bluuebunny/crossref_metadata_2025_split huggingface dataset:
C5b-9 activated, Kv1.3 channel-sensitive Akt pathway mediate oligodendrocyte proliferation (135.7)
Authors: Tegla Cosmin A , Cudrici Cornelia , Singh Anil , Rozycka Monica , Rus Violeta , Judge Susan I.V. , Rus Horea 
Published: January 2023

Abstract
Voltage-gated Potassium (Kv) channels are involved in regulation of growth factors induced cell proliferation. Previously we showed that C5b-9 induced cell cycle activation in oligodendrocytes (OLG) but had yet to investigate the role of Kv channels. Here, we examined the effect of C5b-9 on Kv channel expression and function. OLG Kv channels were identified by recording whole-cell currents and their expression confirmed by western blotting. OLG exposure to C5b-9 significantly increased Kv1.3 and Kv1.4 expression while C5b6 had no effect. However, C5b-9 induced expression of OLG Kv1.3 and Kv1.4 was blocked by 4-aminopyridine, while pretreatment with the recombinant scorpion toxin rOsK-1, a specific Kv1.3 inhibitor blocked the expression of Kv1.3 alone. Since both ERK and Akt pathways are known to be induced by C5b-9 we assessed the possible role of Kv1.3 in activation of these kinases. rOsK1 inhibited Akt phosphorylation and activation by C5b-9 but had no effect on ERK1 activation. These data strongly suggest a role for Kv1.3 in controlling Akt activation induced by C5b-9. rOsK-1 significantly inhibited DNA synthesis induced by C5b-9 in OLG, indicating that Kv1.3 plays an important role in C5b-9 induced cell cycle. In conclusion, our data suggest that Kv1.3 channel plays an important role in controlling cell cycle activation by affecting the activation of Akt induced by C5b-9.
The role of Eri-1 as a small RNA regulator in the immune system (136.43)
Authors: Abdul-Wajid Sarah , Thomas Molly , Ansel K. Mark 
Published: January 2023

Abstract
Mouse Eri-1 is a 3' to 5' exoribonuclease whose C.elegans counterpart has been shown to inhibit RNAi. We have explored a similar role for murine Eri-1 in T-cells; siRNA efficacy and miRNA levels are increased in Eri-1 deficient T-cells. Reconstitution with wild-type Eri-1 via a retroviral construct returned miRNA levels to their wild-type quantities. In addition to validating a small RNA regulatory role for mouse Eri-1, we are exploring various immunological effects of Eri-1 deficiency. Eri-1 deficient T-cells exhibit an increased propensity to differentiate into Th2 cells in vitro and germinal center formation is altered in the absence of Eri-1.
CD8+ T cells induce severe thyroid epithelial cell hyperplasia (TEC H/P) and fibrosis (137.10)
Authors: Yu Shiguang , Sharp Gordon C. , Braley-Mullen Helen 
Published: January 2023

Abstract
IFN-γ -/-NOD.H-2h4 mice develop TEC H/P and fibrosis characterized by abnormal proliferation of thyrocytes and infiltration of thyroids by CD4+ and CD8+ T cells. Mice with severe TEC H/P produce anti-mouse thyroglobulin (MTg) autoantibodies, and their splenocytes transfer severe TEC H/P to SCID recipients, indicating that TEC H/P has an autoimmune basis. The goal of this study was to determine what cells transfer severe TEC H/P and fibrosis. Splenocytes from donors with severe TEC H/P cultured 72 hr with MTg consistently transferred more severe TEC H/P than the same number of freshly isolated cells, indicating that cells able to transfer severe TEC H/P can be activated in vitro. To determine what cells are important for transfer of TEC H/P, spleens of IFN-γ -/- mice with severe TEC H/P were activated in vitro, and CD4+ and CD8+ T cells were isolated and transferred to SCID recipients. Recipients of CD8+ cells had more severe TEC H/P and more fibrosis than recipients of CD4+ T cells or unseparated splenocytes. They had few infiltrating CD4+ cells, suggesting that CD8+ T cells are primary effector cells for severe TEC H/P and fibrosis. Recipients of CD4+ T cells developed mild hyperplasia and minimal fibrosis. Studies are in progress to determine what molecules secreted by CD8+ T cells mediate TEC H/P and fibrosis, and if CD4+ T cells are required for activation of CD8+ T cells. (Supported by VA Merit Review and NIH R56 AI074857).
The Adaptor Protein Shc Plays a Key Role in Early B Cell Development (138.22)
Authors: Deusterman Amber Jin , Bender Timothy P , Ravichandran Kodi S 
Published: January 2023

