bluuebunny/crossref_metadata_2025 huggingface dataset:
Non-thermal atmospheric plasma ameliorates imiquimod-induced psoriasis-like skin inflammation in mice through inhibition of immune responses and up-regulation of PD-L1 expression.
Authors: Lee Yun Sang , Lee Myung-Hoon , Kim Hang-Jun , Won Ho-Ryun , Kim Chul-Ho
Published: January 2023

Plasma medicine is emerging as a novel therapeutic field. It has been reported that plasma can kill bacteria, promote wound healing and induce apoptosis of tumor cells. However, the effects of plasma on immune cells and immune related skin diseases have not been well studied. In this study, we demonstrated that non-thermal atmospheric plasma (NTP) treatment can inhibit psoriasis-like skin inflammation in mice. Psoriasis-like skin inflammation was induced through application of imiquimod for 5 days and NTP was treated on days 3 and 4. On day 5 mice were sacrificed for analysis. NTP treatment in imiquimod-applied mouse skin inhibited increases in epithelial cell thickness compared to imiquimod-applied mouse skin without NTP treatment and suppressed immune cell infiltration in the mouse skins. The expression of cytokines and chemokines and NF-κB activation in the skin were also suppressed by the NTP treatment. In addition, differentiation of Th17 cells, an important cell type for the pathogenesis of psoriasis, was inhibited in the NTP-treated mouse lymph nodes. It was also demonstrated that liquid type plasma (LTP), which is also known as indirect plasma, inhibited Th17 cell differentiation in vitro. Other in vitro experiments showed that LTP inhibits bone marrow-derived dendritic cell (BMDC) activation, and inflammation of HaCaT cells. Interestingly, LTP enhanced PD-L1 expression in BMDC and HaCaT cells, suggesting that NTP may inhibit unwanted over-activation of T cells through increased PD-L1 expression in dendritic cells or epithelial cells. Taken together, these results suggest that NTP can be used to treat CD4+ T cell-mediated autoimmune diseases such as psoriasis.
Identification of a novel tolerogenic cell subset in the treatment of autoimmune diseases
Authors: Marinelarena Alejandra , Bhattacharya Palash , Alharshawi Haled , El-Sayed Osaka , Kumar Prabhakaran , Prabhakar Bellur S.
Published: January 2023

We previously reported granulocyte macrophage colony-stimulating factor (GM-CSF) prevented type 1 diabetes in NOD mice through CD11c+CD8− dendritic cell (DCs) mobilization, precipitating regulatory T-cell (Treg) expansion. Furthermore, we found GM-CSF derived bone-marrow dendritic cells (G-BMDCs) can drive Treg expansion in ex-vivo CD4+splenic T-cell co-cultures through surface-bound molecule OX40L. To determine whether GM-CSF may lead to the formation of a distinct, novel CD11c+OX40L+ expressing DC subset, we generated OX40L+G-BMDCs (CD11c+OX40L+) and analyzed these cells through flow cytometry for various cell lineage markers. We determined OX40L+G-BMDCs expressed increased levels of co-stimulatory molecules CD80, CD86, PD-L1 and MHC-II in comparison to the OX40L− counterpart. Additionally, OX40L+G-BMDCs lacked expression of Ly6G, Ly6C, FceR1, surface markers indicative of granulocyte and mast cell lineage, but to our surprise, highly expressed macrophage markers F4/80, MerTK, and CD200R. To elucidate an in vivo physiological component of these OX40L+ G-BMDCs, we performed an unsupervised hierarchical clustering comparing the transcriptome of OX40L+G-BMDCs to that of all myeloid and lymphoid lineages from the Immgen database. We found that OX40L+G-BMDCs clustered most closely with macrophages and dendritic cells. Furthermore, a principal component analysis with the transcriptome of OX40L+G-BMDCs and that of macrophages and dendritic cells from the Immgen database revealed OX40L+G-BMDCs most closely clustered with macrophages. Collectively, these results suggest OX40L+ G-BMDCs may represent a distinct GM-CSF dependent, macrophage-like cell subset involved in physiological Treg homeostasis.
CD38+ cell depletion with TAK-079 reduces arthritis in a cynomolgus collagen-induced arthritis (CIA) model
Authors: Smithson Glennda , Zalevsky Jonathan , Korver Wouter , Roepcke Stefan , McLean Lachy
Published: January 2023

TAK-079 is a fully human, cytolytic IgG1 mAb against human CD38. We tested the ability of TAK-079 to treat arthritis using a monkey CIA model with features of symmetrical small joint polyarthritis resembling human RA. Monkeys were injected with bovine collagen type II in complete Freund’s adjuvant on Day 0 and Day 21 to induce arthritis. 1 monkey was used as a disease-free, age-matched control (naïve). In monkeys with CIA, the prophylactic group (n=8; 3 mg/kg, QW, iv) started treatment on Day 7 and continued for 8 weeks. The therapeutic treatment groups were randomized, after progression to overt disease, to 1 of 3 treatments: Vehicle (n=5; QW, iv), TAK-079 (n=7; 3 mg/kg, QW, iv), or dexamethasone (Dex; n=4; 0.1 mg/kg QD, po) and treated for 5 weeks.

