bluuebunny/crossref_metadata_2025 huggingface dataset:
Mapping the cellular landscape of the maternal-fetal interface in women with preterm birth and chronic chorioamnionitis
Authors: Levenson Dustyn , Xu Yi , Garcia-Flores Valeria , Galaz Jose , Neshek Barbara , Romero Roberto , Gomez-Lopez Nardhy
Published: November 2024

Preterm birth (PTB) affects over 15 million infants yearly. In up to 40% of late PTBs, placental examination reveals chronic chorioamnionitis (CCA), maternal mononuclear cell infiltration in the chorioamniotic membranes; yet, the cellular landscape remains fully unknown. Herein, we first used imaging mass cytometry (IMC) to map the phenotype and spatial distribution of immune cells at the maternal-fetal interface of women with PTB and CCA and non-CCA controls (n = 4 each). Cases exhibited a 5-fold increase in T cells, including effector memory CD4+ (p=0.02) and proliferative CD8+ T cells (p=0.04), and a 7-fold rise in CD163+CD206+ macrophages (p=0.05) compared to controls. NK cells were also present in both groups. Neighborhood analysis revealed that CD8+ T cells are in proximity to macrophages and CD4+Tbet+ T cells, and distanced from NK cells. Next, we performed spatial transcriptomics (STx) in each study group (n = 8 PTB with CCA, n = 7 PTB no CCA). STx highlighted diversity in several clusters of T cells, macrophages, and other immune cells found to be transcriptionally active in CCA. Gene ontology analysis indicated enrichment for CD8+ T cell activation, as well as IL-12 and IFNγ signaling in macrophages in CCA. Ongoing efforts are focused on validation and functional testing of such immune cells in order to unveil the complex interactions at the maternal-fetal interface in CCA, a placenta pathology associated with adverse outcomes in the mother and offspring.
Fungal Chitin Does Not Contribute To Allergic Asthma Severity
Authors: Ellis Diandra , Makullah Mgayya , Jones Mary Jane , Cheung Suki , Willems Marjoleine , Steele Chad
Published: November 2024

Chitin, a polysaccharide found in the fungal cell wall and exoskeletons of house dust mites and cockroaches, has garnered attention as a potential immunoreactive allergen. Mammals have evolved to express chitin-degrading chitinases that may modulate immune responses to chitin. We have previously reported that mice deficient in acidic mammalian chitinase (AMCase, Chia) demonstrated better lung function (i.e. airway hyperresponsiveness, AHR) during allergic fungal asthma. Here, we show that during allergic fungal asthma, mice overexpressing AMCase (SPAM mice) had worse lung function, and chitin-bearing conidia from Aspergillus fumigatus are detectable in the lung. The lung function of mutant mice directly correlated with the level of chitinase activity during allergic fungal asthma (Chia-/- mice, low chitinase activity, lower AHR; SPAM mice, high chitinase activity, higher AHR), suggesting that chitin catabolism promoted AHR. Chronic exposure of normal mice to purified chitin from A. fumigatus resulted in moderate inflammatory changes in the lung, however this was not sufficient to induce AHR. Moreover, chronic exposure of mutant mice to purified chitin did not induce AHR. Collectively, these results indicate that 1) fungal chitin alone is incapable of driving AHR and 2) chitinase-mediated degradation of A. fumigatus chitin may facilitate unmasking and/or liberation of other fungal cell wall components that preferentially drive inflammatory responses which contribute to AHR.
Teaching Immunotherapy at College of Pharmacy-Therapy-based Learning
Authors: Fazal Nadeem , Graefen Bahar
Published: November 2024

Different types of immunotherapies employed as chemotherapy work in different ways. Some immunotherapy treatments help the immune system stop or slow the growth of cancer cells. Immunology course for Pharmacy year 2 students is taught at College of Pharmacy Chicago State University. The perquisites for the course were Medical Physiology, Biochemistry and Pathophysiology. The students had no prior knowledge of basic immunology. The way the course was designed was that they learn about the current available immunotherapy and then were taken back to the basic concepts of immunology to make sense of that module. We divided Pharmacy Immunology course into five modules, i.e., 1) Monoclonal antibodies and immune checkpoint inhibitors, 2) Non-specific immunotherapies, 3) Oncolytic virus therapy, 4) T-cell therapy, 5) Cancer vaccines. Although the students were taught immunotherapies in two-hour long workshops as discussion in teams, they were also tested for their conceptual knowledge of basic immunology. The assessment results consistently showed a higher level of learning according to Bloom’s taxonomy. The exams were designed to test critical thinking and application and synthesis of knowledge of the students. The assessment data suggested that student performance was significantly improved as compared to previous years of Pharmacy year 2 students who were taught similar content without using this teaching methodology.
A novel mouse model of silicosis and silica-induced inflammatory arthritis.
Authors: de Ocampo Caroline , Mayeux Jessica , Kono Dwight , Pollard Michael , Janssen Lisa
Published: November 2024

Introduction: A strong association between silica dust exposure and development of systemic autoimmune diseases, including rheumatoid arthritis (RA), is widely recognized; however, a direct mechanism has yet to be established. To elucidate this connection, we investigated the role of pulmonary silica exposure in promoting silicosis and RA in BXD2/TyJ mice.

Methods: BXD2/TyJ mice exposed to 5 mg crystalline silica or PBS and examined 1-, 2-, 6-, 12-, 20-weeks later. Total immunoglobulin-G and -M, as well as autoantibody levels, measured in serum and bronchioalveolar lavage fluid (BALF), and lung and ankle tissue assessed histologically. Immune cells characterized in lung, spleen, and tracheobronchial lymph node with flow cytometry. B and T cell-containing pulmonary lymphoid accumulations visualized with immunofluorescent staining.

