bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-12p35 suppresses encephalitis and uveitis in mice
Authors: Choi Jin Kyeong , Yu Cheng-Rong , Mary Mattapallil , Lee Hyunsu , Kang Minkyung , Oladipupo Favour , Caspi Rachel R. , Egwuagu Charles E. 
Published: January 2023

Abstract
Interleukin 35 (IL-35) cytokine is comprised of two subunits, IL-12p35 and Ebi3 and both subunits are thought to be required for the expansion of regulatory B cells (Bregs) that suppress autoimmune diseases. Although use of IL-35 to produce Breg cells has substantial promise for the development of cell therapy against autoimmune and neurodegenerative diseases, some significant challenges have yet to be overcome before it can brought to the clinic. For example, the association of the IL-12p35 and Ebi3 subunits is not strong and readily dissociates, making it difficult to ascertain the effective dose of the bioactive IL-12p35:Ebi3 heterodimer administered or required to ameliorate disease. In this study, we show that IL-12p35 alone has anti-inflammatory properties that recapitulated essential therapeutic functions of the heterodimeric IL-35 cytokine. IL-12p35 induced the expansion of Breg cells in vivo and adoptive transfer of ex-vivo-generated Breg cells using IL-12p35 suppressed inflammation and the development of pathology in mouse models of experimental autoimmune uveitis (EAU) or encephalomyedlitis (EAE). The inflammatory immune cells penetrated the retina, brain and spinal cord in the untreated mice whereas these traits were far more reduced in the IL-12p35-treated mice. In conclusion, the recapitulation of essential immunosuppressive activities of IL-35 by IL-12p35, indicates that IL-12p35 may be utilized for in vivo expansion of Breg cells and autologous Breg cell immunotherapy. These and our unpublished observations suggest that IL-12 single chain proteins may constitute a new generation of therapeutic cytokines that can be exploited to treat autoimmune and neurodegenerative diseases.
The role of CD39-expressing regulatory T cells following treatment with αCD52: an effective therapeutic against central nervous system autoimmunity
Authors: Brass Sara Colpitts , Burgess Joseph N. , Begum-Haque Sakhina , Kasper Lloyd H. 
Published: January 2023

Abstract
Alemtuzumab (αCD52), a novel immunotherapy approved for the treatment of relapsing multiple sclerosis (MS), induces the widescale deletion of lymphocytes, including self-reactive T cells. Using the mouse model of MS, experimental autoimmune encephalomyelitis (EAE), we previously found that following reconstitution of the immune compartment, there was an increased frequency of CD39+Foxp3+ regulatory T cells (Tregs) in the spleen and gut-associated lymphoid tissues of αCD52-treated mice. We hypothesized that the CD39-expressing Tregs that emerge following lymphocyte reconstitution would be suppressive against central nervous system autoimmunity. However, when CD39+ Tregs were purified from αCD52-treated mice and adoptively transferred to naïve recipients, they were unable to reduce the severity of EAE disease. We next used Foxp3/GFP reporter mice treated with αCD52 to compare the suppressive capacity of CD39+Foxp3+ versus CD39+Foxp3− Treg subsets and found that CD39+Foxp3+ Tregs, but not CD39+Foxp3− Tregs, were able to significantly reduce the proliferation of activated naïve T cells in vitro. Finally, we sought to determine if expression of CD39 was required for the therapeutic effects of αCD52 in the EAE model. Preliminary studies using a limited number of CD39 KO mice indicated that treatment with αCD52 was equally effective in both control and KO mice. Thus, our studies have identified potential differences in individual subsets of Tregs emerging after αCD52 treatment and suggest that expression of CD39 is dispensable for the short-term effectiveness of αCD52 in mice. Future studies are required to determine if CD39 expression on Tregs contributes to the long-term suppression of MS disease in patients treated with αCD52.
Effects of anti-CTLA-4 and anti-PD-1 on memory T-cell differentiation and resistance to tumor relapse
Authors: Mok Stephen , Duffy Colm R. , Allison James P. 
Published: January 2023

