Cytokine Ratio Research Articles

MolBV reveals immune landscape of bacterial vaginosis and predicts human papillomavirus infection natural history

Bacterial vaginosis (BV) is a highly prevalent condition that is associated with adverse health outcomes. It has been proposed that BV’s role as a pathogenic condition is mediated via bacteria-induced inflammation. However, the complex interplay between vaginal microbes and host immune factors has yet to be clearly elucidated. Here, we develop molBV, a 16 S rRNA gene amplicon-based classification pipeline that generates a molecular score and diagnoses BV with the same accuracy as the current gold standard method (i.e., Nugent score). Using 3 confirmatory cohorts we show that molBV is independent of the 16 S rRNA region and generalizable across populations. We use the score in a cohort without clinical BV states, but with measures of HPV infection history and immune markers, to reveal that BV-associated increases in the IL-1β/IP-10 cytokine ratio directly predicts clearance of incident high-risk HPV infection (HR = 1.86, 95% CI: 1.19-2.9). Furthermore, we identify an alternate inflammatory BV signature characterized by elevated TNF-α/MIP-1β ratio that is prospectively associated with progression of incident infections to CIN2 + (OR = 2.81, 95% CI: 1.62-5.42). Thus, BV is a heterogeneous condition that activates different arms of the immune response, which in turn are independent risk factors for HR-HPV clearance and progression. Clinical Trial registration number: The CVT trial has been registered under: NCT00128661.

Lactobacilli and Cytokine Modifications during Menopause and Their Relation to Vulvar and Vulvovaginal Disorders.

ObjectivesFemale sexual and reproductive health is heavily influenced by the levels and ratios of Lactobacilli species and vaginal cytokines. Menopause marks a profound body change as it shifts to a natural and permanent non-reproductive state. Vulvovaginal diseases encompass a broad variety of sexual health conditions. Furthermore, both menopause and vulvovaginal diseases affect vaginal Lactobacilli and cytokine levels. Thus, this study aimed to investigate the correlation between menopause, vulvovaginal diseases, and vaginal Lactobacilli and cytokine levels.MethodsVaginal swab samples were collected as part of a prospective data bank creation to study vaginal conditions as approved by the Institutional Review Board of Texas Tech University Health Sciences Center, Lubbock, USA. This study utilized 38 samples in this database, which were assigned to the pre-menopausal with no vulvovaginal conditions (n = 20) and post-menopausal with vulvovaginal conditions (n = 18) groups. A real-time polymerase chain reaction was conducted to determine the relative concentration of Lactobacilli species, while cytokine analysis was performed using multiplex enzyme-linked immunosorbent assay immunoassay. The standardized mean difference, multivariate analysis of variance, and permutational unequal variance t test were used for the statistical analysis.ResultsCytokines, interleukin (IL)-6, macrophage inflammatory protein-1α, IL-8, and Lactobacillus iners expression were significantly elevated in the control group compared to the study group (P = 0.03 for the cytokines, P = 0.0194 for Lactobacilli).ConclusionsThe levels of vaginal cytokine and Lactobacillus profile were significantly different between the pre-menopausal and post-menopausal groups.

Significance of serum Th1/Th2 cytokine levels in underlying disease classification of childhood HLH

Goal of this research was to investigate values of serum cytokines in childhood HLH with different triggers, with the expectation to find secretion spectrum of 5 main types of underlying diseases. 118 newly diagnosed HLH were included, and serum concentrations of 6 cytokines were tested before treatment began. Absolute cytokine levels and ratios between them were then studied in the HLH groups collectively and separately RESULTS: In general, IFN-γ, IL-10 and IL-6 showed differences among 5 HLH groups. Specifically, relative levels of these three cytokines to each other were meaningful in distinguishing 4 types of HLH. Level of IL-6 was higher than those of IFN-γ or IL-10 in HLH driven by Systemic auto-inflammatory disorders (SAIDs) or Langerhans Cell Histiocytosis (LCH), while primary HLH and EBV-HLH shared elevated ratio of IL-10 to IL-6. Although more than one distinctive ratios were found in 3 HLH groups, combination of these parameters didn't offer optimal balance between sensitivity and specificity. As a group of easily gained laboratory findings, cytokine levels were reliable in the procedure of roughly classifying HLH cases with the help of patients' clinical phenotype. However, adequate data is still needed to explore the significance of these indicators in identifying one particular underlying disease accurately.

Comparison of Th1 and Th17 Inflammatory Cytokine Profiles Between Chronic Plaque and Acute Guttate Psoriasis.