Results: Severe silicosis was observed, and B and T cell-containing lymphoid accumulations were detected in the lung as early as 2-weeks following silica exposure. Markedly increased levels of a range of autoantibodies were detected beginning 2-weeks after silica exposure in BALF and 6-weeks in serum. Significantly higher ankle synovitis and bone erosion was observed beginning at 12-weeks post silica exposure compared to controls.

Conclusion: Silica exposure accelerates severe inflammatory arthritis in a susceptible background, establishing a model to study the connection between mucosal inflammation and rheumatoid arthritis.
Circadian clock disruption exacerbates noncanonical inflammasome-dependent pyroptosis via Rev-erbα-SAA1 axis
Authors: Choi Bokeum , Shim Do-Wan , Eo Jun-Cheol , Kim Saeyoung , Yu Je-Wook
Published: November 2024

Circadian arrhythmia has been linked to the increased susceptibility of multiple inflammatory diseases. However, it remains elusive how disruption of circadian clock modulates innate immune responses including inflammasome signaling. Here, we examined a potential role of circadian clock on inflammasome-mediated responses through myeloid-specific deletion of BMAL1, a master circadian clock regulator. Intriguingly, Bmal1 deficiency significantly enhanced macrophage pyroptosis and lethality of mice under noncanonical inflammasome-activating conditions, but did not alter canonical inflammasome responses. Transcriptome analysis revealed that Bmal1 deficiency led to a marked reduction in Rev-erbα expression at steady state and a significant increase in serum amyloid A1 (SAA1) expression upon poly(I:C) stimulation. We found that Rev-erbα is critical for poly(I:C)-induced SAA1 production, resulting in the circadian oscillation pattern of SAA1 expression in myeloid cells. Mechanistically, Rev-erbα inhibited poly(I:C)-induced association of C/EBPβ to the promoter region of Saa1 leading to the reduced transcription of Saa1 in macrophages. Bmal1-deficient macrophages exhibited enhanced binding of C/EBPβ to Saa1. Collectively, our data demonstrate a potent suppressive role of circadian clock BMAL1 on noncanonical inflammasome response via Rev-erbα-C/EBPβ-SAA1 axis and further suggest that circadian arrhythmia can enhance the susceptibility to noncanonical inflammasome-mediated toxicity.
Building a neural network model to define DNA sequence specificity in V(D)J recombination
Authors: Harris Justin , Fagg Andrew , Wu Dee , Rodgers Karla
Published: November 2024

The initial formation of antigen receptor repertoires occurs in developing B and T cells through V(D)J recombination, which results in the joining of two-to-three gene segments that encode for the antigen binding sites of immunoglobulin and T cell receptors. For a gene segment to be targeted for recombination it must be flanked by a semi-conserved recombination signal sequence (RSS). During each V(D)J recombination event, the RAG1/2 recombinase creates DNA double stranded breaks at two RSS substrates that are then repaired to yield the recombined receptor gene. Gene segments are utilized nonequally in generation of the repertoires. The local regulatory and chromatin environment surrounding gene segments and their flanking RSSs have helped to partially explain the nonuniform use of RSSs across each antigen receptor locus. However, each of these loci have distinctive environments, complicating the generalization of RSS usage in V(D)J recombination across loci. Using recombination data of partially randomized 12RSSs, obtained in a chromatin-free environment, a neural network (NN) was built that delineates how changes in sequence at certain positions in the RSS can affect recombination level. External validation revealed the specific effects of distal regions of the RSS on recombination level. Our NN model demonstrates key principles of the entire RSS sequence in defining gene segment utilization in V(D)J recombination.
Lung Inflammation and Pathology are Mast Cell Dependent in a Model of Formaldehyde Toxicity.
Authors: Gibb Matthew , Reinert Angela , Bauer Alison , Brown Jared
Published: November 2024

The mucosal surface of the respiratory tract is continually exposed to the external environment, including potential toxicants. Mast cells are located throughout the lung and are capable of rapid activation, degranulation, and cytokine release, leading to allergic inflammation and disease. Formaldehyde, a ubiquitous chemical, activates mast cells contributing to airway hyperresponsiveness with subsequent neutrophil recruitment and activation, but little is known about the contribution of mast cells following acute, high dose exposure to formaldehyde. To assess the role of mast cells to acute formaldehyde exposure, we utilized wild type (C57BL/6) and B6.Cg-KitW-sh/HNihr-JaeBsm (Kitw-sh, mast cell deficient) male mice. C57BL/6 mice exhibited dose-dependent decreases in migratory dendritic cells, and statistically significant increases in alveolar macrophages and mast cells as assessed by flow cytometric analysis of lung tissue. Bronchoalveolar lavage analyzed lipid mediators, inflammatory cytokines, and cell counts with a significant and dose-dependent increase in cell counts in C57BL/6 mice where Kitw-sh mice showed no significant increase, regardless of dose. Lung pathology revealed increases in immune cell populations and alveolar septal wall thickening in C57BL/6 mice not seen in Kitw-sh mice. Results indicate mast cells may play a significant role in cell recruitment, inflammation, and pathology in the lung post-exposure to chemical pollutants such as formaldehyde.
HBV immune checkpoint receptor CD33 (Siglec-3) is a promising therapeutic target for chronic hepatitis B infection
Authors: Sung Pei-Shan , Lee Chieh-Ju , Huang Yi-Hsiang , Wu Chang-Ru , Hsieh Shie Liang
Published: November 2024