Prophylactic treatment with TAK-079 inhibited arthritis induction and therapeutic treatment reduced arthritic disease to a similar extent as Dex. TAK-079 treatment reduced CD38+ cells in the blood and spleen vs. naïve animal. After the first dose, blood monocytes, B and T cells were each reduced ≤ 55%, but reduction of monocyte and T cell counts were transient despite continued dosing. NK cells showed almost complete depletion. Neutrophil and platelet counts, elevated due to active arthritis, appeared normalized by TAK-079 vs. the naïve animal. TAK-079 treatment reduced systemic and local inflammation. Treated animals suffered less weight loss, and had reduced CRP and ALP levels and increased albumin levels. TAK-079 treatment reduced cartilage degradation and bone resorption in the IP and MCP joints based on radiology findings and confirmed by qualitative histological grading. TAK-079 was effective in treating CIA, supporting its continued investigation for the treatment of autoimmune disease.
Molecular mechanisms promoting progression of autoimmune myocarditis to dilated cardiomyopathy.
Authors: Nalawade Saisha Abhay , Bloom Joshua , Sunsong Robin , Rauhut Anja , Thomas Dominic , Al-Abed Yousef , Forsthuber Thomas
Published: January 2023

Myocarditis is an inflammatory disease of the myocardium, described by infiltration of immune cells and cardiomyocyte necrosis. It is a major cause of sudden death in young adults. Patients often progress to a more severe form of the disease termed dilated cardiomyopathy (DCM). It is characterized by extensive fibrosis leading to impaired cardiac function. Immunosuppressive treatments with corticosteroids (CSs) have not been effective in preventing the disease. We hypothesized that macrophage migration inhibitory factor (MIF), being the only known pro-inflammatory cytokine induced by CSs may play a role in resistance to CSs. Also, MIF counter-regulates CS-mediated immunosuppression. Using the experimental autoimmune myocarditis (EAM) animal model, we observed that MIF−/− mice treated with Dexamethasone (Dex) were highly resistant to EAM and progression to DCM. Furthermore, from a translational approach, treatment using small molecule inhibitors of MIF combined with Dex recapitulated this phenotype in wild type mice. We observed that treated mice showed decreased expression of key chemokines and extracellular matrix molecules compared with controls, implicating these molecules in disease progression. Currently, we are investigating these molecules by selectively inhibiting them in order to understand their contribution to disease pathogenesis. We observed that inhibition of CCL2 after, although not prior to establishment of EAM attenuated progression to DCM. Thereby, suggesting that CCL2 is predominantly involved during the fibrotic phase aiding in progression as opposed to the induction phase of the disease. Besides a novel approach to prevent DCM, our studies may provide new insights into the mechanisms driving DCM.
Therapeutic effects of erythroid differentiation regulator 1 (Erdr1) on atopic dermatitis-like skin inflammation in NC/Nga mice
Authors: Kim Kyung Eun , Cho Daeho
Published: January 2023

Atopic Dermatitis (AD) is a representative chronic inflammatory skin disease. It is characterized by a defective skin barrier and dysregulation of the immune system as result of excessive Th2 immune response. IL-18, a well-known pro-inflammatory cytokine, is particularly closely associated with AD severity and IgE level in AD patients. Recent studies have been reported that erythroid differentiation regulator (Erdr1) has a negative correlation with IL-18 and exerts anti-inflammatory effects on inflammatory skin diseases, such as psoriasis, rosacea, and melanoma. In this study, to investigate whether Erdr1 has a preventive effect on AD, NC/Nga mice were used to induce AD-like skin inflammation. Then, recombinant Erdr1 (rErdr1) was administered by intraperitoneal (i.p.) injection. As results, rErdr1 administration significantly decreased the severity of AD including redness, edema, scaling, and excoriation. In addition, IgE and IL-4 production were suppressed by rErdr1 in the AD mouse model. To elucidate the related mechanisms, the expression of Th2 chemoattractants and angiogenic factor was determined. The expression of CCL17 and CCL22 was significantly lower in rErdr1-treated mice than in vehicle control-treated mice. Also, a representative angiogenic factor, VEGF expression was inhibited by rErdr1 administration. To our knowledge, this is the first evidence that rErdr1 has a preventive effect on AD based on its ability to reduce Th2 chemoattractants and angiogenesis in an AD-like mouse model. Therefore, we suggest that supplementation with rErdr1 might be an effective and improved therapeutic strategy for the treatment of AD.
FVIII BAR-CD8 T cells to target antigen-specific antibody secreting cells
Authors: parvathaneni kalpana , Scott David W
Published: January 2023

Hemophilia A (HA) is an autoimmune bleeding disorder due to a variety of mutations in coagulation factor (FVIII). HA is currently treated by administering clotting factor VIII (FVIII) prophylactically or on demand after bleeds. Due to the lack of tolerance to FVIII, many of these patients make an antibody response (called “inhibitor formation”) to the therapeutic FVIII, thus neutralizing the circulating plasma FVIII levels. The current treatment for inhibitor formation is frequent high dose administration of FVIII, which is expensive and often fails. An alternative immune tolerance protocol is an unmet need. Understanding and applying the CAR therapy has been one of the major notable discoveries in the last decade in both oncology and hematology research areas. Modifying this approach to target the antibody secreting cells, we engineered a B cell antibody receptor (BAR), consisting of antigen domains (FVIII A2 or C2, OVA) fused to lymphocyte signaling domains. BAR-CD8 T cells provide the antigen on their surfaces to be recognized by specific-antibody secreting B cells (ASCs) and can target to kill the cells in a non-MHC dependent manner. The BAR-CD8 T cells in the presence of the respective targets FVIII-C2 or OVA specific hybridoma cells are activated and secrete INF-γ. In vitro, FVIII-C2 BAR-CD8 T cells (both human and mouse origin) specifically kill target cells, including FVIII domain specific hybridomas. We suggest that BAR T cells offer a novel approach for the treatment of autoimmune diseases where formation of autoantibodies is life threatening.
Suppression of acute uveitis following treatment with repository corticotropin injection
Authors: Wright Albion D , Sharma Prabha , Zweifel Ben , Oh Luke
Published: January 2023

