Changes In Cytokine Levels Research Articles

Comparative Analysis of Cytokine Profiles in Cerebrospinal Fluid and Blood Serum in Patients with Acute and Subacute Spinal Cord Injury.

Cytokines are actively involved in the regulation of the inflammatory and immune responses and have crucial importance in the outcome of spinal cord injuries (SCIs). Examining more objective and representative indicators of the patient's condition is still required to reveal the fundamental patterns of the abovementioned posttraumatic processes, including the identification of changes in the expression of cytokines. We performed a dynamic (3, 7, and 14 days post-injury (dpi)) extended multiplex analysis of cytokine profiles in both CSF and blood serum of SCI patients with baseline American Spinal Injury Association Impairment Scale grades of A. The data obtained showed a large elevation of IL6 (>58 fold) in CSF and IFN-γ (>14 fold) in blood serum at 3 dpi with a downward trend as the post-traumatic period increases. The level of cytokine CCL26 was significantly elevated in both CSF and blood serum at 3 days post-SCI, while other cytokines did not show the same trend in the different biosamples. The dynamic changes in cytokine levels observed in our study can explore the relationships with the SCI region and injury severity, paving the way for a better understanding of the pathophysiology of SCI and potentially more targeted and personalized therapeutic interventions.

Cytokine, Anti-SARS-CoV-2 Antibody, and Neutralizing Antibody Levels in Conventional Blood Donors Who Have Recovered from COVID-19

Background: At the beginning of the pandemic, COVID-19 convalescent plasma (CCP) containing anti-SARS-CoV-2 antibodies was suggested as a source of therapy. In the last 3 years, many trials have demonstrated the limited usefulness of CCP therapy. This led us to the hypothesis that CCP could contain other elements, along with the desired neutralizing antibodies, which could potentially prevent it from having a therapeutic effect, among them cytokines, chemokines, growth factors, clotting factors, and autoantibodies. Methods: In total, 39 cytokines were analyzed in the plasma of 190 blood donors, and further research focused on the levels of 23 different cytokines in CCP (sCD40L, eotaxin, FGF-2, FLT-3L, ractalkine, GRO-α, IFNα2, IL-1β, IL-1RA, IL-5, IL-6, IL-8, IL-12, IL-13, IL-15, IL-17E, IP-10, MCP-1, MIP-1b, PDGF-AA, TGFα, TNFα, and TRAIL). Anti-SARS-CoV-2 antibodies and neutralizing antibodies were detected in CCP. Results: We found no significant differences between CCP taken within a maximum of 180 days from the onset of the first COVID-19 symptoms and the controls. We also made a comparison of the cytokine levels between the low neutralizing antibodies (<160) group and the high neutralizing antibodies (≥160) group and found there were no differences between the groups. Our research also showed no correlation either to levels of anti-SARS-CoV-2 IgG Ab or to the levels of neutralizing antibodies. There were also no significant changes in cytokine levels based on the period after the start of COVID-19 symptoms. Conclusions: No elements which could potentially be responsible for preventing CCP from having a therapeutic effect were found.

Association of interleukin-6 with suicidal ideation in veterans: a longitudinal perspective.

Studies showing associations between inflammation in suicide are typically cross-sectional. Present study investigated how cytokine levels track with suicidal ideation and severity longitudinally. Veterans with a diagnosis of major depressive disorder (MDD) with or without suicide attempt history (MDD/SA n = 38, MDD/NS n = 41) and non-psychiatric non-attempter controls (HC n = 33) were recruited, MDD/SA and HC groups were followed longitudinally at 3 months and 6 months. Blood plasma was collected and processed using Luminex Immunology Multiplex technology. Significant differences in depression severity (BDI) and suicidal ideation severity (SSI) were observed across all groups at study entry, wherein MDD/SA group had the highest scores followed by MDD/NS and HC, respectively. Cytokines IL-1β, IL-4, TNF-α, IFN-γ, and IL-6 were examined at study entry and longitudinally, with IL6 levels differing significantly across the groups (p = 0.0123) at study entry. Significant differences in changes in cytokine levels between depressed attempters and the control group were detected for IL-6 (interaction F1,91.77 = 5.58, p = 0.0203) and TNF-α (F1,101.73 = 4.69, p = 0.0327). However, only depressed attempters showed a significant change, in IL-6 and TNF-α levels, decreasing over time [IL-6: b = -0.04, 95% CI = (-0.08, -0.01), p = 0.0245 and TNF-α: b = -0.02, 95% CI = (-0.04, -0.01), p = 0.0196]. Although IL-6 levels were not predictive of suicidal ideation presence [OR = 1.34, 95% CI = (0.77, 2.33), p = 0.3067], IL-6 levels were significantly associated with suicidal ideation severity (b = 0.19, p = 0.0422). IL-6 was not associated with presence of suicidal ideation. IL-6 however, was associated with severity of ideation, suggesting that IL-6 may be useful in clinical practice, as an objective marker of heightened suicide risk.

Probiotic for Pancreatic β-Cell Function in Type2 Diabetes: A Randomized, Double-Blinded, Placebo-Controlled Clinical Trial.