Approximately 300 million people globally suffer from chronic hepatitis B (CHB), leading to complications like hepatic fibrosis, cirrhosis, and hepatocellular carcinoma. Despite a significant reduction in new HBV infections through vaccination, a definitive CHB cure remains elusive. Current treatments rely on antiviral nucleos(t)ide analogues, often failing to induce crucial anti-HBsAg antibody (HBsAb) production. Our prior study unveiled myeloid CD33 (Siglec-3) as an immune checkpoint receptor for HBV, contributing to HBV-induced tolerance. Blocking CD33 with anti-hCD33 mAb enhances antigen-presenting molecule expression and activation markers in dendritic cells, restoring TLR7 agonist-induced cytokine production in CHB patient PBMCs. We recently identified a novel CD19+ B cell population expressing abundant CD33 in CHB patient PBMCs (5-20% of CD19+ B cells), contrasting with rare detection in healthy donors (0.5-3%). Anti-hCD33 mAb treatment induces HBsAb production in PBMCs and CD19+ B cells. Furthermore, establishing a CHB mouse model via pAAV-HBV-1.2 hydrodynamic injection in human CD33 transgenic mice (hCD33 Tg) demonstrated prolonged HBsAg persistence compared to wild-type mice, suggesting human CD33 impedes HBV clearance. Remarkably, anti-hCD33 mAb administration accelerates HBsAg clearance in hCD33 Tg mice, highlighting CD33 as a promising CHB therapeutic target, with anti-hCD33 mAb potential to reactivate host immunity for HBV clearance in CHB patients.
Multi-modal single-cell sequencing of circulating mononuclear cells deconvolutes outcome-specific immune responses in trauma patients
Authors: Chen Tianmeng , Hughes Julia , Gregory Alyssa , Cornoy Julia , Chen Wei , Duerr Richard , Billiar Timothy
Published: November 2024

Immune dysfunction plays a key role in contributing to high morbidity and mortality after trauma. To identify the immune cell subsets associated with a specific clinical trajectory (SR vs. FR, slow vs. fast recovery), we isolated the peripheral blood mononuclear cells from 16 trauma patients, including 8 SR patients (ICU days > 7) and 8 FR patients (ICU days < 7) at 72 hours after injury, along with 4 age and sex matched healthy controls. A total of 20 samples were subjected to DOGMA-seq, a tri-modal single-cell assay that measures gene expression, open chromatin accessibility, and surface protein expression simultaneously in the same cells. Integrated analysis revealed patient group-specific changes in T cells and monocytes. SR patients had lower numbers of cytotoxic T cells (CTLs) and these remaining CTLs were less cytotoxic. We identified a subset of monocytes overrepresented in FR patients. We extracted the genes specifically upregulated in this subset and queried a published whole blood transcriptome dataset of 167 trauma patients. These genes were positively correlated with T cell activation pathways, and were also correlated with a quicker recovery in this bulk dataset. Thus, we were able to validate our findings derived from DOGMA-seq from 16 trauma patients in a much larger cohort. Our analysis provides a landscape of immune responses in trauma patients based on clinical trajectories and characterizes the outcome-specific changes across cell types.
USP30 inhibition augments mitophagy to prevent T cell exhaustion.
Authors: Xin Gang , Zhang Ruohan , Li Jianying , Yang Mingchong , Jin Jiacheng , Pich Thera , Sun Nuo , Xin Gang
Published: November 2024

The quality control of mitochondria through mitophagy is crucial for maintaining effective anti-tumor CD8 T cell responses. However, this process is compromised during T cell exhaustion. The specific mechanisms regulating mitophagy remain unclear, hindering efforts to target it for reversing T cell dysfunction. To tackle this challenge, we employed a mitophagy-reporter mouse model and revealed impaired mitophagy activity in exhausted CD8 T cells exhibiting mitochondrial dysfunction. Through a bioinformatic and experimental approach, we identified that USP30, a mitochondrial deubiquitinase inhibiting mitophagy, is upregulated in exhausted CD8 T cells with the decline of mitophagy. Genetic deletion or pharmacological inhibition of USP30 can significantly enhance mitophagy in exhausted CD8 T cells and restore their mitochondrial fitness and effector function, ultimately reinvigorating anti-tumor CD8 T cell responses and enhancing immunity against tumors. Furthermore, our observations indicated that the stimulation of the T-cell receptor amplifies the transcription of USP30 in exhausted CD8 T cells, highlighting its pivotal role in regulating mitophagy. Overall, our findings shed light on the vital role of USP30-regulated mitophagy in controlling mitochondrial quality during T cell exhaustion while also providing valuable insights into developing targeted therapies to restore mitophagy, thereby improving the immune response against tumors.
Opposing roles of STAT1 and STAT3 in COX-2 protein expression in human follicular dendritic cells
Authors: Jeong Jihye , Choe Jongseon
Published: November 2024

Prostaglandins (PGs) are the lipid mediators of inflammatory responses and acknowledged also as the mediators of immune inflammation. We have explored the role of PGs and their production mechanisms using follicular dendritic cell (FDC)-like cells. In the course of our recent study of the regulation mechanisms of PG production, we observed the involvement of JAK-STAT molecules in the IL-1β-induced PG production. In the current study, we aimed to elucidate the role of JAK and STAT proteins in the expression of COX-2 protein, the responsible enzyme for PG production. COX-2 protein level and PG production were assessed using immunoblotting and EIA, respectively. The intracellular target proteins were detected by a confocal microscopy. STAT1 knockdown resulted in an increase of COX-2 expression, while STAT3 knockdown led to a reduction in the COX-2 induction by IL-1β. IL-1β induced phosphorylation of both STAT1 and STAT3 at the S727 sites, rather than at Y701 or Y705. The phosphorylation and nuclear translocation of pSTAT1 and pSTAT3 were confirmed by a confocal microscopy. Based on these findings, we maintain that STAT1 may play an inhibitory role in COX-2 induction, while STAT3 acts as a promoter, via fine tuning of phosphorylation levels. The current study may enhance our understanding of the precise roles of these STAT proteins in COX-2 induction and PG production and suggests that these molecules could be potential targets for the treatment of immune inflammatory disorders.
A novel interaction between complement product C4a and the receptor for advanced glycation end products (RAGE)
Authors: Wang Fang , Wu Yee
Published: November 2024

