bluuebunny/crossref_metadata_2025_split huggingface dataset:
The H1-isoform of the cell surface receptor Slamf6 suppresses expansion of PD-1+ OPN+ CD4+ T Follicular Helper cells in the lupus-prone B6.Sle1b mice. (123.5)
Authors: Keszei Marton , Detre Cynthia , Castro Wilson , Magelky Erica , Terhorst Cox 
Published: January 2023

Abstract
A major role for the Slamf6 gene in the pathogenesis of lupus-related autoimmunity has been revealed by the analysis of a number of congenic mutant mouse strains. Importantly, upon introduction of one copy of a BAC transgene expressing the protein isoform Slamf6-H1 markedly ameliorates T cell-dependent lupus in the Sle1b mouse, which lacks H1. Here we report that a PD-1+ subset of memory CD4+ T cells dramatically expands in Sle1b mice, but not in Sle1b mice, which express Slamf6-H1. Further analysis showed that this PD-1+ population mainly consists of CXCR5+ T follicular helper cells the expansion of which is prevented by Slamf6-H1 as well as by the absence of the adapter SAP. Surprisingly, these T follicular helper cells express the cytokine osteopontin, which is known to drive polyclonal B cell proliferation. As the human osteopontin gene have been linked to the development of SLE, the outcomes of the present studies could suggest novel therapeutic modalities for SLE patients.
Sexually dimorphic control of regulatory T cell function and autoimmunity protection through B7-H1 and estrogen signals (123.50)
Authors: Curiel Tyler , Kious Mark , Daniel Benjamin , Lin Pei , Thibodeaux Suzanne , Vadlamudi Ratna , Wierman Margaret , Turka Laurence , Zhang Ruan , Sharpe Arlene , Francisco Loise , Lishi Sun , Pandaswara Srilakshmi , Tekmal Rajeshwar , Hurez Vincent 
Published: January 2023

Abstract
CD4+CD25hi regulatory T cells (Tregs) mediate peripheral tolerance and defend against autoimmunity. Among immune sexual dimorphisms, autoimmunity is more common in females but underlying mechanism(s) are poorly understood. We show that B7-H1 signals dampen estrogen-driven Treg mTOR activation through estrogen receptor beta, reducing Treg function independent of PTEN. Poorly functional Tregs from B7-H1 deficient hosts appeared phenotypically normal and did not produce IL-17. B7-H1-deficient female mice (but not males or WT of either sex) were prone to anti-CTLA-4-induced autoimmunity. Autoimmune propensity was rescued by estrogen withdrawal or B7-H1-sufficient dendritic cell transfer. B7-H1-mediated effects on estrogen signals were also mediated by B or T lymphocytes or monocytes, and occurred during induced Treg generation, with no effect on differentiated Tregs. CD80 and PD-1 were not ligands for these effects nor was Treg B7-H1 expression required. Estrogen affected male Tregs in vitro and in vivo provided B7-H1 was blocked. Strikingly, B7-H1 and estrogen did not affect thymic Treg differentiation or function, suggesting differential control of natural Tregs and a potential role for induced Tregs in autoimmunity protection. Similar Treg results were seen in BL6 and Balb/c mice. We thus show a mechanistic basis for the female propensity to autoimmunity, and data are consistent with proposed roles for B7-H1 in female-predominant autoimmune disorders, particularly lupus.
A novel method for kinetic measurements of rare immune cell proliferation using Cellometer image-based cytometry (124.1)
Authors: Lin Bo , Chan Leo , Zhong Xuemei , Pirani Alnoor 
Published: January 2023

Abstract
Measuring cell proliferation is important for studying the effects of various treatments on cultured primary cells or cell lines. Current proliferation analysis methods employ flow cytometry for fluorescence detection of CFSE-labeled cells. However, conventional flow cytometers require a considerable amount of cells per reading, which becomes an issue for kinetic measurements with rare cell population due to the lack of samples for flow cytometric analyses at multiple time points during proliferation period. Here we report the development of a novel cell proliferation kinetic detection method for low cell concentration samples using the new Cellometer Vision image-based cytometry system. Since the Cellometer system requires only 20 µl of sample, cell proliferation can be measured at multiple time points over the proliferation period, whereas typically, flow cytometry is only performed at the end of the proliferation period. To validate the detection method, B1 and B2 B cells were treated with a B cell mitogen and proliferation was measured on day 1, 3, 5, and 6. To demonstrate the capability, B1 B cells were treated with a panel of TLR agonists and proliferation was measured on day 2, 4, 6, and 7. Cellometer was able to obtain proliferation results on each day, which were comparable to flow cytometry. This novel method allows for kinetic measurements of the rare cell samples such as B1 B cell, which has the potential to revolutionize kinetic analysis of cell proliferation.
A novel method for autophagy analysis using statistical microscopy (124.4)
Authors: Pugsley Haley , Hall Brian , Mordecai Scott , Friend Sherree , DeMarco Richard , Kong Raymond , George Thaddeus 
Published: January 2023

