bluuebunny/crossref_metadata_2025_split huggingface dataset:
γδ T cells protect TCRα deficient mice from the autoimmunity induced by scurfy lymphocytes (BA13P.118)
Authors: Ujiie Hideyuki , Shevach Ethan 
Published: January 2023

Abstract
Foxp3+ regulatory T cells (Treg) are critical for the maintenance of immunological tolerance, and a mutation of the Foxp3 gene leads to the absence of Treg and a fatal systemic autoimmune disease in Scurfy mice. Transfer of Scurfy lymphocytes to RAG-/- recipients reproduces the inflammatory phenotype of the Scurfy donor including hepatitis and pneumonitis. Here, we show that TCRα-/- recipients, that lack TCRαβ+ T cells, but have TCRγδ+ T cells and B cells, are significantly protected from autoimmunity induced by adoptive transfer of Scurfy lymphocytes. Co-transfer of Scurfy lymphocytes and B cells from TCRα-/- mice into RAG-/- recipients failed to prevent inflammation. Furthermore, transfer of Scurfy lymphocytes to TCRβδ-/- mice, which lack both TCRαβ+ and TCRγδ+ T cells, but have B cells, resulted in more severe inflammation in the liver and lungs than that seen in TCRα-/- recipients. Taken together, these studies demonstrate that TCRγδ+ T cells, but not B cells, in the TCRαβ-/- recipients mediate protection from disease induced by Scurfy T cells. The TCRγδ+ T cells in the TCRα-/- recipients of Scurfy cells markedly expanded and expressed a highly activated (CD44hi CD62Llo) phenotype, but did not produce IL-10. Surprisingly, the Scurfy T cells in these recipients produced more IL-10 and less IL-4 than Scurfy T cells transferred to RAG-/- or TCRβδ-/- recipients, suggesting that TCRγδ+ T cells may protect TCRα-/- recipients by modulating cytokine production by Scurfy T cells.
Aberrant production of IL-13 by T cells promotes exocrinopathy in Id3 knockout mice (BA14P.206)
Authors: Belle Ian , Mahlios Joshua , Zhuang Yuan 
Published: January 2023

Abstract
Elevated cytokine IL-13 in serum has been associated with several autoimmune diseases, including Sjögren’s Syndrome. However, whether IL-13 plays an early and causative role in disease development is not known and cannot be easily studied in humans. Our previous work has shown that levels of the IL-13 are elevated in Id3 knockout mice, which has been established as a model for primary Sjögren’s Syndrome. Here, we utilized an IL-13 reporter to determine the source of the elevated IL-13 levels observed in Id3 knockout mice and assess its contribution to SS symptoms. Our results indicate that T cells, primarily CD4+ αβ T cells and γδ T cells, in Id3 knockout mice produce IL-13 at an elevated frequency well before disease symptoms become apparent. We also show that T cells developing early in life are more predisposed to produce IL-13. Finally, analysis of Id3 and IL-13 double deficient mice demonstrated that IL-13 plays an essential role in the deterioration of gland function. Our study indicates that enhanced IL-13 production by T cells is capable of inducing exocrinopathy in animal models.
IgA but not IgG serum antibodies to maize prolamins in Mexican patients with celiac disease (CD) and irritable bowel syndrome (IBS) (BA15P.226)
Authors: Sánchez Vargas Luis , Remes Troche Jose , Mellado Sánchez Gabriela , Vivanco Cid Héctor 
Published: January 2023

