bluuebunny/crossref_metadata_2025_split huggingface dataset:
Invariant NKT cells promote expansion of novel CXCR3+CCR4+CD8+ T cells in the liver following allogeneic hepatocyte transplant
Authors: Ringwald Bryce Alan , Zimmerer Jason M , Chaudhari Sachi , Han Jing , Peterson Chelsea M , Warren Robert T , Hart Madison , Abdel-Rasoul Mahmoud , Bumgardner Ginny L 
Published: February 2023

Abstract
Hepatocyte transplant (HcTx) is a treatment for metabolic diseases and a bridge to liver transplant but is limited by immunogenicity. Our prior work identified the critical role of CD4-dependent and CD4-independent CD8+ T cells in HcTx rejection. Herein, we evaluated the influence of invariant natural killer T cells (iNKTs), abundant in the liver, upon CD8+ T cell responses in the presence and absence of CD4+ T cells. To investigate this, C57BL/6 (wild-type; WT) or iNKT deficient Jα18 KO (H-2b) received FVB/N (H-2q) HcTx. Cohorts were depleted of CD4+ T cells and/or reconstituted with iNKTs. Recipients were evaluated for cytotoxicity, CD8+ T cell phenotype, and HcTx rejection. By comparing CD4-sufficient WT and Jα18 KO recipients, we found that the presence of iNKTs significantly enhanced in vivo CD8-mediated cytotoxicity (83±3% vs 23±3%; p&lt;0.001) but did not affect survival (10 days vs 14 days; p=ns). However, in CD4-deficient recipients, iNKTs were critical, since without CD4+ T cells and iNKTs, CD8-mediated in vivo cytotoxicity was completely abrogated (31±3% vs 2±1%; p&lt;0.001) and HcTx survival was significantly prolonged (14 days vs 56 days; p&lt;0.001). iNKTs enhanced the quantity of a novel CD8+ T cell subset, CD44+IFN-γ+CXCR3+CCR4+CD8+ T cells, in the liver (2-fold) in all HcTx recipients (p&lt;0.03 for all comparisons). CXCR3+CCR4+CD8+ T cells, when compared to CXCR3+CCR4−CD8+ T cells, express higher IFN-γ (32±3% vs 9±1%; p&lt;0.001) and TNF-α (37±3% vs 6±1%; p&lt;0.001). They also exhibit greater in vivo cytotoxicity (13±1% vs 5±1%; p&lt;0.001) and mediate rapid HcTx rejection in RAG1 KO recipients (5 days vs 16 days; p=0.02). We show that iNKTs enhance the expansion of highly cytotoxic CXCR3+CCR4+CD8+ T cells that mediate rapid rejection of HcTx.
Xenogeneic Sertoli cells express complement inhibitory proteins necessary for their survival of hyperacute rejection
Authors: Washburn Rachel , Kaur Gurvinder , Dufour Jannette 
Published: February 2023

Abstract
Transplantation is used to treat organ failure associated with various diseases, however there is a severe shortage of organs acceptable for transplant. Utilization of pig organs offers an endless supply of transplantable tissue, but xenografts are rapidly destroyed by hyperacute rejection (HAR). The primary humoral mediator of HAR is the activation of complement (C″) by preformed-antibody binding to xenoantigens, causing a proteolytic cascade that results in cell lysis by membrane attack complex (MAC) insertion. Sertoli cells (SCs) are immunoregulatory testicular cells that survive as xenografts for an extended period without the use of immunosuppressive drugs. The goal of this study was to examine the mechanisms for SC survival of C″. Neonatal porcine SCs (NPSCs) transplanted into rats survived as xenografts while porcine islets (NPIs) did not. Immunohistochemistry of the surviving grafts show deposited C3, but not MAC, suggesting that NPSCs inhibit C″ after activation. An in vitro human serum (HS)-C″ cytotoxicity assay confirmed that NPSCs survive activated human C″. Analysis of NPSC expression of complement inhibitory proteins (CIP) revealed higher levels of membrane cofactor protein (MCP) and decay accelerating factor (DAF) as compared to NPIs, which are killed by C″. Knocking down expression of MCP and DAF using shRNA confirmed their importance in NPSC survival of C″. Less than 10% of these cells survived exposure to the HS-C″ cytotoxicity assay, indicating MCP and DAF are important in NPSC xenograft survival in vitro.
Selective expansion of regulatory T cells by kidney-tethered IL2 mutein
Authors: Larkin Bridget M , Visweswaraiah Jyothsna , Li Bilian , Pradhan Komal , Rowe Michael , Rajani Gaurav , Nagy-Domonkos Celesztina , Higginson-Scott Nathan , Viney Joanne L , Otipoby Kevin L , Shaheen Hussam H , Mascanfroni Ivan 
Published: February 2023