Abstract
Autophagy is a process in which normal cellular components that accumulate during growth and differentiation are degraded via the lysosome; it is a survival mechanism that reallocates nutrients from unnecessary processes to more vital processes in the cell. Basal levels of autophagy are usually low but can be up-regulated by numerous stimuli including starvation, physiological stress, pharmacological agents and infections. In addition, autophagy suppression has been associated with cancer, neurodegenerative disorders, infectious diseases and inflammation. During autophagy, cytoplasmic LC3 is processed and recruited to the outer membrane of forming autophagosomes, and cells undergoing autophagy can be identified by visualizing LC3 puncta by immunofluorescence microscopy. While manual microscopy allows visual per-cell identification of autophagy, objective statistical analysis of autophagy can be challenging with this method owing to its time-consuming and generally subjective nature. To overcome these problems, we used the ImgeStreamX imaging cytometry platform, which objectively collects and quantifies statistically large numbers of images per sample, to measure autophagy in K562 cells, a human CML line. In this study, we demonstrate a method for determining the best image-based parameter for quantifying LC3 puncta in these cells, and apply that parameter to large data sets to measure compound-induced autophagy in an objectively and statistically significant manner.
A simple and rapid method for the isolation of untouched human memory CD8 T cells (124.9)
Authors: Fairhurst Maureen , Posarac Vesna , McQueen Karina , Thomas Terry 
Published: January 2023

Abstract
Memory CD8 T cells are long-lived antigen-specific cells that persist after clearance of infection. Functionally, they are distinguished from naïve CD8 T cells by their less extensive signal requirement for activation and their ability to respond quickly to recall antigens and secrete a broad repertoire of cytokines. In addition, the two populations are phenotypically distinct, with naïve cells expressing CD45RA, and memory cells expressing CD45RO, a marker indicative of previous activation. Current protocols for the isolation of memory CD8 T cells are time-consuming and require the use of columns. We have developed a new kit for easy and rapid isolation of memory CD8 T cells from PBMCs by immunomagnetic, column-free cell separation (EasySep™). Non-CD8 T cells and CD45RA positive CD8 T cell subsets are targeted for depletion by bispecific tetrameric antibody complexes crosslinked to dextran-coated magnetic particles. The labeled cells are separated using an EasySep™ magnet and the desired fraction is poured off. The procedure is performed in 30 minutes and can be fully automated using RoboSep™. The mean enrichment purity and recovery of CD45RO+ CD45RA- CD8+ T cells is 86 ± 4, and 23 ± 11, respectively. The kit provides a simple and efficient means of isolating untouched human memory CD8 T cells that are ideal for studies in signal transduction, activation, cytokine expression, and response to infectious disease.
Allergen specific IgG augments Th2 mediated inflammation in the lungs through FcγRIII signaling and upregulation of IL-33 (125.1)
Authors: Tjota Melissa , Williams Jesse , Lu Tiffany , Byrd Tiara , Clay Bryan , Sperling Anne 
Published: January 2023

