bluuebunny/crossref_metadata_2025_split huggingface dataset:
Unique properties of polarized M1 and M2 macrophages from 2′-5′ Oligoadenylate Like 2 (Oasl2) knock out mouse.
Authors: Ghosh Arundhati , Frank David Anthony , Mosch Nora , Sarkar Saumendra N 
Published: January 2023

Abstract
Our immune system works constantly to keep us safe from invading viruses and bacteria. It can even prevent us from developing cancer. Among these immune cells, macrophages can recognize these foreign threats and neutralize them. Macrophages can be differentiated into specific phenotypes such as M1 and M2 with specific signals. M1 and M2 macrophages each secrete specific cytokines and chemokines promote proinflammatory and antiinflammatory responses in host cells respectively. 2′-5′ oligoadenylate like protein (OASL) protein belongs to a well known family of antiviral proteins knows as Oligoadenylate Synthetase. Both human OASL and mouse isoform Oasl2 perform similar functions in inhibiting RNA virus multiplication within host cells which has been characterized in detail. In the present work, we have characterized the properties of Bone marrow derived macrophages (BMDM) of Wild type (Wt) and Oasl2 knock out (KO) mice in terms of their polarization to M1 and M2 phenotypes. Detailed analysis and comparison of polarized M1 and M2 macrophages from Wt and KO mouse revealed different behavior of KO mouse M1 and M2 macrophages compared to Wt. KO M1 macrophages have more induction of IRF1 resulting more production and secretion of IL-1β in to the culture media. On the other hand, KO M2 produced more Arginase I within the cells. Detail studies of properties of macrophages from Oasl2 KO mice will help us understand the function of this protein within body.
In vivo validation of a novel anti-mesothelin nanobody for early detection of mesothelin-expressing cancers
Authors: Scholler Nathalie , Yin Catherine , sharma khushboo , Prantner Andrew M , Kamat kalika , Repellin Claire , Sambucetti Lidia 
Published: January 2023

Abstract
Antibodies and antibody fragments are widely used in oncology for nanotechnology-based diagnostic, therapeutic, and prognostic assays. Recombinant antibodies that recognize mesothelin, a differentially expressed cancer biomarker with limited normal expression that is upregulated in a variety of epithelial tumors, are important for developing next generation diagnostic or therapeutic platforms and immunosensors because of the flexibility to incorporate various tags or functional groups for site-specific and oriented attachment of antibody fragments to surfaces, labeling reagents or therapeutics. We exploited the inherent stability of camelid heavy chain to develop a site-specific biotinylated, flag-tagged nanobody that recognizes human and mouse mesothelin. We previously published the in vitro characterization of the anti-mesothelin nanobody G3a and then we sought to address whether G3a nanobody could detect mesothelin-expressing tumors in syngeneic and xenograft mouse models of ovarian or lung cancers. Tumor cells were injected subcutaneously (A1847), IV (EKVX, H460, A549), IP or intra ovary (Luc-ID8), and detected at different time points by in vivo imaging (IVIS) or by magnetic resonance imaging (MRI). For in vivo imaging, G3a nanobody was coupled to fluorescently-labelled streptavidin (IRB680W); for MRI, G3a was coupled to avidin-coated magnetic beads. We found that G3a nanobody generated robust and highly mesothelin-specific signals that permitted the detection of very low tumor burden barely detectable by luciferin. Our results indicate that G3a is a valuable reagent for early detection of mesothelin-expressing cancers, as well as a promising delivery agent for imaging and therapeutic conjugates.
Expression of adipocyte/macrophage fatty acid binding protein in tumor associated macrophage promotes breast/mammary tumor progression
Authors: Hao Jiaqing , Yan Fei , Zhang Yuwen , Triplett Ashley , Zhang Ying , Schultz Debra A. , Sun Yanwen , Zeng Jun , Silverstein Kevin A.T. , Zheng Qi , Bernlohr David , Cleary Margot P. , Egilmez Nejat K. , Sauter Edward , Liu Shujun , Suttles Jill , Li Bing 
Published: January 2023