Many clinical studies have proved the effectiveness of probiotics in metabolic disorders associated with insulin resistance. However, the impact of probiotic therapy on pancreatic β-cell function is ambiguous. The influence of probiotic supplementation vs. placebo on β-cell function in people with type2 diabetes (T2D) was assessed in a double-blind, single-center, randomized, placebo-controlled trial (RCT). Sixty-eight patients with T2D were selected for participation in the RCT. Patients were randomly allocated to consumption of live multistrain probiotics or a placebo for 8weeks, administered as a sachet formulation in double-blind treatment. The primary main outcome was the assessment of β-cell function as change in C-peptide and HOMA-β (homeostasis model assessment-estimated β-cell function), which was calculated using the HOMA2 calculator (Diabetes Trials Unit, University of Oxford). Secondary outcomes were the changes in glycemic control-related parameters, anthropomorphic variables, and cytokines levels. Analysis of covariance was used to assess the difference between groups. Supplementation with live multiprobiotic was associated with slight significant improvement of β-cell function (HOMA-β increased from 32.48 ± 13.12 to 45.71 ± 25.18; p = 0.003) and reduction of fasting glucose level (13.03 ± 3.46 vs 10.66 ± 2.63mmol/L and 234.63 ± 62.36 vs 192.07 ± 47.46mg/dL; p < 0.001) and HbA1c (8.86 ± 1.28 vs 8.48 ± 1.22; p = 0.043) as compared to placebo. Probiotic therapy significantly affects chronic systemic inflammation in people with T2D by reducing pro-inflammatory cytokine levels. Probiotic therapies modestly improved β-cell function in patients with T2D. Modulating the gut microbiota represents a new diabetes treatment and should be tested in more extensive studies. NCT05765292.

Efficacy and safety of self-administered acupressure on symptoms, quality of life and nasal mucosal function in patients with perennial allergic rhinitis: study protocol for a randomized controlled exploratory trial

IntroductionAllergic rhinitis is a global health problem that can potentially be managed through acupressure. Our clinical observations have identified Allergic Rhinitis Acupressure Therapeutic (ARAT) as a novel acupressure treatment acting on specific acupoints, which may enhance the effectiveness of acupressure. Therefore, we propose a three-arm randomized controlled trial will be conducted to investigate the efficacy and safety of ARAT for perennial allergic rhinitis (PAR).Methods/designIn this trial, eligible 111 participants diagnosed with PAR will be randomly assigned to one of three groups: the ARAT group, the non-specific acupoints group, or the blank control group. The primary outcome will be the change in the total nasal symptom score, and the secondary outcomes will include: 1) changes in the scores of the standard version of Rhinoconjunctivitis Quality of Life Questionnaire (RQLQs); 2) acoustic rhinometry and anterior rhinomanometry; 3) changes in the scores of relief medication usage; 4) incidence of adverse events. Additionally, we will measure and compare the changes in cytokine levels (IL-5, IL-13, IFN-γ, and TSLP) in nasal secretions.The RQLQs and primary outcomes will be assessed at the beginning, middle, and end stages of the treatment period, with monthly follow-ups conducted over a total of three months. The secondary outcomes and biomarkers in nasal secretions will be measured at the beginning and end of the treatment period. Any adverse events or need for rescue medication will be carefully noted and recorded.DiscussionThis study may produce a new acupressure treatment prescription that is easy to learn, more targeted, and adaptable. This trial represents the first clinical investigation comparing ARAT treatment for PAR with the non-specific acupoints group and blank control group. Our data is expected to provide evidence demonstrating the safety and efficacy of ARAT for PAR patients, while also exploring the functional mechanism underlying ARAT treatment, moreover, the results offer valuable insights for healthcare professionals in managing PAR symptoms.Trial registrationChinese Clinical Trial Registry, ChiCTR2300072292. Registered on June 08, 2023.

The immunogenicity of Alum+CpG adjuvant SARS-CoV-2 inactivated vaccine in mice

The ongoing evolution and emergence of SARS-CoV-2 variants have raised concerns regarding the efficacy of existing vaccines and therapeutic agents. This study aimed to investigate the immunogenicity of an aluminum hydroxide (Alum) and CpG adjuvanted inactivated vaccine (IAV) candidate against SARS-CoV-2 in mice. A comparison was made between the immune response of mice vaccinated with the Alum+CpG adjuvant IAV and those vaccinated with the Alum adjuvant IAV. Mice immunized with Alum+CpG adjuvant IAV demonstrated high antibody titers and a durable humoral immune response, as well as a Th1-type cellular immune response. Notably, compared to Alum alone vaccine, the Alum+CpG adjuvant IAV induced significantly higher proportions of GC B cells in the splenocytes of immunized mice. Importantly, the changes in inflammatory cytokine levels in the sera of mice vaccinated with the Alum+CpG adjuvant IAV followed a similar trend to that of the Alum adjuvant IAV, which had been proven safe in clinical trials. Overall, our results demonstrate that Alum+CpG adjuvant has the potential to serve as a novel adjuvant, thereby providing valuable insights into the development of vaccine formulations.

Effect of Plant Extracts Combinations on TNF-α, IL-6 and IL-10 Levels in Serum of Rats Exposed to Acute and Chronic Stress.

