bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigating DAGLβ expression and activity in human monocytes via proteomic and lipidomic profiling
Authors: Murtagh Olivia , McSorley Caitlin , Hsu Ku-Lung
Published: August 2023

Lipid-metabolizing enzymes are important targets for small-molecule inhibitor discovery as the regulation of inflammation responses have been linked to lipid metabolism cascades controlled by these enzymes. Non-steroidal anti-inflammatory drugs inhibit enzymes in the COX pathway preventing conversion of arachidonic acid (AA) into inflammation-inducing prostaglandins. An upstream precursor to AA is the endocannabinoid 2-arachidonoyl-glycerol (2-AG), which is biosynthesized from the cleavage of arachidonate-containing diacylglycerols (DAGs) by diacylglycerol lipase beta (DAGLβ). Previous work to identify DAGLβ’s role in inflammatory function led to the creation of KT109, a DAGLβ-specific 1,2,3-triazole urea inhibitor. Primary mouse macrophages treated with KT109 showed reductions in AA and 2-AG and an increase in arachidonate-containing DAGs, demonstrating that DAGLβ is key in proinflammatory lipid regulation and signaling. These studies focused on the effects of inhibiting mouse DAGLβ, but information regarding KT109 inhibition of human DAGLβ is still lacking. Herein, we applied activity-based protein profiling (ABPP) and quantitative proteomic and lipidomic methods to determine the effectivity of KT109 inhibition in human monocytic cell line THP1. Potency (IC50 values) for DAGLβ inhibition by KT109 in human THP1 cells were comparable to the magnitude of inhibition in mouse cells. Lipidomic analysis of KT109-treated monocytic and macrophage-differentiated THP1s did not demonstrate an accumulation of arachidonate-containing DAGs. These findings show a potential difference in the expression and activity of human DAGLβ when compared to mouse DAGLβ and additional targets of KT109 in human cells.

Funding and disclosures: This work was supported by the National Institute on Drug Abuse grant no. R01 DA043571.
Open Chromatin: The Guide to Inflammatory Mediators in PCOS
Authors: Shabakesaz Pauniz , Banaszewska Beata , Duleba Antoni , Nicholas Dequina
Published: August 2023

Inflammation comorbid with insulin resistance and metabolic dysfunction are major manifestations of Polycystic Ovary Syndrome (PCOS). However, the type of inflammation associated with POCS and its effect remains uncharacterized making it important to identify inflammatory mediators that contribute to PCOS-related metabolic dysfunction. Our data from a mouse model of PCOS shows increased secretion of proinflammatory cytokines from Th17 cells. Th17 inflammation is associated with metabolic dysfunction in type 2 diabetes. Therefore, we hypothesize that Th17 cells predominate PCOS-specific inflammation. We will test this hypothesis using peripheral blood mononuclear cells (PBMCs) collected from a prospective cohort of 20 fully clinically characterized Polish women: 15 with PCOS and 5 without. Combinations of cytokines and transcription factors, specific to different immune cell types, will have enhanced access to inflammation-related loci promoting transcription. We have used assay for transposase-accessible chromatin with sequencing (ATAC-Seq) of nuclei extracted from the PBMCs of our cohort to test whether chromatin loci important for Th17 cells are accessible as reflected by regions of open chromatin in immune cells of patients with PCOS. We expect our ATAC-seq data to distinguish women with PCOS from control by an open chromatin signature that represents distinct cytokine profiles and Th17 specific transcription factor binding sites. This is the first step to generating a multivariable map of inflammation in this metabolic disease, with the long-term objective being the development of targeted anti-inflammatory therapeutics for PCOS and comorbid insulin resistance.

NIH grant#3TR00HD09833004S1 from NICHD

NIH grant#5TR00HD09833004 from NICHD
Adenylate kinase 4 is critical to the function of classically activated macrophage
Authors: Miaw Shi-Chuen , Chin Wei-Yao
Published: August 2023

Macrophage plays a crucial role in the front line of host defense against pathogens. Classically activated macrophages (M1), induced by IFN-g and LPS, highly express inflammatory cytokines and contribute to inflammatory processes. By contrast, alternatively activated macrophages (M2) are induced by IL-4/IL-13, produce IL-10, and display anti-inflammatory activity. Adenylate kinase 4 (Ak4), an enzyme that transfers phosphate group among ATP/GTP, AMP, and ADP, is a key modulator of ATP. Ak4 is involved in maintaining the homeostasis of cellular nucleotides which is essential for cellular function. We observed Ak4 is preferentially expressed in M1 macrophages compared to M2 macrophages. Whether Ak4 is critical for M1 macrophage function remains elusive. Here we demonstrated that Ak4 maintained ATP homeostasis, and was critical for ROS production, glycolysis, and bactericidal ability in M1 macrophages. Moreover, Ak4 promoted the expression of inflammatory genes, including Il1b, Il6, Tnfa, Nos2, Nox2 and Hif1a, in M1 macrophages via Hif1a and AMPK. However, Ak4 deficiency did not affect the development of murine immune cells. Taken together, our data depict a potential mechanism linking nucleotide homeostasis and the function of M1 macrophage.