BackgroundThe phenotypic heterogeneity of psoriasis is suspected to reflect differences in its pathogenesis, but not yet completely elucidated. Studies of the Th1 and Th17 cytokines associated with different phenotypes of psoriasis have yielded inconsistent results.ObjectiveTo investigate the tissue expression levels of Th1 and Th17 cytokines among patients with chronic plaque psoriasis, acute guttate psoriasis, and healthy control.MethodsA total of 20 patients with psoriasis (10 with chronic plaque type and 10 with acute guttate type) and 5 healthy controls were enrolled. The tissue mRNA and protein levels of following cytokines were measured: interleukin (IL)-12, IL-2, interferon (IFN)-γ, IL-23, IL-17A, and IL-22.ResultsThe tissue mRNA levels of IL-12, IFN-γ, IL-23, IL-17A, IL-22 and the protein levels of IL-12, IL-2, IFN-γ, IL-17A, IL-22 were significantly increased in the psoriasis patients compared with the healthy controls. In comparisons of the subtypes, the tissue mRNA level of IFN-γ was increased in acute guttate psoriasis, whereas the protein levels of IL-12 and IL-17A were significantly increased in chronic plaque psoriasis. The cytokine ratios of IL-17A/IL-2 and IL-22/IL-2 were significantly higher in chronic plaque psoriasis than in acute guttate psoriasis.ConclusionWe confirmed that the tissue levels of Th1 and Th17 cytokines were increased in psoriasis patients compared with healthy controls. The increased IFN-γ mRNA level in acute guttate psoriasis and increased IL-12 and IL-17A protein levels in chronic plaque psoriasis suggest that an imbalance between Th1 and Th17 cytokines may play a role in the phenotypic transition of psoriasis.

Dual role of neutrophils in modulating liver injury and fibrosis during development and resolution of diet-induced murine steatohepatitis

Inflammatory changes in the liver represent a key feature of non-alcoholic steatohepatitis (NASH), the progressive form of non-alcoholic fatty liver disease (NAFLD). Innate immune activation including hepatic neutrophilic infiltration acts as an important inflammatory trigger as well as a potential mediator of inflammation resolution. In this study, we dissected the effects of neutrophil depletion via anti-lymphocyte antigen 6 complex locus G6D (Ly6G) antibodies administration during ongoing high fat-fructose-cholesterol (FFC) diet-induced murine NASH and during inflammation resolution by switching into a low-fat control diet. During NASH progression, protective effects were shown as HSC activation, cell infiltration and activation of pro-inflammatory macrophages were ameliorated. Furthermore, these changes were contrasted with the effects observed when neutrophil depletion was performed during the resolution phase. Impaired resolving mechanisms, such as a failure to balance the pro and anti-inflammatory cytokines ratio, deficient macrophage phenotypic switch into a pro-restorative profile, and defective repair and remodeling processes were observed when neutrophils were depleted in this scenario. This study described phase-dependent contrasting roles of neutrophils as triggers and pro-resolutive mediators of liver injury and fibrosis associated with diet-induced NASH in mice. These findings have important translational implications at the time of designing NASH therapeutic strategies.

Modulation of Production of Th1/Th2 Cytokines in Peripheral Blood Mononuclear Cells and Neutrophils by Hepatitis C Virus Infection in Chronically Infected Patients

This study investigated the influence of Hepatitis C virus (HCV) infection on the cytokine production profiles of the peripheral blood monoculear cells (PBMC) and neutrophils in chronically naïve HCV-infected patients. Seventy-five genotype-4 naïve HCV-infected patients (HCV+) and healthy subjects (HCV−) were enrolled. The neutrophils and the PBMC were separated by density gradient sedimentation and stimulated with a mitogen. The culture supernatants were evaluated for levels of IFN-α, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12, and TNF-α using anti-cytokine antibody MACSPlex capture beads. The PBMC cytokine profiles of HCV+ patients showed significantly lower mean values for IFN-γ, IL-2, IL-6, IL-9, and IL-10 (p < 0.0001) as compared to HCV− subjects. In contrast, HCV+ patients showed higher mean levels of PBMC cytokine values for IL-5 and TNF-α (p < 0.0001). As for neutrophils, HCV+ patients showed significantly lower mean levels of IFN-α, IFN-γ, IL-2, IL-4, IL-6, IL-9, and IL-10 (p < 0.0001). In contrast, the neutrophils from HCV+ patients showed higher mean levels of IL-5, IL-12, and TNF-α (p < 0.0001). Th1–Th2 cytokine ratios suggested a lower Th1 bias in HCV+ subjects as compared to HCV− subjects. Our results suggest that chronic HCV infection brings about an immunomodulatory effect not only on neutrophils, but also to a lower extent on PBMCs

Ювенильный артрит: этиология, патогенез, современные аспекты (обзор литературы)

