bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chronic Vaccine Boost Prolongs Malaria Vaccine Immunity and CD4 Effector and Memory T cells
Authors: Gbedande Komi , Ibitokou Samad A , Ong Monique L , Brown Michael G , Degli-Esposti Mariapia A , Stephens Robin 
Published: January 2023

Abstract
Vaccines to persistent parasite infections have been challenging to develop, and current iterations lack long-term protection. Persistent infection generates effector and effector memory T cells (Teff, Tem), and both contribute to protection, as we recently showed for Plasmodium infection. Sterilizing immunity from live P. chabaudi vaccination lasts less than 200 days. While specific antibody levels remain stable after the acute phase of infection, the decay of T cells corresponds with loss of protection from re-infection. Chronic vaccination with CMV has been shown to generate Tem and Teff, and so we tested MCMV for the ability to prolong protection to P. chabaudi. We generated MCMV encoding P. chabaudi MSP-1 epitope B5 (MCMV-B5), and used B5 TCR Transgenic CD4 T cells to follow the MCMV-B5 response. We showed that boosting the live malaria vaccine with MCMV-B5: i) prolonged vaccine protection to heterologous infection at day 200, ii) increased parasite-specific T cells and promoted a highly-differentiated Teff phenotype, including iii) IFN-γ+ and TNF+ T cells. Importantly, chronic MCMV infection itself improved protection from P. chabaudi infection and promoted generation of a T cell phenotype similar to P. chabaudi infection. Investigating the mechanisms of MCMV in prolonged vaccine-induced protection, we found an impressive role for IFN-γ, as its neutralization in vivo during MCMV latency is associated with loss of protection to P. chabaudi challenge. Blocking IFN-γ before challenge induced fewer Teff and less plasma IL-12 upon P. chabaudi challenge. Our findings suggest a mechanism to boost CD4 T cell immunity using chronic vaccines, which may be broadly applicable to infections that require T cell based immunity.
Supported by R01AI135061
Age-Dependent Responses of γδ+ T cells During Bloodstream Infection in Early Life
Authors: Yokanovich Lila T , Greenfield Kara , Knoop Kathryn 
Published: January 2023

Abstract
Late-onset neonatal sepsis (LOS) is a bloodstream bacterial infection that occurs during the first two months of life. Despite improved clinical care, LOS remains a significant contributor to neonatal mortality. Clinical observations suggest neonates exhibit unique immune responses and excessive inflammation during sepsis compared to adults and older children, however, factors contributing to these differences remain poorly understood. Two common routes of pathogen entry for neonates are direct introduction of a pathogen into the bloodstream, or the translocation of a gut-resident pathobiont. Our lab has developed a mouse model of LOS that disrupts goblet cell homeostasis to increase gut permeability and facilitate the translocation of virulent bacterial species such as E. coli. During both intraperitoneal and oral infection, we observe several key features of clinical LOS including increased pro-inflammatory serum cytokines, organ failure, and death in 5 day-old pups, but not 15 day-old pups. Strikingly, we have found robust activation of γδ+ T cells in 5 day-old pups, following infection, independent of pathogen entry route. This γδ+ T cell activation occurred in an age-dependent manner and was absent in 15 day-old pups. γδ+ T cell activation in 5 day-old pups was also associated with a pro-inflammatory cytokine profile that was absent in older pups. Finally, 5 day-old TCRδ−/− pups were protected from mortality during LOS, suggesting that γδ+ T cells may also contribute to mortality in this model. This study sheds light on the unique responses of γδ+ T cells to bloodstream infection during two distinct periods of early life.
Exploring cutaneous neuro-immune networks during helminth infection
Authors: Jean E Evonne , Rossi Heather , Herbert De’Broski 
Published: January 2023

Abstract
Sensory nociceptive neurons serve important roles in regulation of cutaneous inflammation directed against microbial and fungal pathogens, but whether neurons serve any role in protective immunity against skin penetrating helminths remains unknown. In this study, we sought to identify the key factors responsible for blocking parasite entry of host skin. Data show that CD4 T cells were essential for restricting parasite entry of the skin during secondary infection, but this resistance was not dependent upon the key TH2 transcription factor Signal Transducer And Activator Of Transcription 6 (STAT6). Interestingly, STAT6 deficient mice generated high levels of TH17 associated cytokines, the latter of which are known to be induced by skin nociceptors. To determine whether skin neurons could prime host protective immunity, we used an optogenetic approach that allowed pre-activation of neurons at the site of skin inoculation that expressed transient receptor potential cation channel subfamily V member 1 (TRPV1). Strikingly, preemptive activation of TRPV1 neurons led to a significant inhibition of parasite entry. Ongoing studies are designed to understand whether depletion of TRPV1 skin neurons impairs immunity and whether skin protection relies upon TH17 dependent immunity.
Supported by grants from NIH (R01 AI095289 and R01 GM083204) and NSF GFRP (Fellow ID:2020297782)
Treponema pallidum specific CD4 T cell epitope discovery
Authors: Reid Tara , Godornes Charmie , Tong Denise , Koelle David M 
Published: January 2023

