bluuebunny/crossref_metadata_2025_split huggingface dataset:
Translational control of memory CD8 T cell differentiation (LYM4P.752)
Authors: Araki Koichi , Konieczny Bogumila , Morita Masahiro , Sonenberg Nahum , Ahmed Rafi 
Published: January 2023

Abstract
CD8 T cells play an essential role in controlling viral as well as intracellular bacterial and parasitic infections. Memory CD8 T cells provide protective immunity upon re-encounter with the same pathogens. A better understanding of memory CD8 T cell development is critical for the rational design of vaccines. Transcriptome analyses in antigen-specific CD8 T cells have made a significant contribution to characterize global re-programming of gene expression during memory CD8 T cell differentiation. However, there has been minimal emphasis on understanding translational control of gene expression in antigen specific CD8 T cells. To examine mRNA translation during memory CD8 T cell differentiation, we performed longitudinal analyses of polysome profiles in antigen-specific CD8 T cells after acute LCMV infection. We found that mRNA translation was dynamically regulated during CD8 T cell responses; translation activity was significantly enhanced during clonal expansion phase, and it immediately returned to basal level at the peak of CD8 T cell responses. Genome-wide analyses of mRNA translation showed that a significant number of mRNAs were translationally regulated during CD8 T cell responses. Further experiments revealed that translational control of gene expression plays an important role in memory CD8 T cell formation. Thus, our studies provide a framework for understanding translational regulation that occurs during memory T cell differentiation.
Neutrophil elastase regulates both type 1 and type 2 immune responses to enteric infection (MPF1P.800)
Authors: Sun Rex , Smith Allen , Desai Neemesh , Cheung Lumei , Grinchuk Viktoriya , McLean Leon , Wang Anjiang , Urban Joseph , Zhao Aiping , Shea-Donohue Terez 
Published: January 2023

Abstract
Local neutrophil influx occurs in response to enteric bacterial as well as nematode infection. Neutrophilic release of the serine protease, elastase (NE), is implicated in host defense as well as tissue injury. The aim of this study was to determine the contribution of elastase in host defense against enteric pathogens that induce a Th1/Th17 or a Th2 immune response. Elastase deficient (ELA1-/-) or C57BL/6 (WT) mice (n=5-7) were inoculated with Nippostrongylus brasiliensis (Nb) and studied 5 and 9 days later or infected with Citrobacter rodentium (Cr) and studied 14 days later. Gene expression was determined by real-time PCR in samples of small intestine or colon. Nb infection in WT mice induced a 21-fold STAT6-dependent increase in elastase expression. ELA1-/- mice had accelerated Nb expulsion coupled with significantly (p&lt;0.01) augmented elevation in IL-4, IL-13, and M2 marker (e.g arginase-1) expression when compared to WT mice. Both strains showed similar increases in smooth muscle hypercontractility and epithelial permeability. In contrast, ELA1-/- mice had delayed clearance of Cr infection with impaired upregulation of IFNγ and IL-17A expression when compared to WT mice. These data indicate that during nematode infection, the upregulation of elastase negatively regulates the Th2 response and could be a novel target for modulating type 2 immunity. During bacterial infection, elastase is essential for the upregulation of type 1 cytokines and pathogen clearance in the gut.
Extraintestinal helminth infection reduces the development of colitis-associated colorectal cancer (MPF1P.763)
Authors: Terrazas Luis I. , Callejas Blanca , León-Cabrera Sonia , Rodriguez-Sosa Miriam , Perez-Plasencia Carlos , Chirino Yolanda , Gitierrez-Cirlos Emma , Avila-Moreno Federico , Ledesma-Soto Yadira 
Published: January 2023

Abstract
Whereas a causal association between viral and bacterial infections with tumor development has been well established in animal and epidemiological studies, the impact of parasitic infections in cancer development is almost unknown. Evidences indicate that cancer can be triggered by chronic or deregulated inflammatory processes, such as colitis-associated colorectal cancer (CAC). Using a helminth infection model, we have analyzed the role of helminth-induced anti-inflammatory response in determining the outcome of CAC. Following Taenia crassiceps infection and development of CAC, mice displayed inhibited colonic inflammatory responses and reduced circulating and colonic CD11b+Ly6C+CCR2+ monocytes. Instead they increased the expression of IL-4 and alternatively activated macrophages (AAMs) markers at the colonic tissue. Strikingly, T. crassiceps-infected mice developed few tumors and reduced pathology. In contrast, uninfected mice mounted strong inflammatory responses, did not express AAMs markers, developed severe pathology and displayed multiple tumors. Thus, immunomodulatory activities induced by helminth infections may have a role in the resistance and progression of CAC.
HIV-1 alters pathology and bacterial dissemination of Mycobacterium tuberculosis in co-infected humanized mice (MPF1P.765)
Authors: Nusbaum Rebecca , Calderon Veronica , Huante Matthew , Sutjita Putri , Vijayakumar Sudhamanthi , Goez-Rivillas Yenni , Hunter Robert Hunter , Actor Jeffrey , Cirillo Jeffrey , Valbuena Gustavo , Endsley Janice 
Published: January 2023