Taiwan Ministry of Science and Technology (111-2320-B-002-068-MY3)
Leukocyte and Lymphatic Interactions at the Cribriform Lymphatics
Authors: Laaker Collin , Sandor Matyas , Fabry Zsuzsanna
Published: August 2023

There is mounting evidence that the lymphatic vessels surrounding the central nervous system (CNS) play an important role in maintaining fluid homeostasis, antigen drainage, waste clearance, and steady-state immune cell migration from the CNS. Recently we reported in a murine mouse model of multiple sclerosis and stroke that during inflammatory brain disease a large number of dendritic cells (DCs) are recruited to the cribriform lymphatics, near the olfactory bulbs. These brain-derived DCs then bind to LECs at the cribriform plate (cp) and are retained at the vessel instead of trafficking straight to lymph nodes forming measurable DC-LEC aggregates. Here in this study, we attempted to characterize these DC-LEC interactions, to better understand the role of these binding events on neuroinflammatory disease. To accomplish this we performed a series of scRNAseq experiments which revealed that during neuroinflammation lymphatic endothelial cells at the cribriform plate up-regulate several factors related to leukocyte adhesion, antigen processing/presentation, and cytokine signaling. Additionally, scRNAseq data suggests that many of these DC-LEC interactions could be immunoregulatory in nature, modifying local immune cells population via the PD-L1/PD1 axis. Together our data suggest that the cribriform lymphatics represent an immunological niche where the inflammatory immune response is regulated.

Supported by grants from NIH 1R01NS126595-01A1
Bi-directional crosstalk between gut microbiota and NKT cells
Authors: Olkhanud Purevdorj B , Rodrigues Richard , Betrapally Naga , Bagheri Hana , Trinchieri Giorgio , Berzofsky Jay A
Published: August 2023

The gut microbiome interacts with the host, including local and systemic immunity. However, the causal mechanisms are relatively under-explored. In this study, we sought to identify bi-directional crosstalk between gut microbiota and type I and/or type II NKT cells using C57BL/6 WT (wild-type), Traj18-knock out (KO) (lacks only type I NKT cells), or CD1d-KO (lacks both type I and II NKT cells) mice. Mice were co-housed with each other strain in combination from the age of day 30 (9 days after weaning) for 30 days and housed separately again for another 30 days. Fecal pellets collected at the age of days 30, 60, and 90 were sequenced by 16S rRNA (250 bp paired-end, Illumina) sequencing, identifying 1,276 amplicon sequence variants (ASVs) by analysis (QIIME2). Microbes with low frequency (<0.5%) or exhibiting cage effects within a strain at day 30 were filtered out. Comparing the microbial abundances between the different groups (co-housing or not) and time points revealed an interesting phenomenon regarding four ASVs (the closest known species was Muribaculum intestinale strain YL27 (16S pident 86.7–95.6)). They showed differential abundance (limma p-value < 0.05) between strains at day 30 (e.g., an ASV_802 was most abundant in WT mice, then CD1-KO, and lowest in Traj18-KO mice). These closely related bacteria were not transferred to Traj18-KO mice when co-housed with WT or CD1-KO mice. Because the ASV is present in WT and CD1d KO mice, even though the latter also lack type I NKT cells as well as type II, our results suggest the existence of specific microbe(s) that cannot grow well in the presence of unopposed type II NKT cells.
Alterations in Germinal Center (GC) Formation and B Cell Activation during Severe Orientia tsutsugamushi Infection in Mice
Authors: Gonzales Casey A , Liang Yuejin , Thiriot Joseph , Sun Jiaren , Soong Lynn
Published: August 2023

Scrub typhus is a poorly studied, but life-threatening, disease caused by the bacterium Orientia tsutsugamushi (Ot). Humoral immunity in scrub typhus patients wanes as early as one-year post-infection; yet its underlying mechanisms remain unclear. The objective of this study is to delineate mechanisms underlying humoral immune dysregulation and B cell dysfunction during acute Ot infection. Using a lethal scrub typhus C57BL/6 murine model, we evaluated splenic GC responses by immunofluorescent staining. Splenic germinal centers (GL-7+ cells) were formed at D4, but were nearly absent at D8, accompanied by randomly scattered T cells. Both infection days had comparable numbers of GC B cells and T follicular helper (Tfh) cells, indicating that GC loss at D8 was less likely due to cell death, but rather dysregulation in GC formation. To confirm this hypothesis, we performed RNAseq analysis of purified B220+ B cells and revealed a set of differentially expressed genes associated with B cell adhesion (S1PR1, ICAM1, GPR183) and co-stimulation (CD40, ICOS-L, SLAMF1) at D8. Notably, the downregulation of S1PR2 (a GC-specific adhesion gene) was most evident at D8, correlating to disrupted GC formation. Furthermore, transcriptomic data revealed significant downregulation of 71% B cell activation genes on D8, suggesting attenuation of B cell activation. Additional studies with low- and high-virulence Ot strains and comparative flow cytometry revealed significant differences in both B cell subsets (plasmablasts, marginal zone B cells) and T cell subsets (Tfh and Tfr cells). This is the first study showing marked dysregulation of B cell responses during Ot infection, which helps understand the transient immunity associated with scrub typhus.

Supported by grants from NIH (R01 AI132674, R21 AI156536, R21 AI153586, R01 EY028773, T32 AI007526), Department of Defense Threat Reduction Agency grant (HDTRA1-19-1-0043), UTMB IHII Data Acquisition (CIRWH&IHII Pilot/ Phase II Research grant) and UTMB Center for Biodefense and Emerging Infectious Diseases (Pilot grant)
Host colonization with cutaneous commensal induces humoral immunity via formation of dermal tertiary lymphoid organs
Authors: Gribonika Inta , Perez-Chaparro Paula Juliana , Chi Liang , Veres Tibor , Germain Ronald , Belkaid Yasmine
Published: August 2023