Abstract
Regulatory T cells (Treg) play a critical role in maintaining graft tolerance following organ transplantation, and increased numbers of Tregs in solid-tissue grafts such as the kidney are associated with improved graft survival and function. Therapeutic approaches that increase Tregs offer a promising alternative to current standard-of-care treatment consisting of broad-acting immunosuppressants and their attendant side effects. While efforts to expand Tregs ex vivo for infusion into patients have shown promise, they present manufacturing and administration challenges. Low dose (LD) interleukin 2 (IL-2) drives Treg proliferation and function via the heterotrimeric IL-2 receptor expressed on Tregs and has been shown to expand Tregs in vivo, including in the context of transplantation. However, IL-2 can also activate other immune cells including conventional T cells and Natural Killer (NK) cells, which express IL-2Rb/CD122 and IL-2g/CD132, and this could accelerate rejection. We have enhanced IL-2 selectivity for Treg by introducing mutations that increase affinity for CD25 and decrease affinity for CD122/CD132, resulting in Treg specific expansion. Here, we demonstrate that this IL-2 Mutein (IL-2M) can be “tethered” to the kidney via a bifunctional antibody targeting a tubule-specific protein. This kidney-tethered IL-2M maintains Treg selectivity and localizes to the kidney with extended tissue PK. Moreover, in a CD34-NSG mouse model, animals dosed with kidney-tethered IL-2M exhibited prolonged kidney Treg expansion compared to a systemic, non-tethered IL-2M. This approach has the potential to substantially improve kidney graft acceptance while reducing adverse effects and improving patient quality of life.
IL-12 abrogates mechanisms of immune suppression by B-ALL
Authors: Hunter Rae , Henry Curtis J. , Porter Christopher 
Published: February 2023

Abstract
Despite major advance in therapeutic options for treating patients with acute lymphoblastic leukemia (ALL), including the use of immunotherapies, 15–20% of patients still succumb to disease. Given the high mortality rates, particularly in the context of immune-based therapies, there is still a knowledge gap regarding how leukemia cells evade immune surveillance.
In published studies, we have shown that interleukin-12 (IL-12) secreted by B-leukemia cells stimulate T-cell mediated immunity. We found that recombinant IL-12 treated WT and Rag1−/− mice, which lack B- and T-cells but still have innate immune cells, had prolonged survival as compared to untreated mice. These results demonstrate that IL-12 increases the anti-leukemia immunosurveillance capacity of innate and adaptive immune cells.
Mechanistically, we observed when T-cells were stimulated in ALL supernatant in vitro there is a reduction in the surface expression of the T-cell activation marker, CD44, and marker of degranulation, CD107b. Macrophages co-cultured with leukemia cells were also unable to significantly upregulate the activation markers, CD80 and CD86, compared to macrophages in media alone.
We then explored combination therapeutics in vitro to override ALL immune suppression. Considering the potency of IL-12 alone and Blinatumomab, a bispecific T-cell engager (BiTE) antibody used to treat patients with relapsed and refractory disease, we determined how combining these therapies impacted B-ALL cytotoxicity when cultured with T-cells. Impressively, IL-12 augmented the efficacy of Blinatumomab resulting in more T-cell mediated killing of leukemia cells. Our results demonstrate the therapeutic potential of IL-12 as a treatment for ALL.
Robust Anti-SARS-CoV-2 Humoral and Cellular Immunity through cGAMP-loaded Microparticle Adjuvanted Subunit Vaccination
Authors: Sandor Adam , Varma Devika M , Batty Cole J , Baxter Victoria K , Sakar Sanjay , Thompson Michael A , Gao Hao , Liang Kaixin , Chou Wei-Chun , Bachelder Eric M , Heise Mark T. , Ainslie Kristy M , Ting Jenny P 
Published: February 2023

