CD4 Cytokine Research Articles

A multistage protein subunit vaccine as BCG-booster confers protection against Mycobacterium tuberculosis infection in murine models

The eradication of tuberculosis remains a global challenge. Despite being the only licensed vaccine, Bacillus Calmette-Guérin (BCG) confers limited protective efficacy in adults and individuals with latent tuberculosis infections (LTBI). There is an urgent need to develop novel vaccines that can enhance the protective effect of BCG. Protein subunit vaccines have garnered significant research interest due to their safety and plasticity. Based on previous studies, we selected three antigens associated with LTBI (Rv2028c, Rv2029c, Rv3126c) and fused them with an immunodominant antigen Ag85A, resulting in the construction of a multistage protein subunit vaccine named A986. We evaluated the protective effect of recombinant protein A986 adjuvanted with MPL/QS21 as a booster vaccine for BCG against Mycobacterium tuberculosis (Mtb) infection in mice. The A986 + MPL/QS21 induced the secretion of antigen-specific Th1 (IL-2+, IFN-γ+ and TNF-α+) and Th17 (IL-17A+) cytokines in CD4+ and CD8+ T cells within the lung and spleen of mice, while also increased the frequency of central memory and effector memory T cells. Additionally, it also induced the enhanced production of IgG antibodies. Compared to BCG alone, A986 + MPL/QS21 boosting significantly augmented the proliferation of antigen-specific multifunctional T cells and effectively reduced bacterial load in infected mice. Taken together, A986 + MPL/QS21 formulation induced robust antigen-specific immune responses and provided enhanced protection against Mtb infection as a booster of BCG vaccine.

Sodium houttuyfonate Modulates the Lung Th1/Th2 Balance and Gut Microbiota to Protect Against Pathological Changes in Lung of Ovalbumin-induced Asthmatic mice

Objective The gut-lung axis involves microbial and product interactions between the lung and intestine. Antibiotics for chronic asthma can cause intestinal dysbiosis, disrupting this axis. Sodium houttuyfonate (SH) has diverse biological activities, including modifying gut microbiota, antibacterial, and anti-inflammatory. This study aims to explore the relationship between SH, CD4+ T cells, and gut microbiota. Methods Allergic asthma was experimentally induced in mice through injection and inhalation of ovalbumin. After the administration of different amounts of SH, ELISA was utilized to ascertain the levels of inflammatory cytokines in the serum, flow cytometry was used to examine the levels of Th1/Th2 cytokines in CD4+ cells from lung tissues. The expression of T-bet and GATA3 in lung tissue was determined by Western blotting and quantitative real-time PCR assay. Gut microbiota was determined by 16S rRNA gene sequencing. Results The results showed that SH can alleviate pulmonary injury in asthmatic mice, reducing serum levels of IL-4, IL-5, and IL-13 while simultaneously increasing IFN-γ. Furthermore, SH has been observed to modulate the balance of Th1/Th2 cells by up-regulating the mRNA and protein expression of T-bet but down-regulating GATA3 in the lung tissues of asthmatic mice, thereby promoting the differentiation of Th1 cells. Additionally, SH can regulate the variety and composition of gut microbiota especially genus Akkermansia in asthmatic mice. Conclusion SH can alleviate asthma through the regulation of Th1/Th2 cells and gut microbiota.

Novel bispecific nanobody mitigates experimental intestinal inflammation in mice by targeting TNF-α and IL-23p19 bioactivities.

Inflammatory bowel diseases (IBDs) pose significant challenges in terms of treatment non-response, necessitating the development of novel therapeutic approaches. Although biological medicines that target TNF-α (tumour necrosis factor-α) have shown clinical success in some IBD patients, a substantial proportion still fails to respond. We designed bispecific nanobodies (BsNbs) with the ability tosimultaneously target human macrophage-expressed membrane TNF-α (hmTNF-α) and IL-23. Additionally, we fused the constant region of human IgG1 Fc (hIgG1 Fc) to BsNb to create BsNb-Fc. Our study encompassed in vitro and in vivo characterization of BsNb and BsNb-Fc. BsNb-Fc exhibited an improved serum half-life, targeting capability and effector function than BsNb. It's demonstrated that BsNb-Fc exhibited superior anti-inflammatory effects compared to the anti-TNF-α mAb (infliximab, IFX) combined with anti-IL-12/IL-23p40 mAb (ustekinumab, UST) by Transwell co-culture assays. Notably, in murine models of acute colitis brought on by 2,4,6-trinitrobenzene sulfonic acid（TNBS) and dextran sulphate sodium (DSS), BsNb-Fc effectively alleviated colitis severity. Additionally, BsNb-Fc outperformed the IFX&UST combination in TNBS-induced colitis, significantly reducing colon inflammation in mice with colitis produced by TNBS and DSS. These findings highlight an enhanced efficacy and improved biostability of BsNb-Fc, suggesting its potential as a promising therapeutic option for IBD patients with insufficient response to TNF-α inhibition. A bispecific nanobody (BsNb) was created to target TNF-α and IL-23p19, exhibiting high affinity and remarkable stability. BsNb-Fc inhibited the release of cytokines in CD4+T cells during co-culture experiments. BsNb-Fc effectively alleviated colitis severity in mouse model with acute colitis induced by DSS or TNBS, outperforming the IFX&UST combination.

