Reduction Of CD4 Research Articles

Abstract 1856: Pemetrexed-cisplatin-bevacizumab combo in lung cancer models: impact on tumor immunity

Abstract Dual chemotherapy (i.e., Pemetrexed and Cisplatin, CT) plus anti-VEGF Bevacizumab could be associated with anti-PD1 such as Pembrolizumab in NSCLC patients with low PDL1 levels. To support this new combination, to what extent those drugs could reshape tumor immunity remains to be evaluated, prior to establishing optimal scheduling or sequencing with Pembrolizumab. Both Pemetrexed and Cisplatin are expected to trigger immunogenic cell death, whereas Bevacizumab could help T lymphocytes to extravasate and better infiltrate tumor micro-environment. Here, we studied the impact of Cisplatin and Pemetrexed at two dose levels (i.e., low dose and Maximal Tolerated Dose, MTD) and fixed flat-dose Bevacizumab, alone or in combination (i.e., Cisplatin, Pemetrexed, Cisplatin + Pemetrexed, with our without Beva), on the immune repertoire of BalbC mice ectopically grafted with KLN-205 murine lung tumors. CD45, CD4, CD8, FoxP3-CD4 (Tregs), and MDSCs were monitored by FACS analysis in blood, spleen, and tumors on D2, D7, D14, and D21 after treatment. Results were normalized by CD45 counts. All treatments led to an increase in CD4+ T cells in blood as compared with control (i.e., untreated) animals. Little increase was observed in tumors, regardless of the treatments but mice treated with the Beva+CT combo (both low dose and MTD CT) showed an increase in CD4+ in the spleen on D14. Regarding CD8+ T cells, all treatments led to an increase from D2 to D7. Beva + CT low dose led to a continuous increase in CD8+ cells, whereas in Beva+CT MTD mice, after an initial increase, a reduction in CD8+ was observed after D7 eventually. In the spleen, CD8+ proved to be stable apart from the Beva + CT low dose group which showed an increase up to D14. In tumors, an increase in CD8+ cells was observed in all the groups. No change in MDSCs levels was observed, regardless of the drugs, the dosing and the organ. Conversely, Tregs decreased in the blood in the Beva + CT low-dose group on D14, and decreased in spleen and tumors as compared with Control mice, regardless of the drugs and the dosing. Overall, our data suggest that the triple combination between Beva, Pemetrexed, and Cisplatin has a stronger impact on immune cells than each drug used as a single agent. MDSCs were the only cells to be unaffected by the treatments. Dosing could be relevant as well since the reduction in CD8+ T cells observed with MTD CT could come from drug-related lymphopenia. The Beva followed by low dose Pemetrexed + Cisplatin group is thus the more likely to harness tumor immunity, with peaks in CD4+ and CD8+ T cells and drop in Tregs observed on D14. This pilot study suggests therefore that sequencing Beva + CT low dose followed by a two-week lag time prior to administrating anti-PD1 could lead to synergism in lung cancer models. Citation Format: Guillaume Sicard, Sarah Giacometti, Fabrice Barlesi, Raphaelle Fanciullino, Joseph Ciccolini. Pemetrexed-cisplatin-bevacizumab combo in lung cancer models: impact on tumor immunity [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 1856.

Abstract 906: Development of a repeated challenge potency assay for evaluation of immune cell-mediated cytotoxicity in vitro

Abstract The development of immunotherapies relies on the use of in vitro potency assays - which are key for understanding complex interactions between immune (effector) cells and cancer (target) cells - to evaluate the function, specificity, and sensitivity of a product. A variety of in vitro assays are used to characterize the proliferation, cytokine release, and cell-mediated cytotoxicity of engineered immune cells, such as chimeric antigen receptor (CAR) T cells, each representing an important aspect driving clinical efficacy of the cellular product. Importantly, adoptive cell therapies must exhibit potent tumor killing over a prolonged period in vivo. Here, we describe an in vitro potency assay that quantifies immune cell-mediated cytotoxicity in response to repeated challenges with target cells. The repeated challenge assay was performed with two separate protocols, each providing a unique assessment of the effector cell function. In the first method, SKOV3 target cells (5k cells/well) were seeded into a multiwell microplate with embedded electrodes in the substrate. Electrical impedance measurements from the electrodes in each well detected the attachment and proliferation of the target cells. HER-2 specific CAR T effector cells were added 24 hours after the target cells and co-cultured with the target cells at a 1:1 effector to target ratio. Cytolysis of the target cells was calculated by comparing treated wells to no treatment control wells. Additional SKOV3 target cells (5k per well) were added every 24 hours for the repeated challenge conditions, with total tumor cell killing computed from the area under the impedance curve. The repeated challenge condition reached complete killing after the 6th tumor cell challenge (144 hours after first effector addition), whereas the single challenge condition reached complete killing within 96 hours of effector addition. Flow cytometry analysis revealed that the effector cells proliferated significantly more for the repeated challenge condition (4.7 fold increase) than the single challenge condition (1.5 fold increase). In the second method, the CAR T cells were removed after each challenge, counted, and added to a fresh plate of SKOV3 target cells at a 1:1 effector to target cell ratio. Here, the E:T ratio was kept constant for each challenge, allowing a direct comparison of the killing kinetics (KT50) for each subsequent challenge. The KT50 decreased from the first challenge (19.9 hr) to the fourth challenge (83.5 hr). Flow cytometry analysis revealed a reduction in CD8+ cells for the repeated challenge condition, indicating a change in the make-up of the immune cell population. The paradigms described above facilitate a label-free, in vitro workflow to assess potency of immune effector cells across repeated challenges, with the opportunity to incorporate additional assays, like flow cytometry or cytokine release, after each successive challenge. Citation Format: Denise Sullivan, Stacie Chvatal, Daniel Millard. Development of a repeated challenge potency assay for evaluation of immune cell-mediated cytotoxicity in vitro [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 906.

