bluuebunny/crossref_metadata_2025_split huggingface dataset:
NKT cells in lupus prone NZB/W mice are abnormal and expression of Ly108 (SLAMF6) affects proinflammatory cytokine secretion and help for autoreactive B cells (44.46)
Authors: Tang Xiaobin , Strober Samuel 
Published: January 2023

Abstract
Systemic lupus erythematosus is an autoantibody mediated disease that causes immune complex glomerulonephritis as well as injury to multiple organs. Invariant natural killer T cells (iNKT) from non-autoimmune strains of mice regulate and suppress graft versus host disease, transplantation immunity and autoimmunity. We have shown that iNKT cells from lupus prone NZB/W mice are abnormal and promote rather than suppress spontaneous in vitro immunoglobulin and autoantibody secretion of autoreactive B cells. To determine the role of iNKT cells in the induction and augmentation of immune cell activation, we examined cytokine production of purified iNKT cells from NZB/W and control C57BL/6 mice in vitro. We found that upon stimulation of NKT stimulatory ligand, α-galactosylceramide, young NZB/W splenocytes secreted much higher IFNγ, IL-4 and IL-17 compared with C57BL/6 mice. With activation of CD3 and CD28, sorted NZB/W splenic iNKT cells, but not conventional T cells, also produced dramatically higher IFNγ and IL-4 than controls. Interestingly, we found the expression of Ly108, a SLAM family member (SLAMF6), affected cytokine production, and promoted help for immunoglobulin secretion by autoreactive B cells. Sorted NZB/W Ly108+ iNKT cells produced less IL-17 but more efficiently helped B cells secrete IgG but not IgM as compared to Ly108- iNKT cells. In conclusion, expression of the SLAM family member, Ly108, promoted the interaction of NZB/W iNKT cells with autoreactive B cells.
Structural and quantitative differences between membrane μ chains associated with surrogate or conventional light chains (45.1)
Authors: Hollander Nurit , Ben Moshe Natalie , Raviv Yasmin , Haimovich Joseph 
Published: January 2023

Abstract
Although it is well established that pre-BCR signaling governs proliferation and differentiation during B cell development, the components of the pre-BCR that are important for signaling are a matter of controversy. It has been suggested that signaling by the µ heavy chains (μHC) of the pre-BCR induces survival and differentiation, while the surrogate light chains (SLC) are essential for proliferation and clonal expansion. Using two variants of a murine B-cell line that differ only in surface expression of either BCR or pre-BCR, we demonstrated that surface µHC in the pre-BCR are of the high-mannose type only, while those in the BCR are of the complex type. We further demonstrated that unlike the BCR, the high mannose pre-BCR reaches the surface as a complex with calnexin. The amount of surface μHC in the pre-B cells is lower than that in B cells. We demonstrated that µHC in pre-B cells are in large excess compared to SLC. While LC in B cells are produced in enough amounts to assemble with all µHC, SLC are produced in insufficient amounts, leaving most µHC unassembled and therefore degraded. Igα and Igβ in pre-B cells are not a limiting factor as well, since they are produced in excess compared to SLC. Hence, the lower amount of the pre-BCR as compared to the BCR is probably due to insufficient amounts of SLC. In all, the structural and quantitative differences between μHC in the BCR and pre-BCR may account for their distinct signaling functions.
Dendritic cells express possible ligand(s) for CD38 (45.13)
Authors: Vences Catalan Felipe , Cedillo Barron Leticia , Santos Argumedo Leopoldo 
Published: January 2023

Abstract
CD38 is expressed in cells of the immune system, it has been implicated in several biological functions such as; chemotaxis, cell activation, cell proliferation, rescue or induction of apoptosis. It also has enzymatic activity converting NAD to ADPR and cADPR. It is not known how this molecule can perform different functions in different cells. It has been suggested that such biological functions could be mediated by different ligands, which would explain the pleiotropy attributed to CD38. CD31, present on endothelial cells, has been identified as a possible ligand for CD38; however, these results have not been confirmed in mouse. On the other hand, two molecules were identified in mouse dendritic cells as potential ligands but without further characterization. The aim of this work was to identify new possible ligands in a murine model using chimeric proteins of CD38. We designed three soluble chimeric proteins of the extracellular domain of CD38. One fused with CD8α, a second one fused with the Fc from the IgG1 immunoglobulin, both from human, and a third one containing only a V5 epitope and a histidine tag. The proteins were characterized by WB and enzymatic activity assays. All proteins formed soluble dimers and exhibited enzymatic activity. Using these proteins we were able to identify potential ligand(s) present on DCs but not on T and B lymphocytes from WT or CD38 KO immunized mice. This data suggest that DCs may play a role in CD38 function on B and T cells.
Prostaglandin E2 upregulates p53 mRNA and protein within activated human B lymphocytes (45.17)
Authors: Haque Shabirul , Lee Hyunjoo , Chiorazzi Nicholas , Mongini Patricia 
Published: January 2023

