bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of hematopoietic cell subsets in vivo following treatment with an anti-CD200 antibody in a murine autoimmune hemolytic disease (AIHD) model (83.12)
Authors: Yan Yan , Mickle Angela , Michalowski Christian , Hunter Jeffrey , Johnson Krista , Faas Susan 
Published: January 2023

Abstract
CD200 is an immunoregulatory protein, highly expressed on activated T and B cells and on various liquid and solid tumors. We have previously shown that treatment with an antagonist anti-CD200 antibody in vivo enhances anti-tumor immunity in a hematologic cancer model. The present studies evaluated whether in vivo anti-CD200 treatment modulates hematopoietic cell subsets and allo- and auto- antibody responses in an AIHD model. Mice were immunized weekly with rat red blood cells (RBC) to prompt robust anti-rat RBC allo- and cross-reactive auto- antibody responses. Immunized mice were randomized into groups receiving either vehicle alone, an anti-CD200 antibody (OX90mG2a), a non-reactive, isotype-matched control antibody (12B4mG2a), cyclosporine (CsA), or CsA plus OX90mG2a or 12B4G2a antibodies. Allo- and auto- antibodies were measured prior to and at the conclusion of treatment. Administration of OX90mG2a with or without CsA significantly impacted CD200+ cell subsets in treated mice. Decreased frequencies of CD200+ T cells, B cells, plasma cells and progenitor bone marrow cells were observed in OX90mG2a-treated mice relative to control mice. All CD200+ populations returned to normal levels after termination of anti-CD200 treatment. OX90mG2a administration also inhibited allo- and auto- antibody production in this model. These data suggest that anti-CD200 antibody therapy is a potent immunomodulator and may provide a therapeutic strategy for antibody-mediated diseases.
Sex hormone-independent capacities of female hematopoietic cells from lupus-prone mice (83.23)
Authors: Jorgensen Trine , David Alexandria , MacLeod Megan , Johnson Angela , Kappler John , Marrack Philippa 
Published: January 2023

Abstract
Many systemic autoimmune diseases are more commonly found in females than in males. This is evident in Systemic Lupus Erythromatosis (SLE), a prototypic systemic autoimmune disorder, where the ratio of females to males is 9:1. Our current understanding of the etiology of SLE implies important roles for genes and environmental factors such as sex hormones. Analyses of mouse models of SLE have proven good models for both; however, the relative importance of genes versus sex hormones is still largely unresolved. Using the (NZB x NZW)F1 mouse model system of SLE we have created a system in which we can segregate the effects of genes and hormones on the immune system in vivo. Briefly, male or female (NZB x NZW)F1 mice were lethally irradiated and reconstituted by age-matched male or female bone marrow (BM) or fetal liver (FL) cells. The resulting chimera mice were followed for the development of lupus-like disease. We found that female BM and FL cells express an intrinsic capacity to drive accelerated lupus-like disease development even in the presence of male pubertal sex hormones. Interestingly, chimera mice receiving female cells also presented with increased serum cytokine and immunoglobulin levels, suggesting that activation of both T and B cells happened more easily in female than male cells. These data represent a new system in which to investigate intrinsic differences between male and female immune mechanisms during the development of systemic autoimmunity.
Cathelin-Related Antimicrobial Peptide (CRAMP) regulates B cell IgG1 production (84.13)
Authors: Chen Yao , Kin Nicholas , Kearney John 
Published: January 2023

Abstract
CRAMP belongs to the cathelicidin gene family, which has direct antimicrobial activity against a broad spectrum of microorganisms. Previous studies have shown that CRAMP also modifies the local inflammatory response and regulates cellular and adaptive immunity. However, a possible direct effect of CRAMP on B cells has not been studied. Using CRAMP-deficient mice, we investigated whether B cells express and respond to CRAMP. We show that all mouse B cell subsets tested, including marginal zone, follicular, B1a, and B1b cells, produced CRAMP mRNA directly ex vivo and up-regulate CRAMP upon activation. Purified CRAMP-deficient B cells produced less IgG1 in response to CD40L or LPS plus IL-4 when compared to WT B cells, and the addition of exogenous CRAMP restored the level of IgG1. CRAMP-deficiency had no effect on proliferation, survival, or class switch recombination, but resulted in a decrease in the amount of mature IgG1 mRNA and single cell IgG1 production. Surprisingly, immunization with TNP-OVA with alum resulted in an enhanced TNP-specific IgG1 memory response in CRAMP-deficient mice compared to WT B6 mice. ELISpot and PCR analysis revealed increased numbers of TNP-specific IgG1-secreting B cells and increased CD4+ T cell IL-4 expression in CRAMP-deficient mice. Taken together these results show that B cells express and respond to CRAMP positively in vitro while in vivo CRAMP appears to also indirectly alter B cells through modulation of T cell cytokine production.
The CD4 memory T cell repertoire specific for a hemagglutinin influenza A virus peptide in DR1 individuals is clonotypically diverse (85.5)
Authors: Tutaj Monika , Gorski Jack 
Published: January 2023