В статье даны современные представления об этиологии, наследственной предрасположенности, иммунопатогенезе ювенильного ревматоидного артрита (ЮРА), одного из наиболее частых и самых инвалидизирующих ревматических заболеваний, встречающихся у детей. Этиология заболевания в настоящее время окончательно не расшифрована, существует лишь ряд более или менее обоснованных гипотез. Среди предрасполагающих факторов важную роль отводят персистирующим вирусам. Артротропные персистирующие вирусы — вирусы Коксаки, Эпштейна — Барр, цитомегаловирус, вирусы герпеса, парвовирусы, обладающие тропизмом к тканям суставов, способны длительно в них персистировать и вызывать иммунное воспаление на фоне иммунологических дефектов. Также имеются данные о связи ЮРА с инфекцией, вызванной Mycoplasma pneumoniaе и Chlamydоphila pneumoniaе. В возникновении и развитии ЮРА играет роль генетическая предрасположенность и особенности иммунной системы. Имеются сведения об ассоциации генов гистосовместимости HLA с ЮРА в целом и с отдельными вариантами заболевания. HLA А2, DR5, DR8 являются маркерами олигоартрита у девочек, имеющих увеит, с серопозитивностью по антинуклеарному фактору. HLA В27 — маркер риска развития ЮРА у мальчиков с олигоартритом или ограниченным полиартритом, с преимущественным поражением суставов нижних конечностей, с наличием энтезопатий в школьном или подростковом возрасте. HLA DR4, DR5, DRВ1-04 являются маркерами ЮРА у девочек с поражением мелких симметричных суставов кистей, серопозитивных по ревматоидному фактору, с дебютом заболевания в старшем школьном возрасте. Основу патогенеза ЮРА составляют дефекты Т- и В-клеточных иммунных реакций, приводящие к гиперпродукции провоспалительных цитокинов. Гиперпродукция провоспалительных цитокинов лежит в основе повреждения синовиальной оболочки сустава, хряща, а также в основе развития системных проявлений болезни. Среди большого количества провоспалительных цитокинов центральное место в развитии заболевания занимает фактор некроза опухоли α (ФНО-α), интерлейкины — ИЛ-6, ИЛ-1β. ФНО-α приводит к развитию хронического воспаления, деструкции хряща и кости. ИЛ-6 принадлежит исключительная роль в генезе системного варианта заболевания. С его гиперпродукцией связывают такие внесуставные проявления болезни, как лихорадка, гипохромная анемия, тромбоцитоз. ИЛ-6 стимулирует продукцию гепатоцитами острофазовых белков, играет роль в развитии ряда осложнений, таких как остеопороз и амилоидоз. Состояние цитокиновой сети при различных вариантах ЮРА до конца не изучено. Некоторые авторы указывают на максимальное повышение ИЛ-6 и ИЛ-1 при системном варианте заболевания. Значительное повышение ФНО-α выявлено у пациентов с полиартикулярным вариантом ЮРА, особенно с высокими значениями ревматоидного фактора. Уровень провоспалительных цитокинов коррелирует с активностью воспаления и отражает тяжесть заболевания, определяет дальнейший прогноз. В основе развития ревматических заболеваний у детей существенную роль играет не абсолютное количество образования тех или иных цитокинов, а дисбаланс про- и противовоспалительных цитокинов. Данный дисбаланс может возникнуть под действием повреждающего фактора: бактериальной или вирусной инфекции, переохлаждения, инсоляции, профилактических прививок, травмы суставов, неблагоприятных экологических факторов, хронических психоэмоциональных стрессов. Но у большинства детей после ликвидации действия повреждающего фактора восстанавливается нормальное соотношение цитокинов, а у детей с генетической предрасположенностью дисбаланс сохраняется, что ведет к развитию ревматической патологии.

Immunometabolic factors in adolescent chronic disease are associated with Th1 skewing of invariant Natural Killer T cells

Invariant Natural Killer T (iNKT) cells respond to the ligation of lipid antigen-CD1d complexes via their T-cell receptor and are implicated in various immunometabolic diseases. We considered that immunometabolic factors might affect iNKT cell function. To this end, we investigated iNKT cell phenotype and function in a cohort of adolescents with chronic disease and immunometabolic abnormalities. We analyzed peripheral blood iNKT cells of adolescents with cystic fibrosis (CF, n = 24), corrected coarctation of the aorta (CoA, n = 25), juvenile idiopathic arthritis (JIA, n = 20), obesity (OB, n = 20), and corrected atrial septal defect (ASD, n = 25) as controls. To study transcriptional differences, we performed RNA sequencing on a subset of obese patients and controls. Finally, we performed standardized co-culture experiments using patient plasma, to investigate the effect of plasma factors on iNKT cell function. We found comparable iNKT cell numbers across patient groups, except for reduced iNKT cell numbers in JIA patients. Upon ex-vivo activation, we observed enhanced IFN-γ/IL-4 cytokine ratios in iNKT cells of obese adolescents versus controls. The Th1-skewed iNKT cell cytokine profile of obese adolescents was not explained by a distinct transcriptional profile of the iNKT cells. Co-culture experiments with patient plasma revealed that across all patient groups, obesity-associated plasma factors including LDL-cholesterol, leptin, and fatty-acid binding protein 4 (FABP4) coincided with higher IFN-γ production, whereas high HDL-cholesterol and insulin sensitivity (QUICKI) coincided with higher IL-4 production. LDL and HDL supplementation in co-culture studies confirmed the effects of lipoproteins on iNKT cell cytokine production. These results suggest that circulating immunometabolic factors such as lipoproteins may be involved in Th1 skewing of the iNKT cell cytokine response in immunometabolic disease.

