bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combined TLR/CD40 vaccination produces robust T lymphocyte memory by suppressing mTOR. (P4498)
Authors: Cross Eric , Pennock Nathan , Kedl Ross 
Published: January 2023

Abstract
A long-sought goal of vaccine development is to produce robust cytotoxic T cell responses to prevent future infection. This can be accomplished via live-attenuated vaccines, with the caveat of poor stability and pathogenic reversion concerns. However, a combined TLR agonist/CD40 agonist adjuvant mix (TLR/CD40 vaccination) pioneered by our lab produces potent and protective CD8+/CD4+ T cell memory. However, the mechanistic underpinnings of the protective memory are unknown. Recent work suggests that the metabolic state of a T cell plays a crucial role in the cell fate, effector or memory, realized. While glycolysis is primarily utilized in effector T cells, FAO and catabolic processes are extensively used in naïve and memory T cells. Rapamycin and metformin treatment skews T cell fate decision toward memory by inhibiting mTOR and the respiratory chain complex 1 respectively. Interestingly, CD8+ T cells induced in the presence of these agents have a phenotype identical to that generated by TLR/CD40 vaccination suggesting that our vaccination may influence the metabolic pathways utilized by the responding T cells. Consistent with this, phospho-mTOR levels are suppressed in T cells generated by TLR/CD40 vaccination as compared to those responding to Listeria monocytogenes infection. Ongoing work is establishing the means by which TLR/CD40 vaccination and related costimulatory molecules and cytokines (IL-27 and CD27) influence the metabolic processes in T cells post vaccination.
Vaccine-induced salivary IgA reduces SIV viremia in neonatal rhesus macaques. (P4501)
Authors: Jensen Kara , Wilson Robert , Ranganathan Uma , Van Rompay Koen , Jacobs, Jr. William , Fennelly Glenn , Larsen Michelle , Kozlowski Pamela , Abel Kristina 
Published: January 2023

Abstract
Vertical transmission of HIV in breast milk remains a primary route of pediatric infections. While neutralizing antibodies are important for preventing viral entry, the role of mucosal antibodies for virus neutralization is still not well described. Neonates produce less mucosal IgA than adults and are thus more susceptible to oral pathogens. Here we tested whether an oral pediatric SIV vaccine could induce salivary IgA and protect against oral SIV acquisition. Rhesus macaques were immunized orally at birth with a live, attenuated Mycobacterium tuberculosis strain engineered to express SIVgag, pol and env proteins and boosted at 3 and 6 weeks with rMVAgag/pol/env. Starting at week 9, animals were orally challenged using a regimen of low-dose SIVmac251 (5000 TCID50) weekly. Although the vaccine did not prevent infection, vaccinated infants with higher SIV-specific salivary IgA antibodies at the time of challenge had lower peak viremia (p=0.034). A similar inverse correlation was observed between peak viremia and fecal IgA (p=0.0019). Salivary IgA concentrations correlated with fecal IgA (p=0.018) but not with plasma IgA (p=0.911). In fact, both plasma IgA and IgG showed trends towards a positive association with viremia. Thus, mucosal, but not systemic, antibodies efficiently controlled virus replication. Vaccine strategies that promote the development of mucosal IgA antibodies will likely improve the efficacy of pediatric vaccines to prevent oral HIV acquisition.
Hantaviruses employ multiple mechanisms to evade NK cell recognition (P6298)
Authors: Braun Monika , Klingström Jonas , Gupta Shawon , Björkström Niklas , Ljunggren Hans-Gustaf 
Published: January 2023

Abstract
Rodent-borne hantaviruses, such as Andes virus (ANDV) and Hantaan virus (HTNV) cause hantavirus pulmonary syndrome (HPS) and hemorrhagic fever with renal syndrome (HFRS) respectively, two human diseases with high case-fatality rates and no specific treatment. Patients suffering from HPS or HFRS exhibit massive lymphocyte activation, including high numbers of responding CD8 T cells and NK cells, leading to hyperinflammation and vascular leakage. Interestingly, while endothelial cells are the main targets of hantaviruses, no direct cytopathic effects of the virus on the vascular endothelium have been found. Here, using an in vitro infection model with primary human endothelial cells, we show that hantavirus renders infected endothelial cells resistant to NK cell cytotoxicity by two mechanisms: first, by decreasing NK cell degranulation in an HLA class I-dependent manner and second, downstream of cytotoxic granule-release, by specifically inhibiting granzyme B and caspase 3 activity in infected endothelial cells. Although resistant to NK cell cytotoxicity, infected cells promote an highly activated NK cell phenotype and increased functional capacity. As clinical symptoms in HPS/HFRS most likely stem from the massive immune activation, a more complete understanding of how NK cell activation occurs and the role these activated cells may play in the pathogenesis of disease will be crucial to improve the clinical outcome of these, often fatal, infections.
Defining the immune correlates of pathogenicity of Simian Hemorrhagic Fever virus in Rhesus macaques (P6314)
Authors: Asefa Ben , Jett Catherine , Janosko Krisztina , Bernbaum John , Dodd Lori , Johnson Reed 
Published: January 2023