Abstract
Despite curative therapy and multiple public health campaigns to eliminate syphilis in the US, the incidence of Syphilis is rising in diverse groups: men who have sex with men, women, and children. An effective syphilis vaccine will be a key tool in the eradication of this disease. Prior efforts to create a protective syphilis vaccine focused on raising antibodies against rare outer membrane proteins to facilitate antibody mediated opsonophagocytosis of Treponema pallidum (Tp) in infectious lesions. The inclusion of immunodominant T cell antigens in a subunit vaccine can provide CD4 T cell help to B cells to produce opsonic antibodies. We sought to identify human Tp T cell antigens validated to the epitope level as there are none reported in the literature. Tp specific CD4 T cells were isolated from persons with active or treated syphilis infection by CD137 and CD69 activation–based FACS and polyclonal expansion. These cells were then probed with a panel of 39 recombinantly expressed Tp proteins. Preliminary data show that activation-induced-marker-enriched, expanded T cells can be probed to discover novel CD4 T cell antigens and epitopes using proliferation experiments. The first 4 CD4 T cell antigens identified included antibody targets Tp0435 and Tp0574, a flagellar apparatus protein Tp0870, and a metabolite transporter Tp0684. Further refinement of minimal T cell antigens within these proteins and expansion of the Tp protein panel is ongoing. These are the first human Tp specific T cell epitopes described. Incorporating this information into vaccine design may further increase protection against infection.
Supported by T32AI007140-44
Administration of 3,3-dimethyl-1-butanol is immunomodulatory in collagen induced arthritis murine model independent of TMA lyase activity
Authors: Fechtner Sabrina , Allen Brendan , Chriswell Meagan E , Jubair Widian K , Vroljik M Aaron , Holers V Michael , Kuhn Kristine A 
Published: January 2023

Abstract
Studies of rheumatoid arthritis (RA) and murine collagen-induced arthritis (CIA) link the intestinal microbiome with disease and pathogenic autoantibodies. We have previously identified significant alterations in carnitine metabolism in mice with CIA, providing a potential link between bacterial dysbiosis and autoimmunity. Dietary carnitine is metabolized into trimethylamine (TMA) by bacterial enzyme TMA lyase which is oxidized in the host liver to TMAO. Prior studies determined that 3,3-dimethyl-1-butanol (DMB) and fluoromethylcarnitine (FMC) inhibit TMA lyase. Therefore, we tested if TMA lyase inhibition with DMB or FMC could ameliorate CIA. Male DBA1/j mice at 6 weeks of age were injected with bovine type II collagen in complete Freund’s adjuvant at days 0 and 21. On day 21 drinking water was replaced with vehicle, 1% vol/vol DMB, or 100mg/kg FMC. Mice were monitored for arthritis until day 35. DMB-treated mice had a significant (&gt;50%) reduction in arthritis severity, as well as decreased anti-collagen antibodies, serum IFN-γ, and IL-17 compared to FMC and vehicle-treated mice with CIA. However, only FMC significantly reduced intestinal TMA and circulating TMAO, while DMB did not affect TMA/TMAO, suggesting that the effect of DMB is independent of TMA lyase inhibition. Using gas chromatography, we confirmed that DMB is absorbed and metabolized in mice, thereby identifying a novel metabolite, 3,3-dimethyl-1-butyric acid (DMBut). During CIA, 1% vol/vol DMBut significantly reduced disease, suggesting it is an active metabolite of DMB. Altogether our study suggests that DMB and/or its metabolites are immunomodulatory in CIA. Elucidating its target and mechanism of action may provide directions for RA therapeutic development.
Supported by Grants from NIH (U01HL152405, T32AI074491) and from the State of Colorado Office of Economic Development and International Trade Advanced Industries Accelerator Program
Single-cell mRNA analysis of colon tissue during DSS colitis in CX3CR1cre/+EP4fl/fl mice suggests PGE2 signaling via EP4 promotes survival of protective macrophages
Authors: Murray Page , Emanuel Elizabeth , Kang Byunghyun , Kelsall Brian L 
Published: January 2023