Abstract
Prostaglandin E2 (PGE2), a product of COX-2, plays a central role in inflammation and is elevated in pre-neoplastic tissue and malignancies, as well as in activated normal B lymphocytes. PGE2-induced signals via EP receptors modulate cAMP and/or calcium levels and can cross-talk with other signaling pathways (e.g. MAPK and NF-kB). p53 is a tumor suppressor with broad effects that include negative regulation of cell cycle, DNA repair, and induction of apoptosis. This study describes a role for PGE2 in regulating B cell p53 mRNA and protein. Purified human B2 cells from spleen/tonsil were pre-activated with stimuli resembling those found in sites of inflammation, i.e. surrogate C3dg-coated antigen + IL4 + BAFF. At d 4-5, cell density-limiting cultures were pulsed with PGE2 (50 nM) and harvested 1, 3, 8, 24, and 31 h later. p53 mRNA was monitored by both quantitative and qualitative RT-PCR. Exogenous PGE2 significantly enhanced p53 mRNA expression, in a time dependent fashion, over 3-31 h. This was accompanied by elevated mRNA for BID, a pro-apoptotic protein known to be transcriptionally regulated by p53, and linked to PGE2-elevated levels of p53 protein in replicating blasts. The above findings contrast with evidence that PGE2 promotes the expression of pro-survival Mcl-1 within replicating B cells. Taken together, they indicate that PGE2-mediated signals modulate both pro-survival and pro-apoptotic molecules in normal replicating human B lymphocytes.
The role of Erk pathway in the selection of autoreactive and non-autoreactive B cells (45.3)
Authors: Teodorovic Lenka , Rowland Sarah , Leahy Katelyn , Torres Raul , Pelanda Roberta 
Published: January 2023

Abstract
During development in the bone marrow, B cells undergo gene rearrangements, which lead to the expression of the B cell receptor (BCR) on the cell surface. Often times these BCRs have autoreactive specificities, requiring B cells to undergo tolerance mechanisms before undergoing further differentiation. Tonic BCR signaling, mediated by non-autoreactive BCRs, stops immunoglobulin (Ig) gene rearrangements, and allows cells to differentiate from immature into transitional and mature B cells. Immature B cells that receive antigen-mediated BCR signals are arrested in their differentiation and exhibit BCR downregulation and unresponsiveness to BAFF. Previous studies in our lab showed that the tonic BCR signal that drives immature B cell differentiation is mediated by the Ras-MEK-Erk pathway. However, we have not established whether activation of the Erk pathway is also involved in stopping Ig gene rearrangements. In mouse models where immature B cells co-express non-autoreactive (tonic signals) and autoreactive (antigen-mediated signal) BCRs, both tolerance and cell differentiation occur. Additionally, increased levels of pErk have been associated with positive selection of autoreactive B cells from the bone marrow. We hypothesize that tonic BCR signals overcome antigen-mediated signals by activating Ras-MEK-Erk pathway, leading to the inhibition of receptor editing and differentiation of autoreactive B cells. Results that test this hypothesis will be presented.
Intrathymic cortisone-resistant memory cells (46.11)
Authors: Vagida Murad , Pobezinsky Leonid , Pobezinskaya Elena , Kazansky Dmitry 
Published: January 2023

Abstract
It is known that central memory cells CD8+ are found in immunocompetent lymphoid organs, such as spleen and lymph nodes. Previously, we described the approach allowing to see and estimate selective proliferation of memory cells in MLR with heat-silenced (45 degrees, 1 h) allogeneic stimulator splenocytes. We immunized C57BL/6 (H-2b) mice intraperitoneally with mastocytoma P815 cells (H-2d). Two months later, we tested the proliferation of thymocytes in the MLR with heat-silenced syngeneic C57BL/6 (H-2b), allogeneic B10.D2 (R101) (KdIdDb) and third-party FVB (H-2q) splenocytes. Thymocytes of intact mice did not respond in MLR at all. Thymocytes of immune mice displayed small, but significant MLR responses to heat-silenced allogeneic splenocytes, but not syngeneic or third-party splenocytes. To test the hypothesis that thymic memory cells are cortisone-resistant, we treated immune mice with hydrocortisone. Injection of high dose of hydrocortisone (2,5 mg) 2 days before isolation of responder thymocytes resulted in highly elevated MLR responses. This response was antigen-specific because “immune” thymocytes responded only on heat-silenced R101, but not syngeneic or third-party splenocytes. As expected, in proliferating cultures, the ratio of CD4/CD8 was less than 1 due to proliferation of memory CD8+ cells, responding on H-2Kd MHC class I molecule represented on R101 thymocytes. Thus, we have found that the thymus contained memory-like T cells, which are cortisone resistant.
A role for viral dose in SLEC differentiation. (46.12)
Authors: Amoah Samuel , Alexander-Miller Martha 
Published: January 2023