Abstract
Human memory T cells are important for protection against recurrent pathogens. Complex memory T cell repertoires may be indicative of cross-protection against new influenza strains. Unfortunately, the complexity of memory repertoires and the extent to which complex repertoires are the norm in humans is largely unknown. This lack of understanding limits our ability to optimize vaccine strategies. Here, we investigated the CD4 T cell recall response to an HA peptide, conserved in most H3 flu strains, in three middle aged and three older healthy HLA-DR1 blood donors. The HA307-319 peptide was used to stimulate peripheral T cells in short term culture and the repertoires studied by using TCR CDR3β spectratyping and clonotyping. We sorted multimer-labeled and activated CD4 T cells and assessed the advantage of using both methods in repertoire analysis. We found that HA-specific clonotypes used as many as 18 TCR BV families and all BJ genes. The most prominent and shared responses were observed in TCR BV9, 25 and 28 families. Analysis of four thousand sequences revealed essential structural similarities among different individuals including public clonotypes, conserved CDR3 sequences embedded within different BV-J combinations, and several CDR3 amino acid motifs. This study describes the complexity of antigen-specific CD4 T cell response and indicates that only broad population studies can provide a comprehensive knowledge of memory repertoire selection in influenza infection.
Unique populations of lung mast cells are required for antigen-mediated bronchoconstriction (86.11)
Authors: Cyphert Jaime , Kovarova Martina , Koller Beverly 
Published: January 2023

Abstract
Several studies implicate mast cells as important effector cells of anaphylaxis. Much of our understanding of the functions of mast cells emanates from the study of mice lacking theses cells, particularly mice carrying mutations in c-Kit. Definitive evidence for the role of mast cells in these mice requires the normalization of altered immune responses by reconstitution with cultured mast cells (BMMC). While many niches can be restored with BMMC, this has not been demonstrated for mast cells present in the airways of the lung, cells poised to mediate bronchoconstriction during allergic responses. To determine if mast cell reconstitution is an appropriate model for the study of bronchoconstriction associated with allergic responses, KitWsh/Wsh mice were engrafted with either whole bone marrow (WBM) or BMMC and responses to IgE-mediated anaphylaxis determined. Engraftment of KitWsh/Wsh mice with WBM or BMMC results in reconstitution of the central airways with mast cells. While the treatment of both groups results similar systemic changes in a model of passive anaphylaxis, bronchoconstriction was observed only in animals which received WBM. While BMMC can populate the lung, they cannot restore IgE/Ag-mediated bronchoconstriction to mast cell-deficient animals. This suggests the mast cell population mediating this function may be unique, and to fill this niche in the lung cells must undergo a specific developmental program that is no longer available to cultured mast cells.
Expression of the human basophil (BASO) activation marker, CD63, may reflect the extent of degranulation by the AND (anaphylactic degranulation) mechanism. (86.24)
Authors: MacGlashan Donald 
Published: January 2023