The skin is one of the largest barrier sites which harbors a plethora of microorganisms. Whether skin microbiota promotes humoral responses remains unknown. Here we show that colonization with common skin commensal Staphylococcus epidermidis profoundly modulates cutaneous immunity that results in serum responsiveness with antigen-specific IgG antibodies. Such response is uncoupled from inflammatory signals and is associated with the induction of dermal tertiary lymphoid organs that closely resemble classical germinal centers. Of note, such responses occur in a tissue autonomous manner as evidenced by the preserved antibody repertoire in mice deficient in professional lymphoid organs. Our data also uncover a non-redundant role for Langerhan’s cells for the generation of commensal specific antibodies. Further, our work proposes a model by which commensal specific antigens are collected in the hair follicle leading to the generation of T follicular helper cells that further expand the local B cell pool in an IL-21-dependent manner. We also found that commensal induced antibodies can protect the host against subsequent infection with the same microbe demonstrating the importance of this line of defense in maintaining barrier protection. Collectively, our observations highlight the autonomous potential of skin organizational flexibility and for the first time demonstrate that B cells represent an indispensable local cutaneous compartment that actively maintains skin immunity.
STAT5-regulated autocrine IL-6 signaling dictates IL-10-dependent regulatory functions of neonatal B10 cells
Authors: Sakai Jiro , Akkoyunlu Mustafa
Published: August 2023

Neonates have suboptimal immune responses to immunization, resulting in higher susceptibility to microbial infection. B cells have a central role in humoral immunity to immunization, and antigen recognition by BCRs is essential for the activation and differentiation of B cells. Neonatal B cells, however, do not respond to BCR activation as robustly as adult cells do. Our hypothesis is that unique features of neonatal BCR signaling may contribute to suboptimal humoral responses. The aim of this project is to decipher the neonatal BCR signaling pathways and identify those that may be responsible for suboptimal B cell functions.

Purified adult and neonatal B cells were stimulated with anti-IgM. RNA-seq analysis revealed that STAT3 and STAT5 signaling pathways were activated in neonatal B cells. STAT3 and STAT5 were phosphorylated in neonatal B cells. BCR-mediated STAT5 phosphorylation induces IL-6 production, which in turn activates STAT3 in an autocrine/paracrine manner. Moreover, IL-6-induced STAT3 activation led to the production of anti-inflammatory cytokine IL-10. Further underscoring the role of IL-6 in IL-10 production, neonatal B cells from IL-6-KO mice did not secrete IL-10. To assess whether the autocrine IL-6 is essential for IL-10-dependent immunosuppression, we incubated peritoneal macrophages with conditioned medium (CM) from adult, wild-type neonatal, or IL-6-KO neonatal B cells. We found that CM from wild-type neonatal B cells suppressed TNF-α production by macrophages in an IL-10-dependent manner, whereas CM from IL-6-KO neonatal B cells or adult B cells did not.

Our studies unveiled the essential role for STAT5-induced autocrine IL-6 signaling in IL-10-mediated immunosuppressive functions of neonatal B cells.
Identification of Neutralizing Anti-HMGB1 IgM Autoantibody and Its B Cell Producers
Authors: Vattikota Anirudh , Geng Yajun , Palani Sunil , Singh Nehal , Chen Aoshuang
Published: August 2023

Extracellular High Mobility Group Box 1 (HMGB1) is a prototypic damage-associated molecular pattern (DAMP). While a homeostatic level of extracellular HMGB1 may be beneficial for immune defense, tissue repair and tissue regeneration, excessive HMGB1 is linked to chronic inflammation and inflammatory diseases. This prompts an intriguing question: How does a healthy body control the level of extracellular HMGB1? Here we have identified, in the plasma of both healthy humans and healthy mice, an anti-HMGB1 IgM autoantibody that neutralizes extracellular HMGB1 via binding specifically to a 100% conserved epitope, namely HMW4 (HMGB1 98–112). In mice, this anti-HMGB1 IgM (i.e., anti-HMW4 IgM) is produced by peritoneal B-1 cells, and concomitant triggering of their B cell receptor and toll-like receptor 4 by extracellular HMGB1 stimulates the production of anti-HMW4 IgM. The ability of extracellular HMGB1 to induce its own neutralizing antibody suggests a feedback loop limiting the level of this DAMP in a healthy body. Currently, our studies are ongoing to identify and characterize the human counterpart B cells that produce the anti-HMGB1 IgM, by first amplifying HMW4-reactive B cells from human peripheral blood mononuclear cells (PBMCs) in NSG recipient mice, in the presence of B cell-stimulating adjuvants (such as B cell mitogens). These studies serve as the prelude to exploring human anti-HMGB1 IgM down the road for treating patients with inflammatory diseases and provide insight pertaining to the role and regulation of other DAMP molecules.
Tissue Microenvironment Reprograms the Circulating Memory T Cell Differentiation and Enhances Vaccine-Elicited Anti-Metastasis Immunity
Authors: Xu Haoran
Published: August 2023

During tumor ontogeny and viral infection, heterologous subsets of CD8+ T lymphocytes provide immediate clearance and durable protection. We utilized model antigen (HIV-1-Gag) expressing melanoma cell line and parabiosis model to investigate the role of different components of memory T cells, including circulating memory T cells (Tcirm) and tissue-resident memory T cells (TRM), in melanoma lung metastasis protection. We first proved that TRM generated by DNA vaccine intramuscular prime followed by influenza-vectored vaccine intranasal boost immunization regime mediated potent and long-lasting anti-lung metastasis immunity, whereas Tcirm induced in different vaccination combinations was not capable to provide sufficient protection. To further explain the mechanism behind the potent TRM generated by prime-boost immunization, we found that DNA vaccination elicited a large amount of central memory T cells (TCM) in the circulation and was preferentially differentiated into lung TRM after the intranasal booster immunization. Single-cell RNA sequencing profile illustrated the tissue microenvironment post mucosal immunization reshaped the differentiation of circulating TCM elicited by systemic immunization and enhanced the anti-metastasis T cell immunity. Thus, our research highlighted the importance of TRM in lung metastasis protection and provided insight into the mechanism of lung TRM establishment in the prime-boost vaccination.
Novel bio-signature for the topical administration of a suppressor of cytokine signaling-1 mimetic to the equine eye: implications in treatment of Recurrent Uveitis (RU)
Authors: Stafford Lauren Stewart , Vicuna Valeria , Plummer Caryn , Smith W Clay , Larkin Joseph
Published: August 2023