Oxydative stress, anxiety and depression are associated with changes in cytokine levels. Natural products, including individual and combined plant extracts, have the potential to be used in the treatment of neuropsychiatric disorders. The goal of this study is to investigate the effects of two combined plant extracts, rich in flavonoids, on the levels of the cytokines TNF-α, IL-6, and IL-10 in rats subjected to models of acute cold stress and chronic unpredictable stress. The study utilized common medicinal plants such as Valeriana officinalis, Melissa officinalis, Crataegus monogyna, Hypericum perforatum, and Serratula coronata, which were combined in two unique combinations-Antistress I and Antistress II. The compositions of the used extracts were determined by HPLC methods. Pro- and anti-inflammatory cytokines in rats' serum were measured with Enzyme-linked immunosorbent assay. The results from the acute stress model revealed that the individual extract of Crataegus monogyna decreased levels of TNF-α, while Serratula coronata, Hypericum perforatum, and Valeriana officinalis effectively reduced IL-6 levels. Both combinations, Antistress I and Antistress II, were effective in reducing TNF-α and IL-6 levels, with Antistress II also increasing IL-10 levels. In the chronic stress model, Hypericum perforatum extract decreased the levels of the pro-inflammatory cytokines TNF-α and IL-6, whereas extracts of Serratula coronata and Valeriana officinalis only reduced TNF-α levels. The two combined extracts, Antistress I and Antistress II, decreased TNF-α and IL-6 levels, while Antistress I also reduced the levels of the anti-inflammatory cytokine IL-10. The combinations of plant extracts used in our experiment have not been previously studied or documented in the available literature. However, based on our own experimental results, we can draw the conclusion that the combinations exhibit a more pronounced effect in reducing cytokine levels compared to the individual plant extracts.

Changes in Inflammatory Cytokine Levels in Rectal Mucosa Associated With Neisseria gonorrheae and/or Chlamydia trachomatis Infection and Treatment Among Men Who Have Sex With Men in Lima, Peru.

Neisseria gonorrheae and Chlamydia trachomatis are associated with mucosal inflammation and human immunodeficiency virus 1 (HIV-1) transmission. We assessed levels of inflammatory cytokines in men who have sex with men (MSM) with and without rectal gonorrhea and/or chlamydia in Lima, Peru. We screened 605 MSM reporting condomless receptive anal intercourse for rectal N. gonorrheae/C. trachomatis using nucleic acid testing. We identified 101 cases of gonorrhea and/or chlamydia and randomly selected 50 N. gonorrheae/C. trachomatis positive cases and matched 52 negative controls. We measured levels of IL-1β, IL-6, IL-8, and TNF-α in rectal secretions. Tests for HIV-1, rectal N. gonorrheae/C. trachomatis, and mucosal cytokines were repeated after 3 and 6 months. Cytokine levels in cases and uninfected controls were compared using Wilcoxon rank-sum tests and linear regression. MSM with gonorrhea/chlamydia had elevated levels of all cytokines in rectal mucosa compared with matched controls (all P values <.001). Following antibiotic treatment there were no significant differences in cytokine levels at 3- or 6-month follow-up evaluations (all P values >.05). Rectal gonorrhea/chlamydia infection is associated with transient mucosal inflammation and cytokine recruitment. Our data provide proof of concept for rectal sexually transmitted infection screening as an HIV prevention strategy for MSM. Clinical Trials Registration. NCT03010020.

Studies on the hormone and cytokine producing function of human cumulus cells and its interrelation with fertility in polycystic ovarian syndrome

Cumulus cells (CC) are the specialized layer of follicular cells that are in close contact with the oocyte. They are considered as indirect markers of the oocyte quality. Changes of these cells suggest a damage of the ovary. Determination of cytokines in cumulus cell culture may predict the chance for the conception and development of pregnancy. The objective of the present study was to obtain a primary culture of CC from healthy donors and patients with polycystic ovarian syndrome (PCOS), to identify the most significant differences in production of key cytokines in the CC monocultures of patients and healthy individuals in order to predict the results of in vitro fertilization. Materials and methods: the cell culture technique was used, i.e., cumulus cells of healthy donors and patients with polycystic ovary syndrome were obtained by transvaginal puncture of follicles in the in vitro fertilization (IVF) program. This procedure does not affect the rights of embryo, since the CC are not used at the stage of IVF procedure and intracytoplasmic sperm injection (Ethical Committee Protocol No. 9 of May 16, 2022). IL-6, IL-10, IFN and progesteroneparameters were tested in adhesive cultures of CC by ELISA technique on days 1, 3, 7 of in vitro experiments. Results: We revealed a continuous secretion of progesterone, IL-6, IL-10, IFN in adhesive monocultures of CC. In the patients with PCOS, we have found a sharp increase of progesterone level in cultural media (p 0.01) on the 7th day, By contrast, the initially increased progesterone levels proved to be significantly decreased in donors on the 7th day of culture. Moreover, in the culture of CC from patients with PCOS (7th day of the experiment), the values of IL-6, IL-10 increased only two-fold compared with 30-fold increase of these cytokines in healthy donors (p 0.01). At the same term, we have observed a threefold decrease in IFN in the CC cultures of PCOS patients (p1-7 0.01; p 0.05) compared with cultured controls, which showed a 20-fold increase (p1-7 0.01), thus determining differences in total cytokine balance and, probably, influencing the pregnancy prognosis. Conclusion: Significant multidirectional changes of cytokine levels in the culture of cumulus cells of the patients with PCOS and in healthy individuals may be regarded as determining factors in formation of blastocyst and preservation of the embryo. A further in vitro research on the production of cytokines and sex steroids by CC is especially important on day 5 to 7, when the oocytes are selected for entry into the in vitro fertilization cycle. The study of morpho-functional properties of little-studied cumulus cells using the cell culture technique will enable us for a deeper study on the mechanisms of disturbed folliculogenesis in PCOS, and, thereby, improve the reproductive prognosis in this disorder.