Abstract
A variety of studies of suggested that expression of two commonly studied BASO activation markers, CD203c and CD63 (a protein associated with the granule membrane), do not always reflect the extent of histamine release (HR). While exploring the role of PKC in the expression of CD63 and CD203c using the PKC inhibitors Ro-31-8220 (RO) or bisindoylmaleimide II (BIS), it was noted that at concentrations (&gt; 3µM) well above the IC50 for inhibition of PKC (100 nM), there was marked enhancement of CD63 expression with only modest enhancement of CD203c expression or HR. Previous studies have shown that BASO either shuttle granule contents out of the cell via small vesicles (piecemeal degranulation, PMD) or by fusion of granules to each other and the plasma membrane (anaphylactic degranulation, AND), these results suggested the hypothesis that high concentrations of RO or BIS induced a switch from PMD to AND. Morphological imaging of stimulated BASO showed a dramatic increase in the appearance of the degranulation sacs associated with AND in the presence of 3 µM RO. While the cytosolic calcium (Ca) response and degranulation sac formation were correlated, 3 µM RO did not alter the Ca response. Therefore, it was proposed that two processes determine the extent of PMD vs. AND, Ca and an unknown RO-sensitive kinase. It is also proposed that CD63 expression occurs predominantly with AND and that the relative amounts of PMD and AND determine the relationship between CD63 expression and HR.
Estrogen regulates the expression and activity of serine proteases in mouse splenocytes (87.11)
Authors: Dai Rujuan , Khan Deena , Ahmed S. Ansar 
Published: January 2023

Abstract
In addition to digesting proteins, serine proteases also play critical roles in regulation of inflammatory responses, innate and adaptive immunity via proteolytic cleavage of protease-activated receptors (PARs) and/or signaling molecules such as NF-κB. Dysregulation of serine proteases activity may result in the development of variety of human diseases. We and others have shown that estrogen, an important immune modulator, promoted the production of proinflammatory molecules in different types of cell including murine splenocytes and peritoneal macrophages. Our recent report has demonstrated that serine proteases mediated proteolysis play a critical role in regulation of STAT1 and NF-κB activity in splenocytes from estrogen-treated mice. Inhibition of serine protease activity decreased the production of inflammatory molecules IFNγ and nitric oxide in splenocytes from estrogen-treated mice. In this study, we found that estrogen treatment increased the serine proteases activity in splenocytes. Further, we confirmed that neutrophils-derived elastase mRNA expression level is increased, and serine protease inhibitor SerpinB7 is decreased in estrogen-treated splenocytes when compared to placebo control. Together, our data suggest a novel mechanism underlying estrogen-mediated promotion of inflammatory responses. These studies may form the basis for developing new alternative therapies for estrogen-influenced autoimmune diseases by intervening the activity of serine proteases.
Innate-adaptive interactions provide stability in the homeostatic lung environment (87.16)
Authors: Cavanagh Mary , Hussell Tracy 
Published: January 2023

Abstract
The air is full of innocuous, but highly antigenic particles that are inhaled with every breath. In order to avoid the damage associated with an inflammatory response, lung-resident immune cells are kept under strict regulation and fail to respond to most inhaled antigens. Whilst the majority of cells in the airspaces are alveolar macrophages, many other cell types are present in the airspaces and interstitium. Interactions between lung-resident cells provide the basis for the suppression of immune responses; and the cytokines, chemokines and other factors that they secrete provide further levels of regulation. Using sphingosine-1-phosphate receptor agonists to block the entry of T cells into the lungs, we have been investigating the effects of T cell absence on lung immune homeostasis. FTY720 blocks lymphocyte egress from the thymus and lymph nodes by inducing internalisation of the S1P1 receptor which is required for exit into the blood and lymph. We find that exclusion of T cells from the lungs affects macrophage responses to TLR agonists, both in vitro and in vivo as a result of altered alveolar macrophage activation status. By dissecting the mechanisms involved in maintaining lung homeostasis and resolving inflammation, we hope to further our understanding of lung immunity both in terms of homeostatic disruption and immunopathology during or following infectious disease.
Timing of corticosterone treatment and its effects on the CD8+ T cell response to cell-associated antigen (87.18)
Authors: Yorty Jodi 
Published: January 2023

