bluuebunny/crossref_metadata_2025_split huggingface dataset:
Caloric restriction responses can maintain immune responses during aging (47.14)
Authors: Ruby Carl , Jimenez Hugo , Goldufsky Josef , Chisholm Lana , Lukaesko Lisa , Weinberg Andrew , Kaufman Howard 
Published: January 2023

Abstract
Age-related defects accumulate in both the innate and adaptive arms of immunity to generate a state of immune deficiency. Our laboratory has previously shown a decrease in immune responses in aged mice compared to younger mice in the context of OX40 costimulation. This loss of immunity during aging underscores the need for simple approaches to boost immunity in older individuals. We hypothesize that immune responses in the context of OX40 activation or blockade of CTLA-4 in older mice can be restored and/or maintained through caloric restriction (CR) or dietary supplementation with a CR mimetic. Two CR dietary regimens were used to determine if immunity in old immune-compromised mice could be restored and if immunity could be maintained during aging. In addition mice were fed diets that were formulated with resveratrol (100 ppm), a CR mimetic. Tumor immune responses and Ag-specific T cell responses in these mice were then assessed and compared to young mice and the appropriate dietary controls. Our results demonstrate that caloric restriction cannot restore immune function in old mice. However, CR or resveratrol dietary supplementation initiated at a young age can maintain both OX40-mediated anti-tumor immunity and T cell responses as these mice age. Thus, the induction of a caloric restriction response (via traditional CR or by a CR mimetic) appears to maintain immunological fitness and may increase the effectiveness of immune-based cancer therapies in older subjects.
Loss of nuclear Pro-IL-16 imparts resistance to apoptosis in CTCL T cells (47.5)
Authors: Tuzova Marina , Memari Patrick , Mosaheb Munir , Cruikshank William 
Published: January 2023

Abstract
Cutaneous T-cell Lymphoma (CTCL) is a rare type of non-Hodgkin’s lymphomas characterized by increased T cell proliferation and decreased apoptotic responses. We have reported on the loss of nuclear Pro-IL-16 in CTCL T cells due to a mutation restricting nuclear translocation, which results in dysregulation of Skp2 and p27kip1, and promotes hyper-proliferative state. We have now investigated the relationship between the loss of nuclear Pro-IL-16 and apoptosis. RT-PCR analysis of a panel of apoptosis-related genes was conducted using T cells obtained from IL-16 knock out and wild type control mice. The IL-16 null cells demonstrated a significant reduction in pro-apoptotic genes such as Bax, Bcl10, Bcl2110, and caspases 7, 8 and 9, with an increase in expression of anti-apoptotic genes such as Bag1, Tsc22d3 and Pim2. Annexin V staining of these cells indicated a 43% reduction in the apoptotic response to CD3/CD28 stimulation in the knock-out cells. To confirm the anti-apoptotic effect of nuclear Pro-IL-16, human CTCL-derived T cell line (HUT 78) was transfected to express normal full-length IL-16. Based on Annexin V staining, the apoptotic response increased by 35% in cells, expressing normal IL-16. These studies indicate that loss of nuclear Pro-IL-16 in abnormal T cells correlates not only with increased cell proliferation, but with a decrease in apoptosis as well.
Naringenin modulates intracellular signaling pathways and cell surface receptors in Chlamydia trachomatis-infected mouse J774 macrophages to control concomitantly elicited inflammation (48.3)
Authors: Yilma Abebayehu , Singh Shree , Dennis Vida 
Published: January 2023

Abstract
Sexually transmitted Chlamydia trachomatis infection is of public health concern because of its potential devastating reproduction consequences. C. trachomatis invades the mucosal surface of the female genital tract after it is recognized by cell surface receptors. Increasing evidence now suggests that inflammatory mediators, which are produced by infected cells, contribute to the chronic inflammatory conditions. Controlling excess inflammatory mediators during the early stage of an infection may be a viable alternative to combat the disease. Previously we reported that naringenin, a natural flavonone, reduced IL-6 and TNF levels by up to 70% in C. trachomatis-infected mouse J774 macrophages. Here we hypothesize that naringenin reduces inflammatory mediators by inhibiting intracellular signaling pathways and cell surface receptors. Naringenin inhibited the phosphrylation of p38 MAP kinase and NFκB as induced by C. trachomatis-infected J774 macrophages. Flow cytometric analyses further revealed the ability of naringenin to reduce the cell surface receptor expression of TLR2, TLR4 and CD86 but not CD80 on infected macrophages. These findings suggest the potential of naringenin not only to control inflammatory mediators produced by innate immune cells but also those produced by C. trachomatis-activated T cells during the adaptive immune response. Overall our data imply the positive role played by naringenin in controlling an early C. trachomatis-induced inflammation.
Contact time dependent Effect of human mesenchymal stem cells on IFN-r and IL-17 secreted by CD4+ T effector memory cells (48.4)
Authors: Kim Mee Kum , Jeong Hyun Jeong , Lee Young Eun , Ryu Jin Sook , Lee Won Woo , Wee Won Ryang 
Published: January 2023