Repository corticotropin injection (RCI: H.P. Acthar® gel) contains a purified porcine pituitary ACTH-analogue, and is an FDA-approved treatment for several inflammatory eye diseases. ACTH binds to all 5 known melanocortin receptors and may suppress inflammation by steroid-dependent and independent pathways. Endotoxin-induced uveitis (EIU) in rodents is a useful experimental model to investigate mechanism of action and pharmacological efficacy of potential treatments. This study was conducted to investigate the potential anti-inflammatory benefit of RCI in an acute rat model of EIU. EIU is characterized by clinically relevant signs of inflammation, including inflammatory cytokines and cells in the anterior and vitreous chambers. Rats (n=16 group) were treated with dexamethasone (Dex), placebo, or RCI at 160 IU/kg, 400 IU/kg or 800 IU/kg following EIU induction. Eyes were clinically examined at pre-challenge, 6–8, 24, and 48 hours post LPS challenge using the Combined Draize and McDonald – Shadduck Scoring System.

Treatment GroupOcular Clinical Score (mean ± sem)IL-1a (pg/ml) (mean ± sem)MIP2a (pg/ml) (mean ± sem)Placebo15.5 ± 1.2207.1 ± 68.9240 ± 47Acthar 160 IU/kg9.4 ± 1.0130.3 ± 34.6158 ± 43Acthar 400IU/kg2.0 ± 0.3*95.7 ± 20.3114 ± 18*Acthar 800 IU/kg0.9 ± 0.2*50.9 ± 8.3*126 ± 32*Dex2.6 ± 0.5*54.4 ± 10.5*92 ± 10**= p ≤ 0.01

We show that RCI treatment significantly reduced ocular inflammation and inflammatory cytokines in an EIU model of acute uveitis. RCI has the potential opportunity to be therapeutic for a wide range of inflammatory and autoimmune diseases, including uveitis. The mechanism of action of RCI may involve more than the induction of corticosteroids, and will be explored further in future studies.
Easy ways to increase immunology content engagement in and out of the classroom
Authors: Lukin Kara R
Published: January 2023

Most traditional science courses, especially large-format courses, are lecture-based. However, the literature clearly demonstrates that this is one of the least effective methods for teaching complex content and results in poor retention. Curricula that infuse a variety of discipline-specific active strategies as well as time for “brain dumps” are more enjoyable for students and result in better content integration and long-term retention. To create time for these activities, students can be required to view short videos of core concepts prior to class. Students can be held accountable with no, low or high stakes assessments or group peer-pressure. Interestingly, students report increased content contact time because they also view these resources after class and prior to assessments. Here, multiple low-risk, high-engagement activities are presented with classroom-ready resources. Various tools for short (<2 min) self-assessments (with aggregate class data) and content engagement will be shared. In addition, demonstration of and directions for free, easy to use screen capture programs like Jing will be provided for making quick core content videos. All strategies are appropriate for high school, undergraduate and graduate courses of small and large sizes. Starting one session at a time and using their current teaching materials, educators can slowly integrate these engaging strategies into their syllabi to enhance students’ experiences and outcomes.
Implementation and student perceptions of the Just in Time Teaching (JiTT) strategy in an upper level immunology course
Authors: Brown Deborah M , Brazeal Kathleen R. , Couch Brian A.
Published: January 2023

Instructional pedagogy has undergone innovative changes to meet the demands of 21st century science education. One such teaching strategy, known as Just-in-Time Teaching or JiTT, is an inquiry based, active learning approach that evolved to meet those demands and has been shown to enhance student retention of complex concepts. Prior to class, students read assignments and answer specific conceptual questions that involve linking new material or ideas with previously learned concepts. Importantly, an open-ended, reflective question is also asked about what students found most interesting, confusing, or difficult about the particular assignment. Student responses to these conceptual and reflective questions help identify incoming misconceptions and are used during class to modify instruction and facilitate discussion. Students are graded based on effort, rather than complete correctness, as “low stakes” assessments have been shown to increase student participation and encourage inquiry. Some of the most successful JiTTs in my course are based on the Case Studies in Immunology companion book to Janeway’s ImmunoBiology. These case studies introduce topics such as the importance of MHC molecules in the development of specific CD8 or CD4 T cells. Additional examples will be given that demonstrate the breadth and scope of JiTT concepts and questions as well as student responses to JiTT assessments. Finally, results from a student survey will be presented to demonstrate how students perceive the JiTT strategy to support their learning. Overall, the JiTT approach has been well-received for upper level Immunology students and is beneficial to student and instructor to assess understanding of complex biological concepts.
Self-Directed Learning within Postgraduate Immunology Education
Authors: Sika-Paotonu Dianne
Published: January 2023

Immunology education is essential for the understanding of mechanistic and cellular events responsible for specific diseases. To support proper understanding by students, contemporary learning and teaching methods are becoming increasingly focused on student centered practices that include Self-Directed Learning (SDL). SDL is the process where a student determines their own learning aims and undertakes methods of their own choosing to achieve and assess their own learning outcomes. SDL has become an area of emphasis particularly within the nursing profession. Although the importance of SDL may be emphasized to students, they may not necessarily receive instruction or have SDL strategies to facilitate effective learning. This work sought to explore the impact of teaching and promoting SDL strategies for postgraduate nursing, midwifery and health students undertaking Immunology education by distance within a postgraduate Pathophysiology course. In-class Immunology teaching sessions for these students contained instruction that taught and promoted various SDL strategies. Favorable student outcomes were achieved for Immunology assessment components of the Pathophysiology course with in-class feedback related to specific instruction regarding SDL strategies, positive. Formal student evaluation of the Immunological course elements also highlighted favorable student satisfaction scores. These findings support SDL as an effective learning tool for postgraduate Immunology education.
TLR9 activation impairs phagocytosis-induced neutrophil apoptosis and prolongs E. coli-induced acute lung injury
Authors: Filep Janos G , Edner Natalie , de Oliveira Lima Dos Santos Everton , Sekheri Meriem , El Kebir Driss
Published: January 2023