Abstract
In less than one year, the COVID-19 pandemic has infected over 95 million people with greater than two million deaths globally. Vaccines against the SARS-CoV-2 virus offer the best chance at curbing the severity and spread of COVID-19. While initial COVID-19 vaccine candidates have demonstrated great promise and efficacy, it is unknown how broad and durable those responses will be. Furthermore, eliciting strong antiviral immune responses will prove critical against future emerging strains. Previously, we demonstrated that cyclic-AMP-GMP (cGAMP) encapsulated within acetalated dextran microparticles (MPs) adjuvants influenza subunit antigens to strongly protect against influenza viral challenge when used as an intramuscular vaccination in a mouse and ferret model. Based on these studies, we hypothesized that vaccination with the stabilized spike protein of SARS-CoV-2 (S2P) and cGAMP MPs would lead to protective anti-SARS-CoV-2 immune responses. We demonstrate that S2P vaccination with cGAMP MPs lead to increased total anti-SARS-CoV-2 IgG with a strong Th1 skewed IgG2c antibody response compared to other clinically relevant adjuvants. Vaccination using cGAMP MPs also increased the frequency of activated Th1 antigen-specific T cell responses compared to controls and other vaccine adjuvants, as determined by IL-2 and INFg ELISPOTs. Finally, vaccination using cGAMP MPs lead to comparable SARS-CoV-2 neutralizing antibody production and protection against mouse adapted SARS-CoV-2 infection. These studies demonstrate that vaccination with S2P and cGAMP MPs lead to a protective humoral response and improved antigen-specific T cell responses compared to other clinically relevant adjuvants.
Development and Characterization of Potential Neutralizaing Antibodies to SARS-CoV-2 from Phage Display Human Antibody Library System
Authors: Park In Ho , Lee Sang Pil , Yoon Sunha , Shin Ji-Young , Oh Jooyeon , Lee Hoojung , Jeon Eenkyung , Park Bum-Chan , Shin Jeon-Soo 
Published: February 2023

Abstract
Since the outbreak of Wuhan, China, in 2019, SARS-CoV-2 infection has been increasing globally, and as of January 2021, it has exceeded about 85 million people, with about 2 million deaths. Despite Covid-19 vaccines urgently approved for creating population (herd) immunity, the first need for the infected patients is a treatment. In this perspective, neutralizing antibody is the first concern because it is significant for symptom relief and rapid recovery of patients without therapeutic chemicals.
Phage display human antibody library technology is a potential tool screening neutralizing antibodies specific to the receptor-binding domain, RBD of the spike protein of SARS-CoV-2 virus entry to host cells.
Fifteen antibodies specific to RBD were selected through Ymax®-ABL and Patient-Immune Library and characterized through ELISA, immune fluorescence assay, epitope mapping. Some of the selected antibodies were improved in the spike-neutralizing property through in vitro affinity maturation. Here, the COVID-19 therapeutic values of two promising candidates, both of which recognize different epitope sites on RBD, were functionally proved by using both in vitro cell and in vivo mouse models.
Costimulatory blockade modifies germinal center B cell function in NZB/W F1 mice
Authors: Raparia Chirag , Zeumer-Spataro Leilani , Quach Tam D , Choi Seung-Chul , Morel Laurence , Davidson Anne 
Published: February 2023

Abstract
Systemic lupus erythematosus (SLE) is a complex autoimmune disease with significant morbidity and mortality - demanding further examination of disease reducing treatments. Previously, we showed that a 2-week course of combined CTLA4Ig and anti-CD40L costimulatory blockade prevented loss of tolerance to dsDNA in NZB/W mice while maintaining immune function against exogenous antigen, explained by an altered selection of autoreactive B cells in germinal centers (GC). Here, we examined both B and T cells to clarify if restoration of tolerance is due to a change in GC B cell profile or a decrease in T cell help. 3 months after treatment, large germinal centers persisted in the treated group, but due to smaller spleen size total GC B cells and TFH Cells were decreased compared with controls. The metabolic profile of splenic B and T cells showed a decreased spare respiratory capacity in both subsets and a decrease in B cell glycolysis among the treated group. RNA-seq analysis of GC dark zone and light zone B cells from treated mice showed decreased expression of markers of oxidative phosphorylation and cell proliferation and increased expression of signaling molecules downstream of BCR, including PLCγ, compared with untreated controls. GC B cell stimulation assays showed a delay in phosphorylation of PLCγ in response to anti-IgG in treated mice. These data show a long-lasting effect of short-term costimulatory blockade on both B and T cells with metabolic remodeling of GC B cells and an alteration of BCR signaling. Since both a profound decrease in BCR signaling and an increase in B cell glycolysis have been reported to enhance loss of tolerance in lupus prone mice, our findings help to explain the restoration of tolerance observed after treatment.
Correlation of autoantigen-specific Treg frequency with development of spontaneous organ-specific autoimmunity in a mouse model of uveitis
Authors: Yin Mianmian , Hiam Kamir , Proekt Irina , Chan Jackie , Hu Yongmei , Lowell Clifford A , Gould Douglas B , Anderson Mark S , DeFranco Anthony L 
Published: February 2023