A multicenter double-blind, placebo-controlled randomized trial to evaluate the safety and efficacy of bovine colostrum in the treatment of severe alcoholic hepatitis (SAH)

BackgroundSevere alcoholic hepatitis (SAH) is associated with high mortality. Numerous studies and meta-analysis have reported that corticosteroids reduce the 28-day mortality in SAH, but not the 6-month mortality. Therefore, newer treatments for SAH need to be studied. A pilot study from our group had recently treated ten patients with SAH with bovine colostrum (BC) [20 g thrice in a day for 8 weeks] and prednisolone. This therapy improved the biological functions and 3-month mortality. However, as more and more data showed the failure of corticosteroids to improve the 3- and 6-month mortality, especially in patients with high mDF and MELD scores, we planned this trial to study the safety and efficacy of BC (without corticosteroids) in the treatment of SAH.MethodThis is a multicenter, parallel, double-blind, randomized (1:1) placebo-controlled trial, which will enroll 174 patients with SAH from 5 academic centers in the India. Patients will receive freeze-dried BC or placebo by random 1:1 allocation for 4 weeks. The primary outcome measure is survival at 3 months. The secondary outcome measures are survival at 1 month, change in mDF and MELD scores, change in endotoxin and cytokines (alpha TNF, IL6, and IL8) levels, number of episodes of sepsis [pneumonia, spontaneous bacterial peritonitis (SBP), cellulitis, urinary tract infection (UTI)] from baseline to 4 weeks.DiscussionThis study will evaluate the safety and efficacy of bovine colostrum in improving the survival of patients with SAH.Trial registrationClinicalTrials.gov NCT02473341. Prospectively registered on June 16, 2015.

New-onset hypertension is not associated with systemic changes in inflammatory cytokine levels

IntroductionRecent studies have suggested that hypertension develop�ment may be associated with an altered immune system. However, there is a paucity of data evaluating the association between blood pressure values and inflammatory markers in patients with new-onset hypertension.Material and methodsWe evaluated 61 subjects, including 24 healthy indi�viduals and 37 newly diagnosed hypertensive patients (aged 45 ±9.6 vs. 43.8 ±11.9 years; SBP_24hours 114 ±7.1 vs. 134.2 ±9.5 mm Hg; DBP_24hours 71.2 ±4.7 vs. 85.8 ±9.3 mm Hg, respectively) without prior antihypertensive treatment. The diagnosis of hypertension was based on 24-hour ambulatory blood pressure monitoring (ABPM). We analysed the association between blood pressure values and levels of individual inflammatory markers (ITAC, GM-CSF, fractalkine, IFN-g, IL-10, MIP-3a, IL-12, IL-13, IL-17A, IL-1b, IL-2, IL-21, IL-23, IL-5, IL-6, IL-7, IL-8, MIP-1a, MIP-1b, TNF-a, and IL-15) sepa�rately, as well as in clusters of inflammatory mediators (factor 1 – proin�flammatory: IL-1β, IL2, IL-6, IL-7, IL-12, IL-6, IL-21, TNF-α, IFN-γ; and factor 2 – anti-inflammatory: IL-13, IL-5).ResultsOur study did not show any differences in concentrations of inflam�matory markers between patients and controls. Plasma levels of inflamma�tory markers were not associated with 24-hour ambulatory blood pressure values in patients with new-onset hypertension.ConclusionsPatients with new-onset hypertension did not differ from healthy subjects regarding the levels of plasma inflammatory markers. Our findings demonstrate the need for larger, more comprehensive studies on this topic to further elucidate the relationship between hypertension and inflammation.

Evaluation of Serum Levels of Cytokines in Acute Apical Abscess: A Longitudinal Observational Study

IntroductionCytokine levels are related to the aethiopathogenia of acute apical abscesses (AAA); however, the specific cytokine profiles in these cases are unclear. This study aimed to investigate the changes in systemic cytokine levels in patients with AAA and trismus onset, postantibiotic treatment, and postroot canal disinfection. MethodsIn total, 46 AAA patients with trismus and 32 control subjects were included. After seven days of antibiotic therapy, root canal disinfection was performed in the AAA patients. The serum levels of cytokines were evaluated at basal, seven, and 14 days after endodontic treatment. Quantification of cytokines from T helper (Th) 1, Th2, Th17, and regulatory T cells profiles was determined using the BioPlex MagPix system, and the obtained data were analyzed using SPSS statistical software (P < .05). ResultsAAA patients showed higher tumor necrosis factor-alpha (TNF-α), interleukin (IL) -6, and IL-10 levels than control subjects, at basal measurement (P < .05); there were similar levels of interferon gamma, IL-1β, IL-4, and IL-17 between groups (P > .05). IL-6 and IL-10 levels decreased after antibiotic treatment (P < .05), which was also associated with clinical improvement in patients with AAA and trismus. Patients with AAA had a positive correlation with higher serum levels of IL-6 and IL-10. In addition, TNF-α levels decreased only after antibiotic and endodontic treatment. ConclusionsIn conclusion, patients with AAA had increased systemic serum levels of TNF-α, IL-6, and IL-10. Moreover, increased levels of IL-6 and IL-10 are associated with acute inflammatory symptoms. However, IL-6 and IL-10 levels decreased after antibiotic treatment, while TNF-α levels decreased after antibiotic and endodontic treatment.