Abstract
Tumor associated macrophages (TAMs) play a critical role in cancer development and progression. However, due to the heterogeneity of TAMs, it remains a major challenge to identify clinically-relevant markers for pro-tumor TAMs. Here, we report that expression of adipocyte/macrophage fatty acid binding protein (A-FABP) in TAMs promotes breast cancer progression. While upregulation of A-FABP was inversely associated with breast cancer survival, deficiency of A-FABP significantly reduced mammary tumor growth and metastasis. Furthermore, we demonstrated that the pro-tumor effect of A-FABP was mediated by TAMs. More specifically, A-FABP was preferentially upregulated in a subset of TAMs that exhibited a CD11b+F4/80+MHCII−Ly6C− phenotype, which promoted tumor progression through IL-6/STAT3 pathway. Mechanistically, activated NFκB in A-FABP+ TAMs repressed the expression of miR-29b, resulting in enhanced tumor-promoting IL-6 signaling as compared to A-FABP −/− TAMs. Thus, A-FABP represents as a new functional marker for pro-tumor TAMs and a new target for macrophage-based tumor immunotherapy.
Mechanisms of PTEN inactivation and pAKT dependence in ATM-deficient thymic T cell lymphomas
Authors: An Joseph Byung Hoon , Hathcock Karen S , Hodes Richard J 
Published: January 2023

Abstract
Ataxia telangiectasia mutated (ATM) is a serine-threonine kinase that is essential for maintaining genomic integrity in response to DNA double-strand breaks. AT patients and ATM-deficient mice develop lymphoid malignancies at high frequency, establishing ATM’s role as a tumor suppressor. It was previously reported that murine Atm−/− T cell lymphomas have a recurrent loss of the gene encoding phosphatase and tensin homolog (PTEN), a tumor suppressor that plays a critical role in inhibiting the PI3K/AKT/mTOR pathway. Our analyses of Atm−/− T cell lymphomas revealed that 12/13 express high levels of phospho-AKT (pAKT). This upregulation of AKT signaling is consistent with the finding that 11/12 pAKThigh tumors lack PTEN protein. One tumor was PTEN+ but failed to downregulate pAKT. Here, we study the Atm−/− tumor model in order to uncover the mechanism by which PTEN expression is ablated. Interestingly, our analyses found that 9/11 PTEN-deficient cell lines have incurred a variety of alterations including exonal deletions of pten or truncations of its mRNA, providing a basis for PTEN loss-of-function. The remaining three lines, however, possess full-length Pten exomes and transcripts. Sequencing the Pten mRNA of these cell lines is in progress to identify mechanisms responsible for the absence of PTEN protein. We have further observed that in vitro treatment of Atm−/− T cell lymphomas with the pAKT inhibitor MK2206 resulted in substantial impairment of survival, consistent with their dependence on activated AKT in the absence of regulation by PTEN. Taken together, these results demonstrate that the majority of Atm−/− T cell lymphomas lose PTEN function through multiple mechanisms and become dependent on AKT signaling for their survival.
A sand fly salivary chemotactic protein drives neutrophil migration and exacerbates Leishmania infection
Authors: Guimaraes Anderson , Wen Xi , Castro Waldione , Townsend Shannon , Brzostowski Joseph , Meneses Claudio , Kamhawi Shaden , Valenzuela Jesus , Oliveira Fabiano 
Published: January 2023

Abstract
Neutrophils are the first cells recruited to the site of an infected sand fly bite and interact with Leishmania parasites. Here we investigate the effect of sand fly salivary proteins on neutrophil recruitment. We purified bone marrow neutrophils from C57BL/6 mice or from peripheral blood of healthy human or dog donors. Chemotaxis driven by sand fly saliva of vectors of visceral (Lutzomyia longipalpis) or cutaneous (Phlebotomus duboscqi) disease was measured by modified Boyden chamber assay and by imaging with the EZTAXIScan assay. Neutrophils migrated towards saliva glands of the vectors P. duboscqi and L. longipalpis, in a dose-dependent manner. Proteinase K treatment of saliva completely abrogated neutrophil recruitment, suggesting a protein as the chemotactic factor. To unearth the identity of the chemotactic protein, we generated plasmids coding for the 20 most abundant salivary proteins from P. duboscqi and injected them in the ears of C57BL/6 mice. Neutrophil recruitment was analyzed by flow cytometry staining. We identified 2 different salivary protein-coding plasmids with neutrophil recruitment activity. We produced these 2 recombinant proteins in the HEK293 cell line system and we observed that a family of 40kDa salivary proteins acts as chemokines for neutrophils both in our in vitro and in vivo assays. Moreover, the coinjection of Leishmania major parasites with the recombinant proteins in the ear of C57BL/6 mice exacerbated the infection. We plan to test if neutralization of the salivary chemotactic protein can alter Leishmania infection. We put forward the hypothesis that blockage of this protein will disrupt neutrophil migration early in the Leishmania infection possibly altering the disease outcome.
Metabolic reprogramming of CD4 T cells exacerbates Herpes Stromal Keratitis
Authors: Varanasi Siva Karthik , Rouse Barry T 
Published: January 2023

