bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Prime-Boost Dengue Vaccination Strategy Induces Durable Multivalent Memory B Cell Responses
Authors: Hatch Kristin , Friberg Heather , Koren Michael , Keiser Paul , Lin Leyi , Jarman Richard G , Currier Jeffrey R 
Published: January 2023

Abstract
Denguevirus is a mosquito-borne flavivirus that is comprised of four antigenically related but distinct serotypes. A higher risk for the most severe clinical manifestations of dengue infection, dengue fever and dengue hemorrhagic fever, is associated with sequential infections with heterologous serotypes. While there currently is no precisely defined correlate of protection, durable tetravalent immunity is the desired outcome of dengue vaccination. A Phase 1 clinical trial was conducted at the Walter Reed ArmyInstitute of Research (WRAIR), WRAIR #2136, using live attenuated virus (LAV)and purified inactivated virus (PIV) vaccine candidates containing all fourserotypes. The two products were tested in a prime-boost regimen (PIV followed by LAV and vice versa) in different schedules to determine if dosing schedule or order of administration affected neutralizing antibody levels and/or cell mediated responses. Utilizing a three color Fluorospot assay to measure DENV-specific IgG, IgA, and IgM secreting cells, we tested 6-months post second dose memory B cell cultures to determine if the response was tetravalent and if a change in vaccination strategy affected the valency. We also investigated how memory B cell responses correlated to neutralizing antibody data. Results show that a majority of subjects have a robust multivalent DENV-specific IgG memory B cell response at 6 months post-vaccination. A better understanding of memory B cell responses and how they relate to neutralizing antibody will help find a suitable DENV vaccine that provides durable tetravalent immunity.
CHARACTERIZATION OF INTERFERON-γ PRODUCING CELLS IN THE PERIPHERAL BLOOD OF PATIENTS WITH TUBERCULOSIS
Authors: Ruiz-Sánchez Bibiana Patricia , Cruz-Zárate David , Hernández-Solis Alejandro , García-Paredes Víctor Gabriel , Cicero-Sabido Raúl , Serafín-López Jeanet , Estrada-García Iris Citlali Elvira , Wong-Baeza Isabel 
Published: January 2023

Abstract
Tuberculosis (TB) caused 1.4 million deaths in 2015. The infection starts when Mycobacterium tuberculosis is inhaled and interacts with immune cells in the respiratory tract. Alveolar macrophages, neutrophils and dendritic cells (DCs) engulf the mycobacteria; these DCs migrate to the draining lymph node and induce the activation of mycobacteria-specific Th cells. These Th cells migrate back to the lung and produce IFNγ and TNFα, two cytokines essential for mycobacterial control. In addition to Th cells, other cells, such as Tγδ and NK cells, produce IFNγ in response to mycobacteria; innate lymphoid cells (ILCs) are also able to produce IFNγ, but it still unknown if ILCs, which have been detected in human lungs during cancer, produce this cytokine in response to mycobacteria.
We evaluated, by flow cytometry, the frequency and IFNγ production of Th, Tγδ, NK and ILCs (ILC1, ILC2 and ILC3) in the peripheral blood of healthy individuals (negative and positive for the Quantiferon [QTF] test, which detects latent TB), and patients with active pulmonary TB. The numbers of Th, Tγδ, NK cells and ILCs (ILC1, ILC2 and ILC3) were decreased in TB patients. Peripheral blood was stimulated with M. tuberculosis soluble extract for 5 h in the presence of brefeldin A. In healthy, QTF-negative individuals, IFNγ production was increased in Tγδ and ILC2 after stimulation; in healthy, QTF-positive individuals, IFNγ production was increased in Th, Tγδ and ILCs, and in TB patients, IFNγ production was increased in NK cells and ILC2. These results suggest ILCs produce IFNγ during TB infection.
This project was funded by SIP-IPN-México (20170616).
Long Term Impact of Acute Infection on Lymphatic Function
Authors: Zaretsky Arielle Glatman , Han Seong-Ji , da Fonseca Denise Morais , Hand Timothy , Belkaid Yasmine 
Published: January 2023

