bluuebunny/crossref_metadata_2025_split huggingface dataset:
Simultaneous detection of mouse FoxP3, IL-17 and surface markers by multi-color flow cytometry (96.18)
Authors: Li Li , Wu Xiaowei , Wei Aili , Rohrer Jurg 
Published: January 2023

Abstract
Naïve CD4+ T helper cells can be induced to differentiate towards at least four types of committed helper T cells, namely T helper 1 (Th1), Th2, Th17 and regulatory T cells (Treg) according to the local cytokine milieu. The committed cells are characterized by the expression of specific transcription factors and by the cytokines released. Th17 cells are very efficient inducers of tissue inflammation and crucial initiators of organ specific autoimmunity. FoxP3+ Treg cells are necessary and sufficient to prevent autoimmunity. There is a reciprocal relationship between pathogenic Th17 cells and Foxp3+ Treg cells. Therefore the need to simultaneously detect FoxP3 and IL-17 is becoming increasingly important. Since mouse FoxP3 is sensitive to fixation buffer, it is important to use the appropriated buffers and protocols for its detection. In this report, we described the development of a fix/perm buffer system and protocol for simultaneous detection of mouse FoxP3 and IL-17 with cell surface markers CD4 and CD25. Using the optimized buffer and storage conditions, staining can be done on freshly fixed cells or stored fixed cells. In addition, effects of temperature during the fix/perm steps and the sensitivity of certain clones to the fixation process will be discussed.
Acquistion of Effector and Memory Responses is Reciprocally Regulated by IL-12 and IFN-α/β in Human CD8+ T Cells (96.8)
Authors: Ramos Hilario J , Davis Ann M , Cole Alexander G , Farrar J. David 
Published: January 2023

Abstract
The innate cytokines IL-12 and IFN-α/β act in concert to facilitate clearance of intracellular pathogens. However their precise role in promoting human T cell responses is still unclear. In this study, we demonstrate that these cytokines regulate distinct aspects of effector and memory human CD8+ T cell differentiation. We found that IL-12 programs the development of TEM cells which secrete high levels of IFN-γand TNF-α and attain strong cytolytic activity. Programming of TEM cells by IL-12 correlated to the induction of the IL-12 receptor β 2 (IL-2Rβ2) and was associated with cells which had undergone rapid division. Alternatively, IFN- α/β enhanced a population of TCM cells which expressed high levels of the lymph node homing receptor CCR7. This population did not display immediate effector activity however, upon secondary activation, were capable of rapid proliferation and acquisition of both memory and effector phenotypes. TCM development was regulated by IFN-αand proceeded by slowing the progression of cell division in a subpopulation of cells that selectively expressed elevated IFN- α/βreceptor-2 (IFNAR2). We further examined the effects of the strength of TCR signal on TEM and TCM development. In the presence of both IL-12 and IFN- α/β, these cytokine signals were amplified as the strength of the TCR signal was increased, promoting the simultaneous development of both TCM and TEM. Together, our results support a novel model in which IL-12 and IFN- α/βact in a non-redundant manner to regulate the co-linear generation of both effector and memory cells.
This study is supported by a grant from the National Institutes of Health/National Institute of Allergy and Infectious Disease (AI56222) and by a predoctoral fellowship from the National Instititue of Health/Nation Institute of Allergy and Infectious Disease (AI68622).
IL-4 regulates CD8+ T cell homeostasis in BALB/c but not C57BL/6 mice (97.8)
Authors: Morris Suzanne C. , Ruwe Stephanie , Scott Jane , Finkelman Fred D. 
Published: January 2023

