bluuebunny/crossref_metadata_2025_split huggingface dataset:
Primary medulloblastomas express functional CD1d and can be targeted for immunotherapy with NKT cells (156.25)
Authors: Liu Daofeng , Song Liping , Salsman Vita , Gao Xiuhua , Wei Jie , Teo Wanyee , Lau Ching , Ahmed Nabil , Metelitsa Leonid 
Published: January 2023

Abstract
Medulloblastoma (MB) is the most common brain tumor of childhood. Current therapies are not curable for a third of patients and have debilitating long-term toxicity. In the initial screen of potential targets for immunotherapy, we performed gene expression analysis of twenty primary MB tumors and found that nine of them expressed high levels of CD1d RNA. Nearly all tumors cells in the corresponding specimens expressed CD1d protein on the cell surface. Two of five analyzed MB cell lines (DAOY and MHH-MED-8A) also were CD1d-positive. Functional experiments demonstrated that both cell lines effectively presented αGalactosylceramide (αGalCer) to activate NKT-cell cytokine production, proliferation, and cytotoxicity. The effector to target ratio as low as 2:1 was sufficient to kill nearly 100% of MB cells in vitro. The cytotoxicity was CD1d-restricted as it was inhibited by anti-CD1d blocking mAb. A single intracranial injection of 2X106 ex-vivo expanded human NKT cells and αGalCer in NOD/SCID/β2m mice with 6-day established xenografts of luciferase-transduced DAOY cells resulted in rapid tumor regression with 30% cure rate. The therapy was well-tolerated. Therefore, CD1d-positive MB could be targeted for immunotherapy with NKTs that has a curative potential.
Regulatory B lymphocyte production of IL-10 inhibits lymphoma depletion during CD20 immunotherapy in mice (156.27)
Authors: Horikawa Mayuka , Colin-Minard Veronique , Tedder Thomas 
Published: January 2023

Abstract
Non-Hodgkin’s lymphoma (NHL) is a heterogeneous group of malignancies that represents ~4% of all cancers. Rituximab, a chimeric CD20 monoclonal antibody (mAb) that effectively treats half of NHL patients variably depletes non-malignant tissue B cells. Whether endogenous or remnant B cells affect CD20 mAb-induced lymphoma depletion is unknown. To determine whether endogenous B cells can influence in vivo lymphoma therapy in mice, the therapeutic effectiveness of a potent mouse anti-mouse CD20 mAb was compared in wild type and CD20-deficient (CD20-/-) mice given homologous CD20+ primary lymphoma cells. CD20 mAb efficiently depleted endogenous mature B cells and lymphoma cells in wild type mice and prolonged survival significantly. By contrast, endogenous B cells are not depleted in CD20-/- mice, including a rare IL-10-producing regulatory B cell subset (B10 cells). Furthermore, the persistence of endogenous B cells significantly inhibited the anti-tumor effects of CD20 mAb in CD20-/- mice. Even small numbers of adoptively transferred CD20-/- B10 cells dramatically suppressed lymphoma depletion in wild type mice by inhibiting monocyte activation and effector function through IL-10-dependent mechanisms. Thus, endogenous B10 cells are potent negative regulators of innate immunity, with even small numbers of residual B10 cells inhibiting lymphoma depletion by CD20 mAb. This may provide a new explanation for treatment resistance in some lymphoma patients.
Combination of DTIC and peptide-vaccination elicit a renewal of high-avidity late-differentiated T-cell clones in melanoma patients (156.6)
Authors: Palermo Belinda , Del Bello Duilia , Franzese Ornella , Di Donna Cosmo , Gualtieri Novella , Nuzzo Carmen , Proietti Enrico , Catricalà Caterina , Ferraresi Virginia , Nistico Paola 
Published: January 2023