Abstract
B cell development is a highly ordered process that requires immature cells to pass through multiple checkpoints to advance through development. Successful expression and productive signaling via antigen and cytokine receptors are required at specific stages of development. The adaptor protein Shc is phosphorylated on three critical tyrosine residues and facilitates signaling downstream of multiple receptors, including the pre-T cell receptor. However, its role in B cell development is currently unknown.
Here, using a Cre-loxP mediated approach, we inducibly expressed as transgenes either wild type Shc or a mutant form Shc (ShcFFF, where three critical tyrosines have been mutated). Igα-Cre-mediated induction of ShcFFF expression (but not wt Shc) at the common lymphoid progenitor stage resulted in a severe block at the pre-pro-B to pro-B transition, with diminished numbers maintained throughout B development. Expression of ShcFFF at later stages of B cell development (via CD19-Cre) showed no defect in B cell numbers, emphasizing the importance of Shc during early B development. Mechanistically, Shc appears to affect B cell development at the level of cytokine dependent proliferation, as well as pre-BCR signaling. These results suggest a non-redundant role for Shc in early B development.
Ex vivo expansion of B cells from hematopoietic stem cells to supplement cord blood transplants (141.21)
Authors: Beaudette-Zlatanova Britte C , Le Phong T , Zhang Shubin , Stiff Patrick J , Knight Katherine L 
Published: January 2023

Abstract
Successful cord blood (CB) transplantation in adults is hampered by low numbers of hematopoietic stem cells (HSC) and infection due to delayed establishment of immunocompetence. By performing CDR3 spectratype analysis of T and B lymphocytes from CB transplant recipients, we demonstrated that these patients do not develop a diverse repertoire until 6 months or more after transplant. To provide rapidly engrafting antigen-responsive cells to CB recipients and to increase the number of transplanted cells, we developed an in vitro system to expand lymphoid progenitors from CB HSC. CB HSC (CD34+CD45RA+CD38-) are co-cultured for one to two weeks with either HEK 293T cells or a non-tumorigenic human thymic epithelial cell line (TEC) in media supplemented with a cocktail of cytokines and growth factors. In this study, we focused on B-lineage cells (CD79a+) and found, on average, B-lineage cells expanded more than 70 fold in HEK 293T co-cultures and nearly 100 fold in TEC co-cultures. We conclude that B-lineage cells can be expanded effectively in vitro; consequently, we are testing whether these cells are functional after engraftment in NOD/SCID/IL2Rγ-/- mice. We hypothesize that compared to patients receiving HSC alone, patients receiving transplants supplemented with in vitro-generated lymphoid progenitors will become immunocompetent sooner.
Funding by NIH AI068390 (KLK) and AG023809 (PTL)
T cell responses to hub gene knockout in in-vitro propagated Jurkat Cells: Application to Systems Biology (141.50)
Authors: Khanna Ajai , Lwin Wint Wah , Park Ken , Wauson Matthew , Gao Qin , Perkins David 
Published: January 2023