Abstract
Prostaglandin E2 signaling through its receptor EP4 on macrophages (MPs) protects against DSS-induced colitis, however the mechanisms involved are not clear. Here we perform single cell RNA sequencing on the colons of CX3CR1cre/+EP4fl/fl(Cre+) mice and CX3CR1+/+EP4fl/fl(Cre−) littermate controls at days 0, 3 and 7 following DSS treatment. We identified monocyte (MO), MP, DC1, DC2 and pre-DC populations, with MPs clustering into Hes1hi, Mrc1hi  Siglec1(CD169)hi, Retnlahi, and Il1r2hiCd80hi populations. With colitis there was a dramatic increase in the proportions of inflammatory MOs and a loss of all mature MPs, along with an increase in cDC2 but not cDC1 in both mouse strains. However, in Cre+ mice, which develop worse disease with universal GI bleeding, the loss of Mrc1hi  Siglec1hi MPs was accelerated together with an increased proportion of Il1r2hiCd80hi MPs compared to Cre− littermates. Analysis of DEGs showed the Il1r2hiCd80hi MP cluster expresses high levels of leukocyte chemoattractants, suggesting that PGE2 may prevent leukocyte recruitment during inflammation. Cre+ mice also had fewer Retnlahi macrophages than Cre− mice both at days 0 and 7 post DSS. Genes highly expressed in the Retnlahi cluster are associated with wound healing and maintenance of vascular integrity indicating their loss could account for the enhanced GI bleeding in the inflamed Cre+ mice. This is the first detailed scmRNA analysis of immune cell populations involved in DSS colitis. While additional investigation is necessary, the data suggest a role for PGE2 signaling via EP4 on MPs in preventing excessive inflammatory cell infiltration, decreasing vascular permeability, and contributing to tissue repair by maintaining protective MP populations.
Large-batch evaluation of cell counter instrument-to-instrument consistency for cell therapy applications
Authors: Bell Jordan , Huang Yongyang , Chan Leo 
Published: January 2023

Abstract
Cell therapy has been hailed as a medical revolution, and its rise has only increased the importance of accuracy and precision in cell counting. Ensuring the identity, purity, and viability of cell therapy products is critical for increasing efficacy and avoiding potential harmful side effects. The burden of sample analysis often rests on fleets of automated cell counters, leading to the need for confidence that multiple cell counters will give similar results.
In this work we compare groups of two distinct cell counting instruments. The first type is a well-established single-sample counter (Cellometer K2), similar to those often found in R&D laboratories. The second type of instrument (Cellaca MX) is a high-throughput cell counter that is often more suitable for a manufacturing environment than its single-sample counterparts.
We test the consistency of these cell counters using both live cells and fluorescent beads. We use samples of Jurkat cells to simulate immunotherapy products, and prepare the beads in fixed reference samples to allow comparison of multiple instruments over extended periods of time. We use these beads to investigate agreement among 60 Cellometer K2 instruments and 30 Cellaca MX instruments, what we believe is the largest group of cell counting instruments to be involved in a single comparison study.
Our work illustrates experimental approaches suitable for the comparison of multiple cell counting methods. We measure consistency among the cell counters, both within groups of similar instruments and between instrument types. This consistency encourages optimism for “future-proof” assays that can survive the transition from R&D to production with minimal adjustment.
Characterization of a Novel Anti-inflammatory Biogenic Amine
Authors: Santambrogio Laura , Clement Cristina C 
Published: January 2023