Lymphocyte Expression of Intracellular Cytokines and Heat Shock Proteins in Peripheral Blood of Patients with Atopic Dermatitis.

The present study evaluates expression by activated CD4+ T helper1 (Th1) and T helper 2 (Th2) T lymphocytes of pro-inflammatory cytokines and cytoprotective heat shock proteins (HSPs) in peripheral blood of atopic dermatitis (AD) patients. This research represents preliminary work by the authors to identify correlates between critical immune parameters with the potential to serve as guidelines for the development of pharmacological strategies for altering these factors to promote the restoration of healthy immune profiles in persons afflicted with major atopic diseases. The major experimental strategy used in this research assessed immune activation by peripheral blood mononuclear cells (PBMC) from 21 AD patients and 12 age- and gender-matched healthy control subjects cultured with phorbol myristate acetate (PMA) and ionomycin (PMA/I), which are mutagenic immune activators, to induce expression of pro-inflammatory biomarkers in CD4+ T cells differentiated to express Th1 or Th2 cytokines and heme oxygenase-1 (HO-1) intracellularly (i). Evaluations were performed using an FC500 Beckman-Coulter flow cytometer. Elevated CD4+ T cell expression of cytokines, interleukin-4 (iIL-4), interleukin- 5 (iIL-5), interleukin-10 (iIL-10), interferon-gamma (iIFN-g), tumor necrosis factor-alpha (iTNF-α), were observed. Additionally, the heat shock proteins (HSP) iHO-1 and iHSP-70 were evaluated in cells from the blood of AD patients versus the control subjects. The present study demonstrated an elevated expression of both Th1 and Th2-associated cytokines in CD4+ T cells of AD patients, with a significant direct correlation between Th1 and Th2 cell populations, thus yielding insight into the immune features of the AD-associated systemic inflammatory profile. Finally, the observed increased iHO-1 and iHSP-70 expressions likely represent adaptive physiologic countermeasures to AD-associated inflammatory tissue damage, suggesting that HSP inducers are promising candidates for the management of atopic disorders.

Effects of mesenchymal stem cells on Treg cells in rats with colitis.

The aim was to investigate the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSC) on dextran sulfate sodium (DSS) induced colitis in rats and its effect on regulatory T cells (Treg). A model of DSS-induced colitis was established. BM-MSC was isolated and cultured to observe the efficacy of BM-MSC on colitis, including general vital signs, weight changes, colonic length changes, colonic histopathological changes, and colonic tissue MPO activity. The expression of inflammatory factors (IFN-γ, IL-4, IL-17, TGF-β) in colonic tissues was measured by real-time PCR. The amount of CD4 + CD25 + Treg was detected by flow cytometry. Real-time PCR was used to detect Foxp3+mRNA in CD4 + CD25 + Treg, western to detect Foxp3+protein expression in CD4 + CD25 + Treg, and ELISA was used to detect IL-35 and IL-10 cytokines in CD4 + CD25 + Treg culture supernatant. Results show that intravenous injection of BM-MSC significantly improved the clinical manifestations and histopathological changes in rats with experimental DSS colitis; significantly down-regulated the expression of inflammatory factors IFN-γ, IL-4, and IL-17 and up-regulated the expression of TGF-β in colon tissues; BM-MSC also increased the number of CD4+CD25+Foxp3+Treg and enhanced the function of CD4+CD25+Foxp3+Treg in colon tissues, and up-regulated the expression of IL-35. In conclusion, BM-MSC has a certain therapeutic effect on DSS-induced colitis. It can improve the general signs of colitis rats and reduce intestinal injury and inflammatory response. The immunoregulatory effect of BM-MSC is achieved by enhancing the function of CD4+CD25+Foxp3+Treg and up-regulating the secretion of immunosuppressive inflammatory factors.