Abstract
Combination of chemo/immunotherapy to increase the effectiveness of the antitumor immune responses associated with a clinical efficacy is an attractive antitumor strategy. We have recently reported that the administration of dacarbazine (DTIC) one day before peptide (Melan-A and gp100)-vaccination in disease-free melanoma patients increases the number of peptide-specific effector-memory CD8+ lymphocytes. Notably, patients treated with DTIC before vaccination, showing a longer survival, display a progressive enhancement of TCR repertoire diversity of Melan-A+ CD8+ clones, accompanied by the maintenance of high-avidity, whereas patients treated with vaccine alone showed a reduction of TCR repertoire diversity and a decline of their tumor lytic activity. To study the phenotype and the functional lytic activity of CD8+ T-cell clones in association with their renewal rate we have analyzed the expression of CD28 and CD27 co-stimulatory molecules, in both Melan-A- and gp100-specific T-cell clones, isolated before and at different times after the treatments. We found newly recruited clones with high-lytic activity and highly-differentiated phenotype only after the combined therapy. Studies are in progress to define the role of replicative senescence in the continuous renewal induced by the combined therapy, by studying activity and regulation of telomerase in endogenous-induced or vaccine-driven clones, in correlation with their different phenotype and anti-tumor lytic capability.
Presence of NLRP3 in myeloid cells increases sensitivity to membrane damage (157.3)
Authors: Heid Michelle , Chu Jessica , Sun Chengqun , Thomas L. , Keyel Peter , Salter Russell 
Published: January 2023

Abstract
Pore forming toxins, such as tetanolysin O (TLO), as well as the potassium ionophore nigericin, stimulate the NLRP3 inflammasome. We and others have shown that NLRP3 inflammasome induced processing and secretion of mature IL-1 beta is dependent on the activity of caspase-1 as well as potassium efflux. Unexpectedly, we now observe that NLRP3 knockout macrophages are more resistant to membrane damage compared to wild type macrophages exposed to nigericin or TLO, as assessed by a dye uptake assay. We also were able to increase sensitivity to membrane damage in myeloid cells deficient in ASC, an adaptor protein required for formation of the NLRP3 inflammasome, by over-expression of ASC using retroviral transduction. Through the use of both pharmacological inhibitors and caspase-1 knockout mice, we observed this NLRP3 and ASC dependent effect on membrane integrity is independent of the activity of caspase-1, unlike the secretion of mature IL-1 beta. However, the loss of membrane integrity is dependent on potassium efflux, as is mature IL-1 beta secretion. These data suggest a mechanism through which the NLRP3 inflammasome is able to influence the membrane integrity of myeloid cells in a caspase-1 independent manner, distinguishing the loss of membrane integrity from IL-1 beta processing and secretion.
Pyrin-only protein 3: a pseudogene product in the human genome regulating NF-kB activity (157.4)
Authors: Porter Kristen , Nyland Patricia , Harton Jonathan 
Published: January 2023

Abstract
Pyrin-only proteins influence NF-kB and inflammasome signaling pathways. In viruses, POPs act as pathogenic factors by reducing innate immune responses and favoring viral survival within the host. In humans, two known POPs are thought to have a role in regulating innate immunity and inflammation. POP1 interferes with IKKα/β activation and prevents the downstream activation of NF-kB with no known effect on ASC-dependent inflammasomes. POP2 however, inhibits NF-kB signaling at the level of the p65 (RelA) subunit and down-modulates the formation and activation of a variety of pyrin domain/ASC-dependent inflammasomes. Here we describe a third human POP, POP3, the product of an NLRP pseudogene encoded on chromosome X that is expressed in a wide variety of tissues. Like its close genetic relative POP2, POP3 inhibits p65 activation downstream of IKK, but lacks the capacity of inhibit the NLRP3 inflammasome. Our data suggest that the human POP family has three members, all of which may play distinct biological roles during inflammation.
miR-144 is a negative regulator of the host antiviral response (158.10)
Authors: Rosenberger Carrie , Podyminogin Rebecca , Peschon Jacques , Rodriguez David , Gundapuneni Madhumati , Weiss Mitchell , Aderem Alan 
Published: January 2023