Abstract
Our current understanding of effector cell differentiation suggests that inflammation plays an important role in driving an activated cell down the short lived effector cell (SLEC) versus memory precursor effector cell (MPEC) pathway. While IL-12 has been implicated as an important regulator of differentiation, this appears not to be the case for all infections, e.g. vaccinia virus (VV). To evaluate factors that may regulate effector cell differentiation under these circumstances, we investigated the effect of viral dose. We found that mice infected with a low dose of vaccinia virus had more B8R-specific cells at the peak of the response compared to those infected with a high dose of virus. Surprisingly, the observed increase in total number was the result of an increased SLEC population, leading to a response skewed toward SLEC. The increased SLEC in the low dose infected mice was not due to selective increases in proliferation or decreases in death (as measured by active caspase 3) in this population. In exploring this model we found that T regulatory cells were present at higher numbers in high dose infected mice. However, while depletion of Tregs led to an increase in the number of B8R specific cells, it did not result in a selective increase in SLEC. These data support a role for viral burden in effector cell differentiation, with decreased vaccinia virus dose driving the formation of SLEC.
A rapid method for the isolation of untouched naïve CD8 T cells from peripheral blood mononuclear cells (46.9)
Authors: McQueen Karina , Posarac Vesna , Fairhurst Maureen , Thomas Terry 
Published: January 2023

Abstract
Naïve CD8 T cells are, by definition, those that have been released by the thymus into circulation, but have not yet encountered their cognate antigens. Once they become activated they proliferate and generate effector T cells. We describe an easy and rapid method for the isolation of untouched naïve CD8 T cells from peripheral blood mononuclear cells (PBMC). Briefly, PBMCs are isolated by Ficoll-Paque PLUS density gradient sedimentation. Naïve CD8 T cells are then enriched using immunomagnetic, column-free, negative selection (EasySep™). Using EasySep™, unwanted cells are removed using antibody complexes recognizing non-naïve CD8 T cells and dextran-coated magnetic particles. Labeled cells are separated by placing the tube in an EasySep™ magnet. This procedure can be fully automated using RoboSep™. EasySep™ isolated naïve CD8 T cells are assessed by flow cytometry using a combination of antibodies specific for naïve, effector and memory cells. Naïve CD8 T cells are defined as CD8+CD45RA+CCR7+ and CD45RO-CD56-CD57-. Starting with PBMCs isolated from fresh blood and buffy coat, purities of 89±3% (n=8) are obtained (starting cell frequency 1.0-11.2%). EasySep™ isolated naïve CD8 T cells undergo robust proliferation as measured by flow cytometry of CFSE-labeled cells in culture with CD3/28 T cell expander beads. This simple method to isolate naïve CD8 T cells will be invaluable to researchers studying T cell differentiation, autoimmunity and infectious diseases.
The role of the activating versus inhibitory classes of G protein coupled histamine receptors on experimental allergic encephalomyelitis susceptibility. (47.2)
Authors: Saligrama Naresha , Noubade Rajkumar , Del Rio Roxana , Teuscher Cory 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS) augmented by histamine (HA) and histamine receptor (HR) signaling. To help elucidate the mechanism behind HA and HR signaling on MS susceptibility, we utilize the experimental allergic encephalomyelitis (EAE) mouse model for MS and implement the use of traditional gene knockout studies. HA is known to exert its effects through four different G protein coupled receptors (GPCRs) designated as H1R, H2R, H3R and H4R. H1R and H2R couples to the Gαq/11 and Gs class of activating G proteins, respectively, whereas H3R and H4R couples to the Gi/o class of inhibitory G proteins. Each individual HR has been shown to influence EAE pathogenesis. Here we assessed the overall contribution to EAE susceptibility imposed by the two classes of GPCRs by generating C57BL6/J mice deficient for both H1R and H2R (H1H2RKO), as well as mice deficient for both H3R and H4R (H3H4RKO). The results of our study show that H1H2RKO mice exhibit decreased susceptibility to EAE, less severe neuroinflammation with a higher infiltration of T regulatory cells into the CNS, and decreased blood brain barrier permeability compared to H3H4RKO mice. Moreover, we show that splenocytes from immunized H1H2RKO mice produce significantly less IFN-γ and IL-17, which are cytokines known to exacerbate disease. Therefore, the activating class of GPCRs enhances EAE susceptibility compared to the inhibitory class of HRs.
Genetic loci on New Zealand black chromosome 1 lead to increased generation of T follicular helper and TH17 cells (47.23)
Authors: Talaei Nafiseh , Landolt-Marticorena Carolina , Noamani Babak , Pau Evelyn , Chang Nan , Wither Joan 
Published: January 2023