LBA01-08 EFFICACY OF TARGETED MAST CELL INHIBITION THERAPY IN CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME

You have accessJournal of UrologyCME1 Apr 2023LBA01-08 EFFICACY OF TARGETED MAST CELL INHIBITION THERAPY IN CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME Praveen Thumbikat, Goutham Pattabiraman, Catherine Osborn, Stephen Murphy, and Anthony Schaeffer Praveen ThumbikatPraveen Thumbikat More articles by this author , Goutham PattabiramanGoutham Pattabiraman More articles by this author , Catherine OsbornCatherine Osborn More articles by this author , Stephen MurphyStephen Murphy More articles by this author , and Anthony SchaefferAnthony Schaeffer More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003360.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a component of urologic chronic pelvic pain syndrome (UCPPS) and affects men, leading to chronic pain in the pelvic region or genitalia that is often accompanied by increased urinary frequency and urgency as well as sexual dysfunction. CPPS has an estimated prevalence rate of 6-12% of the male population with most diagnoses between the ages of 35-45. We have previously demonstrated that mast cells can be critical mediators of both chronic pelvic pain and lower urinary tract symptoms (LUTS). In this pilot clinical study, we therefore sought to identify subtypes of CP/CPPS characterized by mast cell dysfunction and evaluate the therapeutic efficacy of functional mast cell targeting on mast cell tryptase levels as well as on symptoms of pain and voiding dysfunction. METHODS: Men with CP/CPPS were recruited and evaluated in an open label, interventional uncontrolled trial after therapy with cromolyn sodium and cetirizine hydrochloride. The primary endpoint was a change in mast cell tryptase concentrations after treatment while secondary endpoints were changes in the NIH-CPSI and AUA-SI. Isolated cells from post-prostatic massage urine were evaluated for immune changes using mRNA expression analysis. RESULTS: 31 patients with a diagnoses of Category III CP/CPPS were consented, 25 patients qualified and 20 completed the study after meeting a prespecified threshold for active tryptase in expressed prostatic secretions. After treatment with cromolyn sodium and cetirizine dihydrochloride for 3-weeks, active tryptase concentrations were significantly reduced from 49.03±14.05 ug/ml to 25.49±5.48 ug/ml (p<0.05). The NIH-CPSI total score was reduced with a mean difference of 5.2±1 along with reduction in the pain, urinary and quality of life sub scores (p<0.001). A reduction in the AUA-SI was observed following treatment (p<0.05). NanoString mRNA analysis of isolated cells revealed downregulation of immune related pathways including Th1 and Th17 T cell differentiation and TLR signaling. Marked reduction in CD45+ cells and specifically macrophages and neutrophil abundance was observed. CONCLUSIONS: This pilot study presents a method to identify CP/CPPS patients with mast cell dysfunction and target them for therapeutic intervention using a novel, safe combination therapeutic approach. The resulting subgroup of CP/CPPS patients were shown to have a therapeutic response as measured primarily by a reduction in mast cell tryptase, a potential surrogate biomarker and reduction in CP/CPPS symptoms. These pilot studies will form the basis for future randomized placebo-controlled studies to evaluate these agents as CP/CPPS therapies. Source of Funding: This work was supported by the National Institute of Diabetes and Digestive and Kidney (NIDDK) grant R01DK083609 to Praveen Thumbikat. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of this abstract © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1179 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Praveen Thumbikat More articles by this author Goutham Pattabiraman More articles by this author Catherine Osborn More articles by this author Stephen Murphy More articles by this author Anthony Schaeffer More articles by this author Expand All Advertisement PDF downloadLoading ...

