bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD8 T cell immunosurveillance and elimination of HIV target cells in the female reproductive tract
Authors: Joag Vineet , Mitchell Jason , Fife Brian T , Masopust David 
Published: January 2023

Abstract
HIV infection is thought to establish in a single founder CD4 T cell in the genital mucosa, however, an effective CD8 T cell-based vaccine must enable timely surveillance of all mucosal CD4 T cells within the first few days after HIV exposure. The HVTN 502 (STEP) vaccine trial employed a CD8 T cell based vaccine that while promising in animal studies, did not protect human vaccinees from mucosal HIV acquisition. We hypothesize that the quantity of mucosal HIV-specific CD8 T cells is a critical determinant of tissue immunosurveillance and was insufficient to adequately survey and eliminate HIV target cells in the STEP trial. To test our hypothesis, we applied intravital mucosal imaging to visualize how efficiently CD8 T cells survey CD4 T cells in the murine uterus. Based on CD8-CD4 T cell contact rates, a density of 4 (0.8%) HIV-specific mucosal CD8 T cells/mm2 would be required to survey all mucosal CD4 T cells within 2 days. However, vaccinees in the STEP trial had ELISPOT readings of ≥55 spot-forming units/106 blood leucocytes or approximately 0.09% mucosal HIV-specific CD8 T cells, which is 10 fold lower than the required density. Surveillance efficiency was directly correlated with the number of CD8 T cells and was independent of target cell numbers. Upon antigen recognition in the context of MHC-I on CD4 T cells, CD8 T cells formed kinapses with CD4 T cells in an LFA-1/ICAM-1 dependent manner, facilitating tethering and prolonged tracking. Direct killing of CD4 T cells by single or multiple CD8 T cells occurred by 3h after antigen presentation. We conclude that CD8 T cells are capable of rapid tissue surveillance and killing of CD4 T cell targets, and that sub-optimal numbers of HIV-specific CD8 T cells may have contributed to the failure of the STEP trial.
Dissecting the role of endogenous epithelial IL-10 using a small intestinal organoid model
Authors: Nguyen Huong D. , Stadnyk Andrew W. 
Published: January 2023

Abstract
IL-10 is widely appreciated as a potent anti-inflammatory cytokine, acting on leukocytes in mucosal immunity. Far less attention has been paid to the impact of IL-10 on epithelial cells, which make up the crucial barrier interface between the host mucosa and the external environment. Furthermore, most studies look into the effects of exogenous IL-10, disregarding the possible presence and function of endogenous IL-10 in the epithelium. Using ex vivo organoids we aimed to dissect any role for endogenous epithelial IL-10. We discovered that cells in small intestinal organoids (enteroids) derived from C57BL/6 mice produced IL-10 constitutively throughout development. IL-10 mRNA increased considerably on day 3, the time when enteroids begin “budding” to create crypt-like structures. In addition, a second IL-10 increase was detected at day 6 when enteroids are typically “mature”. Immunofluorescent staining for IL-10 on day 4 enteroids was localized to Paneth cells. As gut epithelial cells are constantly exposed to bacteria, we added LPS at the start of enteroid cultures and observed that IL-10 staining was significantly increased. To further investigate the role of endogenous IL-10, we grew enteroids from IL-10 gene knockout mice. IL-10 deficient enteroids develop to morphologically resemble wild-type enteroids. We are proceeding to examine any difference in permeability, cell lineages and apoptosis between the two enteroid genotypes. In conclusion, IL-10 is present in the developing small intestinal epithelium and while development is IL-10-independent, whether IL-10 impacts subcellular components is under investigation.
Spatial transcriptomics with single cell resolution
Authors: Braubach Oliver 
Published: January 2023