Abstract
The Clinical Laboratory at the Integrated Research Facility investigates correlates of immunity associated with lethality in nonhuman primates challenged with hemorrhagic fever viruses in high containment. In an intramuscular challenge study with simian hemorrhagic fever virus (SHFv) in rhesus macaques (Macaca mulatta) , we evaluated the factors associated with SHFv and opportunistic bacterial infections (e.g., abscesses) resulting from SHFv-induced immunosuppression. Elevated concentrations of the proinflammatory cytokines MCP-1 and IL-6 and severe septicemia were characteristic of most nonsurvivors of SHFv infection. Other factors associated with morbidity were elevated serum concentrations of AST, ALP, and ALT and decreased concentrations of albumin. Electron microscopy indicated that macrophages and endothelial cells were preferentially infected. Follow up in-vitro studies of SHFV-infected dendritic cells by immunophenotyping demonstrated that mature HLA-DR+ CD163+ cells were rapidly ablated within 48 h of infection with SHFV. Infection with SHFV, a biosafety level-2 (BSL-2) agent, is strikingly similar to infection with Ebola virus, a BSL-4 agent, in hemorrhagic presentation, clinical manifestations, and cellular targets. Further investigation is needed to elucidate the mechanisms of pathogenicity of SHFV infection, compare pathogenic pathways of these hemorrhagic fever virus infections, and identify therapeutic modalities for treatment of viral hemorrhagic fevers.
Age-associated B cells as a connection between anti-viral response and autoimmunity. (P4355)
Authors: Rubtsov Anatoly , Rubtsova Kira , Marrack Philippa 
Published: January 2023

Abstract
Recently our group has discovered a subset of B cells, called age-associated B cells (ABCs), which appear in elderly female mice and autoimmune prone mice of either gender. Our experiments suggest that these B cells can produce autoantibodies and contribute to autoimmune diseases. It has been shown that ABCs develop from follicular B cells and their appearance depends on TLR7 signaling. Here we demonstrate that ABCs also appear at the peak of the response to viral infection. These cells secrete antiviral class-switched (IgG) antibodies and are the major contributors in anti-viral humoral response. Appearance of ABCs during viral infection depends on TLR7 and IFN gamma signaling since mice deficient in these genes fail to accumulate ABCs during infection. Overall, our data suggest a new role of ABCs as a major contributor to anti-viral response. Moreover, ABCs can serve as a new connection between anti-viral response and autoimmunity as they are capable to secrete antibodies with broad reactivity against both self and viral antigens.
Type 2 innate lymphoid cells promote Th2 cytokine production by CD4+ T cells and antibody responses by B cells (P4335)
Authors: Drake Li , Kita Hirohito 
Published: January 2023

Abstract
Type 2 innate lymphoid cells (ILC2s) are part of a large family of ILCs that are important effector cells in innate immunity. Recent studies suggest that lung ILC2s play critical roles in host immunity against helminth infection and in maintaining tissue homeostasis after respiratory viral infection. However, little is known as to whether and how these cells influence adaptive immunity. Here we report that lung ILC2s enhanced proliferation of naïve CD4+ T cells and promoted production of T helper type 2 (Th2) cytokines including IL-4, IL-5 and IL-13. ILC2s regulated T cell function by direct contact through OX40/OX40L interaction. Furthermore, IL-4 produced by ILC2s when cultured with CD4+ T cells played an important role in Th2 cytokine upregulation. We further revealed that lung ILC2s promoted IgA, IgE, IgG1 and IgM antibody production by both B1 and B2 cells, and that this stimulatory effect of ILC2s was mediated at least in part by ILC2 production of cytokines IL-5 and IL-13. We conclude that ILC2s promote both T cell- and B cell-mediated immune responses and propose that ILC2s may play roles in regulating adaptive immunity in addition to their established roles in innate immunity.
Activation and proliferation of innate immune NK cells by adaptive immune B cells (P1082)
Authors: Kim Kwanghee , Lim Seon Ah , Kim Tae-Jin , Lee Kyung-Mi 
Published: January 2023