Abstract
Antiviral responses must be regulated to rapidly defend against infection while minimizing inflammatory damage, but the mechanisms for establishing the magnitude of response within an infected cell are not well understood. microRNAs are post-transcriptional regulators that are attractive candidates for fine tuning the magnitude of the immune response. We identify miR-144 as a negative regulator of the host antiviral response in the lung. We observed that miR-144 expression resulted in increased replication of influenza, EMCV, and VSV in primary mouse lung epithelial cells. We employed a systems biology approach to identify the transcriptional network regulated by miR-144 and demonstrate that it controls the TRAF6/IRF7 antiviral response by post-transcriptionally suppressing TRAF6 expression. Infection of IRF7-null mice revealed that IRF7 is necessary for normal expression of the network of genes that is perturbed by miR-144. Ectopic IRF7 expression restored the antiviral capacity of cells over-expressing miR-144 and reducing TRAF6 levels using shRNAs recapitulated the miR-144 phenotype. In vivo ablation of miR-144 resulted in reduced influenza replication within the lung. These data suggest that miR-144 regulates activation of the IRF7 pathway, thereby creating an exquisitely controlled antiviral response.
MicroRNA-155 enhances ocular inflammation after Pseudomonas aeruginosa infection (158.11)
Authors: Huang Xi , Peng Anping , Sun Mingxia , Wu Minhao , Zhang Ping 
Published: January 2023

Abstract
Abstract: As a gene regulator, microRNA (miR)-155 plays an important role in both autoimmune inflammation as well as antiviral innate immunity. However, the role of miR-155 in Pseudomonas aeruginosa (PA) keratitis remains unknown. Our study demonstrated that miR-155 expression levels were significantly up-regulated in PA-infected vs normal human corneas as well as Macrophages derived from PBMC. Furthermore, in vivo studies showed that miR-155 expression was significantly increased in PA-infected corneas of C57BL/6 (B6) mice. Inhibition of miR-155 led to reduced ocular disease after PA infection, by down-regulating pro-inflammatory cytokine (e.g., IL-1β, IFN-γ, IL-6, IL-12) production. In vitro studies also demonstrated that PA challenge induced miR-155 expression in a time- and dose-dependent manner in peritoneal Macrophages and RAW264.7 cells. The induction of miR-155 was dependent on TLR2, TLR4, TLR9, through NF-κB, IKK, JNK and PI3K, but not P38 or ERK pathways. Moreover, overexpression of miR-155 significantly decreased Macrophage-mediated phagocytosis and intracellular bacterial killing of PA, by down-regulating expression of mannose receptor (MR) and complement receptor 3 (CR3). miR-155 overexpression also enhanced the production of pro-inflammatory cytokines including IFN-γ, IL-1β, IL-6, IL-12, IL-17A, but reduced anti-inflammatory cytokine IL-10 expression. Collectively, these data indicated that miR-155 functions as an inflammatory amplifier in PA keratitis.
Haplotype specific activating NK cell receptors provide recognition of virus-infected cells and mediate resistance to infection (158.7)
Authors: Pyzik Michal , Charbonneau Benoit , Gendron-Pontbriand Eve-Marie , Babic Marina , Krmpotic Astrid , Jonjic Stipan , Vidal Silvia 
Published: January 2023