Neutrophil dysfunction, resulting in delayed apoptosis and inefficient bacterial clearance, is a characteristic feature of severe pathologies, including sepsis and cystic fibrosis. Human neutrophils detect and respond to bacterial DNA (CpG DNA) through TLR9. We investigated the impact of CpG DNA on phagocytosis, phagocytosis-induced neutrophil apoptosis and clearance of E coli. Culture of human neutrophils with CpG DNA (0.1–3.2 μg/ml) resulted in decreased phagocytosis of opsonized E. coli or yeast. CpG DNA upregulated C3R (CD11b) expression, downregulated C5aR (CD88) expression and induced release of neutrophil elastase. C5aR cleavage was prevented by a specific neutrophil elastase inhibitor and the broad-spectrum serine protease inhibitor PMSF. Consistently, CpG DNA reduced phagocytosis-induced NADPH oxidase-mediated activation of caspase8 and caspase-3. These actions of CpG DNA were blocked by the telomere-derived TLR9 inhibitory oligonucleotide 5′-TTT AGG GTT AGG GTT AGG G-3′. In mice, CpG DNA impaired pulmonary clearance of E coli, suppressed neutrophil apoptosis and delayed resolution of lung injury evoked by intratracheal instillation of live E. coli. Genetic deletion of TLR9 rendered mice unresponsive to CpG DNA. These results identify a novel mechanism, neutrophil elastase-mediated inactivation of C5aR-mediated phagocytosis, by which CpG DNA could contribute to neutrophil dysfunction and prolongation of tissue injury. Our findings also suggest a therapeutic potential for TLR9 antagonists or neutrophil elastase inhibitors for enhancing clearance of bacterial infections in an environment where bacterial DNA is abundantly present.
Type 2 innate lymphoid cells impede IL-33-mediated tumor suppression
Authors: Long Alan W , Dominguez Donye , Qin Lei , Chen Siqi , Fan Jie , Zhang Minghui , Zhang Bin
Published: January 2023

Although a number of studies have recently explored the contribution of the adaptive immunity in interleukin 33 (IL-33)-mediated antitumor effects, innate immune involvement has been poorly characterized. Utilizing Rag1−/− mice (lacking T and B lymphocytes), we show here that either systemic administration of recombinant IL-33 or ectopic expression of IL-33 in melanoma cells is sufficient to inhibit tumor growth independent of adaptive antitumor immunity. We have demonstrated that IL-33-mediated antitumor effects relate to expansion and activation of NK cells. Interestingly, IL-33 also promoted the expansion of active type 2 innate lymphoid cells (ILC2s) via its receptor, ST2, which in turn inhibited NK activation and cytotoxicity. This IL-33-induced ILC2 activity coincided with greater expression of the immunosuppressive ecto-enzyme CD73. Removal of CD73 from ILC2s in culture with NK cells resulted in markedly increased activation levels in NK cells, offering a potential mechanism by which ILC2s might suppress NK cell-mediated tumor killing. Thus, our data reveal an important contribution of IL-33-induced ILC2 to tumor growth by weakening NK cell activation and tumor killing, regardless of adaptive immunity.
The Roles of Tumor-associated Neutrophils in Colorectal Cancer Treated with Different Combinatorial Immunotherapies
Authors: Tsai I-Ting , Chang Ching-Cheng , Wu Chia-Jen , Wu Ping-Yi , Tao Mi-Hua
Published: January 2023

Cancer immunotherapy can elicit robust tumor-specific immune responses that can generate durable tumor regression or eradication. Immune checkpoint blockade specific to programmed death-1/ligand-1 (PD1/PDL1) have shown durable clinical responses in various cancer types but benefit only a small portion of patients. Interleukin 12 (IL-12) and granulocyte macrophage colony-stimulator factor (GM-CSF) have shown potent antitumor activities through activating innate and adaptive immunities, and reprogramming tumor-promoting myeloid cells from immune-suppressive to immune-active phenotypes. In this study, we showed the effect of radiation, which increased release of tumor antigens so that augment the efficacy of immunotherapy, in combination with IL-12/GM-CSF (RT/12GM) or anti-PDL1 antibody (RT/PDL1) on TAN in advanced 14-day-CT26 colorectoral cancer. We found that both combination treatment significantly suppressed tumor growth, only RT/12GM but not RT/PDL1 can cure mice; radiation or immunotherapy alone single therapies were much less effective. Mechanistic analysis showed that RT/12GM, but not RT/PDL1, greatly increased accumulation of TAN. These TANs were not only impaired in their suppressive activities, but also able to induce T cell proliferation, in accordance with their increased expressions of MHC class I and II, CD40 and CD86, indicating they presented in a status with higher Ag presentation ability. Antibody depletion experiments showed that TAN was required for tumor regression in RT/12GM therapy. Together, our data demonstrate that combination of radiotherapy and IL-12/GM-CSF improved antitumor immunity through enhancing Ag presentation activity and reducing immunosuppressive functions of TAN.
STAT4 is required for protective Innate Lymphoid Cell responses to gastrointestinal infection
Authors: Dulson Sarah J , Harrington Laurie E
Published: January 2023