Abstract
High prevalence of CD has been found in IBS patients. CD is a chronic immune-mediated enteropathy triggered by exposure to dietary gluten in genetically predisposed individuals. Treatment for CD is to follow, a gluten free diet (GFD). However, some CD patients describe persistent symptoms despite the GFD, suggesting a cross reactivity or specific immune responses against other related-prolamins such as zein present in maize. We evaluated the prevalence of IgA and IgG anti-zein antibodies (AZA) in 37 CD, 375 IBS and 310 healthy controls (HC). All of the IBS subjects met the Rome III criteria and were negative to IgA h-tTG, IgA and IgG Gliadin II. IgA and IgG AZA were measured using an in-house ELISA. Elevated levels of IgA AZA were found in CD patients compared with IBS (p&lt;0.001) and HC (p&lt;0.01). CD patients had the highest prevalence (29.7%) compared with IBS (3.5%) and HC (1.3%) (p&lt;0.0001). IgG AZA production was not demonstrated in any patients or HC. To explore if the results in IBS patients and HC can be explained by cross-reaction among gliadins and zeins, positive serum sample to IgA AZA were analyzed for IgA and IgG antibodies against native Gliadin (AGA). Interestingly, 53.8% of IBS patients and 50.0% of HC got negative results for IgA and IgG AGA, suggesting that IgA AZA response could be specific to maize prolamins. These results open the possibility to explore more about the contribution of the humoral responses to zein in the immunopathogenesis to CD and IBS.
The development of humanized mouse models to study immune regulation of autoreactive T cells. (BA15P.229)
Authors: Aryee Ken-Edwin , Brehm Michael 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is the immune mediated destruction of insulin-producing beta cells in the pancreas, characterized by the loss of peripheral tolerance mechanisms needed to prevent autoimmunity. Regulatory T-cells (TREG) are a specialized CD4 T cell subset important for maintaining peripheral tolerance. IL-2 is critical to maintain TREG fitness and function, and deficiencies in the IL-2 receptor and signaling pathways contribute to the development of T1D. The lack of an in vivo system that fully recapitulates human T1D development impairs the ability to study the role of IL-2 signaling in T1D. Here we show that the Hu-PBL-SCID humanized mouse model can be used to study effector T cells and TREG from T1D-individuals. For these studies we injected PBMC from T1D or non-T1D donors into NOD Rag1null IL2rγnull (NRG) mice and followed the engraftment of human T cell populations. Our results show that PBMC from T1D donors engraft NRG mice with similar kinetics as compared to non-T1D donors. Moreover, our preliminary data show that expression of human IL-2 in recipient NRG mice improves T1D TREG frequency and survival. These results indicate that humanized mice are an effective model for studying autoreactive T cells from diabetic individuals
Effects of Rho GTPase CDC42 on Th2 and Th17 differentiation (CCR6P.277)
Authors: Yang Jun-Qi , Guo Fukun , Zheng Yi 
Published: January 2023

Abstract
We have reported that cell-division-cycle 42 (CDC42) of Rho GTPase family was important for T cell development and homeostasis with the use of conditional CDC42-deficient mice (CDC42flox/LckCre), in which CDC42 was deleted in CD4-CD8- double negative stage of thymocytes. Recently, we generated another line of conditional CDC42-deficient mice with selective expression of the Cre recombinase beginning at a late stage of T cell development (after positive selection of CD4+CD8+) using the distal promoter of the Lck (dLck) gene. We found that CDC42-deficiency (CDC42flox/dLckCre) did not affect T cell development, as expected. The development of natural regulatory T cells (nTreg) was also not affected by CDC42-deficiency. However, CDC42-deficiency impaired in vitro activation of both CD4+ and CD8+ T cells. When naïve CD4+ T cells were cultured under non-polarized conditions, CDC42-deficiency significantly reduced Th1 and Th2 cytokine secretion, but slightly increased Th17 cytokine secretion from CD4+ T cells. Under Th1/Th2/Th17-polarized conditions, CDC42-deficiency significantly downregulated Th2-differentiation, but was less effective to Th1 and Th17 differentiation. CDC42 has no significant role on the induction of Treg in vitro. The ongoing studies are trying to explore the underlying molecular mechanisms. Our findings suggest that CDC42 of Rho GTPase is not only crucial for T cell development and activation, but also involved in Th differentiation, especially Th2 cells.
The dysfunction of the E3 ubiquitin ligase TRIM21 in systemic lupus erythematosus (HUM7P.301)
Authors: Kamiyama Reikou , Yoshimi Ryusuke , Tsukahara Toshinori , Ueda Atsuhisa , Takeno Mitsuhiro , Ozato Keiko , Ishigatsubo Yoshiyuki 
Published: January 2023