Abstract
The ability of numerous activating Ly49 receptors to recognize mouse cytomegalovirus (MCMV) infected cells of various H2 background was examined. Reporter cells expressing either Ly49P1 from NOD/Ltj mice, Ly49L from BALB mice or Ly49D2 from PWK/Pas mice were activated upon co-culture with MCMV infected cells provided the presence of an appropriate H2 molecule and the m04/gp34 viral peptide. Such recognition was most frequent in the H2k background, yet also occurred in the H2d and H2f contexts. To demonstrate whether Ly49 activating receptors confer MCMV resistance, H2 congenic BALB mice were infected with MCMV. Neither BALB.By (H2b) nor BALB/c (H2d) mice were able to control MCMV spread in target organs contrary to BALB.K (H2k) mice, which demonstrated delayed NK cell-mediated virus control. In BALB mice, the antibodies YE1/48 and 12A8 delineate a Ly49L+ NK cell population (YE1/48INT12A8-). BALB.K resilience to the infection correlated with the expansion of the specific Ly49L+ NK cell fraction in the spleen and secretion of IFNγ 6 days after MCMV infection. Adoptive transfer of Ly49L+ but not Ly49L- NK cells provided partial protection from death in neonate BALB.K mice following MCMV infection. Thus, we have uncovered three activating Ly49 receptors partaking in the recognition of MCMV infection by NK cells through an H2- and m04-dependent mechanism, which we find to be more common than the direct recognition of m157.
Elevated numbers and frequencies of CD56- CD3- NK cells with high perforin occur in healthy donors (160.14)
Authors: Hudig Dorothy , Barao Isabel , Hunter Kenneth , Diamond W. , Redelman Doug 
Published: January 2023

Abstract
Routinely, human natural killer (NK) lymphocytes are identified as CD3eneg CD56bright or dim. CD3neg CD56neg cells with low perforin are found in viremic patients infected with human immunodeficiency virus and hepatitis C virus (reviewed, N.K. Bjorkstorm et al., Trends Immunol. 31:401, 2010). To our surprise, we found by flow cytometry that CD3neg perforin(PRF1) positive NK cells included as many as 50% CD56neg cells within the blood of 97 healthy subjects. Among these subjects (aged 21 to 62 years), the CD3eneg PRF1positive cells included at least 10% CD56neg cells in 24 of 97 (~25%) subjects. CD56neg perforin high CD3neg cells were over 50 cells/mm3 of blood in 14% of the healthy donors. As anticipated, the CD56dim cells had ~7 fold higher perforin MFI than the CD56bright cells. Notably in the healthy donors with high frequencies of CD56neg cells, these cells had bimodal perforin levels comparable to the levels in the CD56bright and CD56dim subsets. The perforin levels indicate that many CD56neg NK cells may have strong cytotoxicity in healthy donors in contrast to the low perforin and cytotoxicity observed in CD56neg NK cells of the viremic patients. We conclude that CD56neg NK cells now warrant attention as potential cytotoxic cells after receiving considerable attention as high chemokine-secreting cells.
Impact of T cell receptor reactivity and HLA phenotype on naive CD8 T cell frequency in humans (160.16)
Authors: Bonneville Marc , Legoux Francois , Saulquin Xavier 
Published: January 2023

Abstract
The impact of major histocompatibility complex (MHC) phenotype on the shaping of the peripheral naive T cell repertoire in humans remains unknown. Here we exploited a pMHC multimer-based sorting approach to analyze the impact of expression of a given human leukocyte Ag (HLA) class I allele on the frequency and quality of peripheral blood and thymic T cells directed against immunodominant self, viral and tumor antigens presented by a human MHC class I allele (HLA-A*02, referred to as A2) in individuals expressing or not this allele. Naive T cell frequencies varied from one antigen specificity to another but were restrained for a given specificity. While A2-restricted T cells showed similar repertoire features and antigenic avidities in A2+ and A2- donors, A2 expression had either a positive, neutral or negative impact on the frequency of A2-restricted naive CD8 T cells, depending on their fine specificity. We also identified in all donors CD4 T cells specific for A2/peptide complexes, whose frequencies were not affected by MHC class I expression, but nevertheless correlated with those of their naive CD8 T cell counterparts. Therefore both selection by self MHC and inherent TCR reactivity regulate the frequency of human naive T cell precursors. Moreover this study also suggests that T cell repertoire shaping by a given self MHC allele is dispensable for generation of immunodominant T cell responses restricted by this particular allele.
Thymocyte specific interleukin-7 restores T cell development but not homeostasis in IL-7KO mice (160.6)
Authors: Kim Grace , Lucas Philip , Gress Ronald , Park J. 
Published: January 2023