Abstract
Ocular infection with Herpes Simplex Virus type 1 (HSV-1) sets off an inflammatory reaction in the cornea which leads to both virus clearance as well as chronic lesions that are orchestrated by CD4 T cells. Effector CD4+ T cells rely on glycolysis for inflammatory effector functions and Treg rely on mitochondrial metabolism. How these processes contribute to Herpes stromal keratitis (HSK) is unclear. We show that glycolysis is elevated in CD4+ T cells from mice with HSK as compared to non-immunized controls. In addition, glucose uptake by CD4 T cells at the lesion site and draining lymph nodes were higher in animals with HSK. In vitro, T cells activation led to enhanced glycolysis, which was inhibited by inhibitor 2-deoxy-D-glucose (2DG). Th1 generated in vitro displayed enhanced glycolysis rates compared to Treg cells. Hence, inhibition of glycolysis by 2DG inhibited the differentiation and function of interferon-γ (IFN-γ) producing Th1 cells, largely un-affecting Treg cells. In vivo, treatment of animals with 2DG normalized T cell metabolism rebalancing the Treg to Th1 ratio and limited the SK lesion progression. Furthermore, animals infected with HSV-1 displayed enhanced blood glucose levels at time points when the inflammation at draining lymph nodes and lesion site are at its peak, which may both be the cause and effect of the inflammation. These results suggest that normalization of T cell metabolism through inhibition of glycolysis is a promising therapeutic venue for HSK.
Impaired CD8 T cell response to viral infection following acute spinal cord injury
Authors: Norden Diana Maria , Bethea John R , Jiang Jiu 
Published: January 2023

Abstract
Trauma to the spinal cord disrupts essential communication between the central nervous system and the immune system. This is highly important as patients with spinal cord injury (SCI) are at greater risk for infections that increase morbidity and mortality. Previous studies have confirmed immune depression in mouse models of chronic SCI. In mice, injury at the high thoracic level disrupts sympathetic output and leads to sustained elevations of norepinephrine in the spleen. SCI patients, however, are at high risk for infections acutely after SCI while hospitalized. Here, we induced a mid thoracic injury where integrity of the sympathetic response is maintained and investigated the antiviral response to influenza an acute timepoint. Mice received a 70kdyn contusion injury at T9 and were challenged intranasally with influenza virus (HKx31, H3N2) 7 days after injury. Following infection, injured mice had a diminished CD8 T cell response with decreased numbers of virus-specific CD8 cells in the lungs. Effector functions were also impaired in injured mice. Expansion of CD62Llow/CD44high effector CD8 T cells was reduced in injured mice and recruitment of CD8 T cells to the lungs was decreased compared to uninjured mice. In addition, the number of intracellular IFNγ-secreting CD8 T cells after in vitro stimulation with virus-specific peptide was reduced in both spleen and lung of injured mice. Overall, these data show an impaired CD8 T cell response to viral infection after acute SCI and suggest that immune depression following SCI occurs rapidly and may be independent of chronic norepinephrine elevations.
Enhancement of antigen presentation of a vaccine with the use of human anti-rhamnose antibody both in-vitro and in-vivo
Authors: Hossain Md Kamal , Vartak Abhishek , Karmakar Partha , Sucheck Steven J. , Wall Katherine Ann 
Published: January 2023

Abstract
Since anti rhamnose antibody is the most common natural anti-carbohydrate antibody present in human serum, using rhamnose as a targeting moiety for a rhamnose containing vaccine can be a good approach for effective vaccine formulation. Binding rhamnose of a rhamnose-containing vaccine with anti-rhamnose antibody has been proven effective for capturing and processing of antigen by antigen presenting cells (APC) via stimulatory Fc receptors. This was shown using anti-rhamnose antibodies generated in mice. We wished to determine if human anti-rhamnose antibodies isolated from human serum work in a similar manner. To isolate anti-rhamnose antibodies, we prepared a rhamnose affinity column with a rhamonse-2 amino ethyl linker conjugate coupled to CNBr-activated Sepharose. In-vitro, human anti-rhamnose antibodies enhanced uptake of rhamnose-ovalbumin by murine APC as determined by proliferation of ovalbumin-primed T cells. In-vivo, injection of human anti-rhamnose antibodies into mice improved the priming of T cells to rhamnose-ovalbumin. Also, we found increased priming of both CD4+ and CD8+ T cells toward the MUC1-Tn cancer antigen when mice were given human anti-rhamnose antibodies prior to a rhamnose-modified MUC1-Tn-containing cancer vaccine. Other human immunoglobulins did not enhance priming. Therefore, we have confirmed that human anti-rhamnose antibodies can enhance immune responses to rhamnose-containing vaccines through increased antigen uptake and presentation.
New strategies in vaccine design for the induction of CD8 T cell responses using biodegradable iron oxide nanoparticles
Authors: Powles Liam M , Wilson Kirsty , Xiang Sue D , Selomulya Cordelia , Plebanski Magdalena 
Published: January 2023