Innate Lymphoid Cells (ILCs) are a newly discovered subset of cells that serve both protective and pathogenic roles in intestinal health. ILCs consist of three defined subgroups and perform effector functions similar to CD4 T cells. However, ILCs lack a specific antigen receptor and are activated directly by cytokines early during an immune response. Since the initial events in an immune response can dictate the ability of the host to control intestinal infection, it is critical to identify molecules that regulate ILC function. Signal transducer and activator of transcription 4 (STAT4) is important for driving many pro-inflammatory responses and phenotypes in T cells, but its effects on ILC development and function remain unclear. Contrary to its known role in Th1 cell biology, we find STAT4 to be dispensable in Group 1 ILC (ILC1) development. However, we observe that STAT4 is critical for IFN-γ production by ILC1s, thus influencing the capacity of this subgroup to mount an effector response. Whereas STAT4 deficiency did not impact IL-22 or IL-17 production by ILC3s, the transcription factor specifically modulates IFNγ production by a plastic Tbet+ subgroup of ILC3s that is greatly expanded in response to inflammation. Importantly, STAT4 signaling is also critical for survival against a genetically modified strain of Listeria monocytogenes that is able to model human Listeriosis in mice. Furthermore, both ILC1 and ILC3 respond to infection with robust IFNγ production, and when ILCs are depleted mice are unable to mount a protective innate response and succumb earlier to infection. These results indicate that STAT4 signaling influences critical responses by ILCs, thereby contributing to host protection against intestinal infection.
Inhibition of Innate Lymphoid Cell Activation by Interleukin-23 Secreting Dendritic Cells During Bordetella pertussis Infections in Mice
Authors: Santos Daniel Rebelo , Nguyen Alana , Stokes Jennifer , Pavlov Nicole , Abramson Tzvia
Published: January 2023

Bordetella pertussis (B. pertussis), the causative agent of whooping cough, is a highly contagious respiratory tract disease marked by extensive damage to the mucosal lining and a delay in immune response due to the pertussis toxin (PTX). In this study, the focus is on the characteristics and mechanisms of two innate immune cell populations, the IL-23-secreting dendritic cells (DCs) and the IL-22-secreting innate lymphoid cells (ILCs). We hypothesize that infection with B. pertussis interferes with the IL-23/IL-22 axis pathway due to PTX production. We compared these immune responses in mice models of whooping cough, triggered by B. pertussis (prolonged disease) or Bordetella parapertussis (B. parapertussis) – an acute version of the infection, lacking PTX. Flow cytometry studies show an increased in lung DCs at 2dpi with B. parapertussis compared to B. pertussis and uninfected mice. At 2dpi with B. parapertussis, more lung CD11b+ DC at secrete IL-23 compare to B. pertussis. The percentage of ILC3 is equally increased at 2dpi with B. pertussis and B. parapertussis-infected mice. Proliferation analyses suggest that residential ILCs proliferate locally rather then recruited during infection to the lungs. Despite the increase in IL-23-producing DCs, no differential expression of IL-22 by ILC3 was observed between infections and uninfected mice. Further studies would verify the nature of this inhibition and whether it is translated to a reduced level of antimicrobial substances secreted by lung epithelial cells. The outcomes of this study may clarify the inability of these innate immune responses to resolve B. pertussis infection early.
Protection from Staphylococcus aureus sepsis by bacterial exopolysaccharide
Authors: Paik Wonbeom , Paynich Mallory L , Alonzo Francis , Knight Katherine L
Published: January 2023

Sepsis is the most severe presentation of systemic infection with up to 50% mortality rates. The leading etiological agent of sepsis is a Gram-positive bacterium, Staphylococcus aureus. Antibiotics are the only effective therapy for S. aureus sepsis, and widespread resistance to multiple antibiotics is a major concern. Novel therapeutic agents for sepsis are critically needed. We previously showed that exopolysaccharides (EPS) derived from a probiotic bacterium, Bacillus subtilis, protects hosts from inflammatory disease by the induction of anti-inflammatory M2 macrophages. Using a murine systemic infection model, we tested if EPS also has therapeutic potential for S. aureus sepsis. We treated mice with EPS prior to infection with S. aureus and found that weight loss, serum proinflammatory cytokine levels, and bacterial burden in the liver were mitigated compared to vehicle-treated controls. We also found that compared to cells from vehicle-treated mice, peritoneal cells from EPS-treated mice suppress growth of serum-opsonized S. aureus in vitro. We purified peritoneal F4/80-positive macrophages from EPS-treated mice and found that they were responsible for suppressing S. aureus growth by peritoneal cells. We confirmed this by depletion of macrophages from mice using clodronate-loaded liposomes, which mitigated suppression of S. aureus growth by peritoneal cells from EPS-treated mice. We suggest that B. subtilis-derived EPS may be useful for prevention and/or treatment of S. aureus sepsis.
IL-33 signaling regulates innate IL-22 production via suppression of PGE2 during lung fungal infection
Authors: Garth Jaleesa M , Reeder Kristen M , Mackel Joseph , Dunaway Chad , Blackburn Jonathan , Steele Chad
Published: January 2023