Recurrent uveitis (RU) is a leading cause of ocular decrement in both humans and horses, which can lead to blindness. Equine recurrent uveitis (ERU) serves as the only spontaneous model for human RU as the eye destruction in both diseases is guided by a dysregulated cytokine milieu mediated by T lymphocytes. We have recently shown that the topical administration of a peptide (SOCS1-KIR), which mimics a natural regulator of cytokine signaling, was correlated with a reduction in clinical pathology when applied to the eye of horses with ERU. This administration was without any observed toxicity. The purpose of this study is to establish the intraocular presence of topical SOCS1-KIR treatment. As such, we administered SOCS1-KIR to the eye of experimental horses (lacking ocular pathology) to evaluate peptide localization and detection of a biological signature. SOCS1-KIR peptide, that was intravitreally-injected, was readily detectable within the retina by immunofluorescence. Although we were unable to detect topically administered SOCS1-KIR by immunofluorescence, topical administration reduced levels of TNFa and IL-10 in the aqueous humor of treated eyes. Additionally, we found that SOCS1-KIR modulated TNFa, IL-6, IL-10, RANTES, and IL-8 levels within activated healthy equine PBMC. Together these results suggest a novel biological signature that is mediated by topical SOCS1-KIR administration to the equine eye that has implications for translation to the clinical setting.

This work was supported by the Grayson Jockey Club grant
Pre-clinical pharmacologic and tolerability characterization of a novel KIRxCD8 targeting bispecific CD8 Treg modulator
Authors: McMahan Catherine J , Pham Minh , Boster Daniel , Hermansky Gleda , Gardell Jennifer , Bowser Justin , Rachael Fasnacht , Chen Alex , Gilbertson Emily , Morgan Nadine , Childs Monica , Fitzpatrick Kristin , Tan Cong , Mauer Meghan , Meengs Brent , Therriault Jon , Patton Dan , Julien Susan , Crane Courtney , Swiderek Kristine M
Published: August 2023

We have characterized a subset of CD8 T cells (CD8 Treg) with immunosuppressive characteristics in inflammatory disease settings. Here we describe the binding and specificity profiles of a novel bispecific CD8 Treg modulator in vitro, ex vivo, and in vivo, and initial assessment of tolerability and pharmacology in a humanized mouse model.

We evaluated the binding, pharmacokinetics (PK), and early tolerability of the bispecific CD8 Treg modulator targeting CD8 and KIR2DL1/2/3. Despite single arm binding to NK cells via the KIR targeting arm and CD8 T cells by the CD8 targeting arm of the bispecific, we did not detect activation of immune cells or increased pro-inflammatory cytokines in vitro, nor did we observe any contribution of NK cells to the elimination of pathogenic CD4 T cells. In healthy mice, the CD8 Treg modulator had a PK profile consistent with antibody molecules; treatment delayed disease onset in an acute GVHD setting. In humanized CD34-engrafted NSG-IL-15 transgenic mice, supporting engraftment of NK cells, no activation of immune cells in peripheral blood or terminal tissues or induction of proinflammatory cytokines in the serum was observed following a single dose of the CD8 Treg modulator.

The recently described CD8 Treg network appears dysfunctional in autoimmune diseases. The KIRxCD8 targeting bispecific modulator targets this network, engages and activates CD8 Treg and reduces inflammation without increasing unwanted immune cell activation or pro-inflammatory cytokines, both in vitro and in vivo. Our early data suggest that KIR-mediated enhancement of CD8 Treg function using a bispecific modulator is a broadly applicable and a promising CD8 Treg specific therapeutic modality for the treatment of autoimmune disease.
Prevention of Type 1 Diabetes via a Lipid Nanoparticle-Based JAK inhibitor Treatment that Involves Anergy Formation
Authors: Hou Shenda , Zhang Yichuan , Wang Jialu , Calderon-Colon Xiomara , Tiburzi Olivia , Iglesias-Lozano Marcos , Patrone Julia , Raimondi Giorgio
Published: August 2023

Type 1 diabetes is an untreatable autoimmune disease caused by the destruction of pancreatic beta cells by autoreactive T cells. The inhibition of inflammatory cytokines signaling via JAK inhibitors like Tofacitinib (Tofa) has shown promise in the prevention of this disease. To improve on drug bioavailability and reduce deleterious systemic exposure, we identified a lipid nanoparticle (LNp) formulation with favorable Tofa encapsulation efficiency and negligible cellular toxicity. Live animal imaging indicated these particles have the unique property of accumulating in lymphoid tissues, particularly in pancreatic and mesenteric lymph nodes when administrated via oral gavage. Spectral flow analysis revealed that multiple immune cells, like dendritic cells, T cells, and B cells, uptake the particles in these tissues. Short-term administration (5 gavages once every other day) of Tofa-LNp early (3-week-old) or late (10-week-old) in disease prone NOD mice promoted a significant delay of disease onset. Importantly, the therapeutic effect was associated with a profound increase in anergic CD4 T cells in the draining sites of LNp. Anergic cells have been known to form following stimulation in absence of co-stimulation, however, additional in vitro stimulation assays suggest a distinctive path of anergy induction promoted by blocking cytokine signaling. Overall, our results suggest an unexpected and novel immunomodulatory effect of JAK inhibition that warrants further investigation for the development of an effective treatment of type 1 diabetes.
The Molecular Underpinnings of Treg Plasticity in Cancer
Authors: Friedman David J , Osman Abu , Tousinas Georgios , Marquez Daisy , Carapanceanu Nicoleta , Carapanceanu Valentin , Khazaie Khashayarsha , Gounari Fotini
Published: August 2023