Abstract
Because IL-4 potently stimulates murine CD8+ T cell proliferation during an in vivo Th2 response, we evaluated whether IL-4 also contributes to CD8+ T cell homeostasis in naïve mice. Adult, wild-type BALB/c mice have ~2x the number of total and ~5x the number of memory phenotype (CD44highLy6Chigh) splenic CD8+ T cells as IL-4-, IL-4Rα- and Stat6-deficient BALB/c mice. Rare, IL-4-dependent, memory phenotype CD8+ T cells are found in BALB/c thymus by week 2 of life. The effects of IL-4-deficiency on CD8+ T cells can be reproduced by treating pregnant mice and then their offspring with anti-IL-4 mAb for the first 12 days of life, but not by treating adult mice with anti-IL-4 mAb for 2 weeks. In contrast to BALB/c mice, wild-type, IL-4-deficient and IL-4Rα-deficient C57BL/6 mice have similar numbers of memory phenotype CD8+ splenocytes, which are fewer than those found in wild-type BALB/c mice. BALB/c and C57BL/6 adult mice have similar basal IL-4 production and their memory phenotype CD8+ T cells are similar in size and surface markers, except that the C57BL/6 cells express more IL-15Rα and IL-2/15Rβ. At least 3 genes contribute to the BALB/c vs. C57BL/6 difference. Taken together, these observations suggest that IL-4 acts early in BALB/c T cell development to promote memory phenotype CD8+ T cell differentiation and that greater thymic IL-4 production or responsiveness may contribute to the BALB/c Th2 cytokine predisposition.
Inhibition of anti-apoptotic signals in prostate cancer cells by CCL25 blockade enhances the cytotoxic effects of etoposide (98.1)
Authors: Sharma Praveen K , Singh Rajesh , Eaton John W , Lillard, Jr. James W , Novakovic Kristian R , Grizzle William E , Singh Shailesh 
Published: January 2023

Abstract
Despite of current advancement in treatment and management of prostate cancer (PCa); it is still remaining the second leading cause of cancer-related deaths among men in the US. Current chemotherapeutic regimens provided palliative benefit but relatively modest survival advantage for the patients with hormone refractory PCa. This results in an initial response, but always followed by relapse due to chemoresistance and relatively low rate of apoptosis in PCa cells. Therefore, better efficacy of chemotherapeutics can be achieved by combining additional agents, which can block the mechanisms responsible for this low apoptosis rate. We have previously shown that CCR9 is expressed by the PCa cells and support the mechanisms involved in PCa metastasis. In this study, we have demonstrated that CCR9 and its natural ligand; CCL25 interaction up regulates anti-apoptotic proteins and promotes cell survival. Inhibition of these interactions using antibody blocked enhanced the etoposide-induced apoptosis of PCa cells in vitro and reduces tumor burden in vivo. Therefore, targeting this axis with chemotherapeutic agents may enhance the therapeutic outcomes of PCa. [Supported by funds from DOD PCRP Award W81XWH-06-1-0521]
Anti-inflammatory properties of docosahexaenoic acid (DHA) in bone marrow-derived dendritic cells (98.11)
Authors: Kong Weimin , Yen Jui-Hung , Ganea Doina 
Published: January 2023

Abstract
Omega 3 fatty acids have anti-inflammatory effects in chronic inflammatory and autoimmune diseases. Previous studies showed that dendritic cells (DC) differentiated in the presence of DHA express reduced costimulatory molecules and secrete lower levels of cytokines. We report that 24h exposure to DHA of immature DC prevents LPS-induced upregulation of MHCII, CD40, CD80 and CD86. Immature DC are characterized by CCR5 expression and migration to proinflammatory chemokines, whereas mature DC express CCR7 and migrate to lymph nodes in response to CCL19/21. DHA exposure kept CCR5 expression at high level, and prevented CCR7 expression induced by LPS. DHA-treated DC migrated poorly towards CCL19. Proinflammatory cytokines including IL-12p70, IL-23, IL-6 and TNFα produced by mature DC were inhibited by DHA. We found that DHA inhibited IL-12p70 and IL-23 production induced by ligands for TLR4 and 2, and also by ligands for the intracellular TLR3 and 9. In agreement with previous reports showing that omega 3 fatty acids bind to PPARγ, we found that DHA induced nuclear PPARγ translocation, and that a PPARγ antagonist partially reversed the inhibitory effect of DHA on IL-12p70. In conclusion, our studies indicate that exposure to DHA maintains already differentiated DC in an immature state and prevents expression of proinflammatory cytokines and chemokines.
Support NIH 2RO1AI052306.
Interleukin-18 Downregulates Collagen Production in Human Dermal Fibroblasts via ERK pathway (98.14)
Authors: Kim Hee Jung , Song Seok Bean , Lee Jun Young , Cho Baik Kee , Cho Dae Ho , Park Hyun Jeong 
Published: January 2023