Abstract
Asthma is a chronic inflammatory disease of the lungs marked by mast cell activation, eosinophilia, and airway remodeling. It has been seen that primary sensitization to inhaled allergens leads to increased production of allergen specific IgE and IgG1. Several studies have shown a critical role for IgE in allergic lung disease, but it is unclear how IgG1 might contribute to the initiation and perpetuation of this disease. Previous work in our lab identified a key role for FcγRIII on dendritic cells (DC) in the development of optimal Th2-dependent airway inflammation. As FcγRIII is efficiently engaged by IgG1-immune complexes, we hypothesized that sensitized individuals could form allergen-specific immune complexes upon secondary exposure which could contribute to the augmented Th2 responses found in allergic asthma. We used two different models of immune complex mediated Th2 inflammation through either immune complex formation in vivo or local installation of preformed immune complexes. Both models showed that this process was mediated through FcγRIII. Utilizing both in vitro and in vivo approaches, we then identified IL-33 as a gene downstream of FcγRIII signaling on DCs. Notably, immune complex mediated Th2 inflammation in the lungs was decreased in ST2-/- mice with a specific loss of eosinophilia suggesting a critical role for IL-33. Our findings that previous sensitization can direct the response of DCs could provide a new paradigm for the development of Th2 inflammation.
Environmental exposures alter T cell differentiation and aggravate airway disease through the Aryl Hydrocarbon Receptor (125.6)
Authors: Mezrich Joshua , Zhang Xiaoji , Fechner John 
Published: January 2023

Abstract
Environmental exposures can aggravate airway disease. 10% of World Trade Center disaster responders developed asthma. Polycyclic aromatic hydrocarbon (PAH) levels were the highest ever reported. The Aryl Hydrocarbon Receptor (AHR) directs T-cell differentiation between FoxP3+ Regulatory T-cells and Th17 effector cells, and is the primary receptor for PAHs. Exposure to specific inhaled PAHs found in diesel exhaust particles (DEP), urban dust particles (UDP), and coal fly ash (CFA) has a direct effect on T-cell differentiation in an AHR-dependent manner, that enhances Th17 differentiation and hastens the development of asthma. Naïve CD4+ CD25- T cells from C57BL/6 mice were cultured under Th17 conditions. We utilized standard reference materials (SRM) representing PAHs from UDP, DEP, CFA in cultures and mixed lymphocyte cultures. Human epithelial cells were treated, and then examined for Epithelial to Mesenchymal Transition (EMT), in the presence of PBMC and SRMs. Finally we exposed lungs of mice in vivo to UDP intratracheally (IT). SRMs increased IL-2, IL-17, and IL-22 in the supernatant of naïve T-cells by bead array. SRMs increased IL-17 expression in MLCs. Both IL-17 and UDP were able to cause significant EMT. In lungs exposed to UDP, the AHR was activated and IL-17 was upregulated in tissue by qPCR. PAHs lead to a TH17 deviation in the lung through the AHR. This may play a role in aggravating airway disease in susceptible people, and will be a target for future treatment.
Loss of DRAK2 signaling enhances allogeneic transplant survival by altering effector T cell responses and memory generation (126.24)
Authors: Weist Brian , Hernandez Jeniffer , Walsh Craig 
Published: January 2023

Abstract
Drak2 is a serine threonine kinase that has previously been shown to negatively regulate activation of T cells through the T cell receptor. Here we demonstrate that loss of DRAK2 signaling significantly promotes the acceptance of allogeneic engraftment in two separate transplant models without generalized immunosuppression. Drak2-/- T cells failed to reject allogeneic tumors, and were defective in rejecting Balb/C allogeneic skin grafts on C57BL6/J recipients. A significant fraction of alloreactive Drak2-/- T cells underwent apoptosis following activation, whereas expression of Bcl-xL in Drak2-/- T cells restored rejection. Formation of allogeneic memory was also greatly hampered in T cells lacking the Drak2 gene. Adoptive transfer of memory T cells from Drak2-/- mice failed to promote the rejection of allogeneic tumors, and such cells led to significantly delayed rejection of skin allografts in the Balb/C to B6 model. Costimulatory blockade by in vivo administration of CTLA4-Ig synergized with the DRAK2 deficiency and led to long-term allogeneic skin graft acceptance. Overall, these results demonstrate that DRAK2 plays an important role in primary and memory T cell responsiveness to allografts. Since previous studies have demonstrated that a loss of DRAK2 does not negatively impact antiviral immunity, these studies underscore the potential utility of pharmacological blockade of DRAK2 to achieve transplant maintenance without the imposition of generalized immunosuppression.
Autocrine role for HMGB1 and HSP60 in the growth of murine mammary carcinoma (127.13)
Authors: Kurt Robert , Chalmers Samantha , Eidelman Alec , Ewer Jason , Serrano Antonio , Vail Caroline 
Published: January 2023