Complement proteins (C) including their split products play important roles in host defense and homeostasis. Anaphylatoxins C3a and C5a, by binding to their receptors on immune and non-immune cells, mediate cell recruitment and the release of proinflammatory molecules. The receptor and function of C4a remain unclear. C3a, however, in addition to its conventional receptor C3aR, is known to bind to the receptor for advanced glycation end products (RAGE), a receptor implicated in initiating and sustaining inflammatory responses. This interaction can also be mediated by the TLR9 agonist, CpG. C4a is similar to C3a in its structure and net charge. Therefore, we hypothesize that C4a may interact with RAGE. We performed co-immunoprecipitation experiment on 293T cell lysate expressing C4a and RAGE and found that RAGE did not co-immunoprecipitate with C4a, indicating that C4a does not directly interact with RAGE. Using fluorescently labelled C4a protein, we found that there was no significant C4a binding to 293T cells irrespective of RAGE overexpression by flow cytometry. However, by preincubating C4a with CpG, we detected elevated C4a binding to RAGE-expressing 293T cells compared to control cells. These findings suggest that CpG mediates interaction between C4a and RAGE. Our ongoing investigations focus on determining the binding properties and the functional impact on inflammatory response of this novel interaction.
Obesity reprograms macrophage nucleotide metabolism to enable hyperinflammation
Authors: LIU DANHUI , Liang Shuang , Zhong Zhenyu
Published: November 2024

Obesity-induced NLRP3 inflammasome hyperactivation is a major risk factor for human diseases spanning autoimmune, degenerative, metabolic disorders, and cancer. However, its underlying mechanisms remain elusive. Here we show that obesity compromises the function of SAMHD1 – an evolutionarily-conserved deoxyribonucleotide triphosphate (dNTP) hydrolase essential for cellular nucleotide homeostasis. This dysregulation leads to aberrant accumulation of cytosolic dNTPs in macrophages, which are subsequently transported into mitochondria, and directly serve as building blocks for new mitochondrial DNA (mtDNA) synthesis bypassing the CMPK2-mediated nucleotide salvage synthesis pathway. The uncontrolled mtDNA neosythesis then triggers the overproduction of oxidized mtDNA upon NLRP3 activators stimulation, ultimately resulting in hyperactivation of NLRP3 inflammasome. Genetic ablation of mouse Samhd1 leads to overwhelmed NLRP3 inflammasome activation in vivo and exacerbates immunopathology in multiple NLRP3-dependent disease models. Moreover, mice lacking SAMHD1 exhibit heightened circulating IL-1β, insulin resistance, and metabolic dysfunction-associated steatohepatitis (MASH) upon high-fat diet feeding. Overall, our study establishes SAMHD1 as a macrophage-intrinsic valve restraining NLRP3 inflammasome activation, illuminating that obesity, through compromising SAMHD1's function, rewires macrophage nucleotide metabolism to enable hyperinflammation and disease development.
Emergence of novel CD3+ Macrophages in Brown Adipose Tissue Following Adrenergic Stimulation
Authors: Chae Hee-Don , Levi Jelena
Published: November 2024

Macrophages play a vital role in maintaining tissue homeostasis and defending against infections and cancer. Brown adipose tissue (BAT) is essential for expending surplus energy during cold stress, while white adipose tissue functions as the primary energy reservoir. The sympathetic nervous system responds to cold stress by releasing norepinephrine (NE), activating BAT through β3-adrenergic receptor. BAT-infiltrated macrophages exert opposing effects on thermogenesis, regulating the interplay between BAT cells and sympathetic nerves to either inhibit thermogenesis by clearing NE or enhance it by supporting sympathetic innervation of BAT.

In mouse interscapular BAT, we identified a novel subset of CD3+ macrophages (CD45+ CD3+ CD4+ CD8+ CD11b+ F4/80+), whose frequency increased with chronic adrenergic stimulation. Despite the potential for T cell-monocyte doublets in flow cytometry, CD3+ macrophages demonstrated a unique surface expression pattern, lacking T cell markers CD62L and TCRβ. Furthermore, they showed elevated levels of Ly6C, MHCII, F4/80, and CD11b compared to T cells and CD3- macrophages. CD3+ macrophages exhibit heterogeneity in MHCII expression, with subsets expressing low and high levels.