Abstract
Excessive accumulation of collagen contributes to the fibrotic process. Several experimental studies have shown that interferon (IFN)-γ is effective in preventing fibrogenesis. Interleukin-18, formerly called IFN-γ -inducing factor, is a key mediator of inflammation and host defense responses. Here, we investigated the regulatory effect of IL-18 on expression of type I and collagen genes in dermal fibroblasts. Exposure of human dermal fibroblasts (HDF) to IL-18 resulted in a reduction of collagen gene expression and protein production was in a dose-dependent manner using real-time RT-PCR and Sircol assay. Next, we determined the signaling pathways involved by small interfering RNA(siRNA), reporter assay, and Western blotting. Silencing of the Ets-1 reversed inhibitory effects of IL-18. We demonstrated that IL-18 induced phosphorylation of extracellular signal-regulated kinase (ERK) within 10 minutes, and inhibitor of ERK blocked the inhibitory effects of IL-18. These results suggest that L-18 downregulates collagen production in HDF directly via Ets-1-dependent pathway and ERK pathway.
Relationship between Amyloid β (Aβ) Protein and Inflammatory Markers in Down Syndrome (DS) (98.19)
Authors: Mehta Pankaj D. , Patrick Bruce A. , Mehta Sangita P. , Barshatzky Marc , Dalton Arthur J. 
Published: January 2023

Abstract
Brain autopsy data from persons with DS showed that the neuropathology of Alzheimer disease (AD) is always present in DS 40 years of age and older. The core protein of the neuritic plaque is Aβ protein. Studies have suppoted the role of immune abnormalities and inflammation in the pathogenesis of DS. We hypothesized that the levels of TNFα and Cystatin C would be higher in DS than controls, and would correlate with Aβ levels. The study included 40 persons with DS (44±8 years old) and age-matched controls. Plasma levels of Aβ40 and Aβ42 , TNFα and Cystatin C were quantitated by using an ELISA. Aβ40 and Aβ42, TNFα and Cystatin C levels were higher in DS than controls (p&lt;.001). Although there was significant relation between Aβ40 and Aβ42 levels (r=.6; p&lt;.01)in DS, there was no relation between Aβ40 and Aβ42 levels with TNFα or Cystatin C. The higher TNFαand Cystatin C levels indicate a chronic state of immune activation in DS. Because there was no relation between Aβ and TNFα or Cyststin C the data suggests that increases in TNFα and Cystatin C reflect more infections in DS than AD neuropathology.
Analysis of proinflammatory cytokines as biocompatibility markers of emdogain (98.21)
Authors: SOLIS SOTO JUAN MANUEL , NAKAGOSHI CEPEDA SERGIO EDUARDO , GARZA ENRIQUEZ MARIANELA 
Published: January 2023