Changes in the cytokine status in women with chronic liver pathology during reproductive losses in the first trimester

Research objective: to determine the level of serum cytokines in the pregravidar period in women with chronic diseases of the hepatobiliary system and reproductive losses in the first trimester in the anamnesis.Materials and methods. 90 patients of reproductive age with chronic pathology of the hepatobiliary system (liver steatosis and non-alcoholic steatohepatitis) were examined. All patients had spontaneous miscarriages in the early stages of gestation in anamnesis. The research was conducted during the pregravid preparation period. The laboratory research included an additional study of the interleukins (ІL-2, ІL-6, ІL-8, ІL-10) and tumor necrosis factor (TNFα) in the blood serum using enzyme immunoferment assay. 8 female donors were examined to determine the reference norm of cytokines in the blood serum. Results. The cytokine status in patients with chronic pathology of the hepatobiliary system against the background of early miscarriage was characterized by an increase in proinflammatory cytokines (TNFα in 2,29 times more, ІL-2 in 1,46 times, ІL-6 in 1,84 times and ІL-8 in 1,50 times) and anti-inflammatory cytokines (ІL-10 in 1,31 times). The integral indices that are characterizing the oppositional cytokines ratio increased (TNFα/ІL-10, ІL-2/ІL-10 and ІL-6/ІL-10) which indicates an inflammatory process in the body. Minimal changes were registered in patients with signs of steatosis – a moderate increase in proinflammatory cytokines (TNFα is 1,34 times more, ІL-2 in 1,18 times, ІL-6 in 1,33 times and ІL-8 in 1,13 times) and preservation within the reference norm of ІL-10. The maximum imbalance of serum cytokines was observed in the exacerbation of non-alcoholic steatohepatitis – a significant increase in proinflammatory cytokines (TNFα is 3,66 times more, ІL-2 in 1,95 times, ІL-6 in 2,57 times and ІL-8 in 2,18 times) against the background of a moderate increase in ІL-10 (1,30 times). In patients with clinical and laboratory signs of remission of non-alcoholic steatohepatitis a moderate increase in proinflammatory (TNFα is 1,87 times more, ІL-2 in 1,20 times, ІL-6 in 1,61 times and ІL-8 in 1,20 times) and anti-inflammatory (ІL-10 in 1,51 times) cytokines was diagnosed. The analysis of the integral indices of oppositional cytokines showed a tendency to increase their values in patients with liver steatosis and non-alcoholic steatohepatitis in the phase of clinical and laboratory remission. During the period of the non-alcoholic steatohepatitis exacerbation a considerable increase in the studied coefficients was revealed. Conclusions. Changes in the cytokine status are noted in patients with early spontaneous miscarriages in anamnesis and chronic diseases of the hepatobiliary system, which are characterized by the severity of inflammatory reactions and progression of the pathological process in liver. Obtained results can be used to predict miscarriage and be taken into account in the program of drug preparation in the pregravid period.

A potential diagnostic serum immunological marker panel to differentiate between primary and secondary knee osteoarthritis.

Inflammation contributes to knee osteoarthritis (KOA) where many immunological mediators participate in its initiation and progression. Most clinicians manage primary (pKOA) and secondary osteoarthritis (sKOA) alike. Whether immunological profiles of pKOA and sKOA differ remains obscure. Hence, we aimed to differentially identify potential serum immunologic diagnostic markers of pKOA and of sKOA. This case control study used 46 KOA patients (pKOA, n = 30; sKOA, n = 16), and 60 age, gender matched controls (normal healthy, n = 30; systemic lupus erythematosus [SLE] disease controls, n = 30) where serum was assayed for cytokines (TNF-α, IL-1β, IL-6, IL-10) and nitric oxide derivatives (NOx). Sandwich ELISA assessed cytokine levels, while the 'Griess assay' quantified NOx levels. The diagnostic accuracy of optimal marker combinations was evaluated by the CombiROC web tool. Compared with pKOA, sKOA serum displayed significantly elevated levels of pro inflammatory cytokines (TNF-α, IL-1β, IL-6) with a concurrent decrease in the anti-inflammatory cytokine, IL-10 (P<0.05). This was reiterated by significantly higher Th1:Th2 (TNF-α: IL-10) serum cytokine ratio observed in sKOA compared to that of pKOA. The CombiROC curves identified TNF-α, IL-1β, IL-6 and NOx as the best performing panel of potential diagnostic markers to discriminate pKOA from control groups (~97% accuracy, 90% Sensitivity [SE] and 98% specificity [SP]), while TNF-α, IL-1β and IL-6 discriminated sKOA from control groups (~100% accuracy, 100% SE, and 98% SP). The study identified discrete serum immune biomarker panels to differentiate between pKOA (TNF-α, IL-1β, IL-6 and NOx) and sKOA (TNF-α, IL-1β and IL-6). These findings may assist in developing distinct therapeutic agents for the two types of KOA.

Assessment of the impact of age and of blood-derived inflammatory markers in horses with colitis.