Abstract
Mesenchymal stem cells(MSCs) has known to have antiinflammatory properties in various inflammatory disease, however the effect of MSCs on the inflammation is reported to be variable depending on the disease entity, contact time, application time and administration site for the delivery. The study was aimed to analyze the contact time-dependent effect of human mesenchymal stem cells(hMSCs) on IL-17 and INF-r secreted by CD4+ T effector memory cells. Human peripheral blood mononuclear cells(PBMCs) was obtained from the healthy voluntary donors with informed consents(n=5). The study was performed in accordance with the guidelines of the Declaration of Helsinki. CD4+ T cells were purified through MACS separator with the human CD4+ T Cell Isolation Kit II (Miltenyi Biotec). CD4+ T cells were stimulated with anti-CD3/CD28 and cultured in medium supplemented with IL-1β and IL-23. The bone marrow derived hMSCs got in contact with CD4 T cells on 0 day, 3 days, and 5 days after stimulation at a T cell-to-MSC ratio of 10:1. We found that the earlier hMSCs were contacted to CD4+ CD45RO+ T cells, the more effect was suppressive on both IFN-γ and IL-17A secreted by CD4+ effector memory T cells. In fact, later contact of hMSCs to activated T cells dropped most suppressive effect of hMSCs down on effector memory CD4 T cells. It suggested that the earlier time point contact might be the better for hMSCs to down-regulate the functions of effector memory CD4 T cells after activation.
Staphylococcus aureus protein A mediates innate immune responses during nasal colonization in humans (49.25)
Authors: Cole Alexander , Muthukrishnan Gowrishankar , Eade Colleen , Wood Matthew , Lamers Ryan , Ellis Austin , Cole Amy 
Published: January 2023

Abstract
Mucosa of the anterior human nares can harbor Staphylococcus aureus (SA), an etiologic agent that causes severe infections. Nasal carriage of SA is a complex host-pathogen interaction involving bacterial and host mediators that influence innate host defenses. We have revealed that the exoproteome of SA nasal carrier strains contains unique determinants, which enable SA to colonize the nares. In the current study, we assess how one of these bacterial mediators, Staphylococcal protein A (Spa), affects host innate immune response to SA nasal carriage. Functional disruption of spa was performed in an SA nasal carrier strain using Targetron insertion of a group II intron. Subsequent in vitro growth kinetics analysis and ex vivo culture on human nasal epithelial cells (HNECs) revealed no differences between wild type and spa-disrupted (Δspa) SA strains. However, the Δspa mutant induced lower expression of pro-inflammatory cytokines including IL6, IL8 and TNFα from HNECs compared to the levels elicited by the wild type strain. Additionally, IL17, a crucial regulator of mucosal immunity, was induced at significantly lower levels in HNECs inoculated with Δspa SA compared to those inoculated with the wild type SA strain. Furthermore, expression of the chemokines MIP1β and MCP1, as well as HL60 neutrophil migration, were significantly reduced in conditioned media from HNECs inoculated with Δspa mutants. Spa is likely involved in mediating the mucosal response to SA colonizing the nose.
Building a toolkit: An undergraduate teaching laboratory sequence in immunology (50.1)
Authors: Chatterjea Devavani , Linder Grace 
Published: January 2023