Abstract
For vaccines targeting diseases such as malaria and cancer, CD8 T cell responses are vital. Traditional adjuvants, like alum, are incapable of inducing these responses and thus new delivery strategies are required. Previously, we have shown that biocompatible polystyrene nanoparticles (PSNPs) (carboxylate, 40–50 nm) can induce potent cytotoxic T cell and antibody responses when conjugated to antigen and can protect mice against experimental tumours or clear large established tumours after a single immunization. As a biodegradable alternative; we synthesised ~50 nm iron oxide nanoparticles (IONPs) stabilized with the polysaccharide pullulan. The IONPs share similar characteristics (size, charge and shape) with the PSNPs, are stable, non-toxic, rapidly taken up by dendritic cells in vitro and are considered inert as they do not upregulate costimulatory markers.
The adjuvant/carrier capabilities of IONPs when conjugated to model antigen ovalbumin (OVA) were assessed in vivo. IONPs can induce strong antibody responses but failed to boost the T cell response (Th1 and Th2) after two immunisations, despite their similarity to PSNPs. This implies that the composition of a NP can affect its interactions with the immune system and as such its behaviour as a carrier. To produce an IONP delivery system in which CD8 T cells could also be induced in addition to strong antibody responses, IONPs were co-delivered with the TLR9 agonist CpG. This led to &gt; 2-fold increases in CD8 T cell and antibody responses above either IONPs or CpG alone. Therefore, these biodegradable particles have the capacity to synergize with an inflammatory adjuvant and provide a new strategy for vaccine delivery focused on the induction of CD8 T cell responses.
Induction of stem-like memory T cells by cancer vaccine is associated with TCR downregulation and correlates to strong antitumor effect
Authors: He Yukai , Wu Sha , Zhu Wei , Peng Yibing , Wang Lan , Hong Yuan , Fu Ning 
Published: January 2023

Abstract
To understand why vaccine-activated tumor-specific T cells often fail to generate antitumor effects, we have focused on studying the intrinsic attributes of two alpha fetoprotein-specific CD8 T cells (Tet212 and Tet499). We found that while Tet499 survived in vitro re-stimulation and maintained their functions, Tet212 underwent exhaustion and apoptosis. In vivo, Tet499 generated more expansion than Tet212 and stronger antitumor effects. The different antigen responsiveness and antitumor effects of Tet212 and Tet499 correlated to their activation and differentiation state. Compared to Tet212, Tet499 were in a lower activation state and contained more stem-like memory T cells (Tscm) that could undergo more expansion in vivo. Mechanistically, Tet499 TCR was more downregulated than Tet212 TCR, which may be responsible for the lower activation and differentiation state of Tet499. In contrast to the notion that TCR downregulation dampens T cell antigen reactivity, our data show that it may help T cells maintain their antigen reactivity by halting their activation and differentiation at Tscm stage and by protecting T effector cells from antigen-induced exhaustion and apoptosis. These findings indicate that the focus of cancer vaccination should be on eliciting tumor-specific T cells at earlier differentiation stage including Tscm for better antigen reactivity.
Immune checkpoint function of T cell-derived CD70 in inflammatory T cell responses
Authors: ONeill Rachel , Du Wei , Alquassim Emad , Mohammadpour Hemn , Qiu Jingxin , Chen George , McCarthy Philip , Cao Xuefang 
Published: January 2023