Abstract
Exposure of T cells to glucocorticoids alters their normal program of activation and expansion. In vitro work, often with the synthetic analog, dexamethasone, demonstrated a direct effect of glucocorticoids on T cell proliferation after antigen exposure. In vivo studies have also often utilized dexamethasone. In the current study we evaluated the effect of in vivo administration of the natural glucocorticoid, corticosterone (CORT), on the CD8+ T cell response to SV40 large T antigen (Tag)-expressing cells. C57BL/6 mice respond with a hierarchical CD8+ T cell response to Tag in which epitope IV (amino acids 404-411) is dominant and epitope I (206-215) is subdominant. Exposure to CORT over the course of the immune response to Tag, decreased the number of splenocytes by ~83%. The CD8+ T cell response to epitope I and epitope IV was almost completely abolished during the primary response. Tag-specific memory T cells were also severely affected by CORT treatment. Even when CORT was provided after cross-priming had occurred, the primary T cell response to Tag was still largely inhibited. The ratio of epitope I:epitope IV-specific CD8+ T cells was slightly increased in CORT-treated mice, suggesting that epitope IV-specific T cells may be more susceptible to CORT. Further studies will provide insight regarding when CORT exposure is most detrimental to CD8+ T cell responses that rely on both cross- and direct presentation to achieve maximal expansion.
In vitro characterization of novel anti-human CD27 mAbs (87.23)
Authors: Ramakrishna Venky , Vitale Laura , Sundarapandiyan Karuna , Zhao Biwei , Crocker Andrea , Widger Jenifer , O'Neill Tom , Storey James , Marsh Henry , Keler Tibor 
Published: January 2023

Abstract
CD27, a member of the tumor necrosis factor receptor superfamily (TNFRSF7), is constitutively expressed on the majority of mature T cells, memory B cells, and a portion of NK cells. The interaction of CD27 with its ligand CD70 (CD27L) is important in T cell activation, proliferation, survival, maturation of effector and memory functions, in clonal B cell expansion and germinal center formation and in NK-dependent cytotoxicity. To explore the therapeutic potential of this target, a panel of fully human antibodies recognizing human CD27 was generated using human Ig transgenic mice immunized with recombinant human CD27. These human anti-CD27 mAbs showed specific and high affinity binding to CD27. The antibodies were sub-grouped into those that effectively compete with the binding of CD70 to CD27, and non-blocking antibodies. Several of the anti-CD27 mAbs tested induced pro-inflammatory cytokine and chemokine responses in human whole blood or PBMCs, which could be beneficial in the expansion of Th1 T cell compartment. In addition, some anti-CD27-mAbs mediated in vitro killing of CD27-expressing lymphoma cell lines by ADCC or CDC. The ultimate selection of CD27-activating and CD70-blocking mAbs could potentially complement and amplify existing therapeutic interventions in cancer.
Fucosylation regulates cell death in rheumatoid arthritis (87.24)
Authors: Li Jun , Hsu Hui-Chen , Yang Ping-Ar , Wu Qi , Mountz John 
Published: January 2023

Abstract
Abnormal inflammation and apoptosis are major pathogenic features of synovial fibroblasts (SF) in rheumatoid arthritis (RA). Fucosylation, an important type of glycosylation, has been identified in cancer and inflammation conditions. The role of fucosylation in RA was investigated. Real-time PCR indicated that between RASF and osteoarthritis (OA) SF (N=14 each), there was no difference in the expression of apoptosis related molecules, including tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), its receptors DR4, DR5, DR6, DcR1, DcR2, and regulators of apoptosis FLIPL and cIAPs. However, there was a higher level of α1-2-Fucosyltransferase (Fut1, P=0.016), α1-3/1-4-Fucosyltransferase (Fut3, P=0.018) and α1-3-Fucosyltransferase (Fut6, P=0.038), but not α1-6-Fucosyltransferase (Fut8, P=0.910) in RA compared to OA synovial tissues. Interestingly, a strong positive correlation between TNFα and Fut1, Fut3 and Fut6 was identified. Fut1 also correlated with Il17a. Preincubation of RASFs with 2-Deoxy-D-Galactose (15 mM/ml), a FUT1 inhibitor, decreased the sensitivity to TRA-8, an anti-human DR5 antibody, mediated apoptosis and this effect can be reversed by fucose (15 mM). Similar results were also found on human monocyte/macrophage cell lines, HL60 and THP-1. Our studies suggest that fucosylation is strongly associated with abnormal inflammation and apoptosis in RA effector cells. Fucosylation might be a novel target for RA therapy.
Characterization of antitumor cytokine-induced killer cells generated from immature precursors (87.31)
Authors: Ryu Hwa Sun , Kim Yeon Jin , Lee Hong Kyung , Kim Jee Youn , Hong Jin Tae , Kim Youngsoo , Han Sang-Bae 
Published: January 2023