Impact of Covid 19 pandemic on hematopoietic stem cell transplantation activities: Report from a single center

The current COVID-19 pandemic has placed unprecedented stress on the healthcare system, including HSCT. Several international organizations have created guidelines for managing different aspects of HSCT in the context of the pandemic. Comparing 2019 and 2020, our transplant center performed the same number of transplants. In both periods, transplants were mainly for patients with acute leukemia; thus, the urgency criteria was respected in light of pandemic restraints. Transplants by sibling donors and cord blood units remained the same, while transplants by unrelated donors were increased, in particular from European registries, and transplants by haploidentical donors were decreased. This change was made in light of the necessity of cryopreserving all apheresis products. We decided against cryopreserving bone marrow products due to the greater risk of drastic reduction in CD34 + cell count during the process. For urgent cases with only a haploidentical donor available, we opted for the use of PBSC following stimulation with G-CSF. GvHD prophylaxis was performed with PTCY on days + 3 + 5, cyclosporine with tapering from day + 100, and mycophenolic acid until day + 90 post-HSCT. Post-transplant outcomes such as graft failure, sepsis, and GVHD were not affected by the changes implemented. As a result of logistic difficulties, we halted our Car-T program from the start of the lockdown in March 2020 until September 2020. In accord with international guidelines, we were able to continue our HSCT program in the order to ensure a lifesaving treatment for patients with hematologic diseases for whom this procedure cannot be postponed.

Salidroside alleviates cognitive impairment by inhibiting ferroptosis via activation of the Nrf2/GPX4 axis in SAMP8 mice

BackgroundAlzheimer's disease (AD) is a neurogenerative disease and remains no effective method for stopping its progress. Ferroptosis and adaptive immunity have been proven to contribute to AD pathogenesis. Salidroside exhibits neuroprotective and immunomodulatory effects. However, the underlying mechanisms linking salidroside, ferroptosis, and adaptive immunity in AD remain uncertain. PurposeThe objective of this study is to explore the neuroprotective effects and the potential molecular mechanisms of salidroside against neuronal ferroptosis and CD8+ T cell infiltration in senescence-accelerated mouse prone 8 (SAMP8) mice. Study design and methodsSAMP8 mice were employed as an AD model and were treated with salidroside for 12 weeks. Behavioral tests, immunohistochemistry, HE and Nissl staining, immunofluorescence, transmission electron microscopy, quantitative proteomics, bioinformatic analysis, flow cytometry, iron staining, western blotting, and molecular docking were performed. ResultsTreatment with salidroside dose-dependently attenuated cognitive impairment, reduced the accumulation of Aβ plaques and restored neuronal damage. Salidroside also suppressed the infiltration of CD8+T cells, oxidative stress, and inflammatory cytokines, and improved mitochondrial metabolism, iron metabolism, lipid metabolism, and redox in the SAMP8 mice brain. The administration of salidroside decreased iron deposition, reduced TFR1, and ACSL4 protein expression, upregulated SLC7A11, and GPX4 protein expression, and promoted the Nrf2/GPX4 axis activation. ConclusionIn conclusion, neuronal ferroptosis and CD8+T cells are involved in the process of cognitive impairment in SAMP8 mice. Salidroside alleviates cognitive impairment and inhibits neuronal ferroptosis. The underlying mechanisms may involve the Nrf2/GPX4 axis activation and reduction in CD8+T cells infiltration. This study provides some evidence for the roles of salidroside in adaptive immunity and neuronal ferroptosis in SAMP8 mice.

T cell maturation is significantly affected by SARS-CoV-2 infection.

Coronavirus disease 2019 (COVID-19) is a respiratory tract infection caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). An adequate T cell response is essential not only for fighting disease but also for the creation of immune memory. Thus, the present study aims to evaluate the T cells of patients with moderate, severe and critical COVID-19 not only at the time of illness but also 2 months after diagnosis to observe whether changes in this compartment persist. In this study, 166 COVID-19 patients were stratified into moderate/severe and critical disease categories. The maturation and activation of T cells were evaluated through flow cytometry. In addition, Treg cells were analysed. Until 15 days after diagnosis, patients presented a reduction in absolute and relative T lymphocyte counts. After 2 months, in moderate/severe patients, the counts returned to a similar level as that of the control group. In convalescent patients who had a critical illness, absolute T lymphocyte values increased considerably. Patients with active disease did not show differentiation of T cells. Nonetheless, after 2 months, patients with critical COVID-19 showed a significant increase in CD4+ EMRA (CD45RA+ effector memory) T lymphocytes. Furthermore, COVID-19 patients showed delayed T cell activation and reduced CD8+ suppressor T cells even 2 months after diagnosis. A reduction in CD4+ Treg cells was also observed, and their numbers returned to a similar level as that of healthy controls in convalescent patients. The results demonstrate that COVID-19 patients have a delayed activation and differentiation of T cells. In addition, these patients have a great reduction of T cells with a suppressor phenotype.

Accentuating the detection of immunological signatures as a diagnostic tool for monkeypox virus.