Members of the IL-1 family play both protective and regulatory roles in immune defense against the opportunistic mold Aspergillus fumigatus. Here, we investigated the role of the IL-1 family member IL-33 in lung defense after acute A. fumigatus exposure. IL-33 was detected in nanogram amounts in the naïve lung, which further increased after exposure to A. fumigatus in a Dectin-1 independent manner. Mice deficient in the receptor for IL-33 (IL-1RL1) unexpectedly demonstrated enhanced innate lung clearance of A. fumigatus. IL-33 functioned as a negative regulator of IL-22, as IL-22 was elevated in fungal-exposed Il1rl1−/− mice and IL-33 administration to normal mice attenuated fungal-induced IL-22 production. IL-33 mediated regulation of IL-22 did not involve the modulation of IL-23 but rather prostaglandin E2 (PGE2); PGE2 was significantly increased in fungal-exposed Il1rl1−/− mice and normal mice produced less PGE2 after fungal exposure when administered IL-33, suggesting that IL-33 mediated regulation of IL-22 occurred at the level of PGE2. This was confirmed by in vivo cyclooxygenase 2 (COX-2) inhibition and specific inhibition of PGE2 in vitro, both of which attenuated fungal-induced IL-22 in Il1rl1−/− mice. This study establishes novel mechanisms of IL-22 induction via PGE2 and regulation of the PGE2-IL-22 axis via IL-33 signaling during lung fungal exposure.
Dendritic cell-intrinsic TLR signaling regulates polymicrobial sepsis
Authors: Liu Bei , Iwanowycz Stephen , Fan Hongkuan
Published: January 2023

Dendritic cells (DCs) are professional antigen-presenting cells that play a critical role in both innate and adaptive immune response. DCs recognize and respond to bacteria through multiple pattern-recognition receptors, including Toll-like receptors (TLRs). Heat shock protein gp96 is a master chaperone for TLRs in the endoplasmic reticulum and plays critical roles in innate immunity. Sepsis is a clinical syndrome caused by a severe immune response to infection. However, the role of DC-intrinsic TLRs in sepsis has not been studied. Using a unique DC-specific gp96 knockout (KO) mouse model, we observed that gp96-deficient DCs were unable to respond to both surface and intracellular TLR ligands. Our DC-specific gp96 deficient mice present a DC-specific pan TLR KO mouse model. We challenged WT and KO mice with a lethal dose of LPS, and we found that WT mice significantly produced more serum TNF-a than did KO mice. Consistent with the detrimental effect of this cytokine in sepsis, we found that the WT mice were more susceptible to LPS-induced sepsis than the KO mice. Furthermore, using a well-defined sepsis model of cecal ligation and puncture (CLP), we found that KO mice exhibited significant higher survival rates (33%) compared to littermate WT controls (5%, p<0.05). Collectively, our study demonstrates that DC-specific gp96 KO mice were able to mount effective responses against polymicrobial sepsis, underscoring the importance of DC-intrinsic TLR signaling in innate immunity and sepsis outcomes.
A mouse model of age-associated tuberculosis reactivation positions KLRG1 as a modulator of disease progression.
Authors: Turner Joanne , Gautam Shalini , Dwivedi Varun
Published: January 2023

Tuberculosis (TB) in the elderly can arise from reactivation of M.tb infection that was acquired when younger and immune exhaustion or senescence is thought to play a role in this process. KLRG1 is an inhibitory receptor of T cell function and its expression is elevated on T cells with increasing age and T cell differentiation status. We have previously shown using KLRG1-KO mice that the absence of KLRG1 provides a survival advantage to chronic M.tb infected mice, showing proof of concept for an inhibitory role of this receptor during M.tb infection. We hypothesized that KLRG1 would have a more significant role in a model of age-associated reactivation TB where immuno-senescence can also play a role. We infected young wild type (WT) and KLRG1 KO mice with M.tb and then reduced the M.tb CFU to low levels using the modified Cornell model (2–3 months isoniazid/rifampicin in water), allowing mice to live into old age. Mice were sampled every 2–4 months for determination of TB reactivation and immune function. M.tb CFU were moderately increased in lung and spleen of WT mice around 14 months of age, an age that is associated with shifts in T cell phenotype and function. For WT mice M.tb CFU reached levels similar to a chronic M.tb infection approximately 18–20 months of age. A significant increase in total T cell numbers and antigen specific CD4 T cells in the lung also occurred at that age. In contrast, KLRG1 KO mice had a substantial delay in reactivation of M.tb, indicating that KLRG1 has a functional inhibitory role during M.tb infection. Our data suggest that modulating KLRG1 function, or the development of T cells that express KLRG1, can have beneficial outcome on the susceptibility of the elderly to succumb to reactivation TB.
The analysis of a Puerto Rican cohort of Essential Thrombocytemia patients
Authors: Maysonet-Cruz Johanna , Calzada Natalia , Washington Anthony V. , Hunter Robert
Published: January 2023

Essential Thrombocytemia (ET) disorder, part of the group of myeloproliferative disorders, has been related to acquired mutations that constitutively activate the JAK2 pathway, accordingly JAK2V617F is present in the 56% of ET patients. ET decreases life expectancy and its effects are detrimental on the mid-fifties population, therefore, it is important to understand and devise methods to improve its treatment. For this reason, we completed a comparative analysis of 37 different cytokines in a cohort of 23 Puerto Rican ET patients and 11 controls. Using Mann Whitney comparative analysis between the status of the JAK2V617F mutation and cytokine profiles have revealed that JAK2V617F positive patients have a significantly higher median values of IL-19 (ET: 6.48 pg/ml; control: 5.13 pg/ml, p-value: 0.0076), Il-28/IFNλ2 (ET: 3.76 pg/ml; control: 2.79 pg/ml, p-value: 0.0422) and IL-29/IFNλ1 (ET: 14.47 pg/ml; control: 10.3 pg/ml, p-value: 0.0332) when compared with healthy controls. TWEAK TNFSF12 (ET: 40.05 pg/ml; control: 47.77 pg/ml, p-value: 0.0437), however was significantly lower in this population. Also, we have found that JAK2V617F negative patients have significantly higher levels of IFN-α2 (ET: 14.82 pg/ml; control: 12.53 pg/ml p-value: 0.0096), IL-8 (ET: 6.93 pg/ml; control: 6.33 pg/ml, p-value: 0.0316), and IL-22 (ET: 7.865 pg/ml; control: 6.23 pg/ml, p-value: 0.0087) than control patients. Moreover, we have found that ET CALR mutants have significantly lower median levels of MPP-3 and that ET patients have higher TLT-1 levels than control patients. Together these findings could be used to enhance the characterization and improve prognosis and treatment of ET patients.
Cancer Immunotherapy: knowledge translation and outreach education
Authors: Sika-Paotonu Dianne , Laban Luamanuvao Winnie
Published: January 2023