To determine the impact of age on survival in horses with colitis and to elucidate whether a lower type-1/type-2 cytokine ratio or an exaggerated inflammatory state contribute to reduced survival in aged horses. Part 1: Retrospective cohort analysis. Part 2: Analytic observational study. Part 1: One hundred twenty-four adult horses with colitis. Part 2: Twenty-nine adult horses with new diarrhea onset while hospitalized. Part 1: Patient signalment, select clinicopathological data, diagnoses, treatment, hospitalization length, and invoice were compared between survivors (n=101) and nonsurvivors (n=23). Only age and plasma transfusion retained statistical significance in the final multivariate outcome model, with 8.5 times lower odds of survival in transfused horses (95% confidence interval [CI], 2.6-27.2%). Additionally, the likelihood of nonsurvival increased by 11.8% (95% CI, 4-20.2%) for every year the horse aged (P=0.002). Similarly, geriatric horses (≥20 years) were 15.2 times more likely to die than young-adults (2-12 years, P=0.03), independent of financial investment, documented comorbidities, and duration of hospitalization. Part 2: Select cytokine analyses were performed on serum collected from hospitalized horses within 1hour of diarrhea onset (T0) and 6hours later. At T0, all recorded clinicopathological variables were comparable between geriatric and young-adult horses, suggesting a similar degree of systemic illness. The median concentration of type-2 cytokines interleukin-4 and interleukin-10, and type-1 cytokine interferon-γ did not differ between age groups. Inflammatory cytokines interleukin-6 and tumor necrosis factor-α were significantly higher in geriatric compared to young-adult horses at both sampling time points. Outcome of colitis was less favorable in aging horses and patients receiving a plasma transfusion. Although an exaggerated inflammatory state, based on increased interleukin-6 and tumor necrosis factor-α concentrations, in geriatric horses may contribute to reduced survival, a lower type-1/type-2 cytokines ratio was not identified in our geriatric population.

Association of reduced maternal sHLA-G5 isoform levels and elevated TNF-α/IL-4 cytokine ratio with Recurrent Pregnancy Loss: A study on South Indian women.

Inflammation is of critical importance in successful implantation during pregnancy. However, the establishment of maternal immune tolerance towards semi-allograft foetus is more exigent and is achieved predominantly by human leukocyte antigen-G (HLA-G) isoforms with a special emphasis on soluble HLA-G5 (sHLA-G5). Constant inflammation and lack of resolution by anti-inflammatory milieu, due to aberrant expression of critical immunoregulatory molecules such as sHLA-G5 and dysfunctional T helper cells 1 and 2 (Th1-Th2) cytokine shift, can lead to adverse pregnancy outcomes including recurrent pregnancy loss (RPL). Serum samples of 270 pregnant women (135 healthy parous and 135 with a history of RPL) were evaluated for the concentrations of sHLA-G5, interleukin-4 (IL-4) and tumour necrosis factor-alpha (TNF-α) using sandwich enzyme-linked immunosorbent assay (ELISA) and found elevated levels of sHLA-G5 and IL-4 in controls and higher TNF-α levels and TNF-α:IL-4 ratio in patients (P<.05). Stratified data analysis based on the time of sample collection, that is the first and second trimesters exhibited higher sHLA-G5 and IL-4 in both first and second trimesters in controls than patients, while they displayed lower levels concerning TNF-α and TNF-α:IL-4 ratio (P<.05). However, within patients and controls in the first or second trimesters, there was a significant variation concerning sHLA-G5 alone. Further, the outcome of pregnancies studied in the present investigation revealed a significant elevation in sHLA-G5 levels among women with successful pregnancies compared with women who experienced pregnancy loss, therefore, concluding the potential application of sHLA-G5 isoform as a marker in assisting improved pregnancy outcomes.

Plasma Metabolomics Reveals Pathogenesis of Retained Placenta in Dairy Cows

The complex etiology and pathogenesis of retained placenta (RP) bring huge challenges for researchers and clinical veterinarians in investigating the pathogenesis and treatment schedule. This study aims to investigate the pathogenesis of RP in dairy cows by plasma metabolomics. As subjects, 10 dairy cows with RP and 10 healthy dairy cows were enrolled according to strict enrollment criteria. Imbalanced antioxidant capacity, reduced Th1/Th2 cytokine ratio, and deregulation of total bilirubin (T-bil), alkaline phosphatase (ALP), and reproductive hormones were shown in dairy cows with RP by detecting biochemical indicators, oxidation and antioxidant markers, and cytokines in serum. Plasma metabolites were detected and analyzed by a liquid chromatography–mass spectrometry (LC–MS) system coupled with multivariate statistical analysis software. A total of 23 potential biomarkers were uncovered in the plasma of dairy cows with RP. The metabolic pathways involved in these potential biomarkers are interconnected, and the conversion, utilization, and excretion of nitrogen were disturbed in dairy cows with RP. Moreover, these potential biomarkers are involved in the regulation of antioxidant capacity, inflammation, and autocrine or paracrine hormone. All of these findings suggest that an imbalance of these potential biomarkers might be responsible for the imbalanced antioxidant capacity, reduced Th1/Th2 cytokine ratio, and deregulation of reproductive hormones in dairy cows with RP. The regulation of metabolic pathways involved in these potential biomarkers represents a promising therapeutic strategy for RP.

Predictive Immunological, Virological, and Routine Laboratory Markers for Critical COVID-19 on Admission.