Negative impact on bone homeostasis in postmenopausal women with non-metastatic breast cancer during cytotoxic chemotherapy.

The burden and mechanisms of endocrine therapy-related bone loss are well known, while there are limited data on chemotherapy-induced bone resorption. The study aimed to evaluate the effect of cytotoxic chemotherapy on bone homeostasis among postmenopausal women with non-metastatic breast cancer. Early and locally advanced postmenopausal non-metastatic breast cancer patients aged 45 to 65 planned for three cycles of anthracycline and four cycles of taxane chemotherapy administered along with dexamethasone (cumulative dose-256mg) as an antiemetic from June 2018 to December 2021 were included. Bone mineral density (BMD), bone turnover markers, calciotropic hormones, pro-inflammatory cytokines, oxidative stress, and total antioxidant levels (TAS) were measured. We recruited 109 patients, with early 34 (31.2%) and locally advanced breast cancer 75 (68.8%) with median age 53 (45-65) years. There was a significant decrease in the % BMD at the lumbar spine, neck of the femur, and total hip post-chemotherapy. There was a significant increase in serum C-terminal telopeptide of type I collagen (CTX) and procollagen type I N-terminal propeptide (PINP) levels post-chemotherapy. PINP/CTX ratio significantly decreased post-chemotherapy. Serum 25-OH vitamin D was significantly reduced with a compensatory increase in plasma iPTH levels. The change in CTX, PINP/CTX ratio, 25-OH vitamin D, iPTH, and oxidative stress index was more pronounced during anthracycline as taxane chemotherapy. There were no significant changes in pro-inflammatory cytokine levels. Chemotherapy and dexamethasone as antiemetic resulted in significant bone loss, as evidenced by bone turnover markers. Further studies are required to understand the mechanism of chemotherapy-induced bone loss and the need for bone-strengthening agents during chemotherapy.

Outcomes of Prolonged Intermittent Renal Replacement Therapy Combined with Hemoperfusion among Patients Aged &lt;18 Years Old with Severe Dengue

Context: Fluid overload is inevitable in severe dengue shock syndrome despite intensive fluid management. Aims: To compare the outcomes of prolonged intermittent renal replacement therapy (PIRRT) with or without hemoperfusion (HP) in treating severe dengue among patients ≤18 years old. Settings and Design: Retrospective cohort study in the hemodialysis unit of a tertiary hospital in the Philippines. Methods: Demographic and clinical data, kidney replacement therapy prescription variables, hemodynamic status, and outcomes were analyzed retrospectively for pediatric patients who underwent PIRRT with or without HP for severe dengue. The primary outcome was 28-day all-cause mortality, whereas secondary outcomes were changes in percent fluid overload, vasopressor index (VI), mean arterial pressure, and vasopressor dependency. Statistical Analysis Used: Mann–Whitney U, Chi-square, Fisher exact, and Student’s t test. Results: Among 357 severe dengue patients, 27 underwent PIRRT with (n = 18) or without (n = 9) HP. Baseline demographic and clinical characteristics differed significantly only with respect to the degree of fluid overload, time to therapy initiation, proportion with obesity and overweight, and duration of treatment during the first session. There were no significant differences between the two groups in 28-day mortality (PIRRT, n = 7 of 9, vs. PIRRT + HP, n = 10; P = 0.40) or secondary outcomes except an increased VI in PIRRT group (mean 38.2 vs. 12.4; P = 0.04). Conclusions: PIRRT combined with HP is feasible in hemodynamically unstable children with severe dengue in a resource-limited setting, but the outcomes of the combination are similar to those with PIRRT alone. Larger prospective studies should examine the efficacy of the combination of PIRRT with HP in terms of the dialysis dose and changes in cytokine levels or other inflammatory markers.

Lactobacillus murinus: A key factor in suppression of enterogenous Candida albicans infections in Compound Agrimony enteritis capsules-treated mice.

Compound Agrimony (FuFangXianHeCao, FFXHC) Enteritis Capsules is an ethnomedicine that is derived from Yi Nationality Herbal Medicine for enteritis treatment. We found that FFXHC reduced the mortality outcomes in enterogenic Candida albicans infected mice models and increased the abundance of Lactobacillus murinus in the intestines. Lactobacillus murinus exhibited comparable therapeutic effects to those of FFXHC in enterogenic Candida albicans infected mice. This study provides novel perspectives into the pharmacological mechanisms of FFXHC. We investigated the mechanisms via which FFXHC inhibits C. albicans infections and its effects on L. murinus. Enterogenous C. albicans infection mice models were established and various parameters, including survival rate, weight change, number of colonies, treatment effects on intestinal mucosa, microecology, and immune cytokines evaluated. Susceptibility of C. albicans to L. murinus was evaluated in vitro. Treatment with FFXHC reduced the number of colonies, improved the health status, enhanced the survival rates, increased the abundance of L. murinus, reduced damage to the intestinal mucosa, and elevated occludin as well as claudin-1 levels. Interestingly, TNF-α, IFN-γ, IL-10, IL-22, and IL-17A levels were increased while IL-1β levels were suppressed in the intestinal mucosa without any change in peripheral blood cytokine levels. Moreover, FFXHC promoted L. murinus proliferation. This study also confirmed the incubation-dependent anti-C. albicans effects exerted by the metabolic supernatants of L. murinus. FFXHC effectively alleviated intestinal infections of C. albicans in mice and increased the abundance of L. murinus. Supplementation of L. murinus in food can achieve the effects that are comparable to those of FFXHC. Thus, L. murinus maybe essential in FFXHC-based treatment of intestinal C. albicans infections.