Engineered live attenuated bacteria producing 5-hydroxyindole remodels heterogeneous tumor microenvironment and promotes antitumor immunity

Abstract The highly immunosuppressive tumor microenvironment (TME) stimulates cancer cells resistance to immunotherapy. We have previously shown that a live-attenuated Brucella melitensisΔ vjbR(BmΔ vjbR) bacterial strain, breaks cancer cell resistance to immunotherapy by remodeling the TME and facilitating infiltration of cytotoxic CD8 +T cells (CTLs) into the tumor. As a result, there was shrinkage of tumor size and significant improvement in survival of these mice. However, BmΔ vjbRwas efficient in only controlling colorectal tumors, but was rendered ineffective in B16 melanoma. In this study, we sought to improve the efficacy of BmΔ vjbRin controlling different cancers by engineering it to augment the production of 5-hydroxyindole (HI), i.e., BmΔ vjbR-HI. The effect of BmΔ vjbR-HI in modulating the function and activity of CTLs was assessed by flowcytometry in in vitroassays. The tumor modulating activity of the BmΔ vjbR-HI was also assessed by intravenous injection of this bacteria followed by adoptive CTL therapy in colon carcinoma, melanoma and pancreatic orthotropic murine tumors. We found that BmΔ vjbR-HI increased inflammatory cytokines in CD8 +T cells. The granzyme-B production and cytotoxicity of CD8 +T cells was also enhanced on treatment with the bacteria. BmΔ vjbR-HI remodeled the TME and ameliorated MC32-CEA colorectal cancer, B16-OVA melanoma and Panc-02 orthotropic pancreatic tumors. Moreover, the BmΔ vjbR-HI homed into the TME by uptake in myeloid derived suppressor cells. Overall, our findings demonstrate that BmΔ vjbR-HI remodels heterogeneous TMEs of different cancers to promote CTL-mediated antitumor immunity. These findings have significant translational potential in antitumor microbial immunotherapy. Jugal Das and Jianxun Song are recipients of Careers in Immunology Fellowship 2022.

Modulating T cell inflammation in a mouse model of Alzheimer’s disease

AbstractBackgroundRecent single cell transcriptomic studies using peripheral blood and post‐mortem brain samples from Alzheimer’s disease (AD) patients have suggested a significant role for T cells in modulating AD‐related neuroinflammation. Previous findings from our group and others show that T cells in AD patients and aged individuals are clonally expanded and upregulate genes related to inflammation, cytotoxicity, and senescence. We have found that TSC22D3, a putative anti‐inflammatory transcription factor, is upregulated in brain‐infiltrating CD8+ T cells and found in gene co‐expression modules in peripheral blood from AD patients.MethodWe are pursuing the mechanism of TSC22D3 function and its effect on T cell biology and AD neuropathology. We have generated mice that lack TSC22D3 in CD8 T cells on the 5XFAD background, a mouse model of AD. We used flow cytometry to measure populations of various immune cell subtypes, and sort T cell subtypes for downstream sequencing. We are also using immunocytochemistry to quantify markers of amyloid and tau pathology, microglial interaction with amyloid plaque, and neuronal and synaptic loss in the hippocampus. Finally, we are examining the induction of immune reactions to amyloid and tau protein in TSC22D3 conditional knockout mice without AD transgenes.ResultIn human tissue, TSC22D3 is highly expressed in brain‐infiltrating CD8+ T cells along with genes associated with a senescent, late‐differentiation phenotype. Flow cytometry analysis showed that late‐stage AD mouse splenocytes bore significantly higher percentages of dendritic cells and various cytokines in CD4+ T cells than non‐AD or younger AD mice. Most of these differences were independent of TSC22D3 conditional knockout status. Ongoing bulk sequencing may reveal TSC22D3‐dependent changes in T cell gene expression, with effects on key markers of AD progression in the hippocampus.ConclusionTSC22D3 is known to directly suppress NFkB and other pro‐inflammatory mediators. Its co‐expression with more pro‐inflammatory and cytotoxic genes in brain‐infiltrating CD8+ T cells may represent a compensatory mechanism in the face of neuroinflammation related to aging and AD. We expect that knockout of TSC22D3 in CD8+ T cells will increase expression of pro‐inflammatory genes in T cell subsets, and will manifest in advanced AD pathology in older adult mice.

Lck signaling inhibition causes improvement in clinical features of psoriatic inflammation through reduction in inflammatory cytokines in CD4+ T cells in imiquimod mouse model

Psoriasis is a chronic dermal inflammatory disease with a world-wide prevalence in which different immune/non-immune cells, e.g. T cells, macrophages, neutrophils, and keratinocytes play a decisive role. These immune cells interact among themselves by releasing multiple mediators which eventually cause characteristic psoriatic plaques in the skin. T cells are reported to be significant contributors to psoriatic inflammation through release of multiple cytokines which are controlled by several kinases, one of them being Lymphocyte-specific protein tyrosine kinase (Lck). Lck has been reported to be crucial for expression/production of several key inflammatory cytokines though modulation of several other kinases/transcription factors in T cells. Therefore, in this investigation, effect of Lck inhibitor, A-770041 was examined on PLCγ, p38MAPK, NFATc1, NFkB and STAT3, TNF-α, IFN-γ, Foxp3, IL-17A, in CD4+ T cells in imiquimod (IMQ)-induced psoriatic inflammation in mice. Results from the present study exhibit that p-Lck expression is enhanced in CD4+ T cells of IMQ-treated mice which is concomitant with enhanced clinical features of psoriatic inflammation (ear/back skin thickness, MPO activity, acanthosis/leukocytic infiltration) and molecular parameters (enhanced expression of p-Lck, p-PLCγ, p-p38-MAPK, NFATc1, p-NFkB, TNF-α, IFN-γ, p-STAT3, and IL-17A in CD4+ T cells). Inhibition of Lck signaling led to amelioration of clinical features of psoriasis through attenuation of Th1/Th17 immune responses and upregulation of Treg cells in IMQ-treated mice. In summary, current investigations reveal that Lck signaling is a crucial executor of inflammatory signaling in CD4+ T cells in the context of psoriatic inflammation. Therefore, Lck inhibition may be pursued as an effective strategy to counteract psoriatic inflammation.