Abstract
About 50% of AireGW/+Lyn−/− mice developed autoimmune uveitis, and this was accompanied by the expansion of interphotoreceptor retinoid-binding protein (IRBP) specific CD4+T cells. Moreover, the response to IRBP was necessary for disease initiation, as deletion of IRBP prevented uveitis. Mass cytometry (CyTOF) analysis of CD4+T cells in the retina of the mice with uveitis identified activated T cell subsets characterized as Ly6Chi effectors, PD-1hi effectors, and Treg. To study cellular mechanisms that promoted development of uveitis, we did single-cell RNA sequencing (scRNA-seq) of eye-draining lymph node (LN) CD4+T cells that recognize the P2 epitope of IRBP (amino acid 271–290). Uniform manifold approximation and projection (UMAP) analysis showed that these CD4+T cells included distinct subsets that were Ly6c1hi , Pdcd1hi and Treg, with the Treg population being overrepresented in the mice without disease. Pseudotime analysis indicated that the Ly6c1hi T cell subset represented an earlier stage in activation, whereas the Pdcd1hi subset represented a later stage. Similar analysis of P2–specific CD4+T cells from the retina of mice with uveitis identified 6 subsets of P2-specific T cells, and was largely consistent with the CyTOF analysis of total CD4+ T cells from the retina. Pseudotime analysis indicated that toward the earlier time were cells characterized by being proliferative (Mki67hi), followed by a Lag3hi subset and at other end of the pseudotime axis by a Lag3hi / Pdcd1hi subset. Our results are consistent with the hypothesis that the fraction of Treg within the IRBP-specific CD4+ T cells in the draining LN determines whether these genetically susceptible mice develop eye-specific autoimmunity or are protected.
Contribution of a Hybrid Insulin Peptide-reactive T cell to Diabetes progression in the NOD mouse.
Authors: Lin Yong , Kong Yuelin , Igyarto Botond Z , Zurawski Gerard , Zurawski Sandra , Bettini Maria , Bettini Matthew 
Published: February 2023

Abstract
Type 1 Diabetes (T1D) is a T cell-mediated autoimmune disease characterized by the immune destruction of pancreatic β cells. The search for natural equivalents to the mimetopes that can stimulate diabetogenic T cell clones from the Non-Obese Diabetic (NOD) mouse model of T1D led to the realization that hybrid peptides (HP) formed by the joining of β cell granule protein-derived peptide fragments are highly stimulatory to some clones. These hybrid peptides were later identified in vivo and proposed as a new class of post-translationally modified autoantigens mediating this disease. Recent data suggest high concentrations of these proteins in β cell lysosomes during granule turnover can lead to the hybridization of peptide fragments which may then be presented on MHC molecules. In the thymus, medullary thymic epithelial cells (mTECs) mediate T cell tolerance through promiscuous gene expression, but may not mediate HP-reactive T cell tolerance. We hypothesize that HP-reactive T cells preferentially escape the thymus and contribute significantly to T1D progression. We attempted to enhance T cell central tolerance to 2.5HIP, a hybrid insulin peptide formed from Proinsulin and Chromogranin A, the only two antigens known to be critical for T1D progression in the NOD mice, to better understand its impact on T1D incidence. Using an antibody-peptide conjugation system, we targeted 2.5HIP to a subset of thymic resident DCs to enhance presentation of 2.5HIP in early neonatal thymus. In neonatal BDC2.5 TCR-transgenic mice we found that a single dose of antibody-2.5HIP enhanced antigen-specific thymocyte apoptosis and regulatory T cell development. Importantly, a single dose in wild-type neonates resulted in a significant delay of diabetes onset.
Anti-TRIM21 antibody seropositivity is related to B-cell abnormalities in systemic lupus erythematosus
Authors: Kunishita Yosuke , Yoshimi Ryusuke , Komiya Takaaki , Sakurai Natsuki , Sugiyama Yumiko , Takase-Minegishi Kaoru , Kirino Yohei , Nagaoka Shouhei , Nakajima Hideaki 
Published: February 2023

Abstract

Background/Purpose
TRIM21 is one of the autoantigens which react with anti-SS-A antibody (Ab). Previous studies have shown that TRIM21 dysfunction promotes aberrant B-cell differentiation in systemic lupus erythematosus (SLE). Here we examined the relationship between anti-TRIM21 Ab and clinical and immunological characteristics in SLE patients.

Methods
Twenty-seven patients with SLE before immunosuppressive therapies and four healthy controls (HC) were enrolled in the study. Serum anti-TRIM21 Ab levels were measured using enzyme-linked immunosorbent assays. The serum levels of cytokines and immunoglobulins were measured using cytometer beads arrays. The protein expression levels of TRIM21 in peripheral blood mononuclear cells (PBMCs) were evaluated by western blotting.