Abstract
Over a third of the world’s population is infected with Mycobacterium tuberculosis (M.tb). HIV infection significantly increases risk for M.tb infection and reactivation and TB is the most common cause of death among people with HIV. We have developed a humanized mouse model of HIV-1/M.tb co-infection as an investigative tool to study co-infection biology. Humanized mice were infected intravenously with a macrophage-tropic strain of HIV-1 (JR-CSF) or mock (BSA), and 3 weeks later were infected intranasally with M.tb (H37Rv). Animals became productively infected with HIV-1, as confirmed by serum ELISA for HIV-1 p24 capsid protein and detection of viral RNA by using qPCR. We further observed that HIV infected cells were localized to sites of inflammation and bacterial replication in the lung of HIV-1/M.tb co-infected animals as determined by immunohistochemistry staining for p24. The number of CD4+T cells in peripheral blood did not differ among M.tb- and M.tb/HIV-1-infected groups, indicating that the immune impairments due to HIV-1 were independent of peripheral T cell loss at this stage of co-infection. Consistent with clinical observations in co-infected human patients, greater mycobacterial dissemination was observed in M.tb/HIV-1 co-infected mice compared animals infected with M.tb only. These results suggest promise for use of the humanized mouse as a model to understand co-infection pathobiology and as a translational tool to develop vaccines and therapies.
Chronic infection with trypanosoma cruzi alters the phenotypic profile of circulating memory B cells in humans (MPF2P.812)
Authors: Postan Miriam , Quebrada Palacio Luz , Hernandez-Vasquez Yolanda , Fernandez Esteban 
Published: January 2023

Abstract
Trypanosoma cruzi was shown to induce the loss of parasite-specific IgG-producing B cells during the acute infection of mice, which was proposed to hamper B cell responses and favor parasite replication and chronicity (Zuñiga et al. 2002). Herein, we measured the levels of circulating memory and MZ-like B cells in 12 individuals chronically infected with T. cruzi and 9 healthy controls based on the expression of CD19, IgM IgD, IgG, and CD27. T. cruzi-infected individuals had significantly lower percentages of IgM+IgD-CD27+ (IgM-only), IgM-IgD-CD27+ (isotype-switched) and IgG+IgD-CD27+ memory B cells compared to controls (p&lt;0.05). Also, the percentage of IgM+IgD+CD27+ (MZ-like) B cells was reduced in infected individuals while the proportion of IgG+IgD-CD27- memory B cells was increased (p&lt;0.05). The levels of memory B cells were further analyzed according to ECG findings in infected individuals (asymptomatic [ASY] and chronic Chagas heart disease [CHD]). IgG+IgD-CD27+, IgM+IgD-CD27+ and IgM-IgD-CD27+ B cells were significantly reduced in the ASY group compared to controls (p&lt;0.05); CHD individuals had decreased levels of IgM-IgD-CD27+ and IgM+IgD-CD27+ B cells (p&lt;0.05). Altogether, our data indicate that chronic exposure to T. cruzi modulates the phenotypic profile of the peripheral blood CD19+ B cell compartment in humans, and that some of the defects develop early in the infection, prior the onset of heart symptoms. Supported by Argentinian MINCYT/FONCYT PICT 2010-2148
DH reading frame usage influences HIV-1 epitope recognition (MPF2P.804)
Authors: Wang Yuge , Sanchez-Silva Aaron , Liu Cunnre , Haynes Barton , Schroeder Harry 
Published: January 2023