Dear Editor, The WHO declared the recent monkeypox surge as a Public Health Emergency of International Concern on 23 July 2022. Rapid measures are being taken in terms of diagnosis, immunization, treatment and management of the disease to prevent another catastrophe. Although monkeypox disease is not proven to be fatal for majority of the people, early diagnosis and appropriate therapeutic regiment has to be followed to curb the transmission of virus. Currently, the predominant diagnostic test performed is the Centers for Disease Control and Prevention (CDC)’s Nonvariola-Orthopox Virus PCR test using samples of skin lesions1. Such invasive methods pose the risk of infection, contamination and reluctant behaviour from patients and healthcare staff while obtaining the sample. This raises the need for a simple, feasible yet accurate mode of detection such as blood or other body fluid investigations. But there is lack of evidence on the use of blood, saliva or other body secretions for detection and confirmation of virus. This article explores the available literature on the detection of immunological signatures in blood as a probable testing method for diagnosing monkeypox virus. The altered kinetics in antiviral antibody response is evident among the confirmed cases of monkeypox disease indicating a significant association between onset of rashes and pattern of rise of immunoglobulins (Igs) in patients’ sera. The viral culture results and PCR finding were found to be were consistent with the seroconversion of Igs. IgM enzyme-linked immunosorbent assay and IgG enzyme-linked immunosorbent assay were thus efficient as diagnostic indicators and phase-of-illness indicators of monkeypox infection2. In addition to this, comparing the serum signatures between monkeypox infected monkey and human cell lines, a characteristic elevation of cluster of differentiation (CD) 40, plasmin and histamine was seen in monkey cell lines, while interferons macrophages and neutrophil-related signalling pathways were predominant in human cell lines3. However, these findings have not been studied in a larger cohort and hence require more intense clinical research to establish these biological constituents as immunological signatures of monkeypox disease. Monkeypox disease has also been strongly associated with early T-cell response and increased inflammatory mediators. An early elevation of CD8+ T cells and reduction in CD4+ T cells is seen in patients with monkeypox virus. This is combined with high expression of CD38 activation marker. This elevated T-cell pattern was found to be normalized within 12–20 days from the onset of symptoms. Inflammatory markers such as interleukins, particularly IL-6, IL-8, IL-16 and tumour necrosis factor are also found to be elevated in these patients. A specific characteristic finding was the detection of Th-1 biased pox-specific T cells in the serum. This antigen specific finding was almost predominant in all of the patients diagnosed with monkeypox virus4,5. Although these studies have provided information about the use of immune markers in detection of monkeypox, the laboratory method of using these immunological signatures for diagnosing monkeypox and disease progression is still at its nascent stage. The detection of immune markers in patient’s serum to identify monkeypox disease will improve the quality and efficacy of testing when compared with conventional skin lesion sampling. Thus, we emphasize on conducting more clinical trials and evidence-based studies to make such immune signatures reliable and accurate for disease detection. Ethical approval Not applicable. Sources of funding None. Authors’ contribution V.P.V.: conceptualization, data curation, writing – original draft preparation. L.P., S.P., J.N.B., U.M.: data curation, writing – original draft preparation. K.M.S.: conceptualization, study design, data curation, writing – original draft preparation, reviewing and editing, visualization and supervision. Conflicts of interest disclosure The authors declare that they have no financial conflict of interest with regard to the content of this report. Research registration unique identifying number (UIN) Not applicable. Guarantor Krishna Mohan Surapaneni.

Long noncoding RNA HOXA-AS2 ameliorates chronic intermittent hypoxia-induced lung inflammation by regulating miR-17-5p/tipe2 axis.

The purpose is to confirm whether long noncoding RNA HOXA-AS2 relieves chronic intermittent hypoxia (CIH)-induced lung inflammation. Male Sprague Dawley rats were used to establisha CIH rat model. Hematoxylin and Eosin staining was used on the lung tissue injury to determine the successful construction of CIH animal model. Arterial partial pressure of oxygen (PaO2) and carbon dioxide (PaCO2) were measured. HOXA-AS2 was overexpressed to evaluate its role in the progression and development of CIH. T cell differentiation and cytokine production were determined using flow cytometry. Cell apoptosis was determined using terminal deoxynucleotidyl transferase dUTP nick end labelling assay kit. The target of HOXA-AS2 and miR-17-5p was predicted by the Encyclopedia of RNA Interactomes (ENCORI) and confirmed using luciferase assay. HOXA-AS2 was downregulated in CIH rat models. Lung tissue injury was observed in CIH rats, and the injury was attenuated by the overexpression of HOXA-AS2. PaO2 was reduced and PaCO2 was induced in CIH rats, which was reversed by the overexpression of HOXA-AS2. The overexpression of HOXA-AS2 inhibited CIH-induced cell apoptosis. It also reversed alterations in the levels of interferon gamma (IFNγ), interleukin (IL)-2, IL-6, IL-1β, tumor necrosis factor alpha (TNF-α), and transforming growth factor beta1 (TGF-β1) in rats caused by CIH. The overexpression of HOXA-AS2 prevented the induction in CD4+ IFN-γ+ T cells and reduction in CD4+TGF-β1+ T cells. The overexpression of HOXA-AS2 upregulated tumor necrosis factor-alpha-induced protein 8-like 2 (tipe2) key regulator through directly targeting miR-17-5p. Further experiments proved that tipe2 was the direct target of miR-17-5p. This study manifested that HOXA-AS2 acted as an anti-inflammatory regulator and protected lung tissue injury from CIH in the rat model; this was mediated by upregulation of tipe2 through directly targeting miR-17-5p. HOXA-AS2 upregulated the expression of tipe2, providing new understanding and therapeutic target for CIH.