Abstract
T cells are critically dependent on interleukin-7 (IL-7) as IL-7-deficient mice fail to generate and maintain a functional T cell compartment. Importantly, IL-7 is not produced by T cells and it is only provided by dendritic cells, stromal cells and other non-T cells in trans. Using an lck-proximal enhancer/promoter driven IL-7 transgene (IL-7Tg), here we tested whether T-lineage specific expression of IL-7 can restore T cell development in IL-7 deficient mice. We found that T cell development was completely restored in IL-7Tg.IL-7KO mice as thymocyte numbers, thymic selection, and CD4/CD8 lineage differentiation were identical to WT mice. Importantly, however, IL-7Tg.IL-7KO mice were peripheral T lymphopenic with less than 20% of T cell numbers compared to WT controls. This was presumably induced by termination of transgenic IL-7 expression upon positive selection as qPCR analysis showed a rapid downregulation of IL-7 mRNA expression in mature thymocytes. Surprisingly, the few surviving peripheral T cell in IL-7Tg.IL-7KO mice did not show upregulation of activated or memory T cell markers,and we did not observe any autoimmunity. Further characterization of the function and phenotype of these T cells are currently under progress. Collectively, these data show that IL-7 expression and signaling in cis is sufficient for normal T cell development, and they show in a new in vivo model of peripheral T lymphopenia that continuous IL-7 is critical T cell homeostasis.
CD40 engagement ablates CTLA-4 expression on Th40 effector cells: a mechanism for breach of tolerance (160.9)
Authors: Wagner David , Carter Jessica , Vaitaitis Gisela , Waid Dan 
Published: January 2023

Abstract
Biomarkers defining pathogenic effector T cells have been slowly forthcoming. Towards this we identified CD4+ cells that express CD40 (Th40), and defined these cells as pathogenic in the NOD type 1 diabetes (T1D) model. Th40 cells are unique in their ability to adoptively transfer T1D. To study the origin of Th40 cells and disease pathogenesis we employed a dual transgenic model expressing OVA323-339 peptide as a neo-self antigen on islet beta cells and medullary thymic epithelial cells (mTEC) and a transgenic TCR recognizing the OVA323-339 peptide. Th40 cells and Tregs that both recognize the same cognate antigen, rather than being deleted through central tolerance drastically expand. DO11.RIPmOVA mice develop an autoimmune-type diabetes very rapidly. In pancreatic lymph nodes Th40 cells are greatly expanded while Tregs remain equivalent to controls and importantly remain functional throughout the disease process, suggesting another breach of tolerance mechanism. DO11.RIPmOVA Th40 cells do not express CTLA-4, a molecule for cell auto-regulation, while DO11 Th40 cells do. CD3/CD28 engagement induces CTLA-4 cell surface expression on DO11 Th40 cells while CD40 co-engagement ablates that induction. These data suggest a unique means for Th40 cells to thwart tolerance.
Lymphatic pump treatment enhances pulmonary immunity and protects against solid tumor formation in the rat lung. (161.14)
Authors: McCauley Lyndsey , Pedrueza Mayela , Zhang Xin , Jones Harlan , Hodge Lisa 
Published: January 2023

