained transcription of proinflammatory cytokines, especially IL-6 and MCP-1, than pigs infected with either PRCV or B. bronchiseptica alone. Thus, there appears to be a synergistic effect between PRCV and B. bronchiseptica with regards to proinflammatory cytokine transcription that may partially explain the increased severity of pneumonia in coinfected pigs.

197 Vigilance and cognitive impairment in adults with OSAS

Although the effects of infectious diseases on sleep and sleepiness have been known for thousands of years, it is only within the past 20 years that there has been a systematic study of changes on sleep following infection or microbial product-induced cytokine production, primarily in animals. In humans, the first few studies focused on the circadian pattern of the cytokines and showed that proinflammatory cytokines peak during nocturnal sleep. Since 1997, there has been a focus on the potential role of proinflammatory cytokines in sleep loss and sleep disorders, such as sleep apnea and narcolepsy. IL-6 and TNFa appear to be elevated in patients with sleep apnea and narcolepsy and have been postulated to be mediators of excessive daytime sleepiness and fatigue. Also, it has been shown that cytokines are elevated in obese vs. normal-weight subjects and contribute to their frequent complaints of sleepiness and fatigue. Cytokines are produced by fat tissue and more so by visceral fat, which may explain why obesity per se is an independent risk factor for excessive daytime sleepiness. In addition, it has been shown that cytokines are elevated in conditions that their primary pathogenetic mechanism is insulin resistance, i.e., women with PCOS, and that they contribute both to their sleep pathology (increased prevalence of sleep apnea and sleepiness) as well as to the underlying pathogenetic mechanism of obesity and insulin resistance. In healthy, normal-sleeping humans, cytokines are elevated after total or partial sleep loss, suggesting a role in the mediation of experimentally induced sleepiness also. Interestingly, interventions to reduce or neutralize cytokines in humans are associated with improved alertness and performance. For example, a mid-afternoon nap following sleep deprivation improved sleepiness and performance and decreased IL-6 peripheral levels. Also, the use of a TNFa antagonist in obese patients with sleep apnea was associated with a marked decrease in sleepiness and a significant though slight improvement of sleep apnea. These studies collectively suggest that proinflammatory cytokines play a key role in sleep pathology, i.e., sleep apnea, sleepiness, and in health problems frequently associated with sleep disorders/ sleep loss, i.e., cardiovascular events, obesity, and metabolic syndrome. Future research in this area may lead to novel and more effective treatments for disorders that plague a large percentage of the population.

The role of proinflammatory cytokines in the cause of neuropathic osteoarthropathy (acute Charcot foot) in diabetes

The pathogenesis of the acute Charcot foot of diabetes remains unclear. All patients with this condition have evidence of peripheral neuropathy, with loss of protective sensation and abnormal foot biomechanics. However, the acute Charcot foot is also characterised by a pronounced inflammatory reaction and the pathogenic significance of this inflammation has received little attention. We suggest that an initial insult--which may or may not be detected--is sufficient to trigger an inflammatory cascade through increased expression of proinflammatory cytokines, including TNFalpha and interleukin 1beta. This cascade then leads to increased expression of the nuclear transcription factor, NF-kappaB, which results in increased osteoclastogenesis. Osteoclasts cause progressive bone lysis, leading to further fracture, which in turn potentiates the inflammatory process. The potential role of proinflammatory cytokines suggests the possibility of new treatments for this sometimes devastating complication of diabetes.