Abstract
Emdogain are enamel matrix proteins (amelogenins) that are taken from developing teeth in pigs. It is indicated for the treatment of intra-bony defects due to moderate or severe periodontitis, as well as the surgical treatment of gum recessions. However, its physiological effects are not completely characterized. This work analyzes the effects of emdogain on the production of proinflammatory cytokines and mineralization factors. Monocytes, isolated from peripheral blood, were incubated with different concentrations of emdogain for 2 days. Immunocytochemical methods were used to detect IL-1beta, IL-6, IL-12, TNF-alpha, and IFN-gamma. Fibroblasts, isolated from gingival tissue, were incubated with different concentrations of emdogain for 7 days. Immunocytochemical methods were used to detect osteopontin, osteonectin, osteoprotegerin, and ameloblastin. Morphometrical analysis showed significant increased production of osteopontin (p&lt;0.01), osteoprotegerin (p&lt;0.05) and osteonectin (p&lt;0.05). Ameloblastin production was similar to controls. Of the proinflammatory cytokines only IL-12 (p&lt;0.001) and IL-6 (p&lt;0.05) were produced in considerable amounts. Production of IL-1beta, IFN-gamma and TNF-alpha was similar to controls. Emdogain has a good biocompatibility, because it only increases the production of pleiotropic cytokines. Besides, it stimulates the production of factors of mineralization like osteopontin, osteoprotegerin and osteonectin.
Collagen antibody-induced arthritis (CAIA) in mice: Triggering of arthritis by HMGB1, a late stage lethal mediator of LPS (99.22)
Authors: Waritani Takaki , Cutler Dawn , Terato Kuniaki 
Published: January 2023

Abstract
High mobility group box 1 protein (HMGB1), identified as a late stage lethal mediator of endotoxin, is currently considered a pro-inflammatory cytokine involved in severe inflammatory reactions such as rheumatoid arthritis. Pathological roles of HMGB1 in collagen antibody-induced arthritis (CAIA) model in mice were studied. DBA/1 mice were injected with sub-arthritogenic dose of anti-type II collagen monoclonal antibody (mAb) cocktail on day 0, and then LPS (50 μg) or HMGB1 (500 μg) by IP injection on day 3. Mice receiving HMGB1 developed arthritis (score: 6-8 out of 16 on day 7-10), although the severity of arthritis is milder than mice triggered by LPS (score: 14-16). Importantly, an anti-HMGB1 mAb capable of preventing death of mice by absorption of lethal dose of LPS prevented the development of arthritis by HMGB1, but failed to prevent the arthritis triggered by LPS, indicating that LPS stimulates a variety of inflammatory pathways. On the other hand, other clones of anti-HMGB1 mAb failed to prevent the arthritis induced by HMGB1, indicating the importance of epitope specificity for neutralizing the biological activity of HMGB1. This new arthritis model induced by the combination of mAb cocktail and HMGB1 provides a new tool to study mechanisms of inflammatory reactions linked to toll-like receptors (TLR) 2/4 and/or receptor for advanced glycation end-products (RAGE) in autoimmune mediated arthritis.
IL-21 is increased in CD4+ T cells and serum from patients with systemic lupus erythematosus (99.36)
Authors: Nguyen Vinh , Siaton Bernadette , Rus Violeta 
Published: January 2023

Abstract
Studies in animal models of lupus have reported increased production of IL-21 in MRL/lpr or BXSB mice and attenuation of autoimmune features in mice treated with IL-21 blocking agents. Little is known about IL-21 production and function in patients with systemic lupus erythematosus (SLE). We determined IL-21 levels in serum samples from 28 patients with SLE, 8 disease and 16 healthy controls by ELISA. Mean level of IL-21 was significantly higher in SLE patients (440±372 pg/ml) compared to normal controls (114 ± 101 pg/ml; p=0.05) and inversely correlated with C3 levels. Furthermore, patients with active disease (SLEDAI &gt;4) had higher levels of serum IL-21 (1332.7±1084pg/ml) than patients with inactive disease (217.6±184 pg/ml). IL-21 and IL-21R expression was determined on ex vivo and in vitro stimulated CD4+ T cells by RT-PCR and immunostaining. IL-21 mRNA expression was 7 fold higher in ex vivo CD4+ T cells while intracellular levels were higher in stimulated memory CD4+ T cells from lupus patients compared to controls. IL-21R expression was upregulated on in vitro stimulated CD4+ T cells in both patients and controls. CD4+ T cells from lupus patients had equal susceptibility to proliferative effects induced by IL-21 when compared with controls. Similarly, lupus B cells were equally responsive to IL-21 when compared to controls with respect to proliferation, plasma cell differentiation, IgG and IL-10 production. In conclusion, elevated levels of IL-21 in lupus patients may promote auto-reactive CD4+ T cells proliferation as well as B cell proliferation and differentiation, thus contributing to the generation of T cell dependent autoreactive B cell responses.
Burkitt lymphoma-associated molecules impair HLA class II activation of CD4+ T cells (100.20)
Authors: God Jason , Cameron Christine , Amria Shereen , Haque Azizul 
Published: January 2023