IFN-γ promotes TH1 at the expense of TFH differentiation during viral infections

Abstract Although humoral and cellular immunity upon viral infections usually co-exist, sometimes one of the two responses emerges and is responsible for most of the antiviral activity. For example, vescicular stomatitis virus (VSV) infection induces early and potent neutralizing antibody (nAb) responses, whereas lymphocytic choriomeningitis virus (LCMV) infection induces strong cellular responses, but weak nAb responses. Recent work from our laboratory has shown that unbalance is observed also at the level of CD4 T cells responses, with VSV inducing strong TFH polarization that support nAb responses, and LCMV in contrast promoting TH1 differentiation. Analysis of the VSV and LCMV priming niches led to identification of the spatiotemporal regulation of type I IFN expression as a critical regulator of antiviral CD4+ T cell polarization, with early type I IFN sensing leading to TFH polarization. Notably, this mechanism did not explain the strong TH1 differentiation observed during LCMV infection. In this study we aimed to elucidate the role of known TH1-polarizing cytokines in CD4+ T cell differentiation upon LCMV infection. We found that IFN-γ, but not IL-12, plays a key role in early CD4+ T cell differentiation upon LCMV infection, inducing TH1 cell polarization and suppressing TFH cell development, thus resulting in a shift in the equilibrium towards TFH and humoral responses. Future studies will determine the cellular source for this TH1-polarizing cytokine and the mechanism by which IFN-γ exerts its function, possibly unveiling the mechanisms underlying the reduced humoral response in the context of viral infections like LCMV.

Effects of Local Nasal Immunotherapy with FIP-fve Peptide and Denatured Tyrophagus putrescentiae for Storage Mite-Induced Airway Inflammation.

Allergic diseases are affecting public health and have increased over the last decade. Sensitization to mite allergens is a considerable trigger for allergy development. Storage mite-Tyrophagus putrescentiae shows great significance of allergenic potential and clinical relevance. The fungal immunomodulatory peptide FIP-fve has been reported to possess immunomodulatory activity. We aimed to determine whether T. putrescentiae-induced sensitization and airway inflammation in mice could be downregulated by FIP-fve in conjunction with denatured T. putrescentiae (FIP-fve and DN-Tp). Immune responses and physiologic variations in immunoglobulins, leukocyte subpopulations, cytokine productions, pulmonary function, lung pathology, cytokines in CD4+ and Treg cells were evaluated after local nasal immunotherapy (LNIT). After the LNIT with FIP-fve and DN-Tp, levels of specific IgE, IgG1, and IgG2a in the sera and IgA in the bronchoalveolar lavage fluid (BALF) were significantly reduced. Infiltrations of inflammatory leukocytes (eosinophils, neutrophils, and lymphocytes) in the airway decreased significantly. Production of proinflammatory cytokines (IL-5, IL-13, IL-17F and IL-23) and chemokine (IL-8) were significantly reduced, and Th1-cytokine (IL-12) increased in the airway BALF after LNIT. Pulmonary functions of Penh values were significantly decreased after the methacholine challenge, which resulted in a reduction of airway hypersensitivity after LNIT. Bronchus pathology showed a reduction of inflammatory cell infiltration and epithelium damage after LNIT. The IL-4+/CD4+ T cells could be downregulated and the IFN-γ+/CD4+ T cells upregulated. The Treg-related immunity of IL-10 and Foxp3 expressions in CD4+CD25+ cells were both upregulated after LNIT. In conclusion, LNIT with FIP-fve and DN-Tp had an anti-inflammatory effect on mite-induced airway inflammations and possesses potential as an immunomodulatory therapy agent for allergic airway diseases.

ABCB10 Loss Reduces CD4+ T Cell Activation and Memory Formation.