Abstract
Although the increased expression of type I interferon (IFN)-inducible genes, called “IFN signature”, has been suggested to have important roles in pathogenesis of systemic lupus erythematosus (SLE), its mechanism still remains unclear. Recent studies in vitro or with mice suggest that TRIM21, an autoantigen also called Ro52 or SSA1, is involved in the regulation of type I IFN production as an E3 ubiquitin ligase for IFN regulatory factors (IRFs). Here, we investigated the pathological role of TRIM21 in SLE. We collected peripheral blood mononuclear cells (PBMCs) from 20 patients with SLE and 24 healthy controls (HC). The mRNA levels of TRIM21 and other type I IFN-inducible genes were higher in PBMCs from patients with SLE as compared to HC. Thus, TRIM21 mRNA levels correlated positively with other type I IFN-inducible genes. On the other hand, type I IFN mRNA levels showed a negative correlation with TRIM21 mRNA levels in HC but not in patients with SLE. Consistent with higher TRIM21 mRNA levels, TRIM21 protein levels were higher in SLE patients as compared to HC. After treating with MG-132, the protein levels of IRFs were increased in PBMC from HC, but not in the PBMC from patients with SLE. These results suggest that dysregulation of TRIM21 to function as an E3 ubiquitin ligase for IRF family may be associated with the increased expression of type I IFNs in SLE.
Aberant expansion of CXCR5+ memory CD4 T cells in SLE patients (HUM7P.307)
Authors: Choi Jinyoung , Kim Sang Taek , Kang Insoo , Craft Joe 
Published: January 2023

Abstract
Autoreactive B cells in SLE undergo autoantigen selection, suggesting a requirement for germinal center follicular helper T (Tfh) cells in their maturation. However, evidence for dysregulation of Tfh cells in SLE and their potential contribution to disease remains unclear. Recently, blood CXCR5+ CD4 T cells, a heterogeneous pool consisting of functionally distinct Th1-, Th2-, and Th17-like subsets, have been proposed to be the circulating (blood) memory Tfh cells. We hypothesized that expanded CXCR5+ memory cells in the blood of human lupus patients promote B cell helper function reflecting the abnormal T-B cell responses in secondary lymphoid organs. We have characterized such cells in SLE patients by flow cytometry and T-B coculture studies. SLE patients had significant expansion of CXCR5+ICOShiPD-1hi CD4 T cells compared to controls. Such cells were Bcl6-, but robustly expressed IL-21 with a portion Ki-67+, indicating their functional activity. The blood cells were capable of providing blood-born memory B cells with survival and differentiation signals to secrete isotype switched Igs, including anti-nuclear antibodies. We speculate that therapies that alter T-B collaboration in lupus will abrogate their expansion, accompanied by reduced autoantibody titers and improved disease activity in SLE patients. Our results suggest that aberrant T-B collaboration in lupus is critical to disease pathogenesis and its blockade is likely to be important therapeutically.
Use of humanized mouse models for the study of idiopathic CD4 lymphocytopenia (HUM7P.313)
Authors: Perez-Diez Ainhoa , Liu Xiangdong , Sheikh Virginia , Sereti Irini 
Published: January 2023