Abstract
The lymphatic system facilitates critical processes for immunity, including immune cell trafficking, lipid transport, and the maintenance of tissue homeostasis. In the mesentery, these lymphatic vessels (LVs) allow drainage and cell trafficking from the gastrointestinal (GI) tract through the mesenteric adipose tissue (mAT) to the mesenteric lymph nodes (mLNs). During GI infection, this vasculature is altered by inflammation, which can induce lymphatic leakage. Repair of LVs after infection is resolved is necessary to restore lymphatic function. Although preliminary studies have found acute lymphatic leakage is conserved between GI pathogens, in an acute Yersinia pseudotuberculosis infection, mice control bacterial growth, but develop a chronic mesenteric lymphadenopathy and the increased permeability of LVs does not resolve after the infection is cleared. Consequently, the migratory CD103+CD11b+dendritic cells (DP DCs) cannot reach the mLN and instead pass through the dysregulated LVs into the mAT. The absence of DP DCs in the mLN is associated with impaired development of Th17 adaptive immunity to oral vaccination. Little is known about the mechanisms that regulate lymphatic integrity, however these studies reveal a remodeling of the adipose tissue after infection, and are ongoing to understand the long-term increased permeability of LVs and the potential to restore function. Our results support the idea that acute GI infections are associated with permanent changes in DC traffic, which interfere with the activation of immune responses and can have severe implications for GI tract integrity and predisposition to inflammatory diseases and suggest that dysregulation of the mAT may contribute to the prolonged disruption.
Activation-induced chromatin decondensation in the lymphoid lineage
Authors: Rawlings Jason S , Knaub Alynna , McDonald Sierra 
Published: January 2023

Abstract
It has previously been demonstrated that chromatin decondensation is required for αβ T cell proliferation. B cells, γδ T cells, and Natural Killer cells share the same lineage as αβ T cells, originating from hematopoietic stem cells and a common lymphoid progenitor. In this study, we investigated the hypothesis that chromatin decondensation is required for proliferation of these other cells of the lymphoid lineage. We assessed chromatin decondensation by using Western Blot to test for Histone H3 solubility and Flow Cytometry to test Histone H3 accessibility. We show that Lipopolysaccharide, a known activator of B cells, induces the decondensation of chromatin in B cells. To determine if both T and B cells can decondense chromatin at the same rate, both were stimulated with PMA and Ionomycin. B cells and ab T cells decondense chromatin in a similar manner, although there appears to be differences in kinetics between the two. Preliminary data collected on NK cells and γδ T cells suggest that their rate of decondensation is similar to αβ T cells in response to PMA and Ionomycin stimulation. Recently, our lab demonstrated that calcium is required and sufficient to initiate chromatin decondensation in αβ T cells, future studies will investigate if calcium can also regulate the decondensation of B cells, γδ T cells, and NK cells, establishing a conserved mechanism to control proliferation of the lymphoid lineage.
The expression profiles and regulation of PD-L1 in tumor-induced myeloid-derived suppressor cells
Authors: Lu Chunwan , Savage Natasha , Singh Nagendra , Liu Kebin 
Published: January 2023

Abstract
Programmed death-ligand 1 (PD-L1) is an inhibitory ligand that binds to PD-1 to suppress T cell activation. PD-L1 is constitutively expressed and inducible in tumor cells, but the expression profiles and regulatory mechanism of PD-L1 in myeloid-derived suppressor cells (MDSCs) are largely unknown. We report that PD-L1 is abundantly expressed in tumor-infiltrating immune cells in human colon carcinoma, and 25–67% CD11b+CD33+ myeloid-derived suppressor cells (MDSCs) from peripheral blood of human colon cancer patients are PD-L1+. Using a spontaneous colon cancer mouse model, we observed that PD-L1+MDSCs increased significantly in tumor-bearing mice as compared to MDSCs from tumor-free mice. Interestingly, the highest PD-L1+ MDSCs were observed in the tumor microenvironment in which 56–71% tumor-infiltrating MDSCs are PD-L1+ in vivo. In contrast, PD-L1+ MDSCs are significantly less in secondary lymphoid organs and peripheral blood as compared to the tumor tissues, whereas, bone marrow MDSCs are essentially PD-L1− in tumor-bearing mice. IFNg is highly expressed in cells of the tumor tissues and IFNg neutralization significantly decreased PD-L1+ MDSCs in the tumor microenvironment in vivo. However, IFNg-activated pSTAT1 does not bind to the cd274 promoter in CD11b+Gr1+PD-L1+ myeloid cells. Instead, pSTAT1 activates expression of IRF1, IRF5, IRF7 and IRF8 in MDSCs, and only pSTAT1-activated IRF1 binds to an unique IRF-binding sequence element in vitro and chromatin in vivo in the cd274 promoter to activate PD-L1 transcription. Our data determine that PD-L1 is highly expressed in tumor-infiltrating MDSCs and in a less degree in lymphoid organs, and the pSTAT1-IRF1 axis regulates PD-L1 expression in MDSCs.
Evidence that Th1-polarized Tfh cells drive the differentiation of T-bethi atypical memory B cell expansion in human malaria
Authors: Obeng-Adjei Nyamekye , Crompton Peter D 
Published: January 2023