Abstract
Previously we reported that Myd88 contributed to growth, metastasis, and expression of CCL2 and CCL5 by mammary carcinoma implying that constitutive signaling through Myd88 contributes to tumor progression. To begin to decipher what may be inducing this signaling we focused on proteins that could function as damage associated molecular patterns (DAMPs) since DAMP expression has been reported in tumors, and certain DAMPs mediate effects through toll-like receptors. A screen of mammary carcinoma for DAMP expression showed HMGB1 and HSP60 were significantly elevated relative to normal mammary epithelium. Although suppression of either HMGB1 or HSP60 using RNAi reduced growth of the tumor cells, a significant reduction in growth as well as CCL2 expression was only found when both DAMPs were targeted. Since, besides their ability to function as DAMPs, HMGB1 and HSP60 have important roles in normal cellular functions we used blocking antibodies to determine whether these DAMPs were mediating effects in an autocrine manner. Indeed, antibodies to these DAMPs inhibited growth of the tumor cells, and analysis using a Myd88 inhibitory peptide suggested that HMGB1 mediated effects in a Myd88 dependent manner, while HSP60 mediated effects in a Myd88 independent manner. Collectively, these data show that mammary carcinoma overexpress HMGB1 and HSP60 relative to normal mammary epithelium, their expression influences tumor growth, and the effects of HMGB1 are dependent upon Myd88.
Autoimmune mechanisms in antitumor immunity: tumor as an immunoprivileged self (127.20)
Authors: Miska Jason , Devarajan Priya , Chen Zhibin 
Published: January 2023

Abstract
Blocking cytotoxic T lymphocyte antigen 4 (CTLA4), a prototypical inhibitory molecule of antitumor immunity, is emerging as a promising immunotherapy against cancers. Its clinical efficacy, however, is intricately associated with autoimmune toxicity. This association presents a challenge to selective immunosurveillance against tumors versus normal tissues, and suggests a paradigm change whereto autoimmunity is exploited as an effector mechanism for the destruction of tumor cells. Aided with well-defined murine models of autoimmunity, this study demonstrated that autoantigen-specific effector T (Teff) cell clones were indeed effective in eradicating tumor cells. However, a combination of the tumor microenvironment and regulatory T (Treg) cells presented a formidable barrier to the autoimmune effectors. CTLA4 critically regulated autoimmunity-mediated tumor destruction, such that a subtle reduction in CTLA4, engineered by shRNA transgenesis to mimic human CTLA4 genetic variations, substantially enhanced antitumor immunity. Surprisingly, this enhancement was not due to an extrinsic mechanism of crippling Treg-cell suppression, but an intrinsic promotion of Teff cell function. Overall, this study suggests that the tumor microenvironment may represent an “immunoprivileged self” status, wherein physiological mechanisms of immunoregulation evolved to protect nonmalignant “self” could be exploited to an increased potency within a solid tumor.
TGF-β-associated miR-27a Inhibits Dendritic Cell-mediated Differentiation of Th1 and Th17 Cells by Targeting TAB3, p38 MAPK, MAP2K4 and MAP2K7 in the Tumor Microenvironment1 (127.22)
Authors: Yang Rongcun , Min Siping , Li Ling , Zhang Miaomiao 
Published: January 2023

Abstract
Abstract The alterations induced in dendritic cells (DCs) in the cancer microenvironment have not been extensively explored. We found that the tumor-associated factor TGF-β may selectively upregulate the expression of miR-27a via the SP1 transcription factor. Importantly, miR-27a altered the activity of NF-κB and the MAPKs p38, JNK and ERK. It influences the production of proinflammatory cytokines by targeting TAB3, p38 MAPK, MAP2K4 and MAP2K7. As a consequence, miR-27a hampered the DC-mediated differentiation of Th1 and Th17 cells in vitro and in vivo, but it promoted the DC-mediated accumulation of Tr1 (CD4+IL-10+) and Treg (CD4+CD25+Foxp3+) cells in vivo. The repeated infusion of miR-27a-engineered DCs into tumor tissues accelerated tumor growth, indicating that miR-27a is a potential target for tumor immunotherapy. (119 words)
Tumor-infiltrating lymphocytes in breast cancer tissue comprise granzyme B-expressing B cells with cytotoxic potential (127.23)
Authors: Hofmann Stefanie , Brüggemann Sabine , Beyer Thamara , Barth Thomas , Deissler Helmut , Lotfi Ramin , Kreienberg Rolf , Möller Peter , Schrezenmeier Hubert , Jahrsdörfer Bernd 
Published: January 2023