Abstract
Systems biology is unraveling complex gene interconnections. T cell genes are upregulated during rejection. Suppression of hub and node genes in the T cell genome could result in T cell hyporesponsiveness.
Objective: To knockdown hub or node genes via RNA interference in Jurkat cells and study their effect on T cell proliferation.
Methods: A molecular interaction network was built with Cytoscape using microarray data on Jurkat cells to identify upregulated hub genes: ATP5C1, IL6ST, PRKCZ, MYC, and MAPK1 that served as targets for RNAi knockdown. Jurkat cell lines were lipotransfected with siRNAs for the selected hub genes. Cell proliferation was measured by ATP release bioluminescence assay and BRDU labeling.
Results: At 12 hours following incubation with siRNA MAPK1, there was a reduction in ATP production compared to controls. At 24 hours, siRNAs incorporating genes for ATP5C1, IL6ST, PRKCZ, and MYC showed reduction in Jurkat cell proliferation compared to controls. No difference was observed in MAPK1 group and in the group transfected with a combination of siRNAs for all the identified hub genes.
Conclusions: Downregulation of hub genes by siRNA transfection is possible and could open opportunities to target these genes and modify T cell responses. This will help us in integrating systems biology to identify genes that could be potential therapeutic targets.
The role of Mad, Mnt and c-Myc proteins in OX40 stimulated T cells (33.26)
Authors: Vasilevsky Nicole A , Weinberg Andrew 
Published: January 2023

Abstract
We sought to determine the molecular mechanisms by which OX40 mediates T cell function. Gene array analysis of OX40-activated Ag-specific T cells suggested a role for the transcription factor c-Myc and antagonist Mad family proteins. D011.10 T cells were adoptively transferred into BALB/c recipients and immunized with soluble OVA and an agonist anti-OX40 antibody, which enhances Ag-specific T cell proliferation and survival. Western blot analysis of Ag-specific T cells purified ex vivo revealed that Mad4, Mnt and c-Myc proteins were up-regulated after OX40 engagement. Mad4 and Mnt protein levels peaked at days 3-4 and decreased as the cells contracted in size. C-Myc protein levels remained unchanged over time after OX40 stimulation. We hypothesize that the up-regulation of c-Myc in activated T cells drives proliferation and consequently activation-induced cell death, which is counteracted by Mad4 and Mnt proteins after anti-OX40 engagement, allowing T cells to survive the "blast crisis" phase. Indeed, siRNA knockdown of Mad4 and Mnt proteins led to decreased survival of OX40-activated T cells. We also found that OX40 stabilized both Mad4 and Mnt proteins against degradation and we demonstrated the presence of a serine residue in Mad4 that was essential in mediating accelerated protein degradation. These data provide evidence that Mad4 and Mnt play a role to enhance memory T cell survival after OX40 engagement.
Direct and indirect effects of TLR agonists on FasL, TNF-alpha and TRAIL expression in CD4 T cells (33.7)
Authors: Weinlich Ricardo , Sant'Anna Maíra M , Ferra Luciana C , Campopiano Júlia C , Bortoluci Karina R , Amarante-Mendes Gustavo P 
Published: January 2023

Abstract
The lifespan of CD4 T lymphocytes is partially controlled by the engagement of the so-called Death Receptors (DR) on their surface. De-regulation of DR ligands, such as FasL, TNF-alpha and TRAIL, is associated with many pathological processes, including Immunodeficiency and Autoimmunity. We have recently shown that TcR/CD3-mediated FasL upregulation in CD4 T cells is prevented by PGE2 derived from LPS-stimulated antigen-presenting cells (APCs). Here we show that Toll-like receptor (TLR) agonists other than LPS can interfere with the expression of FasL, TNF-alpha and TRAIL. Except TLR7, stimulation of TLRs in APCs resulted in induction of PGE2, thereby indirectly blocked the expression of DR ligands on T cells. On the other hand, direct stimulation of TLR7 on T cells lead to repression of CD3-mediated FasL expression and consequently reduced AICD. Interestingly, this effect was independent of NF-kB. Altogether, these results suggest that pathogen-associated molecular patterns can alter the expression of DR ligands, either by direct stimulation on CD4 T cells or by inducing the release of PGE2 by APCs.
Supported by FAPESP and CNPq.
MAP kinase phosphatase 1 is necessary for T cell activation and function (35.1)
Authors: Zhang Yongliang , Reynolds Joseph , Chung Yeonseok , Dong Chen 
Published: January 2023