Abstract
CO-Detection by indEXing (CODEX®) is ultra-high plex immunohistochemistry that relies on a DNA-based tagging approach, whereby antibodies are labeled with specific oligonucleotide tags (barcodes), and dye-oligonucleotides (reporters) are iteratively hybridized and dehybridized across multiple cycles. CODEX® can detect dozens of antigens simultaneously, in situ and at single cell resolution. Single-cell RNA sequencing, on the other hand, provides information about hundreds to thousands of mRNA transcripts, but without spatial context. The purpose of our study was to enrich spatially resolved CODEX® data with gene expression data obtained from CITE-seq experiments performed on the same tissues. CITE-Seq concurrently measures single-cell antigen and gene expression data, and it is possible to combine these results with CODEX® data by means of Spatially-resolved Transcriptomics via Epitope Anchoring1. We are testing this approach in intact tissues and cell spreads in order to expand the depth our spatial analysis platform. The combination of CODEX® and single cell gene expression data provides an all-in-one solution for deeply multiplexed spatial analyses of protein and gene expression with single cell resolution.
A universal multiplex assay format for ultra-sensitive cytokine quantification in human serum and plasma
Authors: Lambert Jeremy , Tobos Carmen , Rissin David 
Published: January 2023

Abstract
Quantifying prospective biomarkers of immune signaling in biological samples requires high assay sensitivity. The Simoa Planar Array provides greater sensitivity than ELISA, but has a finite assay menu. This study aimed to develop a universal multiplex ‘homebrew’ planar immunoassay format for development of high sensitivity assays for any analyte.
Anchor antibodies specific for a peptide tag were microprinted in microwell plates. The peptide tags were conjugated to analyte-specific capture antibodies via maleimide chemistry, enabling high affinity binding of capture to anchor antibody. Bound sample antigen was sandwiched between the peptide-tagged capture and biotinylated detector antibodies; chemiluminescent detection was used to quantify signal.
This ‘homebrew’ approach successfully detected sub picomolar levels of cytokines and chemokines in human serum and plasma in a multiplex assay measuring IL-5, IL-6, IL-10, IL-22, TNFa. When compared to commercially available planar assays the universal planar assay format provided comparable LLOQ and LOD with sample correlation (R2) &gt;0.97 for all analytes tested. The universal planar assay was used to successfully screen multiple antibody pairs for human and mouse biomarkers and to develop robust assays using mouse, rabbit or goat monoclonal and polyclonal antibodies.
This universal multiplex planar homebrew assay format combines the accuracy, reproducibility and high sensitivity of Simoa Planar Array technology with total analyte flexibility, and provides assay developers a simple and versatile method to rapidly develop robust high-sensitivity multiplex assays for any analyte of their choice.
Cross/Tangential Flow Filtration of Antisera on an Industrial Scale
Authors: Holland Kathleen M , Fuller Cassandra Nicole 
Published: January 2023

Abstract
Purification of antisera, on an industrial scale, to date has been performed by SEC or affinity chromatography, both of which are costly. The method of cross/tangential flow filtration feeds solution across a filter surface rather than pushing product through it. Cross flow membranes are currently used in the manufacturing of serum proteins in order to dialyze and/or concentrate these solutions. Solvent, solutes, and particles smaller than the membrane’s pore size pass through the membrane, while larger molecules are retained and concentrated. This study was designed to apply membrane purification to increase the titers of polyclonal antisera on an industrial scale, as an alternative method to traditional chromatography techniques. Membranes with varying molecular weight cut offs were used to filter and concentrate neat antisera. The reduction of serum fluids and salts increase the concentration of antibodies, and therefore raise the titers. Concentrated antisera titers were then compared to neat antisera titers, to determine if a significant increase or decrease occurred, or if the method had no effect. This method has a marked resemblance to SEC, but without the need for buffers; other characteristics of the antisera remain unchanged. As the current availability of purified polyclonal antisera is scarce, the goal of this study was to produce a more effective product for use in immunology and medical device research and manufacturing across multiple testing platforms, by producing antisera with higher titers. This could potentially result in being able to process antisera to higher titers using lower concentrations of antigen in the host species, which is important from an industrial perspective for reducing manufacturing costs.
Early cancer detection using ctDNA technology – what is the relevance for Pacific Communities?
Authors: Sika-Paotonu Dianne , Anitelea Toni , Cockburn Alana , Guilford Parry 
Published: January 2023