Abstract
Clinical studies have shown that αPD-1 therapy mediates a greater response rate compared to αCTLA-4 according to RECIST criteria. However, it may not be as durable as αCTLA-4. 25% of patients who initially responded to αPD-1 have tumor relapse within 24 months. In contrast, the 10-year survival rate of patients treated with αCTLA-4 is at least ~25% suggesting a durable response. The goal of this study is to understand the relationship between memory T-cell formation in tumor models mediated by checkpoint antibodies and tumor relapse.
Our results have shown that αCTLA-4 could induce a superior memory antitumor response during tumor rechallenge on mice compared to that treated with αPD-1 after 80 days. We also showed that this memory antitumor response was antigen-specific. These antigen-specific memory T-cells in mice that were treated with αCTLA-4 had better proliferative potential with cytokine producing function and more CD62L+CD44+ memory T-cells were converted back to CD62L-CD44+ effectors during rechallenge. We found that αCTLA-4 and αPD-1 facilitated the development of CD4 and CD8 memory T-cells respectively. We also demonstrated that the memory antitumor activity mediated by the combination of αCTLA-4 and αPD-1 overlapped with αCTLA-4 alone suggesting the effect of the combined treatment is dominated by αCTLA-4.
Bifidobacterium can mitigate intestinal immunopathology in the context of immune checkpoint blockade
Authors: Wang Feng , Sun Shan , Yin Qian , Chen Liang , Davis Mark M. 
Published: January 2023

Abstract
Antibodies that attenuate immune tolerance have been used to effectively treat cancer, but they can also trigger severe autoimmunity. To investigate this, we combined anti-CTLA-4 treatment with a standard colitis model to give mice a more severe form of the disease. Pre-treatment with an antibiotic, Vancomycin provoked an even more severe, largely fatal form, suggesting that a gram-positive component of the microbiota had a mitigating effect. We then found that a commonly used probiotic, Bifidobacterium could largely rescue the mice from immunopathology without an apparent effect on anti-tumor immunity, and this effect may be dependent on regulatory T cells.
Autophagy-deficient breast cancer shows early escape from dormancy and recurrence following chemotherapy
Authors: Aqbi Hussein F. , Smith Timothy , Butler Savannah , Idowu Michael , Payne Kyle K. , Manjili Masoud H. 
Published: January 2023

Abstract
Breast cancer patients who respond to cancer therapies often end up with a distant recurrence of the disease. It is not clear why people with the same type of breast cancer respond to treatments differently, some escape from dormancy and relapse earlier than others. In addition, some tumor clones respond to immunotherapy while others do not. Here, we investigated the role of autophagy in expediting or delaying recurrence of neu overexpressing mammary carcinoma (MMC) following adriamycin (ADR) chemotherapy, and in affecting response to immunotherapy. We took two strategies, including a transient blockade of autophagy with chloroquine (CQ) which transiently blocks fusion of autophagosome and lysosome, and a stable knockdown of autophagy, which inhibits the formation of autophagosome in MMC. We found that while CQ prolonged tumor dormancy, a stable knockdown of autophagy resulted in early escape from dormancy and recurrence. Interestingly, MMC with a stable knockdown of autophagy contained an increased frequency of ADR-induced polyploidy cells which rendered MMC resistant to immunotherapy. On the other hand, a transient blockade of autophagy did not affect the sensitivity of MMC to immunotherapy. Our observations suggest that while chemotherapy-induced autophagy may facilitate tumor relapse, cell intrinsic autophagy delays tumor relapse by inhibiting the formation of polyploidy tumor cells. Our findings are consistent with other reports showing that autophagy-deficient cells contain polyploid nuclei because of increased levels of reactive oxygen species. Our results also suggest that responsiveness of breast cancer patients to neoadjuvant therapies might be determined by tumor intrinsic autophagy.
Chemotherapeutic drugs induce expression of CS1 (CD319) in multiple myeloma cells resulting in enhanced NK cell mediated killing.
Authors: Malaer Joseph D. , Mathew Porunelloor A. 
Published: January 2023