Targeting the Inducible T-cell Costimulator (ICOS) in Patients with Relapsed/Refractory T-follicular Helper Phenotype Peripheral T-cell and Angioimmunoblastic T-cell Lymphoma.

Proliferation of T-follicular helper (TFH) CD4+ T cells is a postulated pathogenic mechanism for T-cell non-Hodgkin lymphomas (T-NHL). The inducible T-cell costimulator (ICOS) is highly expressed by TFH, representing a potential target. MEDI-570 is a monoclonal antibody against ICOS, which eliminates ICOS+ cells in preclinical models. We report the safety, pharmacokinetics (PK), pharmacodynamics (PD), and clinical activity of MEDI-570 in T-NHL. NCI-9930 is a phase I, first-in-human study of MEDI-570 in relapsed/refractory malignant T-NHL known to express ICOS. MEDI-570 was administered intravenously every 3 weeks for up to 12 cycles. Primary endpoints were safety, dose-limiting toxicities (DLT), and recommended phase II dose (RP2D). Secondary and exploratory endpoints included efficacy parameters and various correlative studies. This study is supported by the National Cancer Institute (NCT02520791). Twenty-three patients were enrolled and received MEDI-570 at five dose levels (0.01-3 mg/kg). Sixteen (70%) had angioimmunoblastic T-cell lymphoma (AITL); median age was 67 years (29-86) and the median prior lines of therapies was 3 (1-16). Most common grade 3 or 4 adverse events were decreased CD4+ T cells (57%), lymphopenia (22%), anemia (13%), and infusion-related reactions (9%). No DLTs were observed. The RP2D was determined at 3 mg/kg. Analysis of T-cell subsets showed reductions in CD4+ICOS+ T cells reflecting its effects on TFH cells. The response rate in AITL was 44%. MEDI-570 was well tolerated and showed promising clinical activity in refractory AITL. MEDI-570 resulted in sustained reduction of ICOS+ T lymphocytes.

Effects of infliximab against carbon tetrachloride-induced spleen toxicity in rats.

Carbon tetrachloride (CCl4) is a non-polar molecule used in industry in grain curing, insect-killing and especially in the production of chlorofluorocarbons. It is estimated that an average of 70,000 industry workers in Europe are exposed to this toxic compound. Twenty-four male Sprague-Dawley rats were randomly allocated into four groups: control group (saline only, Group I), infliximab (INF) group (Group II), CCl4 group (Group III) and CCl4+INF group (Group IV). While there was an increase in the numerical density of CD3, CD68, and CD200R positive T lymphocytes and macrophages in the CCl4 administration group (p=0.000), this was not the case in the CCl4+INF administration group (p=0.000). TNF-α inhibitors have a protective effect against CCl4-induced spleen toxicity/inflammation as seen by the reduction in CD3, CD68, CD200R positive T lymphocytes and macrophages.

Hemodialysis-Associated Immune Dysregulation in SARS-CoV-2-Infected End-Stage Renal Disease Patients.

Patients on hemodialysis show dysregulated immunity, basal hyperinflammation and a marked vulnerability to COVID-19. We evaluated the immune profile in COVID-19 hemodialysis patients and the changes associated with clinical deterioration after the hemodialysis session. Recruited patients included eight hemodialysis subjects with active, PCR-confirmed SARS-CoV-2 infection, five uninfected hemodialysis patients and five healthy controls. In SARS-CoV-2-infected hemodialysis patients TNF-α, IL-6 and IL-8 were particularly increased. Lymphopenia was mostly due to reduction in CD4+ T, B and central memory CD8+ T cells. There was a predominance of classical and intermediate monocytes with reduced HLA-DR expression and enhanced production of pro-inflammatory molecules. Immune parameters were analysed pre- and post-hemodialysis in three patients with COVID-19 symptoms worsening after the hemodialysis session. There was a higher than 2.5-fold increase in GM-CSF, IFN-γ, IL-1β, IL-2, IL-6, IL-17A and IL-21 in serum, and augmentation of monocytes-derived TNF-α, IL-1β and IL-8 and CXCL10 (p &lt; 0.05). In conclusion, COVID-19 in hemodialysis patients associates with alteration of lymphocyte subsets, increasing of pro-inflammatory cytokines and monocyte activation. The observed worsening during the hemodialysis session in some patients was accompanied by augmentation of particular inflammatory cytokines, which might suggest biomarkers and therapeutic targets to prevent or mitigate the hemodialysis-related deterioration during SARS-CoV-2 infection.