Abstract
A novel cancer detection method known as circulating tumour DNA (ctDNA) analyses the small fragments of tumour DNA found in the blood circulation. Information relating to a cancer’s size and progression can be easily obtained from the DNA fragments found in a blood sample using specific isolation and quantification techniques. This is a relatively simple method for monitoring cancer and is being adapted to support early cancer detection efforts. Pacific Peoples living in New Zealand (NZ) and in the Pacific Region, remain disproportionately affected by some cancers. Given the increasing cancer rates worldwide and the medical situation for cancer patients in NZ and in the Pacific, the burden of cancer remains concerning as significant health inequalities exist for Pacific Peoples in particular. To support meaningful and respectful scientific knowledge translation activity with appropriate engagement promoting dialogue between researchers and Pacific communities, a Pacific Fono (gathering) event was held in the Wellington Region that included discussion of the ctDNA early cancer detection technology. Feedback from members of the various Pacific communities who attended indicated that researchers had engaged respectfully and facilitated discussion appropriately that was helpful, and also supported understanding of the ctDNA research work. Strong support was also expressed for the development of an early detection tool that involved a simple blood test methodology. The importance of taking into account the cultural context, combined with the appropriate scientific communication was shown to build meaningful connections and support respectful dialogue with Pacific communities.
Antibody-Immobilized &lt;wbr&gt;Nanofiber Scaffolds for the Delivery of Human Tumor Antigen-Specific Cytotoxic T Lymphocytes
Authors: LEE EUNJU , Jia Joann Ng Pei , Kim Tae-Jin , Lim Siyoung , Kim Jiyoung , Kim Jungwon , Kim Jeongsoo , Kim Junghee , Park Sunghee , Lee Kyung-mi 
Published: January 2023

Abstract
Adoptive cell therapy (ACT) employing the transfer of ex vivo expanded effector cells is an attractive approach for cancer treatment. The successful clinical translation of ACT, however, is impeded by inadequate effector cell numbers and poor persistence after infusion. To circumvent these limitations, we propose the use of biocompatible poly (lactide-co-glycolide) (PLGA) nanofiber scaffolds as a platform to effectively deliver human tumor antigen-specific cytotoxic T lymphocytes (CTLs) directly to the tumor site. We modified the PLGA nanofiber scaffolds with monoclonal antibodies (anti-CD8 and anti-CD28 Abs) to provide adhesion and co-stimulatory signals to CTLs. We show that CTLs can be attached and gradually released from the nanofiber scaffolds. We found that the nanofiber scaffold-released CTLs are more activated, have higher CD28 expression, and exhibit higher cytotoxicity with elevated IFNg production, compared to CTLs expanded in a conventional rapid expansion protocol. Our data highlight that anti-CD8/anti-CD28 Ab-conjugated PLGA nanofiber scaffolds can provide a biocompatible biodegradable delivery platform of tumor-specific CTLs by providing T cell persistence and tumor cytolytic function in the patients. Therefore, nanofiber scaffolds incorporated with have become potentially be a more successful strategy for ACT.
A comparison of single cell capture methods and amplification strategies to pair alpha and beta
Authors: Brown Brittany E , Pan Wenjing , Byrne-Steele Miranda , Eisenhower Mary , Li Song , Hou Xiaohong , Depinet Mollye M , Han Jian 
Published: January 2023

Abstract
Immune repertoire amplification of T cell receptors coupled to NGS provides detailed, sequence-level insight into the immune system which can be beneficial for the development of personalized therapies. However, information related to the cognate pairing of alpha and beta chains is lost once RNA extraction is performed on the bulk sample. We have previously presented low through-put (iPair) and high throughput methods for single cell pairing of the alpha and beta receptor chain. Here, we compare different methods to amplify and pair alpha and beta from single cells, as well as track CDR3 rankings from the same sample when sorted for iPair, high throughput pairing, and bulk repertoire. Template switch oligo (TSO) with 5′ cell barcoding, 3′ cell barcoding, and iRepertoire’s multiplex primers were used independently to amplify cDNA from the same cells. The cells are sorted CD14-CD4+ with 1% Jurkat cell line spiked in. Eight thousand cells from the same population were loaded onto 2 different platforms to capture single cells: a droplet-based capture and a microwell array capture. The results demonstrate that the dominant CDR3 pairs in single cell data (regardless of capture method) match the dominant CDR3 clones in bulk repertoire. All single cell methods compared accurately called the 1% Jurkat spike-in and the top paired CDR3.
Housing laboratory mice deficient for Nod2 and Atg16l1 in a natural environment uncovers genetic and environmental contributions to immune variation
Authors: Lin Jian-Da , Devlin Joseph C. , Yeung Frank , McCauley Caroline , Leung Jacqueline M. , Chen Ying-Han , Cronkite Alex , Hansen Christina , Drake-Dunn Charlotte , Ruggles Kelly V. , Cadwell Ken , Graham Andrea L. , Loke P’ng 
Published: January 2023