T cells must shift their metabolism to respond to infections and tumors and to undergo memory formation. The ATP-binding cassette transporter ABCB10 localizes to the mitochondrial inner membrane, where it is thought to export a substrate important in heme biosynthesis and metabolism, but its role in T cell development and activation is unknown. In this article, we use a combination of methods to study the effect of ABCB10 loss in primary and malignantly transformed T cells. Although Abcb10 is dispensable for development of both CD4+ and CD8+ T cells, it is required for expression of specific cytokines in CD4+, but not CD8+, T cells activated in vitro. These defects in cytokine expression are magnified on repeated stimulation. In vivo, CD8+ cells lacking ABCB10 expand more in response to viral infection than their control counterparts, while CD4+ cells show reductions in both number and percentage. CD4+ cells lacking ABCB10 show impairment in Ag-specific memory formation and recall responses that become more severe with time. In malignant human CD4+ Jurkat T cells, we find that CRISPR-mediated ABCB10 disruption recapitulates the same cytokine expression defects upon activation as observed in primary mouse T cells. Mechanistically, ABCB10 deletion in Jurkat T cells disrupts the ability to switch to aerobic glycolysis upon activation. Cumulatively, these results show that ABCB10 is selectively required for specific cytokine responses and memory formation in CD4+ T cells, suggesting that targeting this molecule could be used to mitigate aberrant T cell activation.

Human umbilical cord mesenchymal stem cells alleviate the imbalance of CD4+ T cells via protein tyrosine phosphatase non-receptor type 2/signal transducer and activator of transcription 3 signaling in ameliorating experimental autoimmune thyroiditis in rats.

This study aimed to investigate the therapeutic effect of human umbilical cord mesenchymal stem cells (hUCMSCs) on experimental autoimmune thyroiditis (EAT) and the underlying mechanisms by utilizing a porcine thyroglobulin-induced EAT rat model. The rats received four tail vein injections of vehicle or hUCMSCs at an interval of 7 days and were sacrificed on day 28 after the first injection. Hematoxylin and eosin staining and enzyme-linked immunosorbent assays (ELISAs) were used to assess the therapeutic effects of hUCMSCs on EAT. Splenic lymphocytes were isolated from rats, and the proportions of CD4+ T cell subsets were analyzed by flow cytometry. Splenic CD4+ T cells from EAT rats were cocultured with hUCMSCs. A loss-of-function assay for protein tyrosine phosphatase non-receptor type 2 (PTPN2) was performed to explore the involvement of PTPN2/signal transducer and activator of transcription 3 (STAT3) signaling on the therapeutic benefit of hUCMSCs in EAT. hUCMSC treatment significantly alleviated inflammation, reduced serum thyroid antibody levels, and decreased the ratios of IL-17α+/CD25+FOXP3+ cells and serum IFN-γ/IL-4 in EAT rats. Furthermore, hUCMSC treatment upregulated PTPN2 protein expression in splenic lymphocytes of EAT rats as well as enhanced the PTPN2 protein level and attenuated phosphorylation of STAT3 in CD4+ T cells in vitro. Importantly, knockdown of Ptpn2 significantly reversed hUCMSC-mediated suppression of cell proliferation and hUCMSC-induced alterations in the expression of inflammatory cytokines in CD4+ T cells. Thus, hUCMSC treatment alleviates thyroid inflammation and the CD4+ T cell imbalance in EAT via PTPN2/STAT3 signaling, serving as a promising therapeutic approach for autoimmune thyroiditis.

Expert Roundtable: leveraging cutting edge tools to convert I-O data into knowledge