Abstract
We have designed a laboratory curriculum in immunology where students build and use a toolkit of basic techniques. In the first 10 weeks, students learn how to isolate human and mouse immune cells, label cells with antibodies, perform magnetic enrichment, flow cytometry analysis, T cell proliferation, and ELISA assays. We emphasize keen observation, excellent record keeping, and protocol development. We ensure that each team has at least two opportunities to perform each new technique. Students survey the published literature and formulate a novel question about effects of a dietary supplement of choice on in vitro T cell stimulation. Teams develop experimental designs that are critiqued in a group workshop. Independent experiments are carried out, data collected and analyzed by teams, and a detailed, reflective lab notebook entry compiled by each student. Teams present their findings in a culminating mini-symposium. Our iterative emphasis on skill building, critical record-keeping, application of newly learned skills, and teamwork closely simulates the learning that happens in authentic research laboratories. Students encounter and comprehend these basic procedures in classroom discussions, research seminars, and journal articles. According to students surveyed, this plays a critical role in the productive integration of classroom and laboratory learning, and gives students a strong sense of engagement with the professional world of immunology research.
Anti-MS4a4B treatment abrogates MS4a4B-mediated protection in T cells and ameliorates experimental autoimmune encephalomyelitis (51.13)
Authors: Xu Hui , Yan Yaping , Li Zichen , Zhang Guang-Xian , Williams Mark , Carey Gregory , Zhang Jianke , Rostami Abdolmohamad 
Published: January 2023

Abstract
MS4a4B is a membrane protein that highly expressed in T cells. We have demonstrated previously that MS4a4B has a regulatory role in proliferation and apoptosis of T cells. In this study, we further analyzed the mechanisms underlying MS4a4B-mediated anti-apoptotic effects in T cells and tested the feasibility of using anti-MS4a4B antibody as a therapeutic approach for experimental autoimmune encephalomyelitis (EAE). Our data showed that knockdown of MS4a4B expression by MS4a4B-specific siRNA markedly enhanced levels of anti-CD3 stimulation-induced cleavage of caspase 3, caspase 8 and caspase 9 in pre-activated primary T cells. Interestingly, binding of anti-MS4a4B antibodies to T cells induced activated T cells to undergo apoptosis and administration of anti-MS4a4B antibody in vivo caused T cell depletion in the peripheral blood. To test the therapeutic effects of anti-MS4a4B antibody in suppression of T cell-mediated autoimmune responses, we injected anti-MS4a4B antibodies into EAE mice. The results show that anti-MS4a4B treatment ameliorated the severity of EAE, accompanied by decreased Th1 and Th17 cell responses and reduced levels of pro-inflammatory cytokines in the central nervous system (CNS), suggesting that MS4a4B may serve as a target of antibody-based therapy for T cell-mediated diseases. Supported by American Heart Association grant AHA-0830072N (to H.X)
Durable pharmacological responses from the peptide drug ShK-186, a specific Kv1.3 channel inhibitor that suppresses T cell mediators of autoimmune disease (51.5)
Authors: Munoz Ernesto , Tarcha Eric , Londono Luz , Norton Kayla , Banks Amy , Chi Victor , Beeton Christine , Chandy K. , Iadonato Shawn 
Published: January 2023

Abstract
The Potassium Kv1.3 channel on effector memory T cells is a target to treat autoimmune diseases. ShK-186, a specific peptide inhibitor of Kv1.3, has shown promise in animal models of multiple sclerosis and rheumatoid arthritis. Here, we describe the development of methods to establish a pharmacokinetic (PK) - pharmacodynamic (PD) relationship for ShK-186, and test the hypothesis that the drug has a sustained PD effect in autoimmune disease models. The PK of ShK-186 was evaluated in rats and cynomolgus monkeys using HPLC-MS/MS and ELISA methods to measure the free fraction in plasma. These results were compared with SPECT/CT data collected with an 111In-DOTA-conjugate of ShK to assess whole blood PK parameters as well as the drug’s absorption, distribution and excretion. Our data suggest ShK-186 is absorbed slowly from the injection site resulting in blood concentrations above the Kv1.3 channel-blocking Kd for up to 7 days in monkey. PD studies on human peripheral blood mononuclear cells show that transient ShK-186 treatment results in a sustained suppression of cytokine responses and may contribute to prolonged drug effects. Moreover, in rat delayed-type hypersensitivity and chronic relapsing experimental autoimmune encephalomyelitis, a single dose of drug every 3 - 5 days is as effective as daily administration. These data suggest the PD effects of ShK-186 are governed primarily by slow distribution from peripheral tissues and long residence time on the Kv1.3 channel.
Evaluation of a HLA-DR2 Inhibitor for the Treatment of MS Using Human Cellular Immunologic Models (51.6)
Authors: Ji Niannian , Olson Gary , Self Christopher , Forsthuber Thomas 
Published: January 2023