Colorectal cancer (CRC) is one of the most common cancers in the world, yet it remains one of the most difficult to treat. Immunotherapy of CRC using checkpoint inhibitors is only efficacious in approximately 4% of patients with high microsatellite instability (MSI-H). In earlier studies we showed that in patients with inflammatory bowel diseases (IBD) and CRC as well as in the APCΔ468 murine polyposis model, Tregs overexpress β-catenin. Furthermore, we demonstrated that expression of a dominant active β-catenin in Tregs induces RORγt, compromising suppressive activity and promoting TH17-like properties. This results in a scurfy-like phenotype. By contrast loss of TCF-1 upregulated RORγt, but only compromised Treg anti-inflammatory functions. These observations suggested cooperation of β-catenin and TCF-1 in regulating Treg plasticity. We show here that in healthy mice, β-catenin expression is significantly higher in Tregs as compared to conventional CD4+ T-cells. In Tregs that express a dominant active β-catenin, ablation of TCF-1 or HEB provide partial rescue, while ablation of both TCF-1 and HEB recovers Treg numbers and prevents the scurfy-like phenotype. These observations suggest that TCF-1, HEB and β-catenin cooperate to determine Treg functions. In search for additional partners, we show that loss of TCF-1 upregulates SATB-1, a chromatin modifier that is known to determine T-cell development and function. Upregulation of SATB-1 is enhanced with the loss of both TCF-1 and HEB. All three transcription factors upregulate RORγt suggesting that they cooperatively determine Treg function and plasticity. Elucidating this cooperation will enhance our ability to control Treg plasticity in health and disease.

NIH/NIAID - R01 AI108682-0A1
Gut microbiota changed by tryptophan modulates anti-tumor CTL responses against brain tumor
Authors: Kim Hyeon Cheol , Kim Hyun-Jin , Lee Heung Kyu
Published: August 2023

Glioblastoma multiforme (GBM) is one of the most common brain tumors, and both conventional therapies and current immunotherapies are not effective. Thus, a novel approach is required for GBM treatment. Commensal microbiota reside in the body and are involved in both healthy and disease states of the host. To date, research into the role of microbiota has mainly focused on the gastrointestinal tract. It remains unclear how gut microbes may affect brain and how they may play a role in GBM. Here, we observed a distinct change in gut microbial composition and metabolism during GBM progression using 16S rRNA sequencing in two different animal models. The amount of tryptophan was significantly reduced in GBM-bearing mice. Based on these results, we considered tryptophan supplementation. This diet improved the survival in a commensal microbiota-dependent manner. Surprisingly, some microbiota were restored by tryptophan to levels comparable to those of normal mice. Next, we observed improved survival of GBM-bearing mice co-housed with normal mice. Furthermore, we identified a bacterial species that was present at significantly different levels depending co-housing. Colonization with this bacterium led to increased survival. This bacterium required tryptophan for its growth. Cytotoxic T cells showed more potent immune responses in the tumor microenvironment in tryptophan-supplemented mice and mice colonized with the bacterium. In addition, bacterial metabolites increased apoptosis in glioma tumor cell lines. Collectively, we identified a bacterial species that is critical for defense against GBM and demonstrated that this bacterium potentiates cytotoxic T lymphocyte effector functions and induces the apoptosis of tumor cells.
Loss of testosterone leads to impaired T cell function via glucocorticoids in mouse glioblastoma models
Authors: Lee Juyeun , Chung Yoon-Mi , Sharifi Nima , Lathia Justin
Published: August 2023

While sex differences in glioblastoma (GBM) are well established, the contribution of sex hormones on these differences has not been fully explored. Recently, the immunoregulatory effect of testosterone in other type of cancers has been reported. To evaluate the effect of testosterone on GBM, we performed castration or sham surgery on mice, followed by intracranial implantation of syngeneic glioma cells. Surprisingly, the castrated B6 mice had more aggressive tumors compared to sham surgery controls and the castration induced aggressive tumor growth could be rescued by exogenous testosterone administration. This was further confirmed by chemical blockade of androgen receptor signaling using enzalutamide. In contrast, castration in Rag1KO mouse models did not show a survival difference compared to controls, suggesting a role for lymphocytes. Interestingly, upregulated levels of circulating corticosterone in castrated mice was detected by mass spectrometry, and glucocorticoid receptor blocker reversed the survival of castrated mice, suggesting glucocorticoid-mediated systemic immunosuppression upon castration. Flow analysis revealed decreased cytokine production in CD4+ and CD8+ T cells from tumors and lymph nodes in castrated mice. We also found that tumor growth was delayed in castrated mice when tumor cells were subcutaneously implanted in the flank while decreased cytokine production in T cells in tumor and lymph nodes from flank tumor model as well. These data suggest brain-specific effect of steroid hormone on tumor control. Taken together, our findings demonstrate that testosterone regulates T cell function via glucocorticoids, and ultimately affects GBM tumor growth in a site-specific manner.

Supported by grants from NIH (R35 NS127083, P01 CA245705).
Differential expression of immune receptors in pediatric Acute Lymphoblastic Leukemia (ALL)
Authors: Mathew Stephen O , Ahmed Nourhan G , Powers Sheila , Jose Roslin , Mathew Porunelloor A
Published: August 2023