Abstract
Many chronic infections, including malaria and HIV, are associated with a large expansion of CD21−CD27−CXCR3+CD11c+ ‘atypical’ memory B cells (MBCs) that exhibit reduced effector functions. Little is known about the conditions or transcriptional regulators driving atypical MBC differentiation. Here we show that atypical MBCs in malaria-exposed individuals highly express the transcription factor T-bet. Moreover, a longitudinal study of malaria-exposed children demonstrated a positive correlation between the incidence of febrile malaria and the expansion of T-bethi B cells. The Th1-cytokine containing supernatants of malaria-stimulated PBMCs and B cell receptor (BCR) cross- linking induced T-bet expression in peripheral and tonsilar B cells that were blocked by neutralizing IFN-γ. Accordingly, recombinant IFN-γ plus BCR cross-linking drove T-bet expression in peripheral and tonsilar B cells. Consistent with this, Th1-polarized Tfh (Tfh-1) cells more efficiently induced T-bet expression in naïve B cells. These data illuminate the mechanisms underlying atypical MBC differentiation.
Circulating memory CD4 T follicular helper cell subsets correlate with clinical immunity against human malaria
Authors: Furtado Raquel M , Lauvau Gregoire , Daily Johanna P 
Published: January 2023

Abstract
Malaria remains a global burden with over 200 million infections and half a million deaths annually. While studies in human and surrogate mouse models reveal that B cells and CD4 T cells are essential, it is not known if and how sterilizing immunity against malaria may be achieved. Residents of endemic areas develop clinical immunity to Plasmodium falciparum over time yet protection is partial and requires repeated exposures. Herein we conducted a 2 year longitudinal study on patients with mild malaria in a hyperendemic region in Malawi. We categorized patients with a high number and short interval between reinfections (‘susceptible’) and with a low number and longer interval between reinfections (‘protected’). We hypothesized that clinical immunity correlates with the presence of specific immune cell subsets in protected versus susceptible patients. We conducted comprehensive cellular profiling on patient PBMCs, isolated during infection, by mass cytometry and a panel of 33 markers. We examined the frequency and activation of 40 immune cell subsets by unsupervised clustering analysis and discovered an expansion of CXCR3lo memory CD4T follicular helper cells (Tfh) in protected patients compared to susceptible patients. Importantly, with an in vitro antibody dependent opsonization assay we found increased phagocytosis of P. falciparum merozoites with plasma from protected versus susceptible patients, correlating CXCR3lo memory Tfh cell frequency with enhanced humoral responses. We speculate that CXCR3lo memory Tfh cells are critical for lasting protective responses against malaria in humans and may be essential for an efficacious future vaccine.
Contribution of the T cell receptor to the generation and maintenance of EBV epitopespecific CD8 T cell responses following primary infection
Authors: Kamga Larisa , Brody Robin , Gautam Anita , Ghersi Dario , Selin Liisa K , Luzuriaga Katherine 
Published: January 2023