Abstract
Natural killer (NK) cells belong to innate immune system and consist of circulating lymphocytes which provide innate inspection against infectious disease and cancer. NK cells co-exist with adaptive immune cells, such as T and B cells in secondary lymphoid organ, for example, the lymph nodes and spleen. Though NK cells are likely to interact with B cells, the functional consequence of this interaction remains uncertain. To address this question, , we co-cultured primary NK cells with freshly isolated B cells from the spleen and investigated the influence of B cells on NK cells. Here we show that co-presence of B cells in cultures facilitated NK cell expansion after suboptimal stimulation condition. NK cells cultured with B cells exhibited higher expression of CD69, B220, CD25 activation markers, suggesting that B cells drastically facilitated NK cell activation. In addition, In an in vivo sepsis model, a higher death rate was observed in nude and SCID mice compared to wild type mice after poly I:C treatment. Furthermore, expression of CD25 and CD69 activation markers on NK cells were elevated in nude mice compared to SCID and wild-type mice. Furthermore, this synergistic effect of B cells on NK cell functions could also be detected in humans. Given the fact that NK cells can exert a regulatory role in cancer or infection, co-presence of B cells can potentiate the role of NK cells by promoting their proliferation and cytotoxicity.
The 16 day administration of artemisinin for the long-term cure of malaria in Haiti (P3134)
Authors: Thornthwaite Jerry , Shah Hare , Thornthwaite Bonita , Respess Henry 
Published: January 2023

Abstract
Artemisinin is a natural compound that has been advocated by our Institute in cancer treatment and for its intended purpose in the treatment of malaria. Both malaria parasites and tumors accumulate iron, specifically in the mitochondria, which reacts with the Artemisinin to create a free radical complex that destroys the host malarial parasite or cancer cell. To date, over 50 patients in Haiti, have been treated with our protocol, which encompasses using 100mg orally per day for 16 days. Children under 12 years of age were given 50mg/day, while six babies were given 25 mg/day. Many of these malaria victims had previously experienced multiple recurrences with poor performance and side effects from the conventional malaria treatments. None of the patients treated ever had a recurrence with some over six years after the single treatment (n=17).These cures occurred in spite of repeated mosquito bites of 800-1,000 per year (African estimates). The interesting immunological question is why is a single treatment over 16 days so effective? There must be an in vivo vaccination occurring where the parasites are destroyed so that “super antigens”, regardless of the stage of the cell cycle, are produced. The 16 day treatment assures the transition between early IgM to IgG development. We are currently in the process to secure a collaboration to study antibody and even cytokine levels in these patients during and after treatment to attempt to explain these results.
The costimulatory ligand CD86 promotes resolution of disease after influenza virus infection (P6202)
Authors: Moser Emily , Hufford Matthew , Braciale Thomas 
Published: January 2023

Abstract
Influenza A virus (IAV) infection can produce severe pulmonary inflammation and injury. The host immune response to infection mediates virus clearance but also contributes to pulmonary inflammation and injury, marking the immune response as a potential therapeutic target after severe infections. Host factors mediating IAV clearance and lung inflammation are well-studied, however, factors controlling recovery from disease following IAV clearance are ill-defined. The costimulatory ligand CD86 mediates interactions between immune cells via its receptors CD28 and CTLA-4. We observed in infected mice that blockade of CD86 signaling in vivo after IAV clearance delays recovery and promotes accumulation of neutrophils in the respiratory tract. Furthermore, CD86 blockade reduces both IL-2 and T regulatory cells (Treg) in the lungs. Depletion of Tregs using DREG mice or Treg depleting antibody after IAV clearance also delays recovery and leads to pulmonary neutrophilia. These data suggest that Tregs may play a crucial role in regulating pulmonary inflammation after IAV infection and are in turn dependent on CD86 ligand engagement. We are continuing to explore the role of CD86 in recovery and resolution of inflammation after IAV infection with the long-term goal of exploring the therapeutic potential of optimizing CD86 signaling to promote Treg function in recovery from infection.
Inhibition of reactive oxygen species production attenuates highly pathogenic avian influenza H5N1-induced inflammation via up-regulation of suppressor of cytokine signalling 3 (P6188)
Authors: Ye Siying , Lowther Sue , Stambas John 
Published: January 2023