Abstract
Mitogen-activated protein kinases (MAPK) are evolutionary conserved immune regulators. MAPK phosphatases that negatively regulate MAPK activities have emerged as important players in both innate and adaptive immune responses. MKP-1, also known as DUSP1, was previously shown to negatively regulate innate immunity by inhibiting pro-inflammatory cytokine production. Here we found that MKP-1 positively regulate T cell activation, function and negatively regulate apoptosis. MKP1-deficient dendritic cells exhibited significantly reduced ability to activate T cells than wild-type cells. In T cells, MKP-1 deficiency led to enhanced activation of ERK and JNK but not p38, and impaired the activation, proliferation and function of T cells in vitro and in vivo. MKP-1 deficient T cells exhibited enhanced activation induced cell death. Consistently, MKP-1-/- mice were found to be resistant to experimental autoimmune encephalomyelitis but were defective in anti-influenza immunity in vivo. Our results thus demonstrate MKP-1 as a critical positive regulator of T cell activation and function and may be targeted in treatment of immune diseases.
Enhanced humoral immune responses against T-independent antigens in Fcα/μR-deficient mice (39.14)
Authors: Honda Shin-ichiro , Kurita Naoki , Shibuya Akira 
Published: January 2023

Abstract
IgM is an antibody class common to all vertebrates that plays a primary role in host defences against infection. Binding of IgM with an antigen initiates the complement cascade, accelerating cellular and humoral immune responses. However, the functional role of the Fc receptor for IgM in such immune responses remains obscure. Here we show that mice deficient in Fcα/μR, an Fc receptor for IgM expressed on B cells and follicular dendritic cells (FDCs), had enhanced germinal center formation and affinity maturation and memory induction of IgG3+ B cells after immunization with T-independent (TI) antigens. Moreover, Fcα/μR-deficient mice showed prolonged antigen retention by marginal zone B (MZB) cells and FDCs. In vitro studies demonstrated that interaction of the IgM immune complex with Fcα/μR partly suppressed TI antigen retention by MZB cells. We further showed that downregulation of complement receptor (CR)1 and CR2 or complement deprivation by in vivo injection with anti-CR1/2 antibody or cobra venom factor attenuated antigen retention by MZB cells and GC formation after immunization with TI antigens in Fcα/μR-/-_mice. Taken together, these results suggest that Fcα/μR negatively regulates TI antigen retention by MZB cells and FDCs, leading to suppression of humoral immune responses against T-independent antigens.
The influence of commensal bacteria on anti-viral immunity (39.26)
Authors: Abt Michael C , Beiting Daniel , Kobuley Dymtro , Yu Yimin , Zaph Colby , Wherry John , Artis David 
Published: January 2023