<div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Sood Pic copy.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name" style="color:#5D538A;">Anil K Sood</h3><h5 class="author-title" style="color:#5D538A;">Professor, Department of Gynecologic Oncology and Reproductive Medicine, UT MD Anderson Cancer Center</h5> Dr Anil K Sood is Professor in the Department of Gynecologic Oncology and Reproductive Medicine at the UT MD Anderson Cancer Center. He holds a joint appointment in Cancer Biology and is co-director of the Center for RNA Interference and Non-Coding RNA at the M. D. Anderson Cancer Center. He is also Director of the multi-disciplinary Blanton-Davis Ovarian Cancer Research Program and co-leads the Ovarian Cancer Moonshot Program. Dr Sood received his medical degree from the University of North Carolina, Chapel Hill. A major and consistent theme of his scientific research has been on understanding human cancer biology and converting lab discoveries into novel therapeutics. His research group has made several seminal research contributions in the fields of tumor microenvironment, nanomedicine, and neuroendocrine effects on cancer biology. Dr Sood has received recognition for his research accomplishments including the Hunter Award, the Margaret Greenfield/Carmel Cohen Excellence in Ovarian Cancer Research Prize, and the GCF/Claudia Cohen Research Foundation Prize for Outstanding Gynecologic Cancer Researcher. He is an elected member of the American Society for Clinical Investigation (ASCI), the American Association for the Advancement of Science (AAAS), and the Association of American Physicians (AAP). Dr Sood was selected as an American Cancer Society Research Professor in 2017. </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Prabhakaran_Sandhya_1200x800 - Sandhya Prabhakaran copy.jpg" data-image="1"></div><div class="author-det"><h3 class="author-name" style="color:#00A795;"><strong>Sandhya Prabhakaran</strong></h3><h5 class="author-title" style="color:#00A795;">Applied Research Scientist, Integrated Mathematical Oncology Department, Moffitt Cancer Centre</h5> Dr. Sandhya Prabhakaran is an Applied Research Scientist at the Integrated Mathematical Oncology department, Moffitt Cancer Centre, Florida. Before that she was a Research Scientist at Memorial Sloan Kettering Cancer Centre and Columbia University. Her PhD in Computer Science is from University of Basel and her Masters in Intelligent Systems (Robotics) is from University of Edinburgh. Her research deals with developing statistical theory and Bayesian inference models, particularly to problems in Cancer Biology and Computer Vision. Prior to academics, she was an Assembler programmer working with the Mainframe Operating System (z/OS) at IBM Software Laboratories and has developed Mainframe applications. She has completed 4 out of the 6 World Marathon Majors. </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><strong><strong><strong><img src="https://cdn.insights.bio/uploads/Goldmacher photo.png" data-image="1"></strong></strong></strong></div><div class="author-det"><h3 class="author-name" style="color:#007589;"><strong><strong>Gregory Goldmacher</strong></strong></h3><h5 class="author-title" style="color:#007589;">Executive Director, Clinical Research, Head of Clinical Imaging, Merck Research Laboratories</h5> Greg has been at MSD since 2015. With his team of radiologists and imaging scientists, he oversees the clinical imaging in over 200 clinical trials in oncology and other therapeutic areas. He also leads research in radiomics, artificial intelligence, novel response criteria, tumor growth kinetics, and other innovative approaches to clinical trial imaging. Prior to MSD, he was a senior medical director and head of oncology imaging at a global CRO, and before that in academia. His clinical training was in diagnostic radiology, along with a PhD and postdoctoral work in neuroscience, and business training in finance. In his ‘free’ time, he enjoys playing with his kids, running, and training in the martial arts. </div></div><div class="author"><div class="author-img"><strong><strong><strong><img src="https://cdn.insights.bio/uploads/Sanjabi photo copy.png" data-image="1"></strong></strong></strong></div><div class="author-det"><h3 class="author-name" style="color: rgb(125, 203, 120);"><strong><strong>Shomyseh Sanjabi</strong></strong></h3><h5 class="author-title" style="color: rgb(125, 203, 120);">Principal Scientist, Oncology Biomarker Development, Genentech</h5> Shomyseh Sanjabi, PhD is a Principal Scientist in Oncology Biomarker Development (OBD) at Genentech. She obtained her bachelor’s degree in microbiology and molecular genetics from the University of California, Los Angeles (UCLA) and completed her PhD training with Dr Stephen Smale at UCLA in the department of microbiology, immunology, and molecular genetics elucidating the molecular mechanism of family member specificity among the NF-kappa-B family of transcription factors. She then completed a postdoctoral training in immunobiology at Yale University with Dr Richard Flavell studying the role of TGF-beta and common gamma chain cytokines in CD8 T cell development and function in the context of autoimmunity, anti-pathogen immunity, and cancer immunity. In 2010, she joined the faculty at the Gladstone Institute of Virology and Immunology and UCSF department of microbiology and immunology, where she was the recipient of numerous awards including an NIH New Innovator Award. Her independent academic research focused on using model pathogens and emerging human pathogens, together with preclinical genetic mouse models and human primary tissue samples to study how protective innate and adaptive immune responses are elicited and maintained upon mucosal transmission of viral pathogens. Her passion for human immunology and her vision to apply innovative technologies to study the immune response of patients undergoing cancer immunotherapy brought her to Genentech in 2018. She currently leads the biomarker and translational activities for several early-stage cancer immunotherapy programs. </div></div></div>

The Loss of H3K27 Histone Demethylase Utx in T Cells Aggravates Allergic Contact Dermatitis.

The pathogenesis of allergic contact dermatitis (ACD) requires the activation of Ag-specific T cells, including effector and regulatory T cells. The differentiation and function of these T cells is epigenetically regulated through DNA methylation and histone modifications. However, the roles of altered histone H3K27 methylation in T cells in the development of ACD remain unknown. Two types of histone H3K27 demethylases, Utx and Jmjd3, have been reported in mammals. To determine the role of the histone H3K27 demethylase expression of T cells in the development of ACD, we generated T cell-specific, Utx-deficient (Utx KO) mice or Jmjd3-deficient (Jmjd3 KO) mice. Unlike control mice, Utx KO mice had severer symptoms of ACD, whereas Jmjd3 KO mice showed symptoms identical to those in control mice. In Utx KO mice with ACD, the massive infiltration of myeloid cells, including neutrophils and dendritic cells, has been observed. In addition, the expression of proinflammatory cytokines in CD4+ T cells of the draining lymph nodes (LNs) and in CD8+ T cells of the skin was increased in Utx KO mice, whereas the ratio of Foxp3+ regulatory CD4+ T cells to Foxp3- conventional CD4+ T cells was decreased in both the draining LNs and the skin of Utx KO mice with ACD. Furthermore, Foxp3+ regulatory CD4+ T cells of Utx KO mice with ACD expressed a decreased level of CCR4 (a skin-tropic chemokine receptor) in comparison with control. Thus, in CD4+ T cells, Utx could potentially be involved in the regulation of the pathogenesis of ACD.