It is recognised that Pacific peoples living in New Zealand in the Pacific are affected disproportionately by some cancers with the burden of cancer overall remaining a significant health concern. These communities stand to benefit from research that will contribute to improvements in health outcomes. Community engagement activities and knowledge translation efforts that involve research findings of relevance to Pasifika populations are of importance in supporting the dissemination of knowledge and dialogue. Limited attempts however have been made to communicate major scientific research findings to the Pasifika audience in a manner that is culturally appropriate, clear and meaningful. The purpose of this work was to host a knowledge translation event to present research of direct relevance and importance to Pasifika communities that included a major compartment dedicated to increasing clarity and awareness of emerging cancer immunotherapeutic based strategies for treating cancer. This event was well attended by members of the Pasifika community that included health leaders and representatives, families, scientists, students, academics, health researchers, community representatives and other interested parties. Verbal and written feedback highlighted the programme overall was highly evaluated by those who were in attendance. This knowledge translation and science communication event successfully facilitated engagement and open dialogue with Pasifika members of the health workforce community that included raising awareness of immunotherapeutic strategies for cancer.
Phenotypic characterization of fifty surface markers expressed by human M1 and M2a macrophages cultured with different serums and in the presence or absence of polarizing cytokines.
Authors: Bonnevier Jody L , Hammerbeck Christopher , Goetz Christine , Newman Kristal , Aggeler Birte
Published: January 2023

Macrophages are ubiquitously distributed throughout the various tissues of the body and perform many functions. These include inflammatory responses against pathogens by classically activated M1 macrophages and the regulation of wound healing and tissue remodeling by anti-inflammatory alternatively activated M2 macrophages. The responsibility for these pleiotropic functions lies in the expression of a myriad of surface receptors unique to a given subset. Much of what we know about the function of human macrophage subsets has been gleaned by studying in vitro generated macrophages, matured in the presence of GM-CSF or M-CSF, and polarized with different cytokines. Oftentimes, culture conditions (such as the type of serum used, the duration of the culture and the use of polarizing cytokines) vary between studies making direct comparisons difficult. Furthermore, overlap in surface marker expression can make it difficult to distinguish between the different macrophage subsets.

We directly compared the expression of fifty different surface markers on M1 and M2a macrophages cultured in the presence of fetal bovine serum, human AB serum or serum free media and found that the type or presence of serum used affected the expression of several markers such as CD200R1 and CD32. Moreover, we compared the expression of these surface markers on polarized and unpolarized macrophages and determined that polarization was critical to the expression of several of these markers, such as CD38 and SLAM F7. The results of these studies should significantly expand our knowledge of the phenotypic differences between human M1 and M2a macrophages as well as demonstrate the importance of culture conditions in generating these phenotypes.
Cancer Mutations Targeting TNFRSF14 alter Microenvironment Checkpoint Interactions to Limit Tumor Clearance by Cytotoxic Cells
Authors: Sedy John R , Balmert Mary Olivia , Nguyen Jennifer , Ware Brian C , Bjordahl Ryan , Norris Paula S , Miller Brian R , Aivazian Dikran , Ware Carl F
Published: January 2023

The roles of many non-oncogenic mutations in cancer may influence tumor growth, survival, or how tumors interact with their surroundings. Here we characterize the functional relevance of missense mutations within the gene encoding the tumor necrosis receptor family member HVEM (TNFRSF14), a locus frequently targeted within human lymphoma and other cancers. We find that point mutations identified in human lymphoma were localized to the extracellular domain and specifically target ligand binding, resulting in preferential loss of CD160 and BTLA interactions compared to LIGHT (TNFSF14). Missense mutations were associated with alterations in cytotoxic effector cell signatures within tumor biopsies, while deletion mutations were associated with changes in myeloid cell signatures. Finally, we find that mutated HVEM proteins retained the capacity to inhibit T cell signaling through BTLA, while reducing costimulation of cytolysis in NK cells through CD160. Together, these data provide evidence for how immune selective pressures may drive mutation of TNFRSF14 resulting in greater tumor fitness.
The inflamed phenotype in PD-L1 negative melanoma and response to checkpoint inhibitors.
Authors: Morrison Carl , Pabla Sarabjot , Nesline Mary , Gandhi Shipra , Pandey Manu , Conroy Jeffrey , Glenn Sean , Burgher Blake , Andreas Jonathan , Giamo Vincent , Papanicolau-Sengos Antonios , Ernstoff Marc C. , Galluzzi Lorenzo , Gardner Mark , He Ji
Published: January 2023

Background

PD-L1 positive melanoma patients, with a MEL score of 4 or 5 by immunohistochemistry (IHC), exhibited a response rate of >50% to pembrolizumab in the KEYNOTE-001 trial. Across multiple tumor types, evidence supports that response to checkpoint inhibitors (CPIs) is associated with an inflamed phenotype. Currently there is very little known about predicting response to CPIs in PD-L1 negative inflamed and non-inflamed melanomas.