Abstract
Genetic loci on NZB c1 lead to antinuclear antibody (Ab) production and glomerulonephritis (GN). Using subcongenic lines, we found at least 5 genetic loci within a 35-106cM interval modulate disease. Notably, mice with a 70-100cM interval (c1(70-100cM)) develop high titre anti-dsDNA Ab, increased numbers of large germinal centers and severe GN a characteristic feature of lupus-prone mice with abnormalities of T follicular helper (TFH) and Th17 cells. In this study we determined whether variations in disease severity in c1 subcongenic mice are associated with differences in these subsets. The proportion of splenic TFH cells was significantly increased in c1(70-100) mice. Splenocytes from all c1 congenic mouse strains demonstrated increased production of IL-21 and IL-17 following anti-CD3/CD28 crosslinking which was greatest in c1(70-100) mice. The majority of IL-21 producing cells were TFH, many of which also secreted IFN-γ, but not IL-17. Immunization of 8-wk-old pre-autoimmune mice with ovalbumin (OVA) led to increased generation of TFH and Th17 cells in c1 (70-100) mice. This appeared to result in part from altered T cell function, as adoptively transferred OVA-specific TCR transgenic c1(70-100) T cells and naïve c1(70-100) T cells cultured in-vitro, demonstrated enhanced differentiation to TFH and IL-17-producing cells. Thus, genetic loci on NZB c1 promote differentiation of T cells to TFH and Th17 cells and this capacity correlates with the severity of disease.
Analysis of DRB1 sequence feature variant type associations with systemic sclerosis autoantibodies types and racial groups (47.28)
Authors: Marthandan Nishanth , Guidry Paula , Thomson Glenys , Arnett Frank , Karp David , Scheuermann Richard 
Published: January 2023

Abstract
A novel sequence feature variant type (SFVT) approach to genetic association analysis has been previously shown to identify parts of HLA proteins that are associated with the development of a variety of autoimmune diseases. Using the SFVT approach, we analyzed DRB1 typing data in a cohort of patients with Systemic Sclerosis (SSc) and demographically matched controls. We found no allele that was significant at p-value of 0.05 across all the three racial groups (African American, Caucasian and Hispanic) in the dataset. But, we found 23 SFVTs that were significant across the all three racial groups. In the analysis of SFVT association with the presence or absence of different auto-antibodies, we found a large overlap among the top significant sequence features (SFs). In spite of such overlap of SFs the top 100 SFVTs were exclusively associated with the presence of either anti-centromere or anti-topoisomerase but not both, indicating that different DRB1 sequences predispose to the development of different auto-antibody types. In addition, we found that SFVTs that differ by only a few amino acids can give rise to different autoantibodies in SSc patients (e.g. association of the antigen binding motif 67L_70Q_71R with anti-centromere, versus association of 67F_70D_71R with anti-topoisomerase). Tools for the conversion of four-digit HLA typing data into an SFVT data matrix and the calculation of chi-squared association statistics have been implemented in ImmPort (www.immport.org).
The P2Y11 receptor is associated with narcolepsy and with higher susceptibility to ATP induced cell death in T lymphocytes and NK cells. (47.5)
Authors: Kornum Birgitte , Lin Ling , Kassack Matthias , Kuipers Hedwich , Axtell Robert , Jennum Poul , Steinman Lawrence , Mignot Emmanuel 
Published: January 2023