Since MHCII-expressing macrophages play a crucial role in regulating sympathetic nerve-BAT interaction, further characterization of CD3+ macrophages is pivotal for understanding the intricate mechanisms that govern adaptive thermogenesis regulation.
Sex-biased immunogenicity of a mucosal subunit vaccine against SARS-CoV-2 in mice
Authors: Li Jianping , Hsu Kevin , Hoang Tanya , XIA ZHENG , SUI YONGJUN , Berzofsky Jay
Published: November 2024

Current vaccines against COVID-19 administered via parenteral route have limited ability to induce mucosal immunity. There is a need for an effective mucosal vaccine to combat SARS-CoV-2 virus replication in the respiratory mucosa. Moreover, sex differences are known to affect systemic antibody responses against vaccines. However, their role in mucosal cellular responses against a vaccine remains unclear and is underappreciated. We evaluated the mucosal immunogenicity of a booster vaccine regimen that is recombinant protein-based and administered intranasally in mice to explore sex differences in mucosal humoral and cellular responses. Our results showed that vaccinated mice elicited strong systemic antibody (Ab), nasal, and bronchiole alveolar lavage (BAL) IgA responses, and local T cell immune responses in the lung in a sex-biased manner irrespective of mouse genetic background. Monocytes, alveolar macrophages, and CD103+ resident dendritic cells in the lungs are correlated with robust mucosal Ab and T cell responses induced by the mucosal vaccine. Our findings provide novel insights into optimizing next-generation booster vaccines against SARS-CoV-2 by inducing spike-specific lung T cell responses, and a rationale for considering biological sex differences in future vaccine research and vaccination practice.
Interrogating the CD47-SIRP axis in chronic T cell stimulation for the translation of Graft-vs.-Host disease therapeutics.
Authors: Wallisch Patrick , Bourne Christopher , Pierre Stephanie , Kurtz Keifer , Scheinberg David
Published: November 2024

Graft-vs.-Host disease (GvHD) is the major complication associated with hematopoietic stem cell transplants (HSCTs), characterized by severe alloreactive immunotoxicity of the graft towards the immunocompromised patient. GvHD affects 40-50% of HSCT patients with an average 3-year survival of just 43%, underlining the need for novel therapies.

CD47 the 'don't-eat-me' signal, interacts with Signal Regulatory Protein alpha (SIRPa), an inhibitory transmembrane receptor on macrophages, preventing the phagocytosis of the target cell. In cancer therapy, CD47-blockers are investigated as novel immunotherapies promoting phagocytosis of cancer cells. Serendipitously, we found that either CD47 blockade or pan-SIRP blockade effectively alleviates GvHD onset in a murine xeno-GvHD model. Early phagocytosis in vivo was found not to be the driver of GvHD alleviation, leading us to investigate SIRP proteins as potential regulators of chronic T cell stimulation. Particularly SIRP gamma (SIRPg) is exclusively expressed on T cells, and is known to be the only other SIRP to ligate CD47, however it is regarded understudied and its function, signaling, its regulators and downstream effectors are not known. In this work we could show that SIRPg is upregulated in a setting of chronic T cell stimulation, that its expression levels correlate with exhaustion marker expression in clinical scRNA seq data sets, and that primary human SIRPg KO T cells show a delayed onset of GvHD similar to SIRP blockade.
SIGLEC-3 (CD33) serves as an effective target in tumor progression
Authors: Lin Yi-Jyun , Sung Pei-Shan , Hsieh Shie-Liang
Published: November 2024

Human CD33 (Siglec-3) possesses an immune-receptor tyrosine-based inhibition motif (ITIM), which recruits Src homology 2 (SH2) containing phosphatases (SHP-1/2) and inositol phosphatase (SHIP) and exerts inhibitory effects. The structure and downstream signaling of CD33 bear similarity to programmed cell death protein-1 (PD-1), a promising target in cancer immune therapy. Previous studies indicate that CD33 serves as a therapeutic target in acute myeloid leukemia (AML), additionally, CD33 also acts as a prevalent surface marker of human myeloid-derived suppressor cells (MDSCs). According to the inhibitory signaling upon CD33 activation, CD33 may participate in tumor progression and the regulatory role of CD33 in solid tumors is still unclear. To investigate whether CD33 influences cancer progression, we established a melanoma tumor (B16F10) mouse model in monocyte/macrophage-specific human CD33 transgenic mice (hCD33 Tg mice, Cx3cr1cre/-hCD33M+/-). Compared to wild-type mice (WT), tumor growth and mortality were larger and higher in CD33 Tg mice. Moreover, an antagonistic anti-hCD33 monoclonal antibody (clone 4L3-46) treatment successfully attenuated tumor growth in hCD33 Tg mice and led to a restructuring of the infiltrating MDSCs population. These results indicate that human CD33 contributes to the advancement of tumors, and the blockade of CD33 represents a promising strategy in tumor therapy.
Human implantable tertiary lymphoid organs (TLO) for solid tumor therapy: from organ chips to the clinic?
Authors: Goyal Girija , Barck Lucas , Zhai Yunhao , Prabhala Pranav , Paudel Sudip , Ku Min Wen , Patil Aditya , Isaacs Abdul , Ingber Donald
Published: November 2024

We previously created an in vitro human TLO model from patient-derived, circulating immune cells using organ-on-a-chip devices and 3D culture in extracellular matrix (ECM). B cells in these TLOs express activation induced cytidine deaminase, which is only expressed in lymphoid tissues and is required for class switching and somatic hypermutation (SHM). When challenged by vaccines, these TLO undergo SHM and produce antigen-specific antibodies and CD8 T cells. To model cancer associated TLOs, we integrated human pancreatic and lung cancer cell lines into these TLO Chips to understand how the immune context of hot and cold tumors differentially impacts TLO formation. The high PDL1/L2 expressing lung cancer cell line that displayed a “hot” phenotype in published murine studies induced high levels of cytokines and stimulated increased TLO formation in the lymphoid Organ Chip. In contrast, the cold, low PDL1/L2 expressing lung and pancreatic cell lines did not induce this cytokine signature or TLO formation. Importantly, TLO assembly correlated with increased B cell activation, anti-tumor CD8 activity, and tumor cell death. When the same mix of immune cells and ECM was injected in humanized mice bearing subcutaneous tumors from the hot lung cancer cell line, new lymphoid tissue containing dense aggregates of T and B cells was induced, showing for the first time that direct injection of synthetic TLOs at the tumor site may offer an alternative form of therapy for solid tumors.
Longitudinal assessment of humoral immune response against SARS-CoV-2 in solid cancer patients: Implications for vaccination strategies
Authors: Oh Ji Eun , Lee Eunjin , Lee Ki Seong
Published: November 2024