Abstract
Patients with Idiopathic CD4 Lymphocytopenia (ICL) have low circulating CD4 T-cell counts and develop opportunistic diseases and papillomavirus-related dysplasia. The etiology of ICL is unknown but the low numbers of CD4 T-cells could be explained by decreased thymic production, increased cell death, and/or increased migration or retention of the T cells in secondary lymphoid organs or tissues. To distinguish between these three possibilities we set up a humanized mouse model by transferring either peripheral blood mononuclear cells (PBMC), or Hematopoietic Stem Cells (HSC) from ICL patients or healthy controls (HC) into immune-compromised mice. First, we transferred HC cells into three different strains of mice to identify the most efficient and reproducible host for the human cells. We found that NOD-RAGKO-γcKO (NRG) mice represented the best model, as one to four weeks after transferring PBMC into NRG mice, we reproducibly found human T cells in blood, spleen, lymph node and bone marrow with no graft versus host disease. We next transferred PBMC from either HC or ICL patients into NRG mice to compare survival, expansion, phenotype and migration properties of the human cells. Preliminary results show that mice that received ICL cells developed lymphocytopenia comparable to ICL patients. These data encourage further use of this humanized mouse model to identify the etiology and pathogenesis of ICL and perhaps other human immune-deficiencies that affect T-cell homeostasis.
Expression and function of a novel isoform of Sox5 in malignant B cells (HUM7P.314)
Authors: Edwards Shanique , Desai Anand , Liu Yan , Moore Carissa , Xie Ping 
Published: January 2023

Abstract
Using a mouse model with the tumor suppressor TRAF3 deleted from B cells, we identified Sox5 as a gene strikingly up-regulated in B lymphomas. Sox5 proteins were not detected in normal or premalignant TRAF3-/- B cells even after treatment with B cell stimuli. We cloned the full-length cDNA of Sox5 from B lymphomas spontaneously developed in 4 different individual B-TRAF3-/- mice. Surprisingly, we found that the sequence of Sox5 expressed in TRAF3-/- B lymphomas represents a novel isoform of Sox5, which has not been previously reported. This new isoform of Sox5, Sox5-BLM, contains a 35 aa deletion in the N-terminal region in front of the leucine zipper domain. Sox5-BLM protein was localized in the nucleus of TRAF3-/- mouse B lymphomas and transduced human multiple myelomas. Overexpression of Sox5 inhibited cell cycle progression, and up-regulated the protein levels of p27 and β-catenin in human multiple myeloma cells. Taken together, our findings indicate that aberrant expression of Sox5 regulates the proliferation of malignant B cells.
T cell-mediated immunopathologic mechanism of liver injury in acute viral hepatitis (HUM8P.336)
Authors: Kim Jihye , Chang Dong-Yeop , Shin Eui-Cheol 
Published: January 2023

Abstract
CD8+ T cells play an important role in immune responses against intracellular infection. They recognize and lyse infected cells, and secrete anti-viral cytokines to control infection. However, excessive immune responses may lead to tissue damage during infection. In particular, severe CD8+ T cell-mediated liver injury is known as a primary cause of tissue damage in hepatitis A virus (HAV) infection. In this study, we demonstrate that during acute viral hepatitis, CD8+ T cells specific to HAV-unrelated viruses such as influenza virus, Epstein-Barr virus (EBV), or cytomegalovirus (CMV) became activated even in the absence of those viral reactivations, which means that non-HAV-specific CD8+ T cells are able to be activated even without cognate antigen. Interestingly, the severity of liver injury correlated with the activation of these non-HAV-specific T cells, not with activation of HAV-specific T cells. HAV-infected cells produced IL-15, which induced antigen-nonspecific activation of CD8+ T cells. IL-15-activated CD8+ T cells exerted NK-like cytolytic activity by NKG2D dependent manner, even without T cell receptor engagement. HAV-infected hepatocytes overexpressed NKG2D ligands and, thus, could be cytolytic targets during HAV infection. We report a human viral disease in which host injury is attributed to NK-like cytolytic activity of IL-15-activated CD8+ T cells not specific to the infecting virus.
Severity of infectious mononucleosis (IM) correlates with the frequency of crossreactive influenza A virus (IAV)-M1 and Epstein Barr virus (EBV)-BMLF-1-specific CD8 T cells (HUM8P.345)
Authors: Selin Liisa K. , Aslan Nuray , Watkin Levi , Gil Anna , Luzuriaga Katherine 
Published: January 2023