Abstract
The sleep disorder narcolepsy with cataplexy is caused by the loss of hypocretin-neurons in the hypothalamus as the end result of what is hypothesized to be an autoimmune attack. Using a GWAS design, we recently published a significant association between narcolepsy and purinergic P2Y11 receptor polymorphisms. P2RY11 is a pseudogene in rodents and is thus poorly studied, although human data suggests involvement in extracellular ATP (eATP) induced immune cell death and chemotaxis. We found a P2RY11 genotype dependent effect on expression levels in PBMCs, with functional effects on sensitivity to eATP induced cell death. In further studies, we have found that a balance of P2Y11 and P2X7 expression may determine PBMC subtype sensitivity to eATP induced cell death. Notably, P2Y11 expression levels were CD56+&gt;CD8+&gt;CD19+&gt;CD4+&gt;CD14+, whereas P2X7 expression levels were CD14+&gt;CD8+&gt;CD4+&gt;CD19+. In accordance with this, monocytes were more sensitive to eATP induced cell death than lymphocytes. We have also found that eATP-induced cell death is inhibited by P2X7 antagonists, increased by P2Y11 antagonists and decreased by P2Y11 agonists, in support of our hypothesis. Finally, we show that the genotype associated with narcolepsy affects mostly T lymphocytes and NK-cells. These findings suggest that a balance of P2Y11 and P2X7 activity may be important for selection of specific immune cells in the presence of eATP, and that this process may be important for narcolepsy pathophysiology.
CD30L null SJL mice exhibit reduced lymph node and spleen weights but support growth of transplantable SJL lymphoma RCS-X (48.29)
Authors: Tsiagbe Vincent , Podack Eckhard , Li Yu 
Published: January 2023

Abstract
The process by which endogenous retroviral superantigen (vSAg29) expressing SJL lymphomas (RCS) stimulate host CD4+Vβ16+ T cells, in order to elicit help (notably IL-4 and -5) for their growth, has been characterized as “reverse immune surveillance”. This response is facilitated by the high expression of an array of co-stimulating molecules on RCS cells, including B7.1, B7.2, 41BBL, CD40 and CD30. On the basis of their germinal center derivation, SJL lymphomas bear close resemblance to a subpopulation of human germinal center B cell lymphomas, such as CD30+ Hodgkin’s lymphomas. In our earlier studies, growth of transplantable SJL lymphoma (RCS-X) in vivo was inhibited by anti-CD30 mAb. While the evidence shows that this effect is indirectly on the lymphoma-responsive CD4 T cells, the mechanism of action was unclear. To shed light on the role of CD30-CD30L interactions in SJL lymphoma development, CD30L-/- were bred onto SJL background and tested for their support of transplantable lymphoma growth. A significant reduction in lymph node (17.4-54.7%) and spleen weights (21.5-38.2%) was observed for CD30 null mice, compared to wild type SJL mice. Despite this reduction in the lymphoid compartment, no reduction in lymphoma growth was observed. The results suggest that while CD30-CD30L interactions might be important for SJL lymphoma growth, other costimulating molecules might contribute significantly to the process of reverse immune surveillance in RCS lymphoma.
Tumor-associated macrophages suffocate NKT cells: a novel tumor escape mechanism and a target for therapy (48.7)
Authors: Liu Daofeng , Song Liping , Wei Jie , Gao Xiuhua , Marinova Ekaterina , Dotti Gianpietro , Metelitsa Leonid 
Published: January 2023

Abstract
Vα24-invariant Natural Killer T cells (NKTs) are important for antitumor immunity and immunotherapy. We have demonstrated that NKTs inhibit tumor growth indirectly via targeting tumor-associated monocytes/macrophages (TAMs) in neuroblastoma (NB), suggesting that tumor progression requires an escape from NKTs via yet unknown mechanism. Here, we demonstrate that NB cells via membrane-bound TNFα induce CCL20 production in monocytes and this effect is amplified in hypoxia. Anti-CCL20 neutralizing mAb strongly inhibited NKT-cell in vitro migration towards tumor-conditioned hypoxic monocytes and in vivo localization in NB/TAM xenografts in NOD/SCID mice. The immunofluorescent staining of primary NB tumors revealed selective CCL20 accumulation in TAMs. We also found that hypoxia impairs NKT-cell function, suggesting that NKT-cell trafficking toward CCL20-producing TAMs may serve as a hypoxic trap for tumor-infiltrating NKTs and other T cells. IL-15 protected NKTs from hypoxia in vitro and IL-15-transduced NKTs more efficiently inhibited tumor growth in metastatic NB model in humanized NOD/SCID/IL2rgamma(null) mice compared with parental NKTs. Thus, CCL20 production in hypoxic TAMs and consequent inhibition of NKTs reveals what we believe to be a novel mechanism of immune escape that can be reversed by adoptive immunotherapy with IL-15-transduced NKTs.