Abstract
Background: Some HIV-1-specific neutralizing antibodies, such as 2F5 and 4E10, contain a long H chain complementarity determining region 3 (CDR-H3) includes patches of hydrophobic and charged amino acids, but B cells bearing Igs with charged or hydrophobic CDR-H3s are normally culled from mature B cell subsets. Thus, DH influence on neutralizing antibody generation is very worth to explore. Methods: Mice-BALB/c mice cohort limited to the use of single DH gene segments were generated. ΔD-DFL is a human-like repertoire control. ΔD-DµFS promotes the use of hydrophobic amino acid repertoire. The ΔD-iD is charged amino acid enriched repertoire. Immunization-each strain of 10 mice was immunized with HIV-1 JR-FL gp140 protein. Epitope Identification, serum Assay were conducted. Results: As a general rule, the heterogeneity of the anti-JR-FL gp140 response varied by DH genotype with ΔD-DFL&gt;Wild Type (WT)&gt;&gt;ΔD-DμFS, ΔD-iD. Conversely, the intensity of the response was greatest in the ΔD-iD. However, linear response was not identified with the response to natural HIV-1 epitope. Anti-JR-FL gp140 antibodies present in the sera of the DH altered did not block HIV-1 envelope CD4, 2F5, 2G12 binding site. Conclusion: The pattern of epitope recognition and antigen binding in the response to T dependent antigen, in part, on DH gene segment sequence. Restrictions imposed by natural selection of DH sequence may underlie some of the difficulty that patients generate neutralizing antibody.
Factor H binds to Trypanosoma cruzi trypomastigotes via its C-terminus and contributes to parasite resistance against the alternative pathway of complement (MPF2P.809)
Authors: Ferreira Viviana , Molina Claudia , Tapia Valentina , Nieto Vanesa , Galdames Pablo , Cortes Claudio , Ramirez-Toloza Galia 
Published: January 2023

Abstract
Trypanosoma cruzi (T. cruzi), the causal agent of devastating Chagas disease, possesses multifactorial resistance to the alternative pathway (AP) of complement. AP regulatory protein factor H (fH) uses its C-terminal 19-20 domains to recognize polyanions (i.e. sialic acid) and C3b on host cells, facilitating decay of C3 convertases and inactivation of C3b via the N-terminus of fH. We have shown that recombinant C-terminal fH domains 19-20 (rH19-20) compete with fH for binding to C3b and host cell polyanions, inhibiting protection from the AP. T. cruzi tripomastigotes resist the AP by capturing sialic acid from host cells. The specific role that fH plays in protecting T. cruzi from the AP and the fH domains involved remain unknown. Here we assessed the susceptibility of T. cruzi (MF, Y and Brenner strains) to AP-mediated killing, and the ability of fH to bind directly to the parasite by FACS. For determining the fH domains needed for the interaction with T. cruzi, trypomastigotes were incubated with normal human sera in the presence or absence of equimolar amounts of overlapping recombinant fH proteins (2 to 3 domains each) spanning domains 7 to 20. The ability of each protein to increase AP-mediated killing of T. cruzi was assessed. Only rH19-20 effectively competed with serum fH, inducing a significant decrease in parasite survival (p&lt;0.0001), indicating that the C-terminus of fH is essential for fH-mediated recognition and protection of T. cruzi trypomastigotes from the AP.
IFN-γ is required for spontaneous colitis-associated colon cancer in mice with a lineage-restricted deletion of the Smad4 gene in T cells. (MUC2P.821)
Authors: Kim Byung-Gyu , Choi Sung Hee , Letterio John 
Published: January 2023

Abstract
Immune cells and the cytokines they produce are important mediators of the transition from colitis to colon cancer, but the mechanisms mediating this disease progression are poorly understood. We previously reported that selective deletion in T cells of the TGF-β signaling intermediate in mice, Smad4 (Smad4TKO mice) leads to spontaneous gastrointestinal inflammation which progresses to mucosal epithelia cancers. IFN-γ expression is significantly increased in the T cells and colon of Smad4TKO mice, thus we established a ‘double knockout’ (Smad4/IFN-γDKO) mouse model in which a germ line IFN-γ gene deletion (IFN-γKO) is combined with the Smad4TKO. Smad4/IFN-γDKO mice had a significantly reduced incidence and progression of colitis-associated colon cancer, and a decrease in the number of mucosal CD4+ T cells, compared to those of Smad4TKO. We observed impaired in vitro induction of Foxp3+CD4+ T cells (iTregs) by TGF-β in T cells derived from both Smad4TKO and Smad4/IFN-γDKO. This phenotype is associated with a decrease in the mucosal natural Foxp3+CD4+ T cells (nTregs) in vivo. The colon epithelia of Smad4/IFN-γDKO mice exhibited a non-oncogenic signature with a decrease in the expression of iNOS and p-STAT1 and a restoration of the tumor suppressor gene, 15-PGDH, compared to Smad4TKO mice. Thus, our data indicate the critical role of IFN-γ in promoting carcinogenesis associated with spontaneous inflammation in mice with a T cell-restricted deletion of the Smad4 gene.
image cytometry method for measuring concentration and viability of primary cells used in cellular therapy (TECH1P.846)
Authors: Chan Leo , Lee James , Kuksin Dmitry , Qiu Jean 
Published: January 2023