Abstract
Elotuzumab (Empliciti), a humanized monoclonal antibody against CS1, has been demonstrated an effective drug for multiple myeloma (MM). During clinical trials, elotuzumab in combination with chemotherapeutic drugs was more effective than antibody treatment alone. However, the underlying mechanism for this is not yet known. We hypothesize that chemotherapeutic drugs induce the surface expression of CS1 on MM cells making them more susceptible to natural killer (NK) mediated killing and therefore increasing the effectiveness of elotuzumab. NCI-H929 (multiple myeloma cell line) cells were incubated with lenalidomide and anti-CS1 mAb, or doxorubicin and dexamethasone in order to determine if surface and mRNA expression of CS1 changes. Surface expression of CS1 was determined via flow cytometry, while mRNA expression was determined by RT-qPCR. Our results indicate that treatment with anti-CS1 mAb in combination with lenalidomide or treatment with doxorubicin and dexamethasone increased the cell surface expression of CS1; however, these drugs reduced the expression of CS1 mRNA. Additionally, chemotherapeutic drug incubation increased NK cell effector function against NCI-H929 cells. These results, in conjunction with previous clinical trials, may indicate a novel mechanism in which chemotherapeutic drugs increase surface CS1 expression and therefore increase the effectiveness of anti-CS1 mAb therapy.
Approaches to reverse interleukin-10 mediated immunosuppression in chronic lymphocytic leukemia
Authors: Rivas Jacqueline R. , Alhakeem Sara S. , Eckenrode Joseph , Zhang Yinan , Collard James P. , Rangnekar Vivek M. , Muthusamy Natarajan , Thorson Jon S. , Leggas Markos , Bondada Subbarao 
Published: January 2023

Abstract
B cell Chronic Lymphocytic Leukemia (CLL), the most common leukemia in the western world, is characterized by an accumulation of abnormal B cells in the blood and lymphoid organs. Broad immunosuppression is a serious complication for these patients, with infections and secondary cancers recognized as the leading causes of morbidity and mortality. Although the immune suppression is in part due to clonotypic CLL B cells crowding immune organs, immune responses may also be suppressed by the production of soluble mediators. We and others found that CLL cells produce interleukin-10 (IL-10), a cytokine that suppresses T cell effector function. We hypothesized that this IL-10 may reduce anti-tumor T cell responses. In our studies we use human cells as well as the Eμ-Tcl1 mouse model for U-CLL, in which the oncogene Tcl1 is expressed under the immunoglobulin VH promoter and μ-enhancer. Previously, we found that IL-10R−/− T cells control CLL growth better than WT T cells in the Eμ-Tcl1 adoptive transfer model. We also saw that CLL IL-10 production is dependent on BCR signaling and activation of the transcription factor Sp-1. IL-10 production can be blocked by mithramycin (MTM), an Sp-1 inhibitor. However, MTM is not used routinely as an anti-cancer agent due to its short half-life and associated toxicities, so we prepared and tested novel analogues of MTM to reduce these effects. One of these analogues, MTM23, shows promise as it has similar potency to MTM, but does not interfere with T cell gamma-interferon production or viability. This approach is novel as current therapies for CLL do not target IL-10, or address the immunosuppression seen in CLL patients. IL-10 blockade could therefore be a strategy for harnessing anti-tumor T cells to treat CLL in humans.
A clonotype-independent pre-TCR complex and its downstream signals can support transformation of thymic T cells.
Authors: Hathcock Karen S. , An Joseph BH. , Hodes Richard J. 
Published: January 2023

Abstract
The kinase ataxia telangiectasia mutated (ATM) communicates between molecules that sense DNA damage in the form of double stranded breaks (DSB) and effector mechanisms that maintain genomic integrity. Persistent DSBs can lead to transformation and ATM-deficiency is associated with increased lymphoid malignancies in both humans and mice. ATM-deficient mice exclusively develop thymic T cell lymphomas with high penetrance. We explored the requirement for specific components of the TCR and its signaling pathway for the induction of these thymic lymphomas. We generated multiple strains of ATM-deficient mice in which mature T cell are absent by deletion of CD3ɛ, an invariant component of the pre-TCR complex; by deletion of RAG or TCRβ, in which recombining TCR variable components are deleted; or by deletion of LAT, an important component of pre-TCR and TCR signaling. These strains all showed a similar and severe developmental arrest at thymic DN3. Despite this arrest, mice deficient in RAG or TCRβ, exclusively developed thymic T cell lymphomas. In contrast, mice deficient in CD3ɛ or LAT, developed no or few T cell tumors, respectively, but developed B cell lymphomas. These findings suggest the importance of a clonotype-independent TCR complex and its downstream signals in transformation of T cells. Experiments are in progress to ask if survival of ATMKO T cell lymphomas after transformation is similarly dependent on CD3ɛ and LAT signaling.
Multiple defects in NK cells of surgically resectable pancreatic cancer patients can be reversed by ex-vivo stimulation
Authors: Jeon Seunghyun , Lim Seon Ah , Kim Jungwon , Shin Min Hwa , Kwon Joonha , Lee Kyung-Mi 
Published: January 2023