Background Early identification of COVID-19 patients at risk of critical illness is a challenging endeavor for clinicians. We aimed to establish immunological, virological, and routine laboratory markers, which, in combination with clinical information, may allow identifying such patients. Methods Blood tests to measure neutrophil/lymphocyte ratio (NLR) and levels of ferritin, CRP, D-dimer, complement components (C3 and C4), cytokines, and lymphocyte subsets, as well as SARS-Cov-2 RT-PCR tests, were performed in COVID-19-confirmed cases within 48 hours of admission. RT-PCR cycle threshold (Ct) values from oropharyngeal or nasopharyngeal swabs were determined on the day of admission. Symptom severity was categorized as mild (grade 1), severe (grade 2), or critical (grade 3). Results Of 120 patients who were included, 49 had mild, 32 severe, and 39 critical COVID-19. Levels of ferritin >370 ng/mL (OR 16.4, 95% CI 5.3–50.8), D-dimer >440 ng/mL (OR 5.45, 95% CI 2.36–12.61), CRP >7.65 mg/dL (OR 11.54, 95% CI 4.3–30.8), NLR >3.77 (OR 13.4, 95% CI 4.3–41.1), IL-6 >142.5 pg/mL (OR 8.76, 95% CI 3.56–21.54), IL-10 >10.8 pg/mL (OR 16.45, 95% CI 5.32–50.81), sIL-2rα (sCD25) >804.5 pg/mL (OR 14.06, 95% CI 4.56–43.28), IL-1Ra >88.4 pg/mL (OR 4.54, 95% CI 2.03–10.17), and IL-18 >144 pg/mL (OR 17.85, 95% CI 6.54–48.78) were associated with critical COVID-19 in the univariate age-adjusted analysis. This association was confirmed in the multivariate age-adjusted analysis only for ferritin, CRP, NLR, IL-10, sIL-2rα, and IL-18. T, B, and NK cells were significantly decreased in critical patients. SARS-CoV-2 was not detected in blood except in 3 patients who had indeterminate results. RT-PCR Ct values from oropharyngeal or nasopharyngeal swabs on admission were not related to symptom severity. Conclusion Ferritin, D-dimer, CRP, NLR, cytokine (IL-18 and IL-10), and cytokine receptor (IL-6, IL1-Ra, and sCD25) test results combined with clinical data can contribute to the early identification of critical COVID-19 patients.

High Granulocyte-Macrophage Colony Stimulating Factor to Interleukin 10 Ratio and Marked Antioxidant Enzyme Activities Predominate in Symptomatic Cattle Naturally Infected with Candidatus Mycoplasma haemobos, Theileria orientalis, Theileria sinensis and Trypanosoma evansi.

Simple SummaryThe aim of this study was to evaluate the proinflammatory (interleukin 12, GM-CSF, interferon-γ) to anti-inflammatory (interleukins 10, 4) cytokine ratios, oxidant level (malondialdehyde) and antioxidant enzyme (superoxide dismutase, glutathione peroxidase) activities in sera from symptomatic and asymptomatic cattle infected with blood parasites relative to clinically healthy cattle. Elevations in serum GM-CSF level was recorded in both symptomatic and asymptomatic cattle infected with blood parasites. Cattle naturally infected with C. M. haemobos and T. orientalis but without clinical signs had anti-inflammatory cytokine immune responses characterized by low proinflammatory to anti-inflammatory cytokine ratios which favor parasite persistence and resistance in the host. Serum activities of antioxidant enzymes (superoxide dismutase and glutathione peroxidase) and malondialdehyde concentration were elevated in all the symptomatic and asymptomatic blood parasite infected cattle groups. Therefore, a high GM-CSF:IL10 response alongside marked antioxidative enzyme responses were predominant findings in cattle with clinical signs of natural blood parasite infections.The aim of this study was to measure the serum proinflammatory (IL-12, GM-CSF & IFN-γ) to anti-inflammatory (IL-10, IL-4) cytokine ratio, oxidant (MDA) level and antioxidant enzyme (SOD; GPx) activities after blood parasite infections. The blood and serum samples were obtained from 130 cattle and screened for identity of the infecting blood parasites by conventional PCR. The following blood parasite species were detected: Candidatus Mycoplasma haemobos (70/130); Theileria orientalis (65/130); Theileria sinensis (32/130); Anaplasma marginale (49/130); Anaplasma platys (7/130); and Trypanosoma evansi (4/130). The GM-CSF/IL-10 ratio showed significantly higher values in all the symptomatic blood parasite infected cattle groups except for symptomatic A. platys infected cattle groups. Anti-inflammatory cytokine immune responses were notable findings in symptomatic and asymptomatic cattle infected with C. M. haemobos and T. orientalis characterized by low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 (p < 0.05). Therefore, high serum GM-CSF:IL:10 in the symptomatic blood parasite infected cattle, low serum IL-12:IL-10, IFN-γ:IL-10, IL-12:IL-4 and IFN-γ:IL-4 ratios in asymptomatic cattle, high MDA level, and increased antioxidant enzyme activities could be useful predictive tools for outcome of natural blood parasite infections in cattle.

Lycoperoside H, a steroidal alkaloid saponin in tomato seeds, ameliorates atopic dermatitis-like symptoms in IL-33 transgenic mice.