Abstract
Burkitt Lymphoma (BL) is a very fast growing form of non-Hodgkin's lymphoma. The common characteristic of all BL is the translocation and constitutive activation of the c-myc oncogene. Our present study suggests that BL and Epstein-Barr Virus-transformed B-lymphoblastoid cell lines (B-LCL) constitutively express HLA class II proteins and bind peptide ligands. However, unlike B-LCL, BL cells were not capable of activating CD4+ T cells. While BL and B-LCL possessed similar levels of HLA class II, invariant chain (Ii) and HLA-DM, BL cells expressed lower levels of co-stimulatory molecules (CD80/CD86). Biochemical and functional assays showed that addition of external co-stimulatory signals to BL cells failed to functionally deliver HLA class II antigens to CD4+ T cells. Although both BL and B-LCL were capable of binding peptides at neutral and acidic pH, BL cells were only able to activate T cells at low pH (5.5), suggesting that BL-associated inhibitory molecules were eluted at the acidic pH. BL conditioned medium, as well as acid eluent from BL cells and primary B-cell tumors, inhibited functional class II presentation by both B-LCL and mature dendritic cells, while those eluent from B-LCL and primary B-cells did not. No characteristic inhibitory cytokines were identified in the acid eluent from BL or primary B-cell tumors. Taken together, these results suggest that BL-associated molecules may inhibit HLA class II activation of CD4+ T cells.
Selective depletion of CD4+CD25+Foxp3+ regulatory T cells by low-dose cyclophosphamide is explained by reduced intracellular ATP levels (100.23)
Authors: Huang Bo 
Published: January 2023

Abstract
Adenosine-5'-triphosphate (ATP), the energy molecule and signal element, significantly affects nearly all major biological processes of T cells. Recently, CD4+CD25+Foxp3+ regulatory T (Treg) cells have been demonstrated to play important roles in mediating cancer development and viral infections. Although Cyclophosphamide (CY) has shown promise as a drug to selectively target Treg cells with low-dose in vivo, the underlying molecular mechanism of this selective effect remains unclear. In this report, we provide evidence that the low intracellular ATP concentration accounts for the selective depletion of Treg cells by low-dose CY. Compared to conventional T cells, much lower ATP levels were found in Treg cells. This was due to low expression of one microRNA, miR-142-3p, leading to high adenylyl cyclase 9 to convert more ATP to cAMP, and CD39 on the surface of Treg cells to hydrolyze ATP. As a result, the low level of ATP attenuates the synthesis of glutathione, leading to the decrease of CY detoxification, thus increasing the sensitivity of Treg cells to low-dose CY. Thus, we here identify a molecular pathway through which low-dose cyclophosphamide selectively ablates Treg cells. Our findings also imply that low levels of ATP are probably related to Treg cell function.
Tumor microenvironments direct the recruitment and expansion of human Th17 cells (100.28)
Authors: Peng Guangyong , Su Xinming , Ye Jian , Hsueh Eddy , Zhang Yanping , Hoft Daniel 
Published: January 2023