Abstract
Tumor infiltrating lymphocytes (TILs) are found in advanced pancreatic ductal adenocarcinoma (PDAC) patients and shown to correlate with overall survival (OS) rate. Although majority of TILs consisted of CD8+/CD4+T cells, the presence of NK cells and their role in the pathogenesis of PDAC remains elusive. We performed comprehensive analyses of TIL and PBMC to explore the role of NK cells in the newly developed PDAC patients. Multiple defects in cell migration and effector function were observed in NK cells of patients’ peripheral blood. Extremely low frequencies of NK cells (&lt;0.5%) were found in TILs of PDAC patients which was attributable not to low expression of tumor chemokines, but rather lack of the CXCR2 chemokine receptor. Forced expression of CXCR2 on patients’ NK cells rendered them capable of trafficking PDAC tumors. Furthermore, patients’ NK cells exhibited impaired cell-mediated killing against autologous PDAC tumor cells and failed to proliferate within hypoxic tumor microenvironment. Importantly, these defects can be overcome by ex-vivo activation of NK cells from PDAC patients. When the proliferative capacity of NK cells in vitro was used to stratify patients on the basis of cell expansion, a clear difference in DFS and OS was observed. Patients whose NK cells proliferated &lt;250 fold experienced significantly lower DFS and OS than those expanded ≥250 fold. Multiple defects are observed in NK cells of PDAC patients. Ex-vivo activation of NK cells restores tumor reactivity and provides a therapeutic modality while their fold expansion can be a potentially significant prognostic indicator of OS and DFS in PDAC patients.
Notch-mediated crosstalk of antitumor CD8+T and NK cells
Authors: Thomas Portia , Uzhachenko Roman , Shanker Anil 
Published: January 2023

Abstract
A large majority of immune-oncology research efforts focus on T-cell immunotherapy. However, limited clinical efficacy has been achieved with adoptive T-cell effectors alone. In addition, research has shown that the sole use of adoptive T-cells leads to the development of tumor escape variants and “occult cancer”. Conversely, tumor-infiltrating natural killer (NK) cells have been shown to cause elimination of these escape variants. These observations indicate that the interaction between the innate and adaptive immune effectors is indispensable for effective tumor immunity. Notch is a major signaling system important for communication between neighboring cells. Our previous studies show that it plays a crucial role in the differentiation and antitumor function of CD8+T cells. However, its role in NK cell activation and antitumor function is not clear. Thus, we analyzed Notch receptor-ligand system on NK cells and in the context of their crosstalk with CD8+ T cells. Our data indicate that inhibiting Notch signaling in NK cells decreases NK cell activation. In addition, Notch intracellular domain (NICD) expression is increased upon NK activation, suggesting that the Notch signaling pathway may be involved in NK activation. Our data also show that activated CD8+T cells activate naïve NK cells through direct physical contact. Transmembrane notch receptor expression is decreased during this interaction suggesting that the Notch receptors may be involved in CD8+T cell – NK cell crosstalk. Experiments are underway to further understand the relevance of Notch signaling pathway in innate-adaptive immune crosstalk. The findings will help develop novel strategies for adoptive cell immunotherapies, specifically against solid tumors.
Memory CD4+ T cells guide memory B cell adaptability to drifting influenza vaccination
Authors: Gage Emily , Van Hoeven Neal , Coler Rhea 
Published: January 2023