Acetylcorynoline inhibits microglia activation by regulating EGFR/MAPK signaling to promote functional recovery of injured mouse spinal cord

To investigate the effect of acetylcorynoline (Ace) for promoting functional recovery of injured spinal cord in rats and explore the underlying mechanism. Rat models of spinal cord injury (SCI) were treated with intraperitoneal injection of different concentrations of Ace, with the sham-operated rats as the control group. After the treatment, the changes in motor function of the rats and the area of spinal cord injury were assessed with BBB score and HE staining, and the changes in pro-inflammatory cytokine levels and microglial activation were determined using PCR, ELISA and immunofluorescence staining. In a lipopolysaccharide (LPS)-treated BV2 cell model, the effects of different concentrations of Ace or DMSO on microglial activation and inflammatory cytokine production were observed. Network pharmacology analysis was performed to predict the target protein and signaling mechanism that mediated the inhibitory effect of Ace on microglia activation, and AutoDock software was used for molecular docking between Ace and the target protein. A signaling pathway blocker (Osimertinib) was used to verify the signaling mechanism in rat models of SCI and LPS-treated BV2 cell model. In rat models of SCI, Ace treatment significantly increased the BBB score, reduced the area of spinal cord injury, and lowered the number of activated microglia cells and the levels of pro-inflammatory cytokines (P < 0.05). The cell experiments showed that Ace treatment significantly lower the level of cell activation and the production of inflammatory cytokines in LPS-treated BV2 cells (P < 0.05). Network pharmacology analysis suggested that EGFR was the main target of Ace, and they bound to each other via hydrogen bonds as shown by molecular docking. Western blotting confirmed that Ace inhibited the activation of the EGFR/MAPK signaling pathway in injured mouse spinal cord tissue and in LPS-treated BV2 cells, and its inhibitory effect was comparable to that of Osimertinib. In rat models of SCI, treatment with Ace can inhibit microglia-mediated inflammatory response by regulating the EGFR/MAPK pathway, thus promoting tissue repair and motor function recovery.

17th International Conference on Malignant Lymphoma, Palazzo dei Congressi, Lugano, Switzerland, 13 - 17 June, 2023.

Introduction: Anbal-cel is a novel autologous anti-CD19 CAR-T cell therapy which has been knocked-down for PD-1 and TIGIT expression using OVIS platform. It demonstrated superior T-cell functionality compared to other marketed CD19 CAR-T cells at preclinical studies. Phase 1 dose escalation study confirmed anbal-cel is safe and potentially effective. Methods: Patients with relapsed or refractory LBCL were enrolled from six Korean investigator sites to receive anbal-cel at a dose of 2 × 106 CAR-T cells/kg. Primary endpoint is objective response rate (ORR) assessed by Independent Review Committee (IRC). Secondary endpoints are progression-free survival (PFS), duration of response (DOR), pharmacokinetics (PK) and safety profiles. Results: As of 28 February 2023, a total of 41 patients were infused and 34 patients completed at least one tumor response evaluation at day 28. With the median age of 63 (range 37–82), 66% of the patients (27/41) were double expressor. Non-GCB type was 76% (31/41) and 42% (17/41) were primary refractory patients. Median follow-up period was 4.6 months. ORR (CR/PR) assessed by investigators was 85.3% and CR rate at day 28 & month 3 were 73.5% (25/34) and 71.4% (22/31) respectively. IRC assessment result was similar and will be presented at the meeting. Among the 41 patients, grade 3 CRS and ICANS were reported from 6 patients (14.6%) and 2 patients (4.9%) respectively. No grade 4 CRS and ICANS was observed. Prolonged neutropenia was observed in 11 patients (26.8%) and thrombocytopenia was observed in 12 patients (29.3%). Three (7.3%) serious infections (1 bacteria and 2 COVID-19 infections) were reported during primary follow-up period. Responder group (CR) reported higher incidence and severity of CRS compared to non-responder group (Non-CR), but it was insignificant. Incidence rate, severity of ICANS and prolonged cytopenia were comparable between the two groups. Among CAR+ T-cells in the anbal-cel product, median CD4:CD8 ratio was 3.6 and CCR7+ memory phenotype CAR T-cells account for 65.8% of CAR-T cells. Median knock-down rates of PD-1 and TIGIT were 65% and 96% respectively. The responder group showed a significantly increased level of CCR7+ memory phenotype (68.6% vs. 44.9%) and PD-1 knock-down rate (71.8% vs. 59.3%) compared to the non-responder group, which showed correlation with increased levels of AUC0-28days and Cmax. Anbal-cel expansion was not correlated to tumor volume measured by SPD and TMTV. In addition, the responder group had notable changes in cytokine levels of IL-2, IL-10, IFN-γ, granzyme, GM-CSF and peak levels of various cytokines were proportional to severity of CRS. Patients with higher levels of IL-15 and MCP-1 at baseline had tendency to experience ICANS. Conclusions: Anbal-cel demonstrated unique characteristics in product profile which demonstrated potential to correlate with treatment outcomes and warrant to confirm with more data. The research was funded by: Curocell Inc. Keywords: aggressive B-cell non-Hodgkin lymphoma, cellular therapies, immunotherapy No conflicts of interests pertinent to the abstract.