Abstract
Cytokine-induced killer (CIK) cells have been explored for active immunotherapy of cancer. CIK cells gave strong anti-tumor effects and their cytotoxicity is MHC-unrestricted. In most clinical trials, CIK cells were generated from human PBMCs, mostly containing mature T cell precursors. However, whether immature T cells could be used as precursors of CIK cells was not clarified yet. The possible usage of immature T cells as sources of CIK cells was evaluated in this study. CIK cells were generated from immature T cells of bone marrow and thymus, and their functions were compared with CIK cells generated from mature T cells of spleen. Interferon gamma, anti-CD3 monoclonal antibody, and interleukin 2 (IL-2) were timely added to cultures to generate CIK cells. The absolute number of splenocytes, bone marrow cells, and thymocytes increased by 249, 54, and 14-fold, and CD3+NK1.1+ cells were 30%, 30%, and 33%, respectively. These three different cell populations showed similar profiles in cytokine gene expression, in that they highly expressed IFN-gamma and TNF-alpha. And they showed similar gene expression profiles of granzynme A/B, perforin, and FasL. In addition, they showed similar antitumor activity against B16F10 melanoma. In summary, although CIK cells generated from immature or mature T cells showed similar profiles in phenotypes, cytokine expression, and antitumor activity in vivo, clinical application of immature T cells might be impeded by low ex vivo expansion.
Phytol derivatives as novel immunoadjuvants (87.33)
Authors: Aachoui Youssef , Fitch Richard , Ghosh Swapan 
Published: January 2023

Abstract
Adjuvants are widely used to influence the course of immune response. Improved adjuvants tailored to immunogens are much sought after. In order to achieve this, we are developing new adjuvants based on chemically modified phytol (J Immune Based Ther. Vaccines 2006, 5:4). We developed hydrogenated/nitrated or glycosylated derivatives of phytol (US patent pending). These compounds exhibit excellent adjuvanticity, and in some respects seem superior to Alum or Freunds’ adjuvants. In this study, we show using cytokine microarray how they influence host microenvironment in the peritoneal lavages of BALB/c mice after injection of phytol adjuvants alone or in combination with different antigens. Our results indicate that 1) within 2 hr after injection there was increase in levels of chemotactic factors MCP-1, KC, MIP-1, LIX and IL-6. 2) The levels of these cytokines were more pronounced with PHIS-01 compared to PHIS-02 and PHIS-03. At 24 hours after injection, the inflammatory cascade was amplified stimulating the production of other cytokines that could involve cells of acquired immunity such as B-lymphocyte chemo-attractant (BLC), T-cell activation-3 (TCA), IL-12 , TIMP-1; in addition, they induced innate immunity-related chemotactic factors. Unlike alum, there was no induction of IL-1β and IL-4 suggesting a different mode of action. We also noted that the cytokine profiles significantly changed when protein antigens were incorporated with the phytol-derived adjuvants.
Identification of variant isoforms of CD44 and their functions in murine immune cells (87.6)
Authors: Guan Hongbing , Singh Narendra , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
CD44 is known to be implicated in a variety of immunological processes such as T cell migration, activation and differentiation. CD44 comprises a standard form and a family of isoforms that vary by their degree of post-translational modifications and due to alternative splicing of their primary amino acid sequence encoded by up to 10 variant exons. The isoforms actually determine the differential functions of CD44 in various types of immune cells and immune responses. Upon activation, lymphocytes transiently express certain CD44 splice variants. However, the precise nature of CD44 variant isoforms expressed by various cells types of the immune system has not been studied thus far. In this report, we screened variant exons from exon 1 to 10 using Southern blot hybridization and identified CD44 isoforms on CD4, CD8, FoxP3+ Treg and dendritic cells, either naïve or activated with SRBC, OVA, LPS, anti-CD3 plus anti-CD28, or IL-2 respectively. The results showed distinct isoforms for each cell type that also varied based on naïve or activated status. In experimental autoimmune encephalomyelitis (EAE), targeted deletion of full CD44 sequence prevented EAE whereas sole deletion of variant 7 induced a progressive and non-remitting type of EAE. Our study, has for the first time, identified the unique CD44 isoforms expressed on murine CD4, CD8, Treg and dendritic cells and shown the differential role of CD44 variant isoforms in the regulation of autoimmune disease.