Abstract
Alterations in the composition of intestinal commensal bacteria in humans are associated with enhanced susceptibility to multiple inflammatory diseases suggesting that signals derived from commensal bacteria may influence the development and/or function of the immune system. Supporting this, germ-free or gnotobiotic mice exhibit reduced numbers of lymphocytes in the intestinal intraepithelial compartment. However, whether alterations in the acquisition or composition of commensal bacteria influence immunity to infection remains poorly defined. To test this, mice housed under conventional or gnotobiotic conditions were infected i.p. with Lymphocytic Choriomeningitis Virus (LCMV). At day 7 post-infection, gnotobiotic mice exhibited a significant reduction in the frequency and number of LCMV-specific CD8+ T cells in multiple tissues including the spleen and intraepithelial compartment. Furthermore, LCMV-specific CD8+ T cells from gnotobiotic mice were less capable of producing IFN-γ following LCMV peptide stimulation. In addition, at day 36 post-infection, when conventionally-housed mice have established a LCMV-specific CD8+ memory T cell population, gnotobiotic mice had a significantly impaired population of LCMV-specific CD8+ memory T cells. Diminished LCMV-specific CD8+ effector and memory T cell responses were not the result of inherent defects in gnotobiotic mice as oral administration of a cocktail of antibiotics to conventionally-housed mice also displayed a defective LCMV-specific CD8+ T cell response following infection. Collectively, these data suggest an integral role of commensal bacteria in influencing virus specific CD8+ T cell effector and memory responses.
Response of human epithelial cells and monocytes to UV radiation in a co-culture model of inflammatory acne (39.47)
Authors: Dapul Gina E , Brown Paula , Yaping E. , Lee Wei-Li , Shalita Alan R. , Nowakowski Maja 
Published: January 2023

Abstract
Earlier studies in our laboratories showed that ultraviolet (UV) light has anti-inflammatory properties, as demonstrated by inhibition of IL-1 and ICAM-1 expression by human keratinocytes (hTERT, human telomerase reverse transcriptase - transformed HaCaT cells). The goal of the study was to construct an improved model system that involved a modified co-culture of more than one cell line, exposure to UVA and UVB, and measurement of nitric oxide production as a marker for inflammation. The ability of SC cells to produce nitric oxide in culture was confirmed by incubation in the presence of phorbol myristic acetate (PMA, Sigma) (40-160 nM) for 48 hrs, which resulted in 1.6 fold increase in accumulation of nitric oxide. Human epithelial cells (A549, ATCC) were seeded in a 24-well tissue culture plate (0.1 x 106 cells/well) 48 hrs before use. Cells were exposed to UVA (from 0 to 40J/m2), UVB (from 0 to 200mJ/cm2), and bacterial lipopolysaccharide (LPS, Sigma) (0 to 500ng/ml) and incubated for 48 hours. Supernatants of the irradiated and control A549 cells were co-cultured with human monocytes (SC, ATCC) which were seeded at 0.2 x 106 cells per well in a 24-well tissue culture plate. The modified co-culture was incubated for 48 hours. The supernatants were collected and nitric oxide content was measured (modified Griess assay). Under these conditions, neither LPS nor UV radiation triggered nitric oxide production (&lt;10 μM). The modified co-culture system is a useful model for studying cell-cell interactions and responses to inflammatory stimuli and inhibitory agents or treatments. Supported in part by New York State Dept. of Health, ECRIP Fellowship to G.E.P.
Comparison of the Expression Profile of JunB, c-Jun, and S100A8 (Calgranulin A) in Psoriasis Vulgaris and Guttate Psoriasis (39.48)
Authors: Lee Kyung Ho , Park Chul Chong , Chun Ji Hoon , Woo So-Youn 
Published: January 2023

Abstract
Background: Psoriasis is a chronic, inflammatory, immune-mediated skin disease. Recently, several psoriasis-linked genetic loci have been reported; PSORS4 contains S100A8 (calgranulin A), and the PSOR6 (19p13) locus harbors JunB (19p13.2). S100A8 is considered to be a marker of inflammation in a variety of diseases. The expression of JunB and c-Jun have been reported to be reduced in psoriatic lesions.
Objective: We attempted to assess the role and correlation of S100A8, JunB, and c-Jun in the pathogenesis of guttate psoriasis and psoriasis vulgaris by studying whether any difference of immunohistochemical expression existed.
Methods: Skin biopsy specimens from patients with psoriasis vulgaris (n=37) and guttate psoriasis (n=17), and a normal skin controls (n=9) were utilized in the study. Formalin-fixed and paraffin-embedded tissue sections were prepared and JunB, c-Jun, and calgranulin A were immunohistochemically stained in order to compare the expression of those three proteins in each group.
Results: Reduced JunB expression was observed in patients with psoriasis vulgaris and guttate psoriasis, as compared to patients in the control group; however, c-Jun expression was reduced only in the psoriasis vulgaris group. The expression of S100A8 increased in the psoriasis groups as compared to the control group. In addition, the expression of S100A8 was different between the psoriasis vulgaris and guttate psoriasis groups; S100A8 was expressed more profoundly in the guttate psoriasis group (p&lt;0.05).
Conclusion: Our results indicate that S100A8 contributes to the pathogenesis of guttate psoriasis, and it may be a good target for therapy for guttate psoriasis provoked by microorganisms.
Natural Killer T (NKT) cells mediate anti-tumor activity via killing of tumor-associated macrophages (40.19)
Authors: Metelitsa Leonid S , Song Liping , Salo Jill , Engell Kelly , Wu Hong-wei , Sposto Richard , Seeger Robert C , Asgharzadeh Shahab 
Published: January 2023