Involucrin Modulates Vitamin D Receptor Activity in the Epidermis

Terminally differentiated keratinocytes are critical for epidermal function and are surrounded by involucrin (IVL). Increased IVL expression is associated with a near-selective sweep in European populations compared with those in Africa. This positive selection for increased IVL in the epidermis identifies human adaptation outside of Africa. The functional significance is unclear. We hypothesize that IVL modulates the environmentally sensitive vitamin D receptor (VDR) in the epidermis. We investigated VDR activity in Ivl‒/‒ and wild-type mice using vitamin D agonist (MC903) treatment and comprehensively determined the inflammatory response using single-cell RNA sequencing and associated skin microbiome changes using 16S bacterial phylotyping. VDR activity and target gene expression were reduced in Ivl‒/‒ mouse skin, with decreased MC903-mediated skin inflammation and significant reductions in CD4+ T cells, basophils, macrophages, monocytes, and type II basal keratinocytes and an increase in suprabasal keratinocytes. Coinciding with the dampened MC903-mediated inflammation, the skin microbiota of Ivl‒/‒ mice was more stable than that of the wild-type mice, which exhibited an MC903-responsive increase in Bacteroidetes and a decrease in Firmicutes. Together, our studies in Ivl‒/‒ mice identify a functional role for IVL to positively impact VDR activity and suggest an emerging IVL/VDR paradigm for adaptation in the human epidermis.

Immune system cells modulation in patients with reproductive issues: A systematic review approach.

The aim of this study was to carry out a systematic literature review to investigate the main immune cells responsible for implantation failures. We selected papers from PubMed, Embase and Virtual Health Library databases. Eligible articles included publications between January 1, 2010 and April 24, 2022. Inclusion criteria were: observational and case-control studies; and the exclusion criteria were: review papers, letters to the editor, abstracts, animal studies and case reports. We extracted the following information: day of collection, number of patients, control group, age of patients, type of sample used, immune cells and cytokines. As main findings in our mapping, we found that in peripheral blood, CD3+, CD4+, CD8+, CD16+, CD56+, CD57+, CD69+, CD154+, CD158a+, NKp46 cells were increased and the CD4+, CD45+, Foxp3 and NKp46 markers were reduced. From the endometrial biopsies, there was an increase in CD3+, CD4+, CD5+, CD8+, CD16+, CD25+, CD45+, CD56+, CD57+, CD68+, CD127+ and a reduction in CD45+, CD56+, NKp46 and FoxP3 cells. Cytokines found increased in peripheral blood included IL-6, IL-10, IL-17, INF-γ, TGF-ß, TNF-α; while IL-4, IL-6, IL-10, IL-35, FoxP3, TGF-ß, SOCS3 were reduced. As for the biopsies, there was an increase in IL-2, IL-6, IL-17, IL-22, IL-23, INF-A1, INF-B1, INF-γ, TNF-R and a reduction in IL-6, IL-10, INF-γ, TGFß, TNF-α. We concluded that immune cells can be modulated during pregnancy failure, but further studies are needed to elucidate the modulating effect of the immune system on the endometrium of these patients.

Extract of Laurus nobilis attenuates inflammation and epithelial ulcerations in an experimental model of inflammatory bowel disease

Inflammatory bowel diseases (IBD), including Crohn's disease and ulcerative colitis, are classified as chronic inflammatory disorders and typically require anti-inflammatory drug therapies, such as glucocorticoid regimens, non-steroidal anti-inflammatory drugs, and biologics, aimed at reducing inflammation in the bowel wall. However, each of these therapies is accompanied by a list of possible serious side effects. Because of this, there remains an urgent need to identify new pharmacologic options to reduce or prevent the pro-inflammatory events of IBD while minimizing adverse side effects, and to make available more cost-effective treatment modalities. We have previously identified several herbal extracts that demonstrate potent bio-inhibitory activity of the innate immune response. In particular, Laurus nobilis (LN), or more commonly called bay laurel, demonstrated significant anti-inflammatory function by inhibiting nuclear factor-κB activation. Based upon our original in vitro findings, we have now examined the effects of this herbal extract on a murine dextran sodium sulfate (DSS) model of IBD. Hematoxylin and eosin-stained paraffin sections prepared from DSS treated animals show clear epithelial damage, including ulcerations, extensive neutrophil infiltration into the mucosal layer, and granuloma formation. Tissue from DSS treated animals that also received LN extract showed improved tissue morphology more closely resembling that from control animals. In addition, DSS treated mice with co-administration of LN extract showed a significant reduction in CD4+ antibody staining within the mucosal layer in colonic sections indicating reduced lymphocyte infiltration. Based on these findings, we believe that administration of LN extracts may be effective in reducing the intestinal epithelial damage seen in human IBD and warrants further investigation through clinical trials. Lay Summary: Inflammatory bowel diseases (IBD), such as Crohn's disease (CD) and ulcerative colitis (UC), manifest as chronic inflammation and ulceration of tissues lining the digestive tract. CD involves inflammation of the deeper layers of the digestive tract, including both the small and large intestines, and less commonly, the upper digestive tract. UC involves inflammation along the lining of the colon and rectum. Steroid or biologic treatments for IBD are common, however, are limited due to significant side effects and/or prohibitive cost. In the present study, we provide evidence for use of the natural product, Laurus nobilis (bay leaf), as a safe and effective anti-inflammatory therapy for IBD.