Abstract
Certain subtypes of breast cancer are infiltrated by lymphocytes including B cells, correlating them with a favorable prognosis. Recently, we found that B cells can differentiate into granzyme B (GrB)-secreting cells with cytotoxic potential towards certain tumors. After screening paraffin-embedded tissue sections of various tumors, we show here for the first time that breast carcinoma is infiltrated by CD19+ B cells which express significant amounts of GrB. Testing a panel of different cytokines described to be associated with anti-tumor immune responses revealed interleukin (IL-) 21 as a highly potent stimulus to induce GrB in B cells. Induction of GrB by IL-21 was strongly enhanced in the presence of B cell receptor engagement, and was associated with activation of JAK1/3 and STAT1/3, signaling molecules also activated in NK cells and CTL. Given these parallels with other cytotoxic cells, we tested whether or not GrB-secreting B cells are able to directly induce apoptosis in breast cancer cells. Indeed, we found that IL-21-activated, GrB-secreting B cells showed significant cytotoxic activity against breast cancer cells in-vitro, whereas unstimulated B cells had no such effects. The data presented here support the hypothesis that GrB-secreting B cells may be directly involved in anti-cancer immunity. Our findings suggest, that B cell plasticity is higher than previously thought and that B cells may play an immunological role beyond antibody and cytokine production.
Universal tumor-specific T cells: Redirecting T cells to tagged antigens in vivo (127.30)
Authors: Davila Eduardo , Geng Degui , Srivastava Ratika , Bansal Navneeta , Li Chaoyang 
Published: January 2023

Abstract
The development of a universal yet adaptable gene therapy platform that confers immune cells specificity to various cancer types has valuable clinical implications. T cells engineered to express a chimeric antigen receptor (CAR), that bind tumor-associated antigens, show promise in treating patients however several limitations preclude the widespread use of this form of therapy. We improve the existing CAR technology by developing a versatile system which grants T cells specificity to fluorescein isothiocyanate (FITC) molecules. We report that human antiFITC-CAR T cells recognize various cancer types when bound with FITC-labeled antibodies resulting in efficient target lysis, T cell proliferation, and cytokine/chemokine production. Treatment with antiFITC-CAR T cells plus FITC-cetuximab (FITC-Ctx) delayed colon cancer tumor establishment but unexpectedly led to the outgrowth of epidermal growth factor receptor (EGFR)-negative cells. Using pancreatic cancer cells with uniform EGFR expression, antiFITC-CAR T cells plus FITC-Ctx eradicated an established tumor and prevented tumor growth. These studies are the first to describe the generation of a universal and adaptable CAR system which confers T cells specificity to tagged antibodies which when bound to various cancer types mediate tumor destruction. We consider this platform to be an ‘off-the-shelf’ system that considerably advances the existing CAR technology through its potential to target an assortment of tagged proteins.
Meningiomas include an antigen-experienced B cell infiltrate that produces antibodies directed to tumor antigens (127.36)
Authors: OConnor Kevin , Fang Liangjuan , Meizlish Matt , Anderson Richard , Bruce Jeffrey , Huttner Anita , Lovato Laura 
Published: January 2023

Abstract
Meningiomas are among the most common adult neoplasms of the central nervous system. Like several other tumor types, these intracranial tumors often harbor an immune cell infiltrate that can include substantial numbers of B cells. Characteristic details of the B cell infiltrate remain undefined. To gain a deeper understanding of both the B cell repertoire and insight into the role they play in this tumor, we characterized the immune infiltrate of resected meningiomas. Immunohistochemistry revealed a conspicuous B cell infiltrate in the microenvironment of most tumors examined. Flow cytometry of tissue homogenate showed that the infiltrate included naïve and memory B cells and differentiated plasma cells. Molecular characterization of the immunoglobulin variable regions expressed by tumor-infiltrating B cells revealed clear evidence of antigen experience, in that the cardinal features of an antigen-driven B cell response were present: significant somatic mutation, isotype switching and codon insertion/deletion. This characterization also revealed the presence of B cell clonal expansion and intraclonal variation. Testing the specificity of recombinant immunoglobulin derived from single, dissected, intra-tumoral B cells revealed that tumor antigens were recognized. These data indicate that an adaptive and specific humoral immune response exists within meningiomas, further suggesting that tumor antigens may drive B cell maturation within the tumor microenvironment.