Abstract
We have investigated the potential effect of TCRαβ usage on the maintenance and transcriptome of Epstein-Barr virus (EBV)-specific CD8T cells over the course of primary infection. Blood samples were collected over the course of a year from patients presenting with primary EBV infection. Peptide-MHC tetramers and FACS were used to sort CD8 T cells specific for two HLA-A*02-restricted EBV epitopes: BMLF1 (GLCTLVAML; GLC) and BRLF1 (YVLDHLIVV; YVL); single-cell sequencing and transcriptomics were used to evaluate TCRαβ usage with the expression of 47 immune genes. In 4 donors studied, we identified 16 and 4 public GLC- and YVL-specific CDR3αs; and 17 and 2 public GLC- and YVL-specific CDR3βs. The Morisita-Horn index was used to assess the degree of similarity of the TCRαβ repertoire between primary infection and convalescence. GLC-specific cells tended to have a higher similarity index than YVL-specific cells (mean similarity index: 0.55 vs. 0.27), suggesting that the TCRαβ repertoire GLC-specific cells tended to be more stable than that of YVL-specific cells. The similarity index correlated with the frequency of public CDR3α/β clones (Spearman r: 0.83; p = 0.0198), suggesting there may be a survival advantage to public CDR3αβ usage leading to T-cell persistence. 11 and 10 degenerate GLC- and YVL-specific CDR3αs; and 15 and 12 degenerate GLC- and YVL-specific CDR3βs were identified. In a donor, one of these CDR3αs was paired with up to 9 unique CDR3β. Future analyses will focus on single-cell gene expression to investigate whether there are specific gene patterns associated with public or degenerate TCR and T-cell persistence.
High specificity of human antibody response to nonstructural protein NS1 elicited by Zika virus infection.
Authors: Gao Xiujie , Wen Yingfen , Li Chunlin , Wang Jian , Zhang Fuchun , Yu Lei 
Published: January 2023

Abstract
The newly emerged Zika virus (ZIKV) belonging to the flavivirus family, may appear in the area dengue virus (DENV) infection occurred, raising the issue of diagnosis. The secreted NS1 protein from infected cell is not only an important diagnostic marker but also highly immunogenic. Here we investigated the cross-reaction of NS1 Ab responses between the two close related viruses. First we studied the binding of sera to the recombinant full length NS1 from three ZIKV infected patients (Samples collected at day 4, 7, 15, 106, 188 and 321 for patient 1; day 6, 12, and 66 for patient 2; day 2 for patient 3) by capture ELISA. ZIKV NS1 binding IgG markedly increased in convalescent sera from patient 1 and patient 2 with less cross-reaction to DENV 1–4 NS1. The serum from patient 3 did not bind to ZIKV NS1 and that may be explained by the early time point of serum collected on day 2 from the symptom onset. Specificity of ZIKV NS1 Ab response was evidenced by lack of reactivity to DENV 1–4 NS1proteins of memory B cell derived mAbs (4/5) isolated from the two ZIKV-infected. Only one mAb (1/5) cross-reacted to DENV 2 and DENV 4 NS1. Strikingly NS1 Ab responses of DENV1 infected (six samples) which serotype frequently circulates in our area, Guangdong, China with outbreak in 2014 showed strong cross-reactivity between DENV serotypes with mild or strong reaction to ZIKV NS1. Our data indicated that NS1 Ab response elicited by Zika virus infection was ZIKV-specific and the ZIKV specific mAb can be used for diagnostic tool.
Gut dysbiosis breaks immunological tolerance toward the central nervous system during young adulthood.
Authors: Yadav Sudhir Kumar , Boppana Sridhar , Ito Naoko , Mindur John E , Mathay Martin T , Patel Ankoor , Dhib-Jalbut Suhayl , Ito Kouichi 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an immune mediated disease targeting the central nervous system (CNS) mainly in young adults, and a breakage of immune tolerance to CNS antigens has been suggested to initiate CNS autoimmunity. Recent studies have suggested that gut microbiota affects the development of autoimmune diseases and alterations in the gut microbiota, termed gut dysbiosis, are associated with MS. However, it is still unknown how gut dysbiosis affects the onset and/or progression of CNS autoimmunity. In this study, we investigated the effects of gut dysbiosis on the development of CNS autoimmunity in humanized transgenic mice expressing the MS- associated HLA-DR gene, DR2a, and the T cell receptor (TCR) gene specific for MBP87-99/DR2a that were derived from an MS patient. We show here that the induction of gut dysbiosis triggers the development of spontaneous experimental autoimmune encephalomyelitis (EAE) during adolescence and early adulthood, while increase in immunological tolerance with aging suppresses the disease onset after late young adulthood. Furthermore, gut dysbiosis induces the expression of complement C3 and production of the anaphylatoxin C3a, and down-regulates the expression of the Foxp3 gene in Tregs and anergy-related E3-ubiquitin ligase genes in CD4+CD25− T cells. Consequently, gut dysbiosis was able to trigger the development of encephalitogenic T cells and promote the induction of EAE during the young adulthood age-window. This data suggests that gut dysbiosis increases the risk for CNS autoimmunity in young adulthood through the up-regulation of the complement C3 gene and down-regulation of Foxp3 and E3-ubiquitin ligase genes.
Could Connexin 43 dependent ATP release represent a new therapeutic target for sepsis?
Authors: Dosch Michel Ernest , Salamanca Tamara , Djonova Djulia , Keogh Adrian , Stroka Deborah , Candinas Daniel , Beldi Guido 
Published: January 2023