Tomato seeds contain steroidal saponins called lycoperosides. However, it currently remains unclear whether lycoperosides exert anti-inflammatory or anti-allergic effects. Therefore, we herein investigated the effects of tomato seed extract (TSE) and lycoperoside H (LH) in Interleukin (IL)-33 transgenic mice. TSE (500mg/kg) or LH (10mg/kg) was orally administered once a day for 101days and then evaluated mouse behavior, skin symptoms, and blood and skin inflammatory cytokines. TSE slightly suppressed scratching behavior, while TSE and LH both increased locomotive activity. LH also significantly suppressed inflammation scores in the limbs, and TSE and LH reduced transepidermal water loss. Epidermal hyperplasia and the accumulation of eosinophils and mast cells were decreased by TSE and LH. Skin Th2/Th1 cytokine ratio and serum IgE concentrations were significantly reduced by TSE and LH. The present results suggest that the oral administration of LH derived from tomato seeds effectively ameliorates the symptoms of atopic dermatitis. PRACTICAL APPLICATIONS: It has been reported that tomato seeds contain steroidal saponins, lycoperosides, though the effects of lycoperosides on anti-inflammatory or anti-allergic have not yet been revealed. In this study, we demonstrated that the oral administration of lycoperoside H derived from tomato seeds suppressed atopic dermatitis symptoms in IL-33 transgenic mice.

Abstract CT107: A PHASE 1B/2 OPEN-LABEL STUDY WITH RANDOMIZATION IN PHASE 2 OF IMU-131 HER2/NEU PEPTIDE VACCINE PLUS STANDARD OF CARE CHEMOTHERAPY IN PATIENTS WITH HER2/NEU OVEREXPRESSING METASTATIC OR ADVANCED ADENOCARCINOMA OF THE STOMACH OR GASTROESOPHAGEAL JUNCTION

Abstract Background: HER2/neu is overexpressed in up to 30% of gastroesophageal adenocarcinoma (GEA) patients and associated with poor prognosis. Recombinant mAbs to treat HER2/neu-amplified malignancies are effective with limitations so alternatives are needed. Therefore, we developed a B-cell epitope immunotherapy vaccine (IMU-131/HER-Vaxx) consisting of 3 fused B-cell epitopes (P467) from the HER2/neu extracellular domain coupled to CRM197 and adjuvanted with Montanide. This open-label multicenter Phase 1b study aimed to evaluate the optimal/safe IMU-131 dose leading to immunogenicity and clinical responses.Patients &amp; Methods: Fourteen patients with HER2/neu overexpressing GEA were enrolled and dose escalation with 10, 30, and 50µg was performed in 3 cohorts. Immunogenicity was evaluated by P467- and HER2-specific Ab levels and cellular responses, and clinical response rates by CT/MRI scan evaluated according to RECISTv1.1. Results: IMU-131 was safe with no vaccine-related significant local or systemic reactions or SAEs. 11/14 patients were evaluable for changes in tumor size and vaccine-specific immune responses. One patient showed complete response, 5 showed partial response and 4 stable disease as their best response. HER2-specific IgG levels were dose-dependent. While in cohort 1 and 2 only 1/3 and 2/5 patients respectively responded with moderate to high Ab titers, all patients in cohort 3 mounted moderate to high HER2-specific IgGs. Additionally, cellular vaccine responses were induced, e. g. Th1-biased cytokine ratios and reduction of Tregs. The clinical response (progression free survival) was significantly prolonged in cohort 3 and showed a clear association with the magnitude of humoral and cellular vaccine responses. Conclusions: IMU-131 was well tolerated and safe. The vaccine-induced HER2-specific Abs and cellular responses were dose-dependent and correlated with clinical responses. The highest dose (50 µg) was recommended for further evaluation in a Phase II trial with two arms, chemotherapy with IMU-131 vaccine or chemotherapy alone, which is currently ongoing. https://clinicaltrials.gov/ct2/show/NCT02795988 Citation Format: Marina Maglakelidze, Dinara Ryspayeva, Iurie Bulat, Zoran Andric, Ivan Nikolic, Tanuj Chawla, Rajnish Nagarkar, Vaibhav Chourdhary, Giri Venkata, Rajesh Kumar Singh, Davorin Radosavljevic, Zoran Petrovic, Ursula Wiedermann, Leslie Chong, Nicholas J. Ede, Rita Laeufle, Anthony Good. A PHASE 1B/2 OPEN-LABEL STUDY WITH RANDOMIZATION IN PHASE 2 OF IMU-131 HER2/NEU PEPTIDE VACCINE PLUS STANDARD OF CARE CHEMOTHERAPY IN PATIENTS WITH HER2/NEU OVEREXPRESSING METASTATIC OR ADVANCED ADENOCARCINOMA OF THE STOMACH OR GASTROESOPHAGEAL JUNCTION [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr CT107.