Abstract
Tryptophan (Trp) is an essential amino acid whose catabolism generates a cascade of over fifty molecules in a cell-specific manner. Three major Trp metabolic pathways have been described: synthesis of serotonin, synthesis of melatonin and generation of kynurenines. In mammalian cells, 90% of Trp is processed through the kynurenine pathway and kynurenine metabolites are regarded as one of the most powerful mechanism for immune regulation. We recently reported a novel biogenic amine, 3HKA, generated through a still uncharacterized lateral pathway of Trp catabolism. This novel biogenic amine is generated by both professional (Dendritic Cells) and non-professional (Lymphatic Endothelial Cells) antigen presenting cells and, differently from all others Trp metabolites, display unique “in vivo” immunosuppressive capabilities. In vitro, 3-HKA exhibited an anti-inflammatory profile by inhibiting the IFN-g-mediated STAT1/NF-kB pathway in both mouse and human dendritic cells (DCs) with a consequent decrease in the release of pro-inflammatory chemokines and cytokines; most notably, TNFa, IL-6, and IL12p70. In vivo, 3-HKA exerted protective effects in an experimental mouse model of Psoriasis by decreasing skin thickness, erythema, scaling and fissuring, reducing TNFa, IL-1b, IFN-g, and IL-17 production, and inhibiting generation of effector CD8+ T cells. Similarly, in a mouse model of nephrotoxic nephritis, besides reducing inflammatory cytokines, 3-HKA improved proteinuria and serum urea nitrogen, overall ameliorating the immune-mediated glomerulonephritis and renal dysfunction. Overall we propose that this novel biological amine is a crucial component of Trp-mediated immune tolerance.
Renal Tubular Cell Ferroptosis: A New Player in Pathogenesis of Lupus Nephritis
Authors: Scindia Yogesh , Ali Abdel , Desai Dhruv , Mehrad Borna , Morel Laurence , Conrad Marcus , Clapp William , Abdelmegeed Ahmed 
Published: January 2023

Abstract

Background
New research indicates that the injury to the renal tubular epithelial cells is a better predictor of renal outcomes in lupus nephritis (LN) as compared to glomerular injury. Iron accumulates mostly in the renal tubular cells of LN patients and murine models of LN. Ferroptosis is an incompletely understood form of cell death involving iron mediated lipid peroxidation, but its role in LN is not well established.

Experiment
LN and control patient biopsies were stained for markers of ferroptosis. Kidneys of female (MRL/lpr) and male (NZW X BXSB) F1, lupus mice were analyzed for markers of ferroptosis. Human proximal tubular cells (PTEC) were treated with LN serum with or without Liporxstatin-2, a novel ferroptosis inhibitor.

Results
Compared to controls, LN patients expressed higher levels of ACSL4 and 4HNE, the ferroptosis core proteins in the renal tubules. Compared to non-nephritic mice, nephritic mice had significantly higher gene expression of Aifm2, Acsl4, and Gpx1 as well as protein levels of Acsl4. Nephritic mice had an impaired renal glutathione synthesis pathway. Glutathione is an essential positive regulator of glutathione peroxidase 4 (Gpx4: ferroptosis inhibitor) which resulted in lower protein expression of Gpx4. Collectively, LN kidneys displayed a ferroptosis signature and was associated with an increase in Ngal and Kim1, PTEC injury markers. LN patients’ serum induced PTEC ferroptosis and associated pathology were significantly reduced by Liproxstatin-2.

Conclusion
Our data identify occurrence ferroptosis in renal tubular cell which can contribute to the pathogenesis of LN. Liproxstatin-2 mitigates human LN serum induced PTEC pathology and holds promise as an adjunct therapy to alleviate LN severity.
Supported by grants from Vifor Pharma
The Role of LAG3 in Antibody Responses To Kidney Transplantation
Authors: Nicosia Michael , Fan Ran , Lee Juyeun , Gorbacheva Victoria , Beavers Ashley , Dvorina Nina , Baldwin William M , Fairchild Robert L , Min Booki , Valujskikh Anna 
Published: January 2023

Abstract
The role of lymphocyte activation gene-3 (LAG3) in T cell functions is well studied, however its role in humoral immune responses remains poorly characterized. The goal of this study was to test the role of recipient LAG3 in a mouse model of renal allograft rejectionNaive B6.LAG3−/− mice have elevated numbers of follicular and memory T cells, and plasma cells compared to WT mice, as well as increased frequencies of memory T cells reactive to H-2Dd, H-2Ds, H-2Dq and H-2Dk alloantigens. When kidneys were transplanted from C3H donors to B6 WT and LAG3−/− mice, all C3H kidney allografts survived for &gt; 60d in WT recipients, whereas LAG3−/− recipients rapidly reject allografts and have elevated serum creatinine levels at d14 posttransplant. Graft histology at rejection revealed minimal T cell infiltration, diffuse C4d staining, atrophic peritubular capillaries, endothelial swelling and edema characteristic of antibody mediated rejection (AMR). Compared to WT, LAG3−/− recipients had elevated frequencies of anti-donor IFNγ producing T cells and increased levels of donor specific antibody (DSA) against MHC-I and MHC-II. Depletion of CD8 T cells in LAG3−/− recipients did not alter rejection kinetics, while B cell depletion significantly extended C3H kidney allograft survival, suggesting the predominant role of alloantibody rather than T cell mediated rejection in these mice. However, neither T nor B cell conditional knockout recipients rejected the allograft demonstrating LAG3 expression on both cell types is necessary to mediate rejection.
These findings demonstrate that LAG3 regulates both T and B cell functions in response to kidney allografts, and is an attractive therapeutic target for the prevention of AMR.
Supported by grants from NIH (P01AI087586-10)
PARP-inhibition with IFNg in the ovarian tumor microenvironment induces immunogenic cancer cell death for sustained anti-tumor immunity.
Authors: Kinjyo Ichiko , Adams Sarah F 
Published: January 2023