Abstract
Although Th17 cells play critical roles in the pathogenesis of many inflammatory and autoimmune diseases, their prevalence among tumor-infiltrating lymphocytes (TILs) and function in human tumor immunity remains largely unknown. We have recently demonstrated high percentages of Th17 cells in TILs from ovarian cancer patients, but the mechanisms of accumulation of these Th17 cells in the tumor microenvironment are still unclear. In this study, we further showed elevated Th17 cell populations in the TILs obtained from melanoma, breast and colon cancers, suggesting that development of tumor-infiltrating CD4+ Th17 cells may be a general feature in cancer patients. We then demonstrated that tumor microenvironmental RANTES and MCP-1 secreted by tumor cells and tumor-derived fibroblasts mediate the recruitment of Th17 cells. In addition, we found that tumor cells and tumor-derived fibroblasts produce a proinflammatory cytokine milieu as well as provide cell-to-cell contact engagement that facilitates the generation and expansion of Th17 cells. We also showed that inflammatory TLR and NOD2 signaling promote the attraction and generation of Th17 cells induced by tumor cells and tumor-derived fibroblasts. These results identify Th17 cells as an important component of human TILs, demonstrate mechanisms involved in the recruitment and regulation of Th17 cells in tumor microenvironments, and provide new insights relevant for the development of novel cancer immunotherapeutic approaches.
Cancer-induced Immunostimulatory CD11b+Gr-1+ cells (100.30)
Authors: Shin Tahiro , Guo Miao , Curiel Tyler , Tomihara Kei 
Published: January 2023

Abstract
Both innate and adaptive immune systems are considered equally important for cancer prevention, immunosurveillance, and control of cancer progression. It is known that while both systems initially eliminate emerging tumor cells efficiently, tumors eventually escape immune attack by a variety of mechanisms including differentiation and recruitment of immunosuppressive CD11b+Gr-1+ myeloid suppressor cells into the tumor microenvironment. However, we show here that CD11b+Gr-1+ cells found in ascites of epithelial ovarian cancer-bearing mice at advanced stages of disease are immunostimulatory rather than being immunosuppressive. These cells consist of a homogenous population of cells that morphologically resemble neutrophils. Moreover, like dendritic cells, immunostimulatory CD11b+Gr-1+ cells can strongly cross-prime, augmenting the proliferation of functional CTLs via signaling through the expression of costimulatory molecule CD80. Adoptive transfer of these immunostimulatory CD11b+Gr-1+ cells from ascites of ovarian cancer-bearing mice results in the significant regression of subcutaneous tumors even without being pulsed with exogenous tumor antigen prior to adoptive transfer. We now show for the first time that adaptive immune responses against cancer can be augmented by these cancer-induced granulocyte-like immunostimulatory myeloid (CD11b+Gr-1+) cells, thereby mediating very effective anti-tumor immunity.
Impact of bortezomib-induced proteasome inhibition in vivo on anti-tumor T cell responses (101.1)
Authors: Shanker Anil , De Kluyver Rachel , Sayers Thomas 
Published: January 2023

Abstract
The perturbation of cellular homeostasis offers a new approach for cancer therapy. Our recent reports showed that proteasome inhibitor bortezomib (Velcade/PS-341) not only sensitizes tumors to death receptor-mediated apoptosis but also increases their susceptibility to NK cell lysis. However, the effects of bortezomib on T cell anti-tumor responses are not known. Thus, in mice established with Renca-HA tumor expressing influenza virus hemagglutinin (HA) as a surrogate tumor antigen, we evaluated the anti-tumor functions of adoptively transferred monoclonal HA-specific CD4 and CD8 T cells following bortezomib administration. Interestingly, no inhibition of T cell proliferation or cytolytic activity in vivo was observed following bortezomib treatments. Nor were there any defects in dendritic cell activation or antigen presentation in bortezomib-treated mice. However, a decrease in total T cell yield and a blockade of cellular homeostatic proliferation was noted in these mice. Thus, bortezomib-inhibitory effects on T cell responses appeared quantitative rather than qualitative. Moreover, combined treatment of mice established with Renca-HA tumor with adoptive T cell transfer and bortezomib resulted in superior anti-tumor responses than either therapy alone. These data demonstrate that, with an appropriate scheduling, it may be possible to combine bortezomib-induced sensitization of tumors to apoptosis with tumor-specific T cell adoptive transfers to improve cancer immunotherapy.