Acute Lymphoblastic Leukemia (ALL) represents the most common pediatric cancer. Significant improvements in chemotherapy regimens caused a leap in the 5-year survival rates to surpass 90%. However, relapse significantly lowers the chances of survival to less than 50% in some cases due to resistance developed by malignant cells to chemotherapy. Natural Killer (NK) cells are a promising immunotherapy alternative for their ability to recognize and kill malignant cells directly through activation or inhibition signals or through cytokine release. We have previously identified 2B4 (SLAMF4), CS1 (SLAMF7) and LLT1 that can activate or inhibit NK cells upon the interaction with their ligands. In this study, the expression of 2B4, CS1, LLT1, NKp30 and NKp46 was determined in pediatric ALL and healthy subjects. Whole blood was collected from 42 pediatric ALL subjects before and after induction chemotherapy and the surface expression of receptors/ligand on T-cells, B-cells, monocytes, and NK-cells was compared with blood samples from 20 healthy subjects. The flow cytometry data showed a significant increase in CS1 and NKp46 expression on monocytes and LLT1 in T cells and monocytes of ALL subjects before treatment. A decrease of LLT1, 2B4, CS1 and NKp46 on T-cells of ALL subjects was also observed after treatment along with elevated expression of LLT1 on ALL subjects’ NK-cells before and after treatment. Furthermore, mRNA data showed altered expression of receptors in ALL subjects pre and post induction chemotherapy treatment. The results indicate that the differential expression of the receptors/ligand may play a role in the T cell and NK-cell mediated immune surveillance of pediatric ALL.

Supported by funds from Cancer Research Foundation of North Texas and Team Connor Childhood Cancer Foundation
Analysis of the protumoral mechanisms of F.nucleatum on growth, epithelial-mesenchymal transition and the expression of immunosuppressive markers in cell line of OSCC
Authors: Munoz Camila Paz , Ferrada Luciano , Riquelme Erick , Zuñiga Felipe , González Wilfredo , Betancur Daniel , Oñate Angel , Novoa Sergio Andrés Sanhueza , Cabrera-Garcia Camilo Daniel , Quiroga Romina Andrea , Cáceres Bárbara Evelyn Antilef , Labarca Gonzalo , Nova-Lamperti Estefania , Fernandez Liliana Lampert
Published: August 2023

Background

Oral squamous cell carcinoma (OSCC) is the most common manifestation of oral cancer. It has been proposed that periodontal pathogens contribute to OSCC progression, and preliminary data from our laboratory has identified Fusobacterium nucleatum (F.n) and Epithelial Mesenchymal Transition (EMT) markers in the secretome of biopsies from patients with OSCC in comparison with healthy biopsies from the oral cavity. However, the mechanisms modulated by the tumoral bacteriome in OSCC remains not fully understood.

Methods

Oral cancer cells (HSC3) were infected with F.n using a MOI of 100 to evaluate the effect of the bacteria on tumor growth, expression of EMT markers and immunomodulatory properties.

Results

After infection, we confirmed the presence of bacteria inside and surrounding the cancer cells by confocal microscopy. Then, a significant increment in the size of F.n-infected tumor spheroids was found at 3-, 6- and 10-days post-infection. Related to EMT markers, MMP-9 transcripts were significantly elevated in infected cells, whereas E-cadherin transcripts were significantly downregulated post infection. In addition, the Human XL Oncology array kit revealed that several cancer proteins such as MMP-3, GM-CSF, FOXO1/FKHR, Cathepsina D-S, Amphiregulin, SerpinB5/Maspin and EGFR were upregulated and others like ICAM-1, DLL-1 and ERB2 were downregulated after F.n-infection. Finally, the expression of Galectin-9 measured by flow cytometry was significantly increased in infected cells than control cancer cells.

Conclusions

The periodontal bacterium F.n promotes tumor progression of OSCC through increased tumor growth, acquisition of ETM-associated markers and Galectin-9 upregulation.

Fondecyt Regular Project 1211480
Prepregnancy Hysteroscopic Image in a Patient in whom Spontaneous Uterine Rupture Occurred in the 27th Week of Pregnancy after Adenomyomectomy
Authors: Matsushima Takashi , Kuroki Mutsumi
Published: August 2023

We describe a preconception hysteroscopic image of a patient with a ruptured uterus at 27 weeks’ gestation. A 40-year-old gravida 2, para 1, underwent open adenomyomectomy because of infertility. Subsequently, hysteroscopy performed at our hospital revealed an endometrial deficit from the uterine fundus to the posterior wall, and an area where the endometrium was missing and composed of yellow tissue was seen. She later achieved pregnancy. Lower abdominal pain occurred on day 1 of the 27th week of pregnancy. She suddenly went into a state of shock. Emergency laparotomy was performed, and a uterine rupture wound of approximately 10 cm in the longitudinal direction was seen in the posterior wall. A 1120-g male infant was stillborn. Total blood loss was 6450 mL. The mother was saved without hysterectomy. After adenomyomectomy, a hysteroscopy should be performed to check for endometrial defects before allowing pregnancy.
Timely Laparoscopic Intervention for Ovarian Tumor-related Autoimmune Encephalitis: A Challenging Pathology at Tu Du Hospital in Vietnam and Literature Review
Authors: Pham Thanh Hai , Nguyen Phuc Nhon , Ho Quang Nhat
Published: August 2023

A previously fit and healthy 39-year-old woman was admitted to our tertiary referral hospital with coexisting autoimmune encephalopathy and ovarian tumor. Due to the presence of anti-N-methyl-d-aspartate receptor (anti-NMDAR) in the cerebrospinal fluid, a diagnosis of anti-NMDAR encephalitis was first suggested after ruling out other etiologies. Thus, a laparoscopy was promptly performed to remove the ovarian tumor. The histological endpoint revealed an ovarian teratoma. Consequently, the patient recovered completely in good health condition after 2 months in a coma status. Herein, we report an uncommon case of anti-NMDAR encephalitis associated with ovarian teratoma at our hospital, thus raising awareness of physicians.
Safety of Laparoscopic Cholecystectomy for Cholecystitis during Pregnancy
Authors: Itaimi Ahmed , Abbassi Imed , Baraket Oussama , Kotti Ahmed , Triki Wissem , Bouchoucha Sami
Published: August 2023

Objectives:

The aim of this study is to evaluate the safety of laparoscopic cholecystectomy to treat acute cholecystitis during pregnancy.