Abstract
Influenza A virus infection causes a rapid influx of inflammatory cells, resulting in increased ROS production and acute lung injury. Pro-inflammatory stimuli induce intracellular ROS by activating NADPH oxidase (NOX). Nox2 is the catalytic subunit of the phagocyte NOX. It has been shown that Nox2-containing NOX is the major source of ROS induced by influenza A virus infection in mice. Apocynin, a NOX inhibitor, reduced virus replication and accumulation of inflammatory cells in the lungs of X31- or PR8- infected mice. As hypercytokinemia is thought to contribute to severe disease pathogenesis of high pathogenicity avian influenza (HPAI) H5N1 infection, it is hypothesized that apocynin could reduce HPAI H5N1-induced cytokine production. A panel of airway epithelial and immune cells from mammals and chickens were infected with low pathogenicity avian influenza H5N3 (A/duck/Victoria/0305-2/2012), HPAI H5N1 (A/chicken/Vietnam/0008/2004), or PR8 H1N1 virus in the absence or presence of apocynin. Quantitative real-time PCR showed that apocynin inhibited influenza-induced cytokines (IL-6, IFN-β, CXCL10 and CCL5) and ROS production, although viral replication was not significantly altered. Interestingly, apocynin treatment significantly increased SOCS3 expression, enhancing negative regulation of cytokine signaling. Therefore, apocynin or its derivatives could be used as a therapeutic agent for reducing hypercytokinemia, resulting in amelioration of disease.
High and low molecular weight hyaluronan use TSG-6 or CD44 signaling pathways respectively in activation of different subsets of dendritic cells thereby affecting the development of encephalitogenic T effector cells and autoimmune encephalomyelitis (P5172)
Authors: Guan Hongbing , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Microglia and astrocytes can overexpress hyaluronan in demyelinated lesion of brain in multiple sclerosis (MS), which affects the remyelination of axons. However, the exact role of hyaluronan in the development and treatment of MS is not clear. This becomes more complicated because the prototype hyaluronan is degraded into fragments of various sizes in MS. Previous studies using high molecule weight hyaluronan (HMW-HA) or low molecule weight hyaluronan (LMW-HA) have so far resulted in confusing or contradictory results. In this report, we found that HMW-HA and LMW-HA used different receptors depending on the subsets of dendritic cells (DC), which affect the activation of DC and subsequent development of effector T cells. HMW-HA (950KDa) bound to tumor necrosis factor-6 (TSG-6) receptor on myeloid DC (mDC) and induced a Th2 biased response. LMW-HA (12Kda) bound to CD44 receptors on plasmacytoid DC (pDC) and lymphoid DC (lDC) and induced a Th17 biased response. Accordingly, systemic administration of HMW-HA inhibited the development of experimental autoimmune encephalomyelitis (EAE), whereas systemic administration of LMW-HA exacerbated the development of EAE. We also found that gamma-interferon-inducible lysosomal thiol-reductase (GILT) was required for the therapeutic activity while HMW-HA and LMW-HA regulated GILT activity differentially. Our study, for the first time, elucidated the distinct role of hyaluronan in the development and treatment of MS.
Targeting CD44 ameliorates autoimmune diabetes by reducing pancreatic infiltrates and improving Treg efficacy (P5180)
Authors: Li Cheng-Rui , Mueller Erin , Bradley Linda 
Published: January 2023

Abstract
In type 1 diabetes (T1D), Th1 cells are a major population orchestrating the disease pathology. We showed previously that signals through CD44 are needed for Th1 cell survival and memory development. Therefore, we hypothesized that blocking CD44 could impact progression to overt diabetes. To test this, we treated recently hyperglycemic NOD mice with a short course of anti-CD44 and found a significant delay in the spontaneous development of TID that was associated with a reduction in islet infiltrates. However, previous studies suggested that targeting CD44 might enhance Treg function. To test whether this could contribute to the anti-CD44-mediated effect, we transferred polyclonal in vitro differentiated adaptive Treg cells (aTregs), which we showed can protect ~50% of newly hyperglycemic mice from overt diabetes, together with anti-CD44 treatment. We found a significantly increased the success rate of diabetes prevention. The islet infiltrates in these mice had a higher frequency of CD4+ cells producing IL-10 and IL-2 as well as of endogenous Tregs that displayed enhanced FoxP3 expression. However, anti-CD44 treatment did not alter cytokine profiles or Foxp3 expression of the transferred aTregs. The data therefore suggest that anti-CD44 treatment can alleviate inflammation in the target organ, and can functionally enhance the endogenous Treg population, thereby improving the establishment of long-term control by aTregs, which form a protective memory population.
In situ detection of autoreactive CD4 T cells in the mouse models of autoimmune diseases (P4550)
Authors: Massilamany Chandirasegaran , Jia Ting , Kang Guobin , Zhou You , Li Qingsheng , Reddy Jay 
Published: January 2023