Abstract
The relative contributions of genetic and environmental factors to variation in immune responses are still poorly understood. Here, we performed a deep phenotypic analysis of immunological parameters of laboratory mice released into an outdoor enclosure, carrying susceptibility genes (Nod2 and Atg16l1) implicated in the development of inflammatory bowel diseases. Variations of immune cell populations were largely driven by environment, whereas cytokine production in response to stimulation was affected more by genetic mutations. Multi-omic models identified transcriptional signatures associated with differences in T cell populations. Subnetworks associated with responses against Clostridium perfringens, Candida albicans and Bacteroides vulgatus were also coupled with rewilding. Hence, exposing laboratory mice carrying different genetic mutations to a natural environment uncovered important contributors to immune variation.
Small molecule prevents inflammation in microglia by targeting the NLRP3 inflammasome
Authors: Kulkarni Amol A , Sajith Ayyiliath M , Zhang Changwen , Hao Jiukuan , Bowen J. Phillip 
Published: January 2023

Abstract
Uncontrolled CNS inflammation forms the pathophysiological basis of numerous disorders, including Alzheimer’s disease (AD). Evidence strongly suggests that the inhibition of the NLRP3 inflammasome is a promising strategy for the treatment of AD. A variety of natural products including curcumin, resveratrol, and isoliquiritigenin have displayed encouraging in vitro NLRP3 inhibitory activity. These natural products, however, are reported to display low chemical and/or enzymatic stability and therefore are not considered as promising leads for drug discovery. Using computer-assisted drug design methods and classical medicinal chemistry approaches, we developed a small library of tertiary sulfonylurea compounds that resembled the 3D-structure of the aforementioned natural products without the moieties responsible for the reported chemical and enzymatic instability. The compounds were synthesized, characterized, and tested for their inflammasome inhibitory activity. Preliminary studies indicated two of our compounds decreased the NLRP3 expression in a dose-dependent manner. These compounds reduced the production of inflammatory markers, such as, IL-1b, TNF-a, and caspase 1. These compounds lower the levels of inducible nitric oxide synthase and NO biosynthesis. MTT assays revealed that the sulfonylurea compounds did not affect N9 cell viability. Using computational chemistry and pharmacophore modeling, our efforts are currently on structural modifications to improve their biological activity and penetration the blood-brain barrier.
Gypenosides attenuate lipopolysaccharide-induced neuroinflammation and anxiety-like behaviors in rats
Authors: LEE BOMBI , SHIM INSOP , LEE HYEJUNG , HAHM DAE-HYUN 
Published: January 2023