In the face of evolving SARS-CoV-2 variants, immunocompromised patients, particularly those with solid cancers, pose unique challenges in vaccine responsiveness. We investigated the humoral immune responses in solid cancer patients (n=64) undergoing anti-cancer treatments, comparing them to healthcare workers (n=23). Over 12 months, we tracked SARS-CoV-2 spike (S) and receptor binding domain (RBD) specific antibodies, memory B cells, and circulating follicular helper T cells (cTfh) post two to five doses of monovalent or bivalent vaccinations and after Omicron variant breakthrough infections (BTI). After the third vaccination, solid cancer patients exhibited antibody levels against wild-type and variants similar to healthy controls. Notably, post-BTI, these patients showed heightened and sustained antibody responses, indicating the efficacy of hybrid immunity. S-specific memory B cells initially mirrored healthy individuals after the third dose but declined faster. The fourth dose had minimal impact, while the fifth significantly boosted classical memory B cells. Post-BTI, durable S-specific memory B cells increased, irrespective of prior vaccination doses. Our findings highlight the resilient and prolonged humoral immunity induced by repeated vaccinations and BTIs in solid cancer patients, offering critical insights for tailored vaccination strategies in immunocompromised populations.
B cell regulation of CD8 T cell phenotype in vaccination and homeostasis
Authors: Manes Cammy , Guerrero-Moreno Miguel , Getahun Andrew , Kedl Ross , Klarquist Jared
Published: November 2024

We have found that B cells are responsible for shaping the phenotype of naïve, effector, and memory CD8 T cells. In MD4 mice, which have a fixed BCR repertoire, we find that naïve T cells exhibit an activated/effector phenotype. This is marked by relatively high pAkt, p70-s6K, and 4e-bp1, as well as low Foxo1 and Eomes. Subunit vaccination in MD4 or B cell-depleted mice leads to a CD8 T cell response that is reduced in numbers and is effector-skewed. Additionally, adoptive transfer of CD8 T cells from C57BL/6 WT mice into MD4 mice demonstrate similar reduction in primary CD8 T cell numbers and effector phenotype upon vaccination, indicating the resulting phenotype is mostly dependent the recipient environment. In contrast, the naïve CD8 T cell development is dependent on the B cell environment where they develop, where we recovered on average 10 times more cells from a WT donor than an MD4 donor. Upon looking at memory formation in a B cell-deficient environment, there is impairment in the MD4 mice. Primary response by CD8 T cells is of the same magnitude in WT and B cell-depleted mice, but effector skewing is still prevalent with observed reductions in CD127+ memory cells. These data indicate B cells play an essential role in the establishment of not only the naïve CD8 T cells repertoire, but additionally the regulation of CD8 T cell phenotypes following subunit vaccination or infection.
Unique immune cells in oral cavity are critical for enteric virus pathogenesis
Authors: Brogden Graham , Labayo Hannah , Salazar Emanuel , Atyeo Natalie , Gribonika Inta , Belkaid Yasmine , Chiorini John , Altan-Bonnet Gregoire , Altan-Bonnet Nihal
Published: November 2024

We recently discovered that salivary glands are a major site of norovirus replication and oral-oral transmission is facilitated by saliva1. However, the cell types infected and the role of saliva in virus dissemination remains to be elucidated. Mice inoculated with norovirus were sampled and cells sorted into immune and epithelial cells. Mice became positive for virus in salivary glands and intestine plus associated lymphatic tissues. The majority of virus was present and replicating in CD45+ immune cells and EpCAM+ epithelial cells. Spatio-temporal analysis of the infection revealed that the earliest and highest viral pools were associated with the mandibular lymph nodes. Furthermore, a unique population of immune cells contained the highest concentration of virus, suggesting that these cells potentially play a sentinel role in disseminating norovirus. Notably, enteric infection of the intestine resulted in high levels of inflammation and cell death, however this was absent in the salivary glands, suggesting a level of immune modulation. Our findings suggest that the immune response in the salivary glands is different from that seen in the intestinal tract, and there is a unique population of immune cells which potentially plays an important role during the infection.

1 Ghosh S, et al. Enteric viruses replicate in salivary glands and infect through saliva. Nature. 2022; 607:345–350. doi: 10.1038/s41586-022-04895-8.
CD8 T cells with an exhaustion-like phenotype remain highly functional in hematologic tumors
Authors: Minnie Simone , Waltner Olivia , Zhang Ping , Takahashi Shuichiro , Nemychenkov Nicole , Ensbey Kathleen , Legg Samuel , Comstock Melissa , Boiko Julie , Bhise Shruti , Wilkens Alec , Koyama Motoko , Dhodapkar Madhav , Chesi Marta , Riddell Stanley , Green Damian , Spencer Andrew
Published: November 2024