Abstract
During EBV-associated IM IAV-specific crossreactive memory T cells are activated and play a role in disease severity. In HLA-A2+ IM patients, influenza M1-58 (IAV-M1)-specific CD8 memory T cell responses crossreacted with two different EBV lytic epitopes, BMLF1-280 (17/29) and BRLF1-109 (19/20). Furthermore, 11/22 IM patients demonstrated some intra-viral crossreactivity between EBV-BRLF1 and -BMLF1 responses. Disease severity of IM directly correlated with significantly increased frequencies of crossreactive IAV-M1/EBV-BMLF1, IAV-M1, and EBV-BMLF1 specific CD8 cells, and with mean viral load over the first 5 weeks of infection. Disease severity did not correlate with BRLF1 or M1/BRLF1 crossreactive responses. When severity of IM was scored and patients were assigned to either mild or severe groups, disease severity correlated with specific TCR Vb usage in IAV-M1 population suggesting that TcR selection is driving disease outcome. Consistent with IAV-M1 and EBV-BMLF1 responses driving increased immunopathology was the observation that patients with severe disease had significantly more IAV-M1 and EBV-BMLF1 cells producing IFNg/MIP1-b in response to antigen as compared to patients with mild disease. These results suggest that T cell crossreactivity impacts T cell selection and function and ultimately disease outcome. Insights on these issues are important for the intelligent design of vaccines and to develop therapeutic interventions for virally induced disease.
Selectivity of Toll-like receptor stimulation in human B cells (HUM8P.350)
Authors: Simchoni Noa , Cunningam-Rundles Charlotte 
Published: January 2023

Abstract
B cells activated by nucleic-acid sensing Toll-like receptor 7 and TLR9 proliferate and secrete immune globulin. Memory B cells are more responsive due to higher TLR expression levels, but further selectivity remains largely unknown. In this study, human B cells stimulated by TLR ligands, with or without IFNα, or by T cell mimic CD40L plus IL-21 were examined to identify differentially responsive subsets, defined phenotypically or through BCR characteristics. Both stimuli induced CD27hiCD38hi plasmablasts, with TLRs inducing more IgM+ plasmablasts and IgM+ CD27hiCD38lo pre-plasmablasts than CD40L/IL-21. TLR stimulation also increased the relative proportion of IgM+ memory B cells and fostered higher IgM secretion, while CD40L/IL-21 expanded IgM- memory cells and fostered higher IgG secretion. Ig heavy chain deep sequencing showed V and J preference among TLR responsive cells, clustering TLR stimulated conditions apart from CD40L/IL-21 and control conditions. TLR stimulation expanded BCRs containing V3 at the expense of V1, and particularly expanded V3 to J4 rearrangements. More somatically hypermutated sequences were seen following TLR stimulation as compared to CD40L/IL-21. Additionally, complementarity determining region 3 segments from TLR activated cultures were shorter, more hydrophilic, and less negatively charged compared to CD40L/IL-21 or control CDR3s. Human B cells show selective sensitivity to TLR stimulation, with distinctive proliferative and genetic signatures.
Functional polymorphisms of TLR8 gene contribute to Mycobacterium tuberculosis infection (INC7P.401)
Authors: Eng Hock-Liew , You Huey-Ling , Wang Chiou-Huey , Su Pei-Yi , Lin Tsun-Mei 
Published: January 2023