Materials and Methods:

We conducted a retrospective multicenter study including pregnant women with acute cholecystitis managed in surgery departments in Tunisia from January 1, 2015, to December 31, 2019.

Results:

Seventeen centers of surgery department participated in this study including 107 cases of acute cholecystitis. The average maternal age was 30.5 years. Nonoperative management was performed in eight patients, whereas 99 other patients had surgery. Postoperative follow-up was uneventful in 93.8% of cases and eventful in 6.2% of cases. There was no mortality as far. A medical complication occurred in two patients with a medical morbidity rate of 1.7%. It was about thromboembolic disease. A surgical complication occurred in two other patients with a surgical morbidity rate of 1.7%. It was about intraperitoneal infection in one case and biliary collection in the other case. In univariate analysis, variables related significantly to maternal complication were: age equal or over 35 years old (P = 0.001), jaundice (P = 0.024), C-reactive protein value equal or over 20 mg/L (P
= 0.05), and biliary peritonitis (P = 0.05). In multivariate analysis, independent variable predictive of maternal complications was age equal or over 35 years old (P = 0.003), jaundice (P = 0.003), and biliary peritonitis (P = 0.011).

Conclusion:

Laparoscopic cholecystectomy for cholecystitis can be safely achieved in pregnant women with low rates of morbidity and mortality. This study showed that independent variable predictive of maternal complications was age equal or over 35 years old, jaundice, and biliary peritonitis.
Hysterectomy through Minimally Invasive Surgery for Cervical High-grade Intraepithelial Neoplasia: Reassessment of the Specimens’ Eligibility for Histological Examination
Authors: Kurihara Shuichi , Hamasaki Yoichiro , Onjo Sachiko , Nishiyama Kenichi , Nishida Makoto
Published: August 2023

Objectives:

The objective was to investigate the microscopic artifacts made in the uterus of cervical high-grade squamous intraepithelial lesion (HSIL) resected by hysterectomy through minimally invasive (H-MI) procedures and to verify whether these specimens are suitable for histopathological assessment.

Materials and Methods:

This single-center retrospective study analyzed 28 patients with cervical HSIL, consisting of 21 premenopausal and seven postmenopausal women, who underwent H-MI. The proportion of the cervical mucosa covered by intact surface epithelium (residual ratio [RR]) was measured on microscopically. Surgical margin’s status was also verified.

Results:

All cases developed detachment of the cervical surface epithelium to a varying extent. The RR was significantly higher in the premenopausal patients (median: 75.5%) than in the postmenopausal patients (median: 37.6%). Among the premenopausal patients, the RR was lower in the cases on whom uterine manipulator (UM) was used (median: 70.5%) than in the cases without UM use (median 92.7%). Among the 21 cases whose resected uterus contained HSIL, the vaginal resection margin was not assessable in three (14.2%) of the seven postmenopausal cases due to the artifact.

Conclusion:

Although transvaginal manipulation of the uterus causes detachment of the cervical surface epithelium, H-MI for cervical HSIL provides an acceptable specimen for histological assessment in premenopausal patients, even if UM is used. In postmenopausal women, H-MI easily develops artifactual loss of cervical surface epithelium, sometimes providing an unfavorable specimen for microscopic assessment.
Dermatological Manifestations in COVID-Related Illness in Children: A Retrospective Observational Study at a Tertiary Care Hospital of Eastern India
Authors: Purkait Radheshyam , Roy Kakali , Manna Nivedita , Samanta Moumita , Mahapatra Tapan Kumar Sinha
Published: August 2023

Objectives:

To identify the spectrum of cutaneous manifestations and to evaluate temporal relationship between each type of dermatological lesions and the severity of COVID-19-related illness in the pediatric population.

Subjects and Methods:

Retrospective observational study on 35 children (upto12 years) from Kolkata and different district areas of West Bengal, admitted to our tertiary care hospital with COVID-related illnesses (COVID-19 and/or multisystem inflammatory syndrome in children [MIS-C]).

Results:

Maculopapular rash was heading the list (n = 18, 51.4%) followed by chilblain-like lesions (n = 12, 34.2%), vasculistic lesion (n = 8, 22.8%), vesicular rash (n = 5, 14.3%), and urticaria (n = 3, 8.5%). In majority of patients (n = 26, 74.2%), dermatologic manifestations proceeded (n = 16, 45.7%) or merged (n = 10, 28.6%) with the onset of systemic symptoms. In rest of the patients (n = 9, 25.7%), skin manifestations occurred after the systemic symptoms, particularly in vascular lesions. 57.1% children required pediatric intensive care unit (PICU) admission. All children with vasculitis (n = 8) required PICU admission and 87.5% (n = 7) of them needed inotrope support. Intravenous immunoglobulin (IVIG) was mostly given in children with chilblains (n = 11, 92%). Methylprednisolone and repeat dose of IVIG was mostly needed in patients with vasculitis (75% and 25% of vasculitis children, respectively). The duration of PICU stay was least in patients with vesicular rash (4.4 ± 2.5 days) and longest in vasculistic lesions (18.75 ± 1 days), highest being 56 days. The overall mortality rate among MIS-C patients with cutaneous manifestations was 25.7% (n = 9) while patients with vasculistic lesions had the highest mortality rate of 50% (n = 4).