Abstract
Cellular therapy has become a major clinical research field that creates tailor-made medical treatments for many human diseases. Primary cells obtained from patients and mouse models often contain nonspecific particles such as red blood cells (RBC), platelets, and cellular debris, which can make the cell sample analysis difficult. To remove nonspecific particles that can interfere with analysis, a ficoll gradient separation or RBC lysis is routinely performed. Measurements of concentration and viability of the cell sample are necessary for clinical researchers to qualify the collected patient samples for research and downstream processing. In this work, we validated a fluorescence-based image cytometry method using acridine orange (AO) and propidium iodide (PI) to rapidly measure concentration and viability without the need for tedious purification protocols. Using the Cellometer Vision instrument we performed the viability and concentration analysis and validated our results using the traditional hemacytometer method for samples including but not limited to peripheral blood mononuclear cells, mononuclear cells, Leuko Pac, bone marrow, cord blood, whole blood, bronchoaveolar lavage, and primary murine samples. This image-based cytometer method can increase clinical research efficiency by eliminating the need for purification steps and for manual counting. Furthermore, it can eliminate the user-to-user variation, thus improving the accuracy of the cell analysis.
New multiplex assay panels for simultaneous quantification of 13 Th cytokines in human and mouse samples (TECH1P.852)
Authors: Sun Binggang , Yao Jingjing , Hu Jiaming , Ji Shaoquan 
Published: January 2023

Abstract
T helper or “Th” cells, play important roles in regulating the responses of the immune system. They secrete cytokines to stimulate various effector cells, such as cytotoxic T cells, B cells and macrophages. Accurate measurement of Th cytokine expression is critical to identify the corresponding Th cell responses and to develop in depth understanding of the immune responses. We have developed two multiplexed assay panels, i.e., the human and mouse Th panels, using fluorescence-encoded beads that are suitable for use on various flow cytometers. Both panels allow simultaneous quantification of 13 human and mouse cytokines including interleukins (IL-2, 4, 5, 6, 9, 10, 13, 17A, 17F, 21, 22), IFN-g and TNF-α, which are collectively secreted by Th1, Th2, Th9, Th17, Th22 or T follicular cells. Each antibody pair was carefully selected for assay specificity, sensitivity, accuracy and reproducibility. The assays provide higher detection sensitivity and broader dynamic ranges than traditional ELISA methods. The panels have been validated by detecting expected changes in biological samples. Further advantages include small sample volume, flexible assay configurations, and time- and cost-effectiveness. The Th cytokine panels can be used for serum, plasma, cell culture supernatant and other sample types, offering useful tools for biomedical research and drug discovery.
Simultaneous detection of IgG antibodies associated with viral hemorrhagic fever by a multiplexed luminex-based immunoassay (TECH1P.855)
Authors: Zhang Shuo , Wu Wei , Qu Jing , Zhang Quanfu , Li Chuan , Li Jiandong , Jin Cong , Liang Mifang , Li Dexin 
Published: January 2023

Abstract
Viral hemorrhagic fevers (VHFs) are worldwide diseases caused by several kinds of viruses. With the emergence of new viruses, advanced diagnostic methods are urgently needed for identification of VHFs. Based on Luminex xMAP technology, a rapid, sensitive, multi-pathogen and high-throughput method which could simultaneously detect hemorrhagic fever viruses (HFVs) specific IgG antibodies was developed. Recombinant antigens of nine HFVs were produced and purified from a prokaryotic expression system and the influence of the coupling amount was investigated. Cross-reactions among antigens and their rabbit immune sera were evaluated. Serum samples collected from 51 laboratory confirmed hemorrhagic fever with renal syndrome (HFRS) patients, 43 confirmed SFTS patients and 88 healthy donors were analyzed. Results showed that recombinant nucleocapsid protein of the five viruses belonging to the genus Hantavirus, had serological cross-reactivity with their corresponding rabbit immune sera, but not apparent with immune sera of other four viruses. Evaluation of this new method with clinical serum samples showed 98.04% diagnostic sensitivity for HFRS, 90.70% for SFTS detection and the specificity was ranging from 66.67% to 100.00%. The multiplexed luminex-based immunoassay has firstly been established in our study, which provides a potentially reliable diagnostic tool for IgG antibody detection of VHFs. Corresponding author: Prof. Dr. Dexin Li; Email: lidx@chinacdc.cn
Close proximity between ICAM-1 and agonist pMHC-I ligands facilitates T cell integrin activation advancing local cytoskeleton remodeling (TECH1P.856)
Authors: Anikeeva Nadia , Sykulev Yuri 
Published: January 2023