Abstract
Innate immunity is important for antimycobacterial defence through senses various pathogen-associated molecular patterns of Mycobacterium tuberculosis (MTB). Davila et al. first reported the evidence of association of TLR8 polymorphisms with tuberculosis (TB) susceptibility in males, but the mechanism through which TLR8 recognizes MTB and intracellular signaling remains unknown. In this study, we demonstrated that TLR8-129C /+1 A allele was present at higher frequency in males TB infected patients as compared to the healthy controls (24.1% vs. 6.8%, p=0.001) in Taiwan. Ex vivo phagocytosis assay of the functional effects of these polymorphisms revealed significantly lower TLR8 expression (p=0.002) but higher phagocytosis ability (p=0.03) in monocytes of male individuals of TLR8 -129C/+1A genotype than those of wild type, TLR8 -129G/+1G. Since Mycobacteria within monocytes can escape the immune destruction, individuals with the TLR8 -129C/+1A might thus be higher susceptible to TB infection due to higher phagocytosis ability. In addition, ex vivo analyzing mRNA expression and cytokine production in whole blood stimulated with inactivated MTB, TLR7 expression (p=0.05) and TNFα production (p=0.03) were significantly downregulated in TLR8 -129C/+1A subjects. In conclusion, these findings provide an evidence of association between TLR8 gene polymorphisms and susceptibility to MTB infection which is related to the effectiveness of MTB phagocytosis and impaired TLR signaling.
Differential cytokine response by macrophages to ManLAM of Mycobacterium tuberculosis under various culture conditions (INC7P.419)
Authors: Bagriacik Emin , Albayrak Nurhan 
Published: January 2023

Abstract
Lipoarabinomannan, a lipoglycan is a major virulence factor for Mycobacterium. Mannosylated form of Lipoarabinomannan (ManLAM) is commanly found in pathogenic Mycobacterium tuberculosis. It has been shown that ManLams inhibit production of TNFα and IL-12 by macrophages under in vitro culture conditions. In this study, we tested cytokine responses of macrophages stimulated with ManLAM under various culture conditions. ManLAM was isolated from M. tuberculosis strain H37Rv (ATCC 25618). Macrophages (J744.1; murine macrophage cell line) were stimulated with ManLAM in three different conditions, either alone or in co-cultures with fibroblasts (3T3; ATCC, CRL1658) in contact-culture and non-contact culture systems. Cytokines were measured using commercially available ELISA kits. In comparison to unstimulated controls, TNFα, IL-6 and IFNγ levels increased significantly in co-culture systems after stimulation with ManLAM while a slight increase occurred for IL-12 in the same cultures. However, we observed no change in TNFα, IL-6 and IL-12 for stimulated macrophages when they were cultured alone in the absence of fibroblasts. These results suggested that macrophage activity to ManLAM changed in the presence of the fibroblast in co-culture systems that might be an in vitro model for granuloma formation in tuberculosis infections.
Neutrophils in granulomas from Mycobacterium tuberculosis-infected macaques express granzyme B in response to mycobacterial products and pro-inflammatory signals (INC7P.406)
Authors: Mattila Joshua , Flynn JoAnne 
Published: January 2023

Abstract
Inflammation is a critical component of tuberculosis (TB), and may contribute to a poor prognosis. The factors responsible for this inflammation are poorly studied. In addition, the role of neutrophils in TB, and whether neutrophils express granzyme B (grzB), a cytotoxic pro-apoptosis enzyme, is controversial. We examined neutrophils in peripheral blood (PB) and granulomas to determine if mycobacterial products or pro-inflammatory factors induce neutrophil grzB expression. PB neutrophils did not express grzB. In contrast, granuloma neutrophils expressed grzB and these cells contained more grzB than T cells. Elevated neutrophil, but not T cell, grzB expression correlated with higher tissue bacterial load. Perforin is required for T cell-mediated cytotoxicity but was not expressed by neutrophils. PB neutrophils upregulated grzB expression when infected with M. tuberculosis or stimulated with M. tuberculosis culture filtrate protein or LPS from E. coli, but not peptide antigens associated with T cell stimulation. Moreover, stimulated PB neutrophils secreted grzB in a perforin-independent process. GrzB was not bactericidal or bacteriostatic, suggesting secreted neutrophil grzB acts on extracellular targets, potentially enhancing neutrophil migration or regulating activation in other cell types. These data indicate M. tuberculosis-specific factors in granulomas upregulate neutrophil grzB expression and suggest a previously unappreciated aspect of neutrophil biology in TB.