The cAMP responsive element modulator (CREM) is a regulator of CD4+ Tcell function.

The cAMP responsive element modulator (CREM) is a transcriptional regulator of different effector cytokines in CD4+ Tcells including IL-2, IL-17, IL-21 but also IL-4 and IL-13 and thus an important determinant of central T helper cell functions. Our review gives an overview over the regulation of CREM in Tcells and the pleiotropic effects of CREM on CD4+ Tcells in health and autoimmune diseases with a particular focus on systemic lupus erythematosus.

LAG3 and PD1 Regulate CD8+ T Cell in Diffuse Large B-cell Lymphoma Patients.

Background Diffuse large B-cell lymphoma (DLBCL) is a clinically and genetically heterogeneous lymphoid malignancy. The unsatisfactory outcome for refractory patients has prompted efforts to explore new therapeutic approaches for DLBCL. However, the mechanisms involved in treatment associated with immune checkpoints remain unclear. This study is aimed at investigating the potential roles of programmed cell death protein 1 (PD1) and lymphocyte activation gene 3 (LAG3) in CD8+ T cells for treatment in DLBCL. Methods Utilizing flow cytometry, we examined the content of T cells, the levels of cytokines, and the expression of PD1 and LAG3 in patients with DLBCL as well as in healthy controls. Levels of cytokines in CD8+ T cells from DLBCL patients before and after treatment were compared by blocking of PD1 and LAG3 in magnetic bead-sorted CD8+ T cells. Results We found that the proportion of CD4+ T cells and CD8+ T cells was increased in DLBCL patients after treatment. The levels of cytokines trended toward those of healthy controls in treatment. PD1 (+), LAG3 (+), or PD1 (+) LAG3 (+) were all expressed in lower amounts in CD4+ T cells and CD8+ T cells after treatment than in untreated DLBCL patients. In addition, blockade of PD1 and LAG3 in sorted CD8+ T cells markedly inhibited cytokine production in response to treatment. Conclusion PD1 and LAG3 in CD8+ T cells may be important targets of therapy and play therapeutic role in patients with DLBCL.

Identification and Characterization of a Master Transcription Factor of Th1 Cells, T-bet, Within Flounder (Paralichthys olivaceus).

T-bet, a T-box family member, is a transcription factor essential for the differentiation of naive CD4+ T cells into Th1 cells that are involved in both innate and adaptive immune responses. In this study, the transcription factor T-bet of flounder (Paralichthys olivaceus) was cloned and characterized, and its expression profile after infection was analyzed. T-bet+ cells were identified in flounder, and the expression and localization of T-bet in T lymphocyte subsets and B lymphocytes were investigated. Finally, the proliferation of T-bet+ cells, T lymphocyte subsets, and B lymphocytes were studied after stimulation with IFN-γ, IL-2, and IL-6, respectively, and the variations of some transcription factors and cytokines in CD4+ T lymphocyte subsets were detected. The results showed that T-bet in flounder consists of 619 aa with a conserved T-box DNA binding domain. T-bet was abundantly expressed in the spleen, head kidney, and heart, and it was significantly upregulated after infection with Vibrio anguillarum, Edwardsiella tarda, and Hirame rhabdovirus, especially in the group of Edwardsiella tarda. A polyclonal antibody against recombinant protein of T-bet was prepared, which specifically recognized the natural T-bet molecule in flounder. T-bet+ cells were found to be distributed in the lymphocytes of peripheral blood, spleen, and head kidney, with the highest proportion in spleen, and the positive signals of T-bet occurred in the cell nucleus. T-bet was also detected in the sorted CD4-1+, CD4-2+, CD8+ T lymphocytes, and IgM+ B lymphocytes. In addition, T-bet+ cells, coordinated with CD4-1+ and CD4-2+ T lymphocytes, were proliferated after stimulation with IFN-γ, IL-2, and IL-6. Especially in sorted CD4-1+ and CD4-2+ T lymphocytes, IFN-γ and IL-2 were able to upregulate the expression of T-bet, forming a positive feedback loop in Th1-type cytokine secretion. These results suggest that T-bet may act as a master transcription factor regulating flounder CD4+ T lymphocytes involved in a Th1-type immune response.