PORTO-001 study: Changes in the oral microbiome and matrix metalloproteinases in patients with oropharyngeal squamous cell carcinoma undergoing definitive chemoradiotherapy.

e18034 Background: Head and neck cancer patients treated with concurrent chemoradiotherapy (CRT) develop oral mucositis, with varying severity. However, knowledge about risk factors, biomarkers, treatment, and prognosis of oral mucositis is still limited. Methods: We designed an exploratory study to examine the changes in the oral microbiome and salivary and plasma levels of matrix metalloproteinases (MMP) in patients with locoregionally advanced oropharyngeal squamous cell carcinoma (OPSCC) undergoing definitive CRT. Patients were followed from screening to week 7 of treatment with CRT, which consisted of 70 Gy/35 fractions of radiation with cisplatin given at 100 mg/m2 q3weeks or weekly 40 mg/m2. Clinical and demographic data were collected at specified timepoints. Samples were collected at screening and on week 4 of treatment with CRT. A tumor swab was collected for microbiome analysis. Blood and saliva samples were collected for measurements of 7 MMPs and 26 other cytokines. The study population was summarized descriptively. The change in the cytokines and the α-diversity of microbiome data (week 4 vs screening) were examined using Wilcoxon signed rank test. Results: A total of 11 patients were included. Nine (82%) were male. 5 patients (45%) were stage III, and there were 2 patients (18%) for each of stages I, II, and IV. A history of smoking was present in 5 (45.4%) patients. p16 was positive in 9 (81.8%) patients. There was a median weight loss of 7.0% (5.1 kg) between screening and week 7 visits. Almost half (45%) of patients developed grade 2 oral mucositis. All patients developed radiation dermatitis, with most cases ranging between grades 2 and 3. There were no statistically significant changes in the salivary levels of cytokines. However, 1 patient developed clinical oral mucositis before all the other patients, at week 1, and this was associated with an upsurge in the salivary levels of IFN-γ, IL-6, TNF-α, MMP-1, MMP-2 and MMP-7. For all patients, there were statistically significant changes (median (95% confidence interval)) in the blood plasma levels of IP-10 (-0.1 fluorescence intensity (FI) (-0.3, -0.04), p = 0.01), MIP-1ß (0.2 FI (0.07, 0.5), p = 0.02), SDF-1α (1.4 pg/ml (0.07, 3.7), p = 0.02), MMP-1 (-0.4 pg/ml (-0.5, -0.09), p = 0.01), MMP-3 (0.4 pg/ml (0.03, 0.5), p = 0.02), and MMP-8 (-0.2 FI (-0.3, -0.02), p = 0.04). There was a 29.1% ((-3.0%, -32.8%), p = 0.005) reduction in α-diversity of microbiome at week 4 from screening. Conclusions: In this small series of patients diagnosed with OPSCC, there was a change in the plasma levels of IP-10, MIP-1ß, SDF-1α, MMP-1, MMP-3, and MMP-8, as well as a reduction in the α-diversity of the oral microbiome. Larger studies are needed to confirm these results. If confirmed, these changes might imply a potential role as predictive or pharmacodynamic biomarkers for oral mucositis.

Assessment of short-chain fatty acid (SCFA) and circulating cytokine trends in a clinical trial of nivolumab/ipilimumab and CBM588 in metastatic renal cell carcinoma (mRCC).

4556 Background: Recent evidence suggests that blood and stool SCFA levels (including acetic acid and butyric acid) are associated with clinical outcomes with immune checkpoint blockade in melanoma (Nomura et al. JAMA Network Open 2020). Herein, we report trends in plasma SCFA and cytokine levels in a randomized phase I clinical trial of CBM588, a live bacterial product containing a butyric acid-producing bacterium, C. butyricum, in patients with mRCC treated with immune checkpoint blockade. Methods: Treatment naïve patients with clear cell and/or sarcomatoid mRCC and intermediate- or high-risk IMDC disease were randomized to nivolumab/ipilimumab or nivolumab/ipilimumab and CBM588 in a 1:2 fashion. Blood samples were collected at baseline and on treatment (week 12 +/- 5 weeks). Quantitative assessment of plasma SCFAs and cytokines was performed by liquid chromatography-mass spectrometry and a Human Cytokine 30-plex protein assay, respectively. Changes in SCFAs and cytokine levels were assessed using paired sample T-tests. Results: A total of 29 patients were randomized to the nivolumab/ipilimumab (n=19) and nivolumab/ipilimumab with CBM588 (n=10) arms. Objective response rates (ORR) with nivolumab/ipilimumab with and without CBM588 were 58% and 20%, respectively. No statistically significant change in SCFA levels was observed between baseline and on-treatment samples in either arm. Notably, however, a numerical increase in levels of acetic acid (414.3 vs 814.0 nmol/g) and isobutyric acid (7.9 vs 9.0 nmol/g) was seen in the nivolumab/ipilimumab and CBM588 arm. In contrast, both factors decreased in the nivolumab/ipilimumab arm (563.1 vs 401.1 nmol/g and 10.0 vs 8.8 nmol/g, respectively). In patients with an uptrend in isobutyric acid levels with nivolumab/ipilimumab and CBM588, ORR was 64% compared to 50% in those without an uptrend in such levels). Similarly, patients with increases in butyric acid with nivolumab/ipilimumab and CBM588 had an ORR of 66%, compared to 43% among those without an increase in butyric acid levels. Correlations between acetic acid and chemokines, including CCL2 and CCL4, were observed with correlation coefficients of 0.83 and 0.82, respectively (p&lt;0.001 for both). Conclusions: A substantial increase in isobutyrate levels in CBM588 responders is indicative of altered amino acid metabolism (specifically, valine and leucine) by modified gut flora in these patients. These observations, combined with alterations in serum cytokines, provide biologic rationale for the clinical activity seen with CBM588 in combination with nivolumab/ipilimumab in this trial. Clinical trial information: NCT03829111 .