Exploratory Evaluation of the Relationship Between iNKT Cells and Systemic Cytokine Profiles of Critically Ill Patients with Neurological Injury

Neurological injury can alter the systemic immune system, modifying the functional capacity of immune cells and causing a dysfunctional balance of cytokines, although mechanisms remain incompletely understood. The objective of this study was to assess the temporal relationship between changes in the activation status of circulating invariant natural killer T (iNKT) cells and the balance of plasma cytokines among critically ill patients with neurological injury. We conducted an exploratory prospective observational study of adult (18years or older) intensive care unit (ICU) patients with acute neurological injury (n = 20) compared with ICU patients without neurological injury (n = 22) and healthy controls (n = 10). Blood samples were collected on days 1, 2, 4, 7, 14, and 28 following ICU admission to analyze the activation status of circulating iNKT cells by flow cytometry and the plasma concentration of inflammation-relevant immune mediators, including T helper 1 (TH1) and T helper 2 (TH2) cytokines, by multiplex bead-based assay. Invariant natural killer T cells were activated in both ICU patient groups compared with healthy controls. Neurological patients had decreased levels of multiple immune mediators, including TH1 cytokines (interferon-γ, tumor necrosis factor-α, and interleukin-12p70), indicative of immunosuppression. This led to a greater than twofold increase in the ratio of TH2/TH1 cytokines early after injury (days 1 - 2) compared with healthy controls, a shift that was also observed for ICU controls. Systemic TH2/TH1 cytokine ratios were positively associated with iNKT cell activation in the neurological patients and negatively associated in ICU controls. These relationships were strongest for the CD4+ iNKT cell subset compared with the CD4- iNKT cell subset. The relationships to individual cytokines similarly differed between patient groups. Forty percent of the neurological patients developed an infection; however, differences for the infection subgroup were not identified. Critically ill patients with neurological injury demonstrated altered systemic immune profiles early after injury, with an association between activated peripheral iNKT cells and elevated systemic TH2/TH1 cytokine ratios. This work provides further support for a brain-immune axis and the ability of neurological injury to have far-reaching effects on the body's immune system.

Effect of Dezocine on the Ratio of Th1/Th2 Cytokines in Patients Receiving Postoperative Analgesia Following Laparoscopic Radical Gastrectomy: A Prospective Randomised Study.

PurposeTo evaluate the effect of dezocine on the postoperative ratio of Th1/Th2 cytokines in patients undergoing laparoscopic radical gastrectomy.Patients and MethodsSixty patients undergoing laparoscopic radical gastrectomy were randomly divided into two groups (n=30): dezocine group (Group D) and sufentanil group (Group S). They received patient-controlled intravenous analgesia (PCIA) after the operation with either dezocine 0.8 mg/kg (Group D) or sufentanil 2 µg/kg (Group S). Both groups also received ondansetron 8 mg diluted to 100 mL with saline. The primary outcome was the Th1/Th2 cytokines ratio at predetermined intervals, 30 min before the induction of general anaesthesia and 0, 12, 24 and 48 h after surgery. The secondary endpoints were patients’ pain scores, measured on a visual analogue scale (VAS) at predetermined intervals (0, 12, 24 and 48 h after surgery), and side effects at follow-up 48 h after surgery.ResultsThe Th1/Th2 cytokines ratio in Group D was significantly higher than Group S (P<0.05) 12, 24 and 48 h after the operation. There were no significant differences in VAS pain scores between groups at 0, 12, 24 and 48 h after surgery (P>0.05). Compared to Group S, the incidence of postoperative nausea, vomiting and lethargy was significantly lower in Group D (P<0.05).ConclusionDezocine increases the ratio of Th1/Th2 cytokines, relieves postoperative pain and causes fewer side effects in patients undergoing laparoscopic radical gastrectomy.

T‐cell mediated responses against alpha‐foetoprotein in hepatocellular carcinoma: Relationship with hepatitis C virus infection, tumour phenotype and patients’ survival

AbstractBackgroundAlpha‐foetoprotein (AFP) is a potential immunotherapeutic target in hepatocellular carcinoma (HCC). However, T‐cell response (TR) to AFP is suppressed in HCC due to immune evasion. It is unknown whether HCV infection may pre‐condition TR against AFP, or whether TR may influence the clinical course of HCC.MethodsWe prospectively enrolled 18 HCV+ treatment‐naïve patients with cirrhosis (CC), 18 HCV+ HCC cases and 17 HCV‐ HCC cases. TR was quantified by ELISPOT using assays specific to interleukin (IL) 2, IL10 and granulocyte‐monocyte colony stimulating factor (GM‐CSF) on ex‐vivo peripheral blood mononuclear cells (PBMC) stimulated in vitro with AFP peptides. Cytokine ratios were compared between groups and with clinicopathological features of HCC, including overall survival (OS).ResultsThe proportion of AFP‐specific responses was not different across the studied groups for any of the assayed cytokines. AFP‐specific IL‐2 responses were increased in larger (P = .02), multifocal tumours (P = .01) and correlated with advanced disease (P = .01). TRs did not correlate with other clinicopathological factors and did not predict for OS.ConclusionTumour stage but not HCV infection is related to the emergence of anti‐AFP TRs. These data enable formulation of a rationale for the further development of anti‐AFP immunotherapy in HCC, facilitating optimal patient selection for future studies.