Abstract
Multiple sclerosis (MS) is the most common human neurological disease affecting approximately 400,000 Americans with no cure available. It is believed to be driven by autoimmune CD4+ T cell responses directed against myelin antigens in the CNS. CD4+ T cell activation and response require antigen presentation by MHC (HLA in human) class II molecules. The HLA class II allele HLA-DRB1*1501 (HLA-DR2) is highly associated with MS, therefore we tested the immunological properties and activity of PV-267, a small molecule DR2 inhibitor in vitro in human cellular immunologic models related to MS using flow cytometry analysis, human cytokine ELISPOT and Bio-Plex assays. In vitro studies of human PBMCs from MS patient or healthy control subjects and testing a myelin basic protein specific T cell line showed that: 1) DR2 inhibitors can inhibit myelin antigen-specific human T cell responses mediated via HLA-DR2; 2) inhibit the production of pro-inflammatory cytokines; 3) treatment with the inhibitors does not have undesired non-specific immune-activating effect on human PBMCs expressing either HLA-DR2 or other HLA-DR molecules. In sum, our preclinical study has established the immunologic efficacy and assessed the safety of DR2 inhibitors in vitro. The results provide a strong rationale for the application of MHC inhibitors for the treatment of MS and potentially other T cell-mediated autoimmune diseases based on the immunological efficacy and good safety profile.
Disparate adjuvant properties among three formulations of “alum” (52.11)
Authors: Cain Derek , Sanders Sergio , Cunningham Michael , Kelsoe Garnett 
Published: January 2023

Abstract
Aluminum salts, commonly referred to as “alum,” are the most common adjuvants in human vaccines and serve as a benchmark in the development of novel adjuvants. Although the mechanisms that promote humoral responses to alum-adsorbed antigens are still enigmatic, alum is thought to induce inflammatory signals that, in turn, promote antibody production. It has been noted that some formulations of alum used in animal studies, notably Imject alum®, are not the chemical equivalents of the aluminum used in human vaccines, raising concerns where Imject serves as the comparator for novel adjuvants. Here, we compared the capacity of Imject alum, Alhydrogel®, and a traditional alum-antigen (Alum-Ag) precipitate to induce humoral responses in mice to the hapten-carrier NP-CGG. Primary humoral responses elicited by Alhydrogel and the Alum-Antigen precipitate were significantly greater than those induced by Imject alum. The three alum formulations elicited comparable responses in terms of acute eosinophil and monocyte infiltration at the immunization site, but neutrophil recruitment by Imject alum was signficantly reduced compared to Alum-Ag precipitate or Alhydrogel. These findings reveal substantial differences in the adjuvant properties and inflammatory effects of distinct alum preparations, an important consideration in the analysis of new adjuvants and mechanistic studies of alum’s adjuvanticity.
Inhibition of mTOR promotes dendritic cell activation and survival and enhances therapeutic autologous vaccination in mice (52.4)
Authors: Amiel Eyal , Everts Bart , Freitas Tori , King Irah , Curtis Jonathan , Pearce Erika , Pearce Edward 
Published: January 2023

Abstract
Dendritic cells (DCs) are potent inducers of T cell immunity and autologous DC vaccination holds promise for the treatment of cancers and chronic infectious diseases. In practice, however, therapeutic vaccines of this type have had mixed success. Here we show that brief exposure to inhibitors of mechanistic Target Of Rapamycin (mTOR) in DCs during the period that they are responding to TLR agonists makes them particularly potent activators of naïve CD8+ T cells, and able to induce enhanced immune responses capable of controlling melanoma in a therapeutic autologous vaccination model in the mouse. The improved performance of DCs in which mTOR has been inhibited is correlated with an extended lifespan following activation and prolonged, increased expression of costimulatory molecules. In this work, we define molecular and metabolic processes that determine DC lifespan and identify that how mTOR inhibition alters these pathways to extend the post-activation lifespan of DCs. These findings define mTOR as a molecular target for augmenting DC survival and activation and document a novel pharmacologic approach for enhancing the efficacy of therapeutic autologous DC vaccination.
Candidate adjuvant restores DC-NK cross-talk and improves immunization outcomes in nicotine-exposed hosts (53.14)
Authors: Nouri-Shirazi Mahyar , Nourishirazi Erika , Lang Kevin , Guinet Elisabeth 
Published: January 2023