Pembrolizumab in combination with epigenetic therapy is safe and active in heavily treated patients with peripheral T‐cell lymphoma and cutaneous T‐cell lymphoma

Introduction: Peripheral T-cell lymphomas are uniquely sensitive to epigenetic modifiers. We have shown that decitabine and 5-azacytidine (Marchi et al.; Br. J. Haematol. 171; 2015) induce the expression of cancer testis antigens in pre-clinical models of PTCL, and that pralatrexate modulates genes involved in cytokine production, viral response and apoptosis. These data suggest a role for the incorporation of the immune-checkpoint inhibitor, pembrolizumab, to epigenetic agents. Herein we report on the clinical activity of this approach in patients with relapsed or refractory (R/R) PTCL and CTCL. Methods: This is a phase 1b study of pembrolizumab combined with pralatrexate (Arm A), pralatrexate + decitabine (Arm B), or decitabine (Arm C) in patients with R/R PTCL and CTCL. A 3+3 dose-escalation/de-escalation is applied in Arms A and C while a DLT-adapted partial order continual reassessment method for dose-finding with combinations of agents is applied in Arm B. Changes in serum cytokine levels were assessed during cycle 1 using a Luminex-based immunoassay. T-cell subpopulation profiling in peripheral blood was performed using a multi-color flow cytometry assay (Cytek® Aurora). Results: Patient characteristics, toxicity data, and response analysis are shown in table 1. Serum levels of three cytokines, TNFα, MIP-3α and IL-10, measured at day 1 pre-treatment were significantly elevated (p = 0.0001, 0.011 and 0.0017, respectively) in trial patients compared to healthy controls. Patients who responded to treatment demonstrated a more consistent decline in levels of TNFα, MIP-3α, and IL-10 during cycle 1, whereas non-responders showed stable to increased levels of these cytokines. T-cell subpopulation analysis (n = 6) showed mean CD4:CD8 ratio of 0.5 in trial patients prior to treatment compared to 1.5 for healthy age and sex-matched controls (p = 0.016). No significant change in the proportion of PD1+CD8+ central memory cells was seen in trial patients between D0 and D28 of treatment (p = 0.12), and between D0 in comparison to healthy controls (p = 0.11). The proportion of PD1+CD8+ effector T cells was lower in trial patients relative to healthy controls both before and after treatment (p = 0.04 and 0.001, respectively). There was no difference in the proportion of PD1+CD8+ effector T cells among trial patients from D0 to D28 of treatment (p = 0.19). Patients who responded to treatment (n = 3), had a lower proportion of PD1+ CD8+ effector cells at the start of treatment relative to non-responders (p = 0.002). Conclusions: These clinical data suggest that the integration of pembrolizumab on an epigenetic backbone is safe and demonstrates encouraging responses in heavily treated patients with PTCL and CTCL. These correlative data suggest that specific cytokines (TNFα, MIP-3α, and IL-10) and changes in circulating T-cell subpopulations may have predictive value as biomarkers of disease response. The research was funded by: Merck and Co. Ongoing Trial Keywords: Aggressive T-cell non-Hodgkin lymphoma, Combination Therapies, Ongoing Trials Conflicts of interests pertinent to the abstract. N. N. Benani Consultant or advisory role: Vividion Therapeutics, Kymera, Secura Bio, Affirmed GmbH, Astellas Pharma, Acrotech Biopharma LLC M. E. Williams Consultant or advisory role: Astra Zeneca, Gilead, Janssen, Kite Pharma, Kymera, TG Therapeutics Honoraria: International Oncology Network, Research to Practice Research funding: Janssen, Kymera, Pharmacyclics J. S. Manavalan Research funding: Astex O. A. O'Connor Employment or leadership position: TG Therapeutics Stock ownership: TG Therapeutics E. Marchi Employment or leadership position: Myeloid Therapeutics Consultant or advisory role: Myeloid Therapeutics Honoraria: Kyowa Kirin Research funding: Merck and Co., Celgene/BMS, Astex, NomoCan Pharmaceuticals Other remuneration: University of Virginia (Patents & Royalties: 3062/170 PROV), Daiichi Sankyo (advisory board)