Loss of immunosurveillance in solid tumors is associated with CD8 T cell dysfunction/exhaustion (TEX). Immunotherapies target TEX differentiation, which may be modified by differences in microenvironments across cancer types and tissue sites. We characterized TEX differentiation in bone marrow (BM), where liquid tumors reside, using mouse models of myeloma and B cell leukemia progressing after autologous stem cell transplantation and CD19-directed CAR T cells respectively. We used flow cytometry and concurrent analysis of RNA expression and DNA accessibility in single cells. We identified a novel CD8 T cell subset with a terminally exhausted phenotype that retained functional capacity, including secretion of cytotoxic molecules and IFNγ. These cells are referred to as interferon-Gamma-secreting, Phenotypically Exhausted (TG-PhEX). Expression of functional genes in TG-PhEX correlated with Batf expression and accessibility within the BATF motif. Chromatin remodeling in stemness associated genes indicated a loss of self-renewal capacity. Indeed, disease progression was associated with a lack of TG-PhEX expansion and not dysfunction as TG-PhEX effectively killed myeloma with comparable activity to CX3CR1+ TEX. Importantly, we observed analogous cells in myeloma and lymphoma patients. TG-PhEX are a clinically relevant target for immunotherapy of hematologic cancers and demonstrate differences in TEX differentiation across tumor sites and cancer subtypes.
Heterogeneity in tumor-intrinsic sensitivity to interferon gamma stimulation
Authors: Merritt Elliot , Granados Genesis Lara , Monga Isha , Carvallo-Fleming Julie-Ann , Pham Giang , Sritharan Ramja , Farias Richard , Zuluaga Laura , Sia Daniela , Sfakianos John , Badani Ketan , Horowitz Amir , Hirsch Fred , Brody Rachel , Tsankov Alexander , Hopkins Benjamin , Tocheva Anna
Published: November 2024

Interferon gamma (IFNg) drives resistance to PD1-targeting immunotherapies by altering the tumor-intrinsic immune phenome associated with the upregulation of inhibitory ligands and immunosuppressive mediators. Major bottleneck to improving the efficacy of IFNg-inducing immunotherapies is our lack of knowledge as it pertains to tumor-intrinsic features that drive tumor-specific differences in IFNg sensitivity. There is a critical need to define the cellular and molecular mechanisms underlying tumor-intrinsic sensitivity to IFNg in pre-clinical models that reproduce the oncogenic programs of human tumors.

We leveraged patient-derived tumor organoids (PDO) with high-dimensional immunophenotyping and RNA-sequencing to determine IFNg sensitivity across different tumors. Our approach identified significant differences in IFNg sensitivity between malignant cells of the same tumor associated with the expression of therapeutically relevant immune checkpoints and secreted proteins. These differences were driven by unique malignant programs underlying patient-specific intratumoral divergence in IFNg sensitivity.

PDO model intra-tumoral and inter-patient heterogeneity in IFNg sensitivity. This is the first study to reveal how the underlying oncogenic pathways intersect with sensitivity to IFNg stimulation and uncover the breadth of IFNg-induced molecular programs in patient tumors.
FOXP1 and KLF2 Modulate the Transition from Stem-like to Effector in CAR T Cell
Authors: Zhu Ziang , Wu Tuoqi , Yao Chen
Published: November 2024

T cells are a pivotal component of the immune system in combating infections and cancer. In cancer, the presence of self-renewing, stem-like CD8+ T cell subpopulation is crucial for limiting tumor growth and providing sustained immunity. However, prolonged antigen stimulation can induce differentiation of some CD8+ T cells into exhausted T cells, characterized by a loss of self-renewing capacity, impaired function and upregulation of immune checkpoints. Exhaustion poses challenges for CAR-T therapy efficacy. Transcription factors are central to the control of T cell differentiation. Utilizing scATAC+RNA-seq analysis of CAR-T cells, we identified transcriptional regulators of CAR-T cell differentiation. FOXP1, a central element in the stem-like gene regulatory network, facilitates the expansion and preservation of stemness in CAR T cells while restraining excessive effector differentiation. In addition, transcription factor KLF2 plays a key role in promoting the differentiation of effector CAR CD8+ T cells while averting their terminal exhaustion. Thus, our study has demonstrated FOXP1 and KLF2 as crucial regulators of CAR T cells and highlighted their potential as targets for enhancing effectiveness and longevity of CAR T cells in cancer immunotherapy.
COMPOUND 2 is a first-in-class small molecule cGAMP competitive oral STING antagonist that reduces neuroinflammation in mouse models
Authors: Banerjee Monali , Shrivastava Ritesh , Middya Sandip , Basu Sourav , Middya Anindita , Shinde Vaibhav , Ghosh Rajib , Padaliya Manisha , Bano Shahnaj , Mansuri Rubeena , Sharma Priti , Roy Sneha , Soram Thanilsana , Yadav Dharmendra , Puniya Kavita , Gautam Anuj , Pryde David
Published: November 2024

Background: Aberrant activation of the cGAS/STING pathway has been implicated in chronic inflammatory conditions such as neuroinflammation. Emerging evidence reveals that the cGAS-STING-mediated Type I interferon (IFN) signaling axis contributes to neuroinflammation and dopaminergic neurodegeneration in Parkinson's Disease (PD). COMPOUND 2 is a novel cGAMP competitive synthetic small molecule STING antagonist discovered at Curadev. Materials and Methods: Competitive binding was established using radioligand binding assays. The neuroprotective effects of COMPOUND 2 were demonstrated in MPTP treated mice. PK and plasma to brain partitioning of COMPOUND 2 was estimated after oral dosing in mice. Results: COMPOUND 2 inhibits the binding of cGAMP to STING and causes reductions in the level of Type I interferons and pro-inflammatory cytokines from immune cells as well as in mice treated with a STING agonist. There is significant partitioning of COMPOUND 2 into the brain after oral dosing. Once a day oral administration of COMPOUND 2 provided protection from neurodegeneration, decreased striatal tyrosine hydroxylase expression and led to a recovery in dopamine levels in MPTP treated mice. Conclusions: COMPOUND 2 is an orally administered novel first-in-class cGAMP competitive STING antagonist with high levels of brain exposure and is a promising candidate for the treatment of neuroinflammatory conditions.
Identification of immune modulating strategies to promote cartilage regeneration.
Authors: Choi Eirene , Smith Carly , O'Donnell Lauren , Collins John , Freeman Theresa
Published: November 2024

The immune response is a key modulator of regeneration. Apoptosis Signal-Regulating Kinase 1 (ASK1) loss can promote cartilage regeneration, thus we investigated how ASK1 loss modulates the immune response.