Abstract
We report here the utility of major histocompatibility complex (MHC) class II dextramers for detecting self-reactive CD4 T cells in the target tissues using two mouse models of autoimmunity namely, experimental autoimmune encephalomyelitis and experimental autoimmune myocarditis. We determined the conditions for dextramer staining using brain sections obtained from SJL mice immunized with proteolipid protein (PLP) 139-151 by staining the sections with a cocktail of IAs/PLP 139-151 (specific) or Theiler’s murine encephalomyelitis virus (TMEV) 70-86 (control) dextramers conjugated with the fluorescent dyes, and CD4 antibody. The sections were examined by Olympus Fluoview laser scanning microscope, and the analysis revealed PLP 139-151 dextramer-positive cells to be colocalized to CD4-expressing cells, but such reactivity was lacking with TMEV dextramers, suggesting that the staining obtained with PLP dextramers was specific. We next extended these observations to detect cardiac myosin-specific T cells by deriving IAk dextramers for cardiac myosin heavy chain-α (Myhc) 334-352 and ribonuclease 43-56 (control). Expectedly, heart sections prepared from A/J mice immunized with Myhc 334-352 on day 21 postimmunization showed infiltrations of Myhc-specific CD4 T cells. The data suggest that MHC class II dextramers can be potentially used to enumerate the precursor frequencies of antigen-specific CD4 T cells in situ directly, without having to amplify the fluorescent signals.
Growth hormone effect on type 1 diabetes development (P4546)
Authors: Kakabadse Schmidt Dimitri , Villares Ricardo , Gomez Lucio , Martinez Carlos , Mellado Mario 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune disease that affects approximately 34 million people around the world. T1D is caused by the destruction of insulin-producing pancreatic β-cells by autoreactive T cells that lack effective immune regulation. One of the most common animal models for diabetes study is the NOD (non-obese diabetic) mouse strain. Using female NOD mice, which develop spontaneous T1D, we observed that growth hormone (GH) induces a protective effect on insulin-producing cells and as a result, inhibits T1D development. We generated a transgenic NOD mouse that expresses bovine GH and showed almost complete protection against diabetes development. Histochemical analysis demonstrated an increase in the number of pancreatic endocrine progenitor cells, overgrowth of islets of Langerhans, and although these mice showed peri-insulitis, infiltration was much reduced. The mechanism involved an increase in regulatory T cell number and activity, a polarization phenotype toward anti-inflammatory macrophages, and altered B cell responses with reduced antibody production. Our results identify a regulatory role for GH in immune cells and pinpoint a new target for therapeutic intervention on type 1 diabetes.
CD40L at the interface of the innate and adaptive immune responses. (P5206)
Authors: Ragheb Jack , Bushar Nicholas 
Published: January 2023

Abstract
Induction of CD40L expression on CD4+ T cells is a critical event in the generation of cellular and humoral immune responses. CD40L interacts with B cells via its cognate receptor to trigger immunoglobulin isotype class switching and promote Ig secretion and the differentiation of memory B cells. Through its induction of IL-12 and upregulation of costimulatory molecules on antigen presenting cells (APC), CD40L is also vital to the generation of Th1 cellular responses. We have previously demonstrated that a combination of cell-contact dependent and soluble signals act in concert to orchestrate the biphasic pattern of CD40L expression that underlie its complex role in promoting the adaptive immune response. We now report that CD40L likely plays an equally important role in the innate response. We show for the first time that TCR-dependent expression of CD40L on primary human T cells can be induced by interaction of β1 integrins on CD4+ T cells with extracellular matrix (ECM) in a monocyte and IL-2 independent fashion. We provide evidence in a model of vascular wall injury that ECM mediated induction of CD40L on CD4+ T cells plays a role in wound healing. These findings establish a pivotal role for CD40L at the interface of the innate and adaptive immune responses and suggest that therapeutic modalities directed towards CD40L may have benefit in modulating vascular pathophysiology.