Extract of Laurus nobilis attenuates inflammation and epithelial ulcerations in an experimental model of inflammatory bowel disease

Inflammatory bowel diseases (IBD), including Crohn's disease and ulcerative colitis, are classified as chronic inflammatory disorders and typically require anti-inflammatory drug therapies, such as glucocorticoid regimens, non-steroidal anti-inflammatory drugs, and biologics, aimed at reducing inflammation in the bowel wall. However, each of these therapies is accompanied by a list of possible serious side effects. Because of this, there remains an urgent need to identify new pharmacologic options to reduce or prevent the pro-inflammatory events of IBD while minimizing adverse side effects and making available more cost-effective treatment modalities. We have previously identified several herbal extracts that demonstrate the potent bio-inhibitory activity of the innate immune response. In particular, Laurus nobilis (LN), or more commonly called bay laurel, demonstrated significant anti-inflammatory function by inhibiting nuclear factor-κB activation. Based upon our original in vitro findings, we have now examined the effects of this herbal extract on a murine dextran sodium sulfate (DSS) model of IBD. Hematoxylin and eosin-stained paraffin sections prepared from DSS-treated animals show clear epithelial damage, including ulcerations, extensive neutrophil infiltration into the mucosal layer, and granuloma formation. Tissue from DSS-treated animals that also received LN extract showed improved tissue morphology more closely resembling that from control animals. In addition, DSS-treated mice with co-administration of LN extract showed a significant reduction in CD4+ antibody staining within the mucosal layer in colonic sections indicating reduced lymphocyte infiltration. Based on these findings, we believe that the administration of LN extracts may be effective in reducing the intestinal epithelial damage seen in human IBD and warrants further investigation through clinical trials. Lay Summary: Inflammatory bowel diseases (IBD), such as Crohn's disease (CD) and ulcerative colitis (UC), manifest as chronic inflammation and ulceration of tissues lining the digestive tract. CD involves inflammation of the deeper layers of the digestive tract, including both the small and large intestines, and less commonly, the upper digestive tract. UC involves inflammation along the lining of the colon and rectum. Steroid or biological treatments for IBD are common, however, they are limited due to significant side effects and/or prohibitive costs. In the present study, we provide evidence for use of the natural product, Laurus nobilis (bay leaf), as a safe and effective anti-inflammatory therapy for IBD.

CD34+ progenitor cells as diagnostic and therapeutic targets in Alzheimer's disease.

CD34+ progenitor cells as diagnostic and therapeutic targets in Alzheimer's disease.

Impaired adaptive immune response in COVID-19 convalescent patients with hematological malignancies.

The immune dysregulation during SARS-CoV-2 has the potential to worsen immune homeostasis after recovery. Patients with hematological malignancies with COVID-19 have changes both in the innate and adaptive immune responses. Little is known about the severity of immune dysfunction following recovery from COVID-19 in hematological patients. Here, we performed a comprehensive analysis of the lymphocyte subsets in peripheral blood mononuclear cells by FACS Canto II in 55 patients, including 42 with hematological malignancies 4-6 weeks after COVID-19. Hematological COVID-19 convalescents had deep reduction in CD3+ T cells, including helper T cells (CD3 + CD4+), naïve helper T cells (CD3 + CD4 + CD45RA+), and memory CD4+ T cells among with extremely low levels of Treg cells and decreased expression of both TCRα/β and TCRγ/δ. Severe immune dysregulation in hematological convalescents was expressed by increased activation of T lymphocytes, both as elevated levels of activated T cells (CD3 + HLA-DR+) and activated cytotoxic T cells (CD3 + CD8 + HLA-DR+). Our findings showed a profound impairment of the adaptive immune response in hematological convalescents which might be a result of persistent activation of T cells. Convalescents with lymphoid malignancies showed more pronounced depletion of key T lymphocytes subpopulations in creating an effective adaptive response and immune memory.

Antitumor Effects of Ononin by Modulation of Apoptosis in Non-Small-Cell Lung Cancer through Inhibiting PI3K/Akt/mTOR Pathway.