Abstract
The CD27-CD70 pathway is known to provide a costimulatory signal with CD70 expressed on antigen-presenting cells while CD27 functioning on T cells. Although CD70 is also expressed on activated T cells, it remains unclear how T cell-derived CD70 affects T cell function. Therefore, we have assessed the role of T cell intrinsic CD70 using multiple models including autoimmune inflammatory bowel disease (IBD) and allogeneic graft-versus-host disease (GVHD). Compared with WT T cells, CD70−/− T cells surprisingly caused more severe IBD and GVHD and produced higher levels of inflammatory cytokines. Mechanistic analyses reveal that IFNg induces CD70 expression in T cells, and CD70 limits T cell expansion via a regulatory T cell-independent mechanism that involves caspase-dependent T cell apoptosis and upregulation of other immune checkpoint molecules. Overall, our findings demonstrate that T cell-derived CD70 plays a novel immune checkpoint role in inhibiting inflammatory T cell responses.
Vio®Dyes meet REAfinity - High Resolution Antibody-Fluorochrome Conjugates for Multi-Parameter Flow Cytometry Analysis
Authors: Evaristo Cesar , Vahldieck Marco , Krauthaeuser Susanne , Jansen Nicole , Richter Anne , Dose Christian 
Published: January 2023

Abstract
State-of-the-art sensitivity and target specificity of fluorochrome-conjugated antibodies limit applications in flow cytometry. Identification of cell subsets can be challenging due to low expression levels of surface and intracellular markers. The combination of Vio®Dyes and REAfinity™ antibodies provides researchers with the most powerful and reliable tools for flow cytometric cells analysis.
REAfinity™ are recombinant antibodies of human IgG1 isotype with a mutated region within the Fc part, which prevents Fcγ receptor interactions. They are characterized by high antigen specificity and consistent quality compared to conventional hybridoma-derived monoclonal antibodies. Vio®Dyes, e.g. Vio®515, VioBright®515, Vio®667, VioBright®667, and PE-Vio®615, allow due to their excellent fluorescent properties the detection of low expressed markers and provide new choices for multi-parameter analysis. To demonstrate the potential of the combination of REAfinity™ antibodies and Vio®Dyes, we developed detection panels and protocols for the characterization of FOXP3+, HELIOS+, and CD45RA+ in CD4+ CD25+ CD127low/− human regulatory T cells (Tregs). Tregs were detected and enumerated in whole blood, PBMC, and in samples of magnetically enriched Tregs utilizing the newly developed reagents. In addition, we set-up new and fast staining protocols using 96-well plates or magnetic columns, which allow improved identification of Treg subsets.
In summary, the portfolio of Vio®Dyes conjugated REAfinity and special protocols open up new ways for designing and implementing multicolor panels towards highly sensitive, reproducible, and rapid cell profiling approaches.
Human Th17 activity profile in oral chronic Graft-versus-Host Disease
Authors: Dodge Joshua , da Silva Ana C Costa , Dhamala Susan , Rose Jeremy J , Pavletic Steven Z , Hakim Frances T , Mays Jacqueline W. 
Published: January 2023

Abstract
Immunotherapy using allogeneic hematopoietic stem cell transplantation (alloHSCT) is highly effective; however, its use is limited by the incidence of chronic graft-versus-host disease (cGVHD). The oral cavity, which is affected by cGVHD, is a highly accessible mucosal site with its own local biofluid – saliva – but little is known about the pathogenic progression of cGVHD in oral tissues. This study examined the relationship between salivary gland gene expression, salivary proteins and the immune cell network within oral tissues at onset of oral cGVHD.
Patients enrolled in NIH cGVHD trials (NCT00331968, NCT00520130, NCT01851382) were evaluated after cGVHD onset. Whole saliva from individual oral cGVHD patients (n=58) and healthy controls (n=10) was analyzed for a 13-plex panel of immune-related analytes and revealed elevated expression of tissue remodeling factors and IFN- and IL-17-induced chemokines that correlated with clinical scoring of cGVHD severity in a multivariate analysis. Microarray of patient MSG indicated canonical pathway expression including innate immune pathways, and IFN, IL-17 and IL-22-induced processes. Confocal microscopy confirmed the presence of immune cells in oral tissues. Extending our laboratory’s prior observations of Th1 effectors in BM, Th17 cells (CD4+/CD161+/IL-17+) and Type I IFN-induced factors were identified in the majority of available BM and MSG of cGVHD patients, but not in unaffected post-transplant controls.
Current analyses in cGVHD patient samples implicate activation of IFN pathways and presence of Th17 cells in oral tissues in the pathogenesis of oral cGVHD, though the specific sequence and role of these cells in salivary gland pathogenesis during cGVHD remain to be determined.