Methods

86 melanomas were tested using PD-L1 IHC and a custom NGS immune gene expression assay. 72 of these patients were treated with one or more CPIs. A PD-L1 MEL score of 4 or 5 was considered positive for IHC, and over expression of a majority of 43 genes related to T-cell activation and 11 genes associated with tumor infiltrating lymphocytes were considered positive for the inflamed phenotype. RECIST v1.1 was used to assess patient response

Results

8 of the 86 samples were PD-L1 IHC positive, 10 of the 86 were PD-L1 IHC negative, and all 18 samples in this group showed an inflamed phenotype. Of the 72 patients that could be evaluated for response to CPIs, 8 demonstrated an inflamed phenotype while 64 were non-inflamed.

Conclusion

A positive PD-L1 IHC MEL score of 4 or 5 fails to identify most melanoma patients with response to CPIs. The inflamed phenotype also fails. Most responders demonstrate a PD-L1 IHC negative/non-inflamed phenotype. Additional studies of the immune phenotype in melanoma are required to evaluate prediction of response to CPIs.
Secretion of IL-1β from imatinib-resistant chronic myeloid leukemia cells contributes to BCR–ABL mutation-independent imatinib resistance
Authors: Lee Wongeun , Lee Cho-Rong , Kang Jung-Ah , Yang Taewoo , Park Sung-Gyoo
Published: January 2023

Some cases of chronic myelogenous leukemia are resistant to tyrosine kinase inhibitors (TKIs) independently of mutation in BCR–ABL, but the detailed mechanism underlying this resistance has not yet been elucidated. In this study, we generated a TKI-resistant CML cell line, K562R, that lacks a mutation in BCR–ABL. Interleukin-1β(IL-1β) was upregulated in imatinib-resistant K562R than in the parental cell line K562S, and higher levels of IL-1β contributed to the imatinib resistance of K562R and to cell survival. In addition, IL-1β secreted by K562R cells could stimulate a bone marrow stromal cell line to produce chemokines, such as CXCL11, that lead to the migration of the CML cells into the stroma. Thus, elevated IL-1β production from TKI-resistant K562R cells may contribute to TKI resistance by increasing cell viability and promoting cell migration.
Activator of G-Protein Signaling 3 Modulates Prostate Cancer Development and Progression
Authors: Richardson Micheler , Adekoya Timothy , Smith Nikia , Mowart Tonelia A
Published: January 2023

Prostate cancer (PCa) is a leading cause of cancer death among men, with greater prevalence of the disease among the African American population in the US. AGS3, a receptor-independent activator of G-protein signaling has been shown to affect different cellular processes and cell cycle activity. The aim of this study is to assess the role of AGS3 on the development and progression of prostate cancer as well as to understand the molecular dynamics involved. Preliminary results indicate a differential in the level of expression of AGS3 in PCa cells with the African American cell line (MDA PCa 2b) expressing high amount of AGS3 while the Caucasian cell lines (PC3 and LNCAP) do not. Overexpression of AGS3 in PC3 augmented tumor development in nude mice xenografts whereas depletion of AGS3 in MDA PCa 2b decreased tumor size. Interestingly, a xenograft model of Lewis Lung Carcinoma (LLC) cells in AGS3 conditional knockout (AGS3−/−) mice resulted in a significant increase in tumor progression when compared with the wild-type (AGS3+/+) mice. Taken together, these results indicate that AGS3 expression modulates tumor development and progression and may play a role in PCa health disparity.
EFhd2/ Swiprosin-1 regulates the cytoskeleton of B lymphocytes and impacts germinal center reaction
Authors: Reimer Dorothea , Heyland Edmar , Wiesmann Veit , Wittenberg Thomas , Brachs Sebastian , Voehringer David , Hauser Anja , Niesner Raluca , Mielenz Dirk
Published: January 2023

The germinal center (GC) reaction is a decisive checkpoint for efficient humoral immunity. Most GC associated processes, e.g. cell migration and cell adhesion, are strongly dependent on cytoskeletal dynamics and B cell receptor (BCR) signaling. However, the molecular connection between downstream BCR molecules and cytoskeletal proteins is not yet fully elucidated.

The adaptor protein EFhd2/ Swiprosin-1 co-localizes with the BCR at the plasma membrane. It controls F-Actin bundling as well as Actin remodeling through Cofilin activity and is involved in spreading, lamellipodia formation and migration in non-lymphoid cells. We showed that constitutively EFhd2 deficient mice display enhanced GC responses upon challenge with the helminth Nippostrongylus brasiliensis in a B cell intrinsic manner in mixed bone marrow chimeras. Moreover, B cell specific deletion of EFhd2 leads to enhanced GC B cell numbers already in steady state. We therefore hypothesize that EFhd2 controls the GC reaction by regulating cytoskeletal processes. In vitro transwell migration assays and in vivo studies using two-photon microscopy revealed decreased directed chemotaxis but increased random chemokinesis of EFhd2−/− B cells. Moreover, activated B cells spread faster in the absence of EFhd2 on anti-BCR antibodies which correlates with accelerated Cofilin activity. Finally, also BCR capping was increased in activated EFhd2 deficient B cells.

Taken together, we suggest that EFhd2 controls Actin dynamics through Cofilin in B cells, thereby, controlling chemotaxis of B cells and BCR mediated B cell spreading. Hence, EFhd2 links proper coordination of the B cell cytoskeleton with the GC checkpoint and impacts TH2 response.