Abstract
Poly(ADP-ribose) polymerase (PARP) inhibitors have been a remarkable breakthrough in the treatment of ovarian cancers with BRCA1/2 mutations. We have previously reported that PARP-inhibition with immune checkpoint blockade extended survival of BRCA1-deficient ovarian cancer (ov ca) preclinical models. A significant increase in IFNg producing effector T cells in the tumor environment promoted tumor cytotoxicity in a caspase-independent manner.
In this study, we hypothesized that in the presence of IFNg, PARPi may induce immunogenic cell death (ICD) rather than apoptosis and promote immune-mediated tumor control. In vitro, PARPi+IFNg treatment induces hallmarks of ICD in BRCAmt ov ca; such as cell surface translocation of calreticulin, ATP and HMGB-1 release but not by PARPi or IFNg single treatment. Additionally, the PARPi+IFNg treated Ov ca cells induced maturation and activation of dendritic cells with enhanced antigen-presentation to stimulate anti-tumor T cells. Evidence of immunogenicity was confirmed by the benefit of vaccination with PARPi+IFNg treated Ov ca cells in vivo. The vaccination established the long-lasting protective anti-tumor immune responses to clear sequential tumor inoculations. T cell-depletion and adoptive transfer experiments showed the ICD-induced tumor immunity was surprisingly, both CD4 and CD8 T cell-mediated. Currently, we are addressing whether anti-tumor antibodies contribute to tumor control.
Our results identified a novel mechanism by which PARP-inhibition with IFNg induces ICD in the ovarian tumor environment. These effects can be leveraged to enhance tumor clearance in the combination with immune therapies to modulate TME by stimulating tumor-immune cycles.
Supported by grants from NCI (R37 CA229221)
Dendritic cells to prevent cancer: Immune responses against neoantigens after dendritic cell vaccination of Lynch Syndrome patients
Authors: Abidi Asima , Westdorp Harm , Gorris Mark A J , Scheijen Blanca , Boller Anna-Lena , Irusquieta Cristina Berganza , Hoogerbrugge Nicoline , Schreibelt Gerty , de Vries Jolanda I J M 
Published: January 2023

Abstract
Lynch syndrome (LS) is an autosomal dominantly inherited syndrome caused by monoallelic germline aberrations affecting a DNA mismatch repair (MMR) gene, which leads to the development of microsatellite instability. Individuals with LS are 60–80% more prone to colorectal cancer (CRC). MMR deficiency in tumor DNA causes shifts in the translational reading frame resulting in the production of altered peptides called neoantigens. In a phase I/II clinical trial we vaccinated 20 healthy LS mutation carriers and 3 LS patients with surgically resected CRC with monocyte-derived dendritic cells (DC) loaded with neoantigens of TGF-βRII and caspase-5, and CEA.
The objective was to analyze the presence and functionality of antigen specific CD8+ T-cells in peripheral blood and biopsies of post-treatment skin tests. Cell lines stably expressing the (neo)antigen were generated and used in calcein based killing assays with expanded (neo)antigen specific patient CD8+ T cells.
The vaccination induced antigen specific T cell responses in 20/23 patients. Multifunctional antigen specific T-cells producing multiple cytokines and high amounts of interferon-γ upon peptide stimulation were detectable in peripheral blood. Moreover patient T cells of 3/4 patients tested demonstrated cytotoxicity against tumor cells presenting TGFβRII neoantigen, the T cell receptor repertoires of which had shared motifs in the clonotypes between patients as identified by Illumina MiSeq sequencing.
Conclusion-DC vaccination with (neo)antigens is well tolerated in LS patients and induces functional (neo)antigen specific T cells capable of eliciting a cytotoxic immune response thus able to enhance antitumor immunity in a prophylactic and therapeutic setting.