Abstract
Vaccines are aimed at all target populations including smokers, who constitute one billion people worldwide. There have been several studies published on the link between reduced vaccine effectiveness and cigarette smoke. However, there have been no studies targeted at developing vaccine formulas to increase the effectiveness of vaccines in this population. Recently, we reported that exposure to nicotine significantly diminishes the development of antigen-specific effector memory Th1 cells and antibody production, leading to poor host response to an otherwise protective and therapeutic vaccine. We also reported that defects in the immunobiology of the DC system in vivo underlie this poor outcome (Accepted in The Journal of Immunology). In this study, we further demonstrate that among TLR agonists tested, a candidate adjuvant overcame the impact of nicotine and restored DC-NK cross-talk in vitro. Furthermore, the addition of this adjuvant to vaccine formulation significantly improved vaccination outcome in nicotine-exposed mice as evidenced by clonal expansion and differentiation of adoptively transferred OVA-specific T cells into IFNg-secreting effector Th1 cells in the spleen and lymph nodes of both control and nicotine-exposed mice. In summary, our research has revealed the physiological changes triggered by nicotine on host immunity and opened the possibility of developing a vaccine formulation that works optimally in all target populations.
Immunostimulatory properties of dynamic oligonucleotide micelles (53.9)
Authors: Liu Haipeng , Irvine Darrell 
Published: January 2023

Abstract
Synthetic immunostimulatory oligonucleotides such as double-stranded RNA or unmethylated cytosine-guanosine motifs (CpG-ODNs) mimic molecular signatures of pathogens and trigger an immunostimulatory cascade; as a result, these synthetic ODNs have been extensively studied as therapeutic agents for cancer and as vaccine adjuvants. Using a self-assembly approach, we designed lipo-CpG-ODN molecules composed of CpG oligos conjugated with diacyl lipid tails. These dynamic amphiphiles form compact micelles ~20 nm in diam. in water but can also disassemble and insert their acyl tails into the plasma membrane when incubated with cells. These two key elements of the CpG micelle-ultra-small physical size and cell membrane insertion capability- combine to greatly enhance the adjuvant properties of CpG or other co-associated TLR agonists. Injection of CpG micelles s.c. resulted in a 5-15-fold enhancement of draining lymph node (dLN) retention compared with free CpG or stabilized CpG-micelles incapable of inserting into membranes. Immunization with soluble antigen mixed with CpG micelles induced 10-fold increased frequencies of antigen-specific T-cells in blood and elicited higher antibody production than equivalent doses of soluble CpG, and also elicited stronger responses than CpG-micelles lacking membrane insertion potential. Thus, dynamic micellar TLR agonists may be of interest as novel vaccine adjuvants.
In vivo induced deletion of the mitochondrial protein stomatin-like protein-2 is associated with development of an inflammatory motor deficit disorder (54.10)
Authors: Mitsopoulos Panagiotis , Christie Darah , Welch Ian , Madrenas Joaquin 
Published: January 2023

Abstract
Stomatin-like protein (SLP)-2 is a mainly mitochondrial protein identified in proteome analysis of glycolipid-enriched membrane microdomains of mammalian cells, including lymphocytes. Our preliminary data suggest SLP-2 recruits prohibitins to cardiolipin-enriched domains in the mitochondrial inner membrane to form specialized membrane microdomains, required for optimal cellular respiration. We hypothesized that deletion of the SLP-2 gene will cause defective mitochondrial function that may result in organ-selective disease. We found that a conventional SLP-2-deficient mouse was embryonic lethal at the pre-implantation stage. Thus we generated viable tamoxifen-inducible systemic SLP-2 knockout mice. Administration of tamoxifen (3mg/40g; 10 days; i.p.) resulted in a motor phenotype characterized by poorer SHIRPA scores, weight loss and death (68% at day 28, p&lt;0.05). This phenotype correlated with SLP-2 gene deletion and decreased protein levels in select tissues. Histological analysis revealed multifocal myofibrillar degeneration of the diaphragm with a mononuclear inflammatory infiltrate. In addition, apoptosis throughout the exocrine pancreas and pancreatic atrophy due to loss of normal acinar tissue were observed. In conclusion, in vivo SLP-2 deletion induces a mitochondrial disease-like phenotype, suggesting this protein is required for optimal metabolic function. SLP-2-deficient mice may be useful to assess the function of SLP-2 in vivo and to study mitochondrial diseases.