Abstract
Tumor infiltration with CD1d-reactive Vα24-invariant NKT cells associates with favorable outcome in neuroblastoma and other cancers. However, the function of NKTs in tumor immunity remains largely unknown. We demonstrate that CD68+ tumor-associated monocytes/macrophages (TAMs) represent the majority of CD1d-expressing cells in primary neuroblastomas. TAMs in vitro and in vivo stimulated neuroblastoma growth via IL-6 production and were the source of IL-6 in primary tumors and metastatic bone marrows in patients. Gene expression analysis with TaqMan® low-density arrays of 129 primary MYCN non-amplified neuroblastomas revealed that high level expression of TAM genes (CD14, CD16, IL-6, IL-6R, and TGFβ) was associated with poor 5-year event-free survival. While NKTs were not cytotoxic against neuroblastoma cells, they effectively killed monocytes pulsed with tumor cell lysate. The killing was CD1d-restricted since it was inhibited by anti-CD1d mAb. Co-transfer of human monocytes and NKTs to tumor-bearing NOD/SCID mice decreased monocyte number at the tumor site and delayed tumor growth compared to mice transferred with monocytes alone. Thus, killing of TAMs reveals a novel mechanism of NKT cell anti-tumor activity that relates to the disease outcome and should be considered for NKT-based therapeutic strategies. Supported by NIH grants: RO1 CA116548 (LSM) and PO1 CA81403 (RCS).
SYNERGY BETWEEN MOLECULAR TARGETS OF COSTIMULATION IN PROMOTING T CELL PERSISTENCE (40.31)
Authors: Song Jim , Croft Michael 
Published: January 2023

Abstract
Costimulation regulates multiple cellular processes including proliferation, expansion and survival of T cells, and molecular targets of costimulation might be useful to augment T cell activities. Two defined targets of costimulatory signals in CD4 T cells are the anti-apoptotic bcl-2 family molecule bcl-xL, and survivin, an Inhibitor of Apoptosis family member that might regulate cell division and survival. However, the relative importance and relationship between these molecules in CD8 T cells is not clear. To understand whether they have overlapping or synergistic functions, we used retrovirus-mediated transduction to introduce bcl-xL and survivin separately, or together linked by a 2A picornavirus self-cleaving peptide, into antigen (Ag)-responding CD8 T cells. In vitro, we found that CD8 effector T cells expressing both bcl-xL and survivin strongly expanded at an early stage and had a long-term survival advantage. In vivo, with response to tumor-expressed Ag following adoptive T cell transfer, Ag-reactive CD8 T cells expressing both bcl-xL and survivin displayed a greatly enhanced tumor protective activity compared CD8 T cells expressing either molecule introduced separately. These results indicate that bcl-xL and survivin can critically contribute in a synergistic, non-redundant, manner to augment the accumulation and persistence of CD8 T cells following encounter with Ag. The data provide new insights into why costimulatory signals might need to be sustained over time or provided periodically via ligation of multiple receptors.