Abstract
To mimic clustering of ICAM-1 and pMHC-I molecules, we assembled pMHC-I and ICAM-1 proteins on fluorescent nanoparticles (NP) and compared the binding of ICAM-1/NP, pMHC/NP and ICAM-1/pMHC-I/NP conjugates to live T cells. The binding of the ICAM-1/NP was poor, but it has been significantly enhanced in the presence of Mg2+ that induce high-affinity conformation of LFA-1. The addition of agonist pMHC-I/NP or anti-CD3 antibody did not influence the binding of the ICAM-1/NP to the T cells. However, when both agonist pMHC-I and ICAM-1 were assembled on NP, a profound increase in the binding of the ICAM-1/pMHC-I/NP was observed. The amplification effect was non-additive suggesting that ICAM-1 and pMHC-I displayed on the NP cooperate in their binding to TCR and LFA-1 on T cells. In control experiment, NP bearing ICAM-1 and null pMHC did not reveal the cooperative binding to the T cells. We conclude that the close proximity of agonist pMHC-I and ICAM-1 facilitates conversion of LFA-1 to a high affinity form upon productive TCR engagement. We propose that this cooperative affect advances initial local cytoskeleton remodeling enhancing the sensitivity and kinetics of TCR signaling.
Using quantitative imaging to link immune profile and tumor grade in breast cancer. (TECH1P.861)
Authors: Blenman Kim , Andersen Emily , Lee Peter 
Published: January 2023

Abstract
Tumor grade determines how quickly a tumor proliferates and spreads. It is one of the factors used to determine the best treatment approach for cancer patients. The Nottingham Histological Score is used to determine tumor grade in breast cancer. Three factors comprise this score: amount of gland formation, nuclear features, and mitotic activity. These factors tell how close the cancer cells and the tumor tissue structure resemble normal cells and tissue. Well-differentiated (grade 1) tumors closely resemble normal cells and tissue and grow and spread slowly. Moderately (grade 2) - and poorly (grade 3) - differentiated tumors have abnormal cell appearance, lack normal tissue structures, and grow and spread faster. In this pilot project, we were interested in determining if we could link tumor microenvironment immune profiles to tumor grade. We used our quantitative imaging process including multiplexed immunohistochemistry, multispectral imaging, and quantitative analysis software to generate immune profiles of CD3+ T cells, CD20+ B cells, CD1a+ dendritic cells, and cytokeratin+ cancer cells in primary tumor and tumor draining lymph nodes of breast cancer patients. Tumor grades were collected from pathology reports. Our preliminary data suggests that we can link immune profiles and tumor grade. Our next steps are to determine if combination of immune profiles and tumor grade can improve selection of treatment approach for breast cancer patients versus tumor grade alone.
New multiplex assay panels for simultaneous quantification of major proinflammatory chemokines in human and mouse samples (TECH1P.866)
Authors: Sun Binggang , Hu Jiaming , Yao Jingjing , Ji Shaoquan 
Published: January 2023

Abstract
Chemotactic cytokines or chemokines play pivotal roles in various processes such as immune surveillance, organ development, angiogenesis, and immune responses. Expression profiling of chemokines, especially those involved in inflammation and immune disorders, is important in achieving a deeper understanding of disease states. Using fluorescence-encoded beads, we have developed two multiplex immunoassay panels to simultaneously detect 13 human or mouse chemokines, respectively. Both human and mouse panels include MCP-1 (CCL2), RANTES (CCL5), IP-10 (CXCL10), Eotaxin (CCL11), TARC (CCL17), MIP-1α (CCL3), MIP-1β (CCL4), MIG (CXCL9), and MIP-3α (CCL20). In addition, the human panel contains ENA-78 (CXCL5), GROα (CXCL1), I-TAC (CXCL11) and IL-8 (CXCL8), and the mouse panel contains LIX (CXCL5), KC (CXCL1), BLC (CXCL13) and MDC (CCL22). Each antibody pair was selected for sensitivity, specificity, accuracy and reproducibility. Suitable for use on various flow cytometers, the assays have higher sensitivity and wider dynamic range than typical ELISA methods. Further advantages include small sample volume requirement, flexible assay configurations and time- and cost-effectiveness. In addition, the assays have been validated with expected changes in relevant biological samples. The chemokine panels can be used for serum, plasma, cell culture supernatant, and other sample types, offering useful tools for biomedical research and drug discovery.