Two mm biopsy punches of ear tissue samples were taken from WT and ASK1 KO mice (n=3) on day 0, 4 and 7 after injury. RNA was extracted and gene expression analyzed using the NanoString nCounter Immunology Gene Panel. Functional enrichments were analyzed using Gene Set Enrichment (GSEA) and Variation (GSVA) Analysis.

GSVA showed general apoptosis and p53 pathways were significantly decreased in all KO samples, as expected. The Nanostring curated gene set across all time points and samples showed TLR, NFkB and TNF signaling down regulation. Conversely, MHC class II and cell adhesion were upregulated. GSEA indicated significant differences between cell types involved in the innate and acquired immune response. The most increased GSVA results from the Hallmark gene set was apical junction and most significant decrease was the epithelial-mesenchymal transition (EMT) and IL-6-JAK-STAT3 signaling pathway. GSEA-generated heatmaps of genes that significantly contributed to these pathways give additional insight into how ASK1 loss modulates immune signaling pathways to promote a regenerative environment.

This analysis identified signaling pathways that modulate the immune response to support a regenerative environment and insights into how to clinically modulate this process.
Understanding the role of IL-17 signaling pathway in regulating the immune response against Mtb infection.
Authors: Chauhan Kuldeep , Das Shibali , Dela Peña Noreen , Ahmed Mushtaq , Khader Shabaana
Published: November 2024

Tuberculosis (TB) remains a significant global health concern, with a quarter of the world's population infected. Understanding the immune response against Mycobacterium tuberculosis (Mtb) is crucial for developing effective strategies to control TB. Previous work from various groups, including our lab, has indicated that the induction of IL-17 plays a protective role during Mtb disease progression. However, the downstream cells that sense and respond to IL-17 and how they shape the induction of protective immune responses remain unclear. The current study focuses on understanding the role of IL-17RA-mediated signaling in regulating the immune response against Mtb infection. The study aims to investigate the role of IL-17 signaling in specific innate myeloid immune cell populations and its effect on TB disease progression. Using Cre-Flox based conditional knockout mice lacking IL-17RA, we explored the role of IL-17 signaling in different cellular components of myeloid immune cells, regulating disease progression and initiating the immune response. Our work elucidated the involvement of LysM-expressing cells as the primary responder to IL-17RA-mediated signaling in regulating the protective immune response against TB during the early stages of infection. By understanding the role of IL-17 signaling in the immune response against Mtb, this study aims to provide insights into potential strategies for controlling TB and improving vaccine design.
Regulation of macrophage inflammatory response by potato-derived bioactive peptide
Authors: Okeke Emeka , Udenigwe Chibuike , Ventura Esmeiry
Published: November 2024

The normal immune response to infection requires a well-regulated mechanism of leukocyte activation and recruitment, a process generally termed inflammation. Dysregulation of this inflammatory response is a major problem in modern medicine and is associated with several disease conditions including autoimmune diseases, cancer and even COVID-19.

Food-derived peptides with anti-inflammatory properties have gained popularity due to their availability in the daily diet and limited side effects. Previous reports have shown that the potato protein patatin has anti-inflammatory property. However, the sequence of amino acids in patatin that is critical for anti-inflammatory activity is not well established. In this work, we investigated the ability of the potato protein-derived peptide DIKTNKPVIF to down-regulate the inflammatory response of monocyte-derived macrophages (MDMs) activated with lipopolysaccharide (LPS). We found that DIKTNKPVIF inhibited the inflammatory response of MDMs to LPS as evidenced by decreased production of pro-inflammatory cytokines. Importantly, using computational and experimental screening techniques, we identified TNKPVI as the bioaccessible and biostable pharmacophore necessary for the activity of DIKTNKPVIF. Our results highlight the potential of potato-derived bioactive peptides to regulate the inflammatory response.
Maresin-1 attenuates neuroinflammation in EAE via metabolic reprogramming of disease-associated cell types
Authors: Zahoor Insha , Nematullah Mohammad , Ahmed Mohammad Ejaz , Fatma Mena , Mir Sajad , Rattan Ramandeep , Giri Shailendra
Published: November 2024

Inflammation resolution is dysregulated in people affected with multiple sclerosis (MS) due to impaired metabolism of docosahexaenoic acid (DHA). We hypothesize that supplementing its downstream metabolite maresin 1 (MaR1) will alleviate inflammation and demyelination in a preclinical mouse model of MS; experimental allergic encephalomyelitis (EAE). Administration of the resolution agonist MaR1 mediated resolution and improved neurological outcome in the relapsing-remitting (RR) model of EAE. MaR1 induced metabolic changes in CD4, macrophages, microglia, and oligodendrocytes. It modulated the phenotype of disease-associated cell types by regulating their effector functions. It restored the impaired efferocytosis in EAE, promoting clearance of damaged myelin and dead cells; thereby lowering the disability with disease course. MaR1 is a potential interventional candidate to attenuate dysregulated inflammation to restore neurological deficits in EAE and other autoimmune diseases.