Adjuvant nivolumab following salvage resection in head and neck squamous cell carcinoma patients previously treated with definitive therapy: A single-arm phase II multi-institutional study.

6031 Background: Salvage surgery for locally recurrent head and neck squamous cell carcinoma (rHNSCC) results in local control rates of 33-50% but only 20-40% of patients achieve long-term survival necessitating additional therapy (Haque et al., Oral Oncol. 2019). Many patients are ineligible for re-irradiation and chemotherapy alone after salvage surgeryhas shown no survival benefit. The clinical activity and tolerability of immune checkpoint inhibitors has been demonstrated in metastatic HNSCC, but the benefit after salvage surgery (SS) has not been studied. Here we report the results of a multi-center phase II investigation of nivolumab, a PD-1 inhibitor, after SS in recurrent HNSCC (NCT03355560). Methods: HNSCC patients undergoing curative-intent SS were enrolled to receive 6 months of nivolumab beginning 4-11 weeks after surgery. All received radiation with or without chemotherapy as prior definitive therapy and had no other curative treatment options at the time of surgery. Key exclusion criteria included: distant metastatic disease, gross residual disease, or a history of immunodeficiency, autoimmunity, or pneumonitis. The primary endpoint was 2-year disease-free survival (DFS) measured by Kaplan Meier curves. Safety was evaluated by CTCAE v5.0. Results: 39 patients were enrolled. Median age was 68 years (range, 49-85). 12/39 (31%) were female. 34/39 (87%) were white. Disease sites included oropharynx 9/39 (23%), oral cavity 14/39 (36%), and larynx 16/39 (41%). P16 status was 26% (+), 48% (-), and 26% (unknown). 17/39 (44%) had high risk pathologic features (positive margins or extranodal spread) at time of SS. 28/39 (72%) patients experienced treatment-related adverse events (TRAE), the most common of which were fatigue (26%), hypothyroidism (10%) and acneiform rash (13%). Grade 3-4 TRAEs were rare, occurring in 3/39 (8%) patients and included diarrhea, oral pain, neck pain, productive cough, stridor, and COPD exacerbation. 3/39 (8%) required treatment discontinuation and there were no grade 5 events. The 2-year DFS was 60% (95%CI 0.39-0.91). 2-year overall survival was 74% (95% CI 0.54-1). In single-cell multiplex cytokine analysis, patients who relapsed following adjuvant nivolumab had a significantly higher proportion of peripheral blood CD8 T cells which displayed a polyfunctional cytokine profile. IFN-γ and Granzyme were the dominant CD8 cytokines in both responders and non-responders, however CD8 expression of MIP1a and TNF-α were significantly higher in patients who ultimately relapsed. Conclusions: Nivolumab after salvage surgery in rHNSCC is well tolerated and shows promising antitumor activity in this high-risk patient population with unmet need. Immunotherapy after salvage surgery should be studied in randomized clinical trials. Clinical trial information: NCT03355560.

Etanercept attenuates immune-mediated hepatitis induced by concanavalin A via differential regulation of the key effector cytokines of CD4+ T cells

AimsThe current study aims to investigate the role of the key effector cytokines produced by CD4+T cells in the pathogenesis of Con A-induced liver injury in mice and testing whether etanercept can be repurposed to differentially regulate these cytokines. Main methodsFour groups of mice were used: group I: control group, group II: mice received 15 mg/kg Con A i.v, group III: mice received 15 mg/kg etanercept i.p, group IV: mice received both Con A and etanercept as described. Hepatic injury and necroinflammation were assessed. Infiltration of CD4+ T cells and neutrophils were evaluated. Hepatic levels of TNF-α, IL-4, IL-10, and MDA were assigned and expression of NF-κB as well. Key findingsA significant decrease in ALT, AST, and LDH levels occurred when etanercept was injected before Con A. Hepatic necrosis and infiltration of CD4+ T cells and neutrophils were reduced by etanercept. Levels of TNF-α, IL-4, and MDA were significantly decreased in group IV compared to group II while that of IL-10 was increased. Also, number of NF-κB positive cells was significantly low in group IV. SignificanceThe study elucidates an interplay between the two effector cytokines of CD4+ T cells, TNF-α and IL-4, and their key role in Con A-induced liver injury. Additionally, our results showed that etanercept could be repurposed to differentially regulate effector cytokines produced by CD4+ T cells. Not only TNF-α, but also IL-4 signaling pathways, through which it exerts immunomodulatory, anti-inflammatory, and anti-oxidant effects leading to attenuation of Con A-induced liver injury.

Single-cell RNA Sequencing Deciphers Immune Landscape of Human Recurrent Miscarriage

Single-cell RNA Sequencing Deciphers Immune Landscape of Human Recurrent Miscarriage