Lung cancer is a leading global cause of cancer-related death in both males and females. Non-small-cell lung cancer (NSCLC) is the most commonly diagnosed cancer type that can be difficult to control with conventional chemotherapeutic and surgical approaches resulting in a poor prognosis. Paclitaxel (PTX) is a commonly used chemotherapeutic drug for NSCLC, which can cause tissue injury in healthy cells and affect the quality of life in patients with cancer. In order to treat NSCLC, alternative medications with minimal or no side effects are highly needed. Ononin is an isoflavone glycoside extracted from Astragali Radix (AR) that has various pharmacological activities. Therefore, this study investigated whether ononin inhibits NSCLC progression and promotes apoptosis synergistically with PTX both in vitro and in vivo. Antitumorigenic properties of ononin were determined by MTT assay, colony formation assay, migratory capacity, and apoptotic marker expression in A549 and HCC827 cells. The combination of ononin with PTX increased the expression of apoptotic markers and ROS generation and inhibited cell proliferation through the PI3K/Akt/mTOR signaling pathways. Furthermore, ononin prevented the translocation of NF-κB from cytosol to the nucleus. Also, we used the xenograft NSCLC mice model to confirm the in vivo antitumorigenic efficacies of ononin by reduction of CD34 and Ki67 expressions. Based on the histological analysis, the cotreatment of PTX and ononin reduced PTX-induced liver and kidney damage. Overall, our findings suggested that the therapeutic index of PTX-based chemotherapy could be improved by reducing toxicity with increasing antitumor capabilities when combined with ononin.

Utility of HLA‐DR in screening panel for inborn errors of immunity

AbstractA comprehensive evaluation of immune parameters while screening for inborn errors of immunity (IEIs) provides valuable information about the underlying defect. In 2019, the ICMR‐National Institute of Immunohaematology adopted a screening panel modified from the Euroflow consortium‐recommended PIDOT tube, the difference being the addition of HLA‐DR. In the present study, we evaluate the utility of HLA‐DR in our screening panel. The HLA‐DR expression on CD3 + T, its CD4+ and CD8+ subsets, natural killer (NK) and double‐negative T cells (DNT) was compared between patients and controls. HLA‐DR expression on CD3+ T cells or its subsets was significantly elevated in SCID and Omenn's phenotype (P &lt;.001), combined immunodeficiencies (P &lt; .05) including DOCK8 and WAS, diseases of immune dysregulation including FHL (P &lt; .0001) and ALPS (P &lt; .05) and CVID with complications (P &lt; .05). An absence of HLA‐DR marker can help in identifying cases of MHC II deficiency at the time of screening including atypical cases which present without reduction in CD4+ T cell counts and reversal of CD4+/CD8+ ratio. ROC analysis revealed a cut‐off of 14.62%, 19.11% and 1.5% for percentage HLA‐DR expression on CD3+ and CD8+ T cells and HLA‐DR on CD8/HLA‐DR on CD4, respectively, for FHL. Random forest analysis identified HLA‐DR (%) on NK cells, ratio of HLA‐DR CD8+/HLA‐DR CD4+, NK cell (%), T cell (%), HLA‐DR+ CD8(%) and CD45pos NK cell (%) as important predictors for FHL. Evaluation of HLA‐DR expression during screening can provide corroborative clues to the diagnosis of underlying IEI along with other immunological abnormalities.

Free Light Chains, High Mobility Group Box 1, and Mortality in Hemodialysis Patients.

Background: Uremic toxins are associated with immune dysfunction and inflammation. The inadequate removal by hemodialysis (HD) of serum free light chains (FLCs) determines their accumulation. This study evaluated FLCs in HD patients, analyzing their relations with other biomarkers, such as serum high mobility group box 1 (HMGB1). Methods: FLC and HMGB1 were evaluated in a cohort of 119 HD patients. κFLC and λFLC were summated to give a combined (c) FLC concentration. Patients were followed prospectively until the end of the observation period of four years, or until the endpoint: the patient’s death. Results: cFLC values in HD patients were 244.4 (197.9−273.5) mg/L. We detected a significant reduction in CD8+ cells and a decreased CD4+/CD8+ ratio. HMGB1 levels were 94.5 (55−302) pg/mL. After multivariate analysis, cFLCs correlated with β2-microglobulin and the CD4+/CD8+ ratio. Subjects with cFLC values above 263 mg/L and with sHMGB1 values < 80 pg/mL experienced a significantly faster evolution to the endpoint (mean follow-up time to progression of 27.5 and 28.5 months, respectively; p < 0.001). After an adjusted multivariate Cox analysis, cFLCs were associated with 11% increased risk of death, whereas low sHMGB1 increased this risk by 5%. Conclusions: cFLCs and HMGB1 reflect the inflammation and immune dysfunction in HD patients representing two strong and independent risk markers of mortality.