Abstract
Lymphatic pump treatments (LPT) are used by osteopathic physicians to treat edema and infection. In animals, LPT increases lymph flow and leukocyte numbers, which may enhance protection against infection and cancer. To determine if LPT would enhance anti-tumor immunity and reduce tumor formation in the lungs, F344 rats were injected intravenously with MADB106. Rats received no treatment (control), 4 min of light touch under anesthesia (sham), or 4 min of LPT under anesthesia, for 7 days. At day 8, tissues were measured for tumors and leukocyte numbers/activities. Serum was measured for vascular endothelial growth factor C (VEGF-C). LPT reduced solid tumors and increased B cell, CD4+ T cell, CD8+ T cell, NK cell and macrophage numbers in the lungs. In addition, Leukocytes isolated from the lungs of LPT rats secreted more IL-2, IL-10 and IFN-γ but did not enhance tumor lysis in vitro. Previously we demonstrated LPT mobilizes gut-derived lymphocytes into lymphatic circulation; therefore, we stained pulmonary lymphocytes for the gut marker α4β7. LPT increased α4+β7+ leukocytes in the lungs, suggesting LPT mobilizes gut-derived leukocytes into the lungs. Finally, there were no differences in the levels of VEGF-C between treatment groups, suggesting that LPT does not promote lymphangiogenesis in this model. Our data suggests LPT inhibits pulmonary tumor formation by enhancing the numbers of leukocytes with anti-tumor activities that migrate into the lungs.
Defining the phenotype, migration, and longevity of true tissue lung CD8 T cells. (161.15)
Authors: Anderson Kristin , Skon Cara , Vezys Vaiva , Masopust David 
Published: January 2023

Abstract
Thus far, CD8 T cell responses to respiratory infections have been restricted to broad tissue analyses, which may not accurately represent the true nature of the tissue, nor the response to infection. In order to thoroughly explore the dynamics of the CD8 T cell response, we used respiratory and systemic infection models to perform a detailed analysis of the anatomic location, phenotype, migration capacity, and longevity of CD8s in the lung. Interestingly, we found that as many as 98% of the T cells that would commonly be perceived to be in the tissue were actually in the capillaries. We show that the phenotype of cells in the vasculature is distinct from cells in the tissue and the airway, and that migration to the tissue is chemokine dependent. Additionally, local infections generate a memory CD8 population with a tissue distribution different than a systemic infection. These findings have implications for establishing and maintaining CD8 T cell dependent immunity within the lung.
Interleukin-33 and its receptor ST2 contribute to inflammation in the lungs of patients with idiopathic pulmonary fibrosis (161.16)
Authors: Pickering Edward , Todd Nevins , Lockatell Virginia , Atamas Sergei , Luzina Irina 
Published: January 2023

Abstract
Interleukin (IL)-33 and its receptor ST2 have been recently implicated as critical regulators of responses to injury and of ensuing inflammation. The roles of IL-33 and ST2 in pulmonary inflammation and scarring remain unclear. We analyzed bronchoalveolar lavage (BAL) from patients with idiopathic pulmonary fibrosis (IPF), a disease characterized by inflammation, with known elevated levels of IL-8 and excessive collagen deposition in the lungs. IL-33 was not found in BAL from 5 controls but was found in 7 out of 11 IPF samples, at 10.0 ± 3.2 pg/ml. Soluble ST2 was found in BAL from healthy controls at 93.2 ± 82.1 pg/ml, and in all IPF samples at 352.1 ± 135.2 pg/ml (p &lt; 0.05, Mann-Whitney U test). The elevation of IL-33 in BAL suggested that this cytokine may act directly on pulmonary epithelial cells and through this action contribute to inflammation. Cultured human primary pulmonary epithelial cells were stimulated with rhIL-33, and the levels of pro-inflammatory cytokines IL-6, IL-8, MIP-1α, and TNF-α in cell culture media were analyzed by ELISA. Of the analyzed cytokines, only the levels of IL-8 were increased by 2.0 ± 0.2 fold at 24 h and 1.7 ± 0.2 fold at 48 h. In conclusion, levels of IL-33 and ST2 are elevated in the lungs of IPF patients; IL-33 may contribute to inflammation and fibrosis by activating IL-8 production in pulmonary epithelium.