Abstract
Influenza A annually infects 5–10% of the world’s human population resulting in an estimated one million deaths. Unlike other infectious agents, where either infection or vaccination against the disease confers long-term immunity, influenza causes annual epidemics and re-infects previously exposed individuals as a result of antigenic drift in the surface glycoprotein hemagglutinin (HA). Due to antigenic drift, the human immune system is simultaneously exposed to both novel and conserved parts of the influenza virus through vaccination and/or infection multiple times throughout life. Preexisting immunity by infection or vaccination influences subsequent neutralizing antibody responses, the correlate of protection for influenza. To understand how preexisting immunity augments future responses to drifted influenza immunization, we established mouse models, sequentially infecting mice with a H1N1 strain and then immunizing with a second drifted H1N1 strain. Mice previously infected with A/CA have increased neutralizing antibody response (nAb) to A/PR upon immunization with A/PR HA. This increase in nAbs was dependent on CD4+ T cell and memory B cell, correlating with CD4+ T cell reactivity conserved across both influenza HA’s. We also found increased germinal center PR8-specific B and T follicular helper cells post vaccination in the draining lymph node. These results suggest conserved MHC Class II restricted epitopes within HA are critical for B cells to adapt to drifting influenza and could be leveraged to boost out subsequent neutralizing antibody responses. Understanding the mechanism by which preexisting immune responses shape future responses is essential to optimize and leverage vaccination strategies.
Pattern recognition receptor agonists enhance protection elicited by a single, low dose protein vaccine in an immunization route-specific manner
Authors: Lampe Anna T. , Hain Jacey L. , Farris Eric J. , Pannier Angela K. , Brown Deborah M. 
Published: January 2023

Abstract
Despite advances to increase vaccine efficacy, influenza A virus (IAV) remains a significant health risk. Current vaccines against IAV rely on the generation of strain specific antibodies that fail to provide protection against heterosubtypic IAV. Vaccination may be improved through the use of vaccine adjuvants to amplify antigen presenting cell (APC) activation and enhance T and B cell priming. Combined adjuvants that trigger multiple pattern recognition receptors (PRR) are a promising strategy to better mimic live infection. In this study, the efficacy of combining CpG and MPLA, which activate TLR9 and TLR4 respectively, as vaccine adjuvants was examined. It was observed that single, low dose hemagglutinin (HA) protein vaccines containing combination CpG and MPLA delivered intramuscularly (i.m.) or intranasally (i.n.) elicit superior protection against lethal challenge compared to vaccines containing single agonists, as indicated by lower mortality and significantly lower morbidity. Early systemic cytokine responses are being investigated to elucidate the mechanism behind this improved vaccine efficacy. However, it was observed that i.m. administration of combination agonist vaccines induced higher IgG2c antibody responses compared to single agonist vaccines, while minimal levels of antibodies were observed in mice vaccinated i.n. In addition, T cell analysis following lethal infection indicated that i.n. vaccination induces distinct activation profiles compared to i.m. Taken together a comparison of the antibody and T cell profiles of mice vaccinated i.m. and i.n with combined TLR9 and TLR4 agonists suggests that route of vaccination can dictate immune response and may modify effective correlates of protection.
Antibody levels to Influenza A are greater in adults with Trisomy 21 but are less functional compared to disomic adults.
Authors: King Scott , Kusher Samantha , Jacobs Brennan , James Stephanie F. 
Published: January 2023

Abstract
Adults with Trisomy 21 (Down syndrome; DS) are particularly susceptible to respiratory infections which is a primary cause of death in adults with DS. Due to medical advances there has been a rapid increase in the lifespan of people with DS, creating a critical gap in our understanding of immunity in these adults. Although no studies to date have identified the pathogens responsible for the majority of respiratory infections observed in adults with DS, data from previous influenza epidemics suggest that adults with DS have significantly higher rates of morbidity and mortality as a result of influenza infection. Their high rates of pneumonia and subsequent death may result from an impaired ability of B cells to generate antibodies of sufficient quantity and function to control infections. Data evaluating the effectiveness of immunizations in adult individuals with DS is inconclusive, although it has been established that those with DS have decreased B cell counts. Herein, we hypothesized that adults with DS will have an attenuated antibody response following immunization compared to typical adults. Antibody concentrations to influenza A were evaluated using a standard ELISA, and antibody titer was measured using a hemagglutination inhibition assay. Our results show adults with DS have significantly increased IgG antibody concentration to Influenza A compared to age matched controls (p=0.005). However, when evaluated by a hemagglutination inhibition assay (to A/HongKong) adults with DS had lower levels of inhibition compared to typical adults (p=0.02). These data indicate that although adults with DS can develop an antibody response to influenza, this response may not be as protective against disease as in typical, disomic individuals.