Conclusion:

Our study finding reveals that COVID-related disease severity is highly depends on the type of skin lesions but not just on mere occurrence of skin manifestations. Lesions such as maculopapular, chilblains, and vesicular rash had good prognosis and urticaria and vasculitis had poor prognosis.
Acute-onset Alopecia in a Child: A Surrogate Marker of Azathioprine-induced Myelotoxicity
Authors: Mendiratta Vibhu , Agarwal Akanksha , Singh Smita
Published: August 2023

Azathioprine (AZA) is a widely used immunosuppressive which when not monitored cautiously can lead to life-threatening complications like myelotoxicity. In resource-limited countries, acute hair loss starting as early as 2 weeks after drug initiation can be used as a surrogate marker of impending AZA-induced myelotoxicity.
Giloy (Tinospora Cordifolia): A COVID-19 Herbal Drug Causing Erythroderma in a Pediatric Patient
Authors: Gupta Apoorva , Bansal Mansi , Gulati Sanjeev , Gupta Vatsala , Sharma Vinod Kumar
Published: August 2023

Erythroderma or exfoliative dermatitis is characterized by erythema and scaling affecting more than 90% of the body surface area. Drug-induced erythrodermas are relatively uncommon in children but can be potentially life-threatening. We hereby report an unusual case of erythroderma due to oral intake of Giloy (Tinospora
cordifolia) in a 10-year-old child.
Unilateral Eyelid Angioedema: An Atypical Manifestation in Childhood IgA Vasculitis
Authors: Deepashree R. , Prasad Arakali Lakshminarayana Shyam
Published: August 2023

Immunoglobulin A (IgA) vasculitis (Henoch–Schönlein purpura) is an immune-mediated small vessel vasculitis, in which edema of the nondependent areas of the body, including the face and scalp is a rare cutaneous finding, found mostly in children <2 years of age. We report a case of a 7-year-old girl presenting with unilateral eyelid angioedema as a manifestation of IgA vasculitis. Early diagnosis and treatment are necessary, as the risk of renal complications increases with the incidence of angioedema.
Clinicoradiological parameters predicting operative difficulty in laparoscopic partial nephrectomy for renal tumors
Authors: Pirzada Faisal Masood , Sood Rajeev , Taneja Anil , Sharma Umesh , Goel Hemant Kumar , Gahlawat Sumit
Published: August 2023

Introduction:

The number of renal tumors amenable to laparoscopic surgery is rising, both, due early detection by improved imaging techniques and due to progressive improvements in minimal access surgery. Conversion to open surgery, which is a significant event, can be minimized by proper case selection. We assessed the pre-operative factors that can predict the operative difficulty and can help in case selection, thus avoiding complications and reducing the chances of conversion to open.

Methods:

One hundred and sixteen patients (73 males and 43 females) with the mean age of 50.78 ± 14.2 years, meeting the inclusion criteria underwent transperitoneal laparoscopic partial nephrectomy (LPN). Various clinical, anthropometric, radiological, and pathological parameters were recorded. Intraoperative difficulty was assessed and graded on a scale of 1 (easiest) to 4 (most difficult or open conversion) by an independent observer to calculate the difficulty score, which along with the other parameters of operative difficulty, was used to calculate the difficulty scale. Significant parameters on the univariate analysis, were subjected to a multivariate analysis, to find parameters that can predict the operative difficulty.

Results:

The mean age was 52 ± 14.29 years, mean size was 4 ± 1.04 cm, male:female ratio was 1.6:1, most of the tumors were exophytic (60%) and anteriorly located (62%) and had a mean perinephric fat surface density (PnFSD) of 6446.026 ± 2244 surface density pixel units (SDPU). On the univariate analysis, age >60 years, Eastern Cooperative Oncology Group performance score >1, presence of perinephric fat stranding, increased PnFSD (>10,000 SDPU), large tumor size (>4 cm), hilar/posterior location, endophytic tumors and higher clinical stage were significantly associated with intraoperative difficulty. However, on the multivariate analysis, no single factor could independently predict intraoperative difficulty in LPN for Renal tumors.

Conclusion:

It is difficult to predict the intra-operative difficulty during LPN. Feasibility of LPN should be based on multiple factors rather than a single factor.
Intervertebral disc degeneration and vertebral end plate damage in acute lumbar disc herniation
Authors: Nehru Arul P , Kanna Rishi M , Shetty Ajoy P , Shanmuganathan Rajasekaran
Published: August 2023

Background:

Although microtrauma, gene polymorphisms, or subclinical infection could initiate lumbar disc herniation (LDH), the final pathway culminating in herniation can involve two possible trajectories—degeneration of nucleus pulposus or vertebral end plate (EP) damage. We performed a retrospective case–control study to understand the relative role played by disc degeneration (DD) and EP changes in acute LDH.

Materials and Methods:

MRI of 300 consecutive patients with acute LDH (<6 weeks) was evaluated for the type of LDH, severity of DD by Pfirrmann’s grading, and EP Modic changes (MC) for its type, distribution, and location in EPs. Among the1500 discs evaluated in 300 patients, 308 had disc herniation (cases) and 1192 acted as controls.

Results:

98.8% of LDH had grade ≥3 DD (P < 0.001). The mean DD was higher in herniated discs (P < 0.05). From a mean 1.98 in normal discs, it increased to 3.31 in disc bulges, 3.73 in disc protrusions/extrusions, and 3.83 in sequestrations. Herniated discs had more MCs than normal discs (38.3%, 118/308) (P < 0.001). Although in normal discs, only 7.7% had MC, herniated disc subtypes showed a progressive increase in the incidence of MC (26.2% in disc bulges, 37.5% in disc protrusion/extrusions, and 58.3% in sequestrations). MC on both EP was strongly associated with LDH (odds ratio = 9.76). Posterior corner MC had a significant association with LDH (72.72%, P < 0.001).

Conclusion:

DD seems to be a common thread in all patients (98.8%) with LDH, whereas EP damage seems to be specific pathway in a subset (38.3%). The study showed a significant degeneration of nucleus pulposus in most (98.8%) herniated discs. Compared with nonherniated discs, vertebral MC had a higher incidence in herniated discs (38%), and interestingly these were of subacute fatty type 2.