Results
Nineteen and 8 patients showed seronegativity and seropositivity for anti-TRIM21 Ab, respectively. There were no significant differences in the background parameters, including SLE activity and laboratory data. The serum levels of interferon (IFN)-α2 were significantly higher in patients with anti-TRIM21 Ab as compared with those without anti-TRIM21 Ab (p = 0.03). The levels of IgG1, IgG2, and IgA were significantly higher in patients with anti-TRIM21 Ab as compared with those without anti-TRIM21 Ab (p = 0.005, 0.04 and 0.01, respectively). The PBMCs of patients with anti-TRIM21 Ab showed a significantly higher expression of TRIM21 protein as compared with those of patients without anti-TRIM21 Ab.

Conclusion
Anti-TRIM21 Ab seropositivity is related to B-cell abnormalities and type I IFN overproduction in SLE patients, which may be due to the inhibitory effect of anti-TRIM21 Ab on TRIM21 functions.
Pertussis toxin inhibits encephalitogenic T cell infiltration and promotes a B cell driven disease during TH17-EAE
Authors: Maria Zahra , Turner Emma , Axtell Robert C. 
Published: February 2023

Abstract
Pertussis toxin (PTX) is required as a co-adjuvant to induce disease in active Experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis. The objective of this study was to investigate the role of PTX during adoptive transfer TH17-EAE and observe alterations in the disease phenotype. TH17-EAE was induced by transferring myelin-specific TH17 cells, harvested from MOG35-55 immunized wild type (WT) donor mice, into WT or B cell deficient (μMT) recipient mice. The recipient mice also received PTX or PBS on days 0 and 2 and monitored for paralysis. In WT mice, PTX significantly reduced disease severity and delayed onset of paralysis. PTX reduced accumulation of pathogenic T helper cells and myeloid cells in the CNS, which we found to be associated with PTX inhibition of chemokine receptor dependent trafficking. Strikingly, our data shows that PTX promotes the accumulation of B cells in the CNS and elevates the capacity of B cells to present antigen to T cells in the periphery, suggesting an altered disease toward a B cell-dependent pathology. We found that without PTX treatment, the severity of disease was equivalent between WT and μMT mice. In contrast, with PTX treatment, the μMT mice had significantly less disease compared to the WT mice. In conclusion, our data suggests that during TH17-EAE, PTX restricts the trafficking of pathogenic T helper cells and myeloid cells into the CNS and thereby delays the disease onset. However, not only PTX does not alter trafficking of B cells into the CNS it also stimulates a more inflammatory B cell phenotype. This study provides novel insights to interactions between TH17 and B cells in mice and has defined a new model to study the pathologic functions of B cells during multiple sclerosis.
Gene Edited Human CD4+ T Cells Reveal that Loss of PTPN22 Function Enhances Mildly Autoreactive T Cell Responses
Authors: Anderson Warren R , Barahmand-pour-Whitman Farbia , Linsley Peter S , Cerosaletti Karen , Rawlings David 
Published: February 2023

Abstract
A genetic variant in the gene PTPN22 (R620W, rs2476601) is strongly associated with increased risk for multiple autoimmune diseases. PTPN22 is a non-receptor tyrosine phosphatase that negatively regulates proximal T cell receptor (TCR) signaling. PTPN22 deficiency increases TCR signaling in mouse and human T cells. Mouse models of the variant similarly exhibit increased TCR signaling, while primary T cells from human carriers of the variant exhibit decreased signaling. Here, we utilize gene editing in primary human T cells to address these discordant findings and explore how the risk variant impacts TCR signaling. Using Crispr/Cas9 nucleases and DNA repair templates, we established a robust platform for precise homology-directed-repair based gene editing of PTPN22, allowing the generation of PTPN22 risk edited, control edited (silent modification), and knock-out T cells from the same donor. In contrast to control edited cells, PTPN22 risk edited T cells exhibit increased activation in response to TCR engagement with higher expression of activation markers including CD40L and CD25, mimicking PTPN22 KO cells. Further, using lentiviral delivery of transgenic TCRs (derived from autoreactive T cells of T1D patients), we demonstrate that loss of PTPN22 leads to enhanced TCR signaling in weakly autoreactive, but not strongly autoreactive T cells, causing increased phosphorylation of S6 kinase, enhanced proliferation, and greater Th1 skewing. Taken together, these findings suggest that direct effects of the PTPN22 rs2476601 variant represents a loss of function in T cells, increasing TCR signaling in mildly autoreactive T cells, potentially expanding the pool of autoreactive T cells and skewing them toward an inflammatory phenotype.