Abstract
Neuroinflammation is considered a major factor in several neuropsychiatric disorders. Gypenosides (GPS) have pharmacological properties with multiple beneficial effects including anti-inflammatory and protective properties. In the present study, whether GPS could anxiolytic-like effects and chronic inflammation induced by injecting lipopolysaccharide (LPS) into rat hippocampus was investigated. Effects of GPS on inflammatory factors in the hippocampus and the mechanisms of these effects were also examined. Induction of LPS into the lateral ventricle caused inflammatory reactions, anxiety-like symptoms on the rats. Every day treatment of GPS for 21 consecutive days significantly increased the time spent and number of visits to the open arm in the elevated plus maze test, and significantly increased the number of central zone crossings in the open field test. GPS administration significantly reduced the freezing response to contextual fear conditioning. GPS administration significantly decreased pro-inflammatory mediators such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and nuclear factor-kappaB (NF-κB) levels in the brain. Furthermore, GPS reduced LPS-induced elevated levels of toll-like receptor 4 (TLR4) mRNA, and inhibition of brain-derived neurotrophic factor (BDNF) mRNA level. Collectively, these results showed that GPS may anxiolytic-like effects and provide a potential therapy for anxiety-like behaviors caused by neuroinflammation.
Human neural stem cells induce central nervous system specific regulatory T cells from the exTreg pool and promote repair in models of multiple sclerosis
Authors: Greilach Scott A , McIntyre Laura L , Hasselmann Jonathan , Othy Shivashankar , Ngyuen Quy , Sears-Kraxberger Ilse , Steward Oswald , Kessenbrock Kai , Cahalan Michael D , Blurton-Jones Mathew , Lane Thomas E , Walsh Craig M 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is a chronic inflammatory autoimmune disease affecting the central nervous system (CNS) and for which there is no cure. Current treatments focus on suppression of the immune system but fail to repair the resulting damage to the CNS. Neural stem cell (NSC) transplantation is a promising therapeutic strategy for treating neurodegenerative diseases through cell replacement and repair however it is unclear how these cells would mediate repair in MS. We report that human NSCs promote CNS specific T regulatory cells (Tregs) which activate endogenous repair pathways and promote remyelination in a murine model of MS. We observed remyelination, decreased inflammation and an increase in (CNS)-infiltrating CD4+CD25+FoxP3+ Tregs in EAE mice receiving an intra-spinal transplant of hNSCs. Recovery was not a result of cell replacement, as hNSCs underwent xenograft rejection, and was Treg dependent, as ablation of Tregs abrogated histopathological improvement. Treg expansion is antigen driven as hNSCs expanded CD25+FoxP3+ Tregs in-vitro when cultured with neural antigen restricted RAG2−/−2D2+ (R2D2) splenocytes but not RAG2−/−OT-II+ splenocytes. When co-cultured with B6 splenocytes, hNSCs drove the expansion of unique TCRs when compared to controls. Additionally, hNSC-Tregs also appear to derive from the exTreg pool suggesting both antigen specific expansion and antigen dependent maintenance of FOXP3 in CNS-specific Tregs. hNSC Tregs also have a unique expression profile and express transglutimase-2 which is implicated in oligodendrocyte dependent repair in the CNS.
IL-10 signaling in T cells is essential for induction of transplant tolerance
Authors: Lozano Marcos Iglesias , Bibicheff Darrel , Chicco Maria , Brandacher Gerald , Raimondi Giorgio 
Published: January 2023

Abstract
Induction of long term transplant survival by “costimulation blockade” (CoB) regimens is impaired by inflammatory responses. We and others recently revealed new interrelations between inflammatory cytokines and interleukin 10 (IL-10) that alter the signaling outcome of the latter. In this study we investigated the role of IL-10 during transplant tolerance induction.
Using a murine full-mismatch skin transplantation model, we showed that IL-10 has a fundamental role in the protective effect of CoB. This tolerizing regimen also failed to be effective in recipient mice with T cell specific unresponsiveness to IL-10, revealing an unexpected important direct modulation of T cells by this cytokine. Accelerated rejection in these transgenic mice correlated with increased accumulation of T cells producing TNF-α, and IFN-γ, and a doubling of IL-17A secreting lymphocytes. We then investigated the mechanism behind modulation of T cells physiology by IL-10. Using in vitro generated rested-effector cells, we show that IL-10 did not reduce their signaling threshold nor their effector functions (cytokine secretion). Moreover, IL-10 did not promote the expression of inhibitory receptors. Instead, we discovered that IL-10 can completely neutralize the TLR-mediated (CD28 independent) costimulation of memory and effector T cells (a largely unexplored phenomenon). We have an ongoing RNAseq analysis to decipher this novel mechanism at the molecular level.
Overall, these results highlight the importance of IL-10 signaling for the therapeutic efficacy of CoB. A better understanding of the effects of this cytokine on T cells will reveal targets of strategies to sustain and potentially improve the clinical efficacy of CoB.