Abstract
During sepsis, ATP is released into the extracellular space where it modulates immune response via specific receptors and ectonucleotidases present on various immune cells. ATP can be released by hemichannel Connexin 43 (Cx43). Here, we examine the importance of Cx43 mediated ATP release in macrophages and the role of this pathway in modulating innate immune responses in the context of sepsis.
For murine sepsis model, caecal ligation and puncture (CLP) was used. Mice were treated with Gap27, a Cx43 inhibitor. Liver, lung, spleen, bone marrow, peritoneal fluid and blood cells expressing Cx43 were characterized (IHC, FACS). Macrophages were isolated from peritoneum and liver of wild-type mice and stimulated with PAMPs. ATP was quantified using luciferin-luciferase assay. Cytokine levels (ELISA), gene expression (qPCR) and protein expression (immunoblot) were assessed.
Blocking of Cx43 during CLP prolonged survival up to 48 hours in comparison with 24 hours for controls. In addition, it decreased systemic levels of inflammatory cytokines (TNFa, IL1b and IL6) and chemokines (CCL2) and lowered bacterial load (CFU/ml) in the blood and peritoneal fluid. Cx43 was not constitutively expressed in the liver, but was induced following CLP. Cells expressing Cx43 in septic livers were identified as infiltrating M1 macrophages and neutrophils. Ex vivo, Cx43 expression was upregulated in primary macrophages upon stimulation with LPS. Macrophages were actively releasing ATP in response to TLR 4 and 2 agonists, and Cx43 inhibition reduced extracellular ATP levels.
We observed improved survival to sepsis by blocking Cx43. By mediating release of ATP, and potentially other DAMP, Cx43 might overactivate macrophages and exacerbate the immune response.
TCR recognition is a mediator in the control of HIV infection by CD8 T cells
Authors: Nicodemos Rodrigo A Matus , Casazza Joseph , Price David , Koup Richard , Douek Daniel 
Published: January 2023

Abstract
CD8 T cell responses to HIV-specific MHC class I epitopes are critical for the control of HIV infection. The precise mechanisms that govern the control of HIV by CD8 T cells are largely unknown. Certain protective HLA alleles, such as, B*5701, are associated with long-term control of HIV-1. However, other mechanisms by CD8 T cells might be contributing to the control of HIV, because non-controllers with these protective alleles have been reported.
Therefore, to determine the molecular mechanisms that underlie HIV control by CD8 T cells we assembled a cohort of HIV positive individuals, which consisted of controllers (viral load less than 2000 copies per mL), ART-naive non-controllers, and non-controllers on ART-therapy. Using PBMCs from our cohort we first stained the cells with a panel of thirteen T cell markers and then performed polychromatic single cell index sorting of CD3+CD8+ MHC class I tetramer-binding cells. Our tetramers consisted of HIV epitopes bound to HLA-B*5701, A*0201, B*0702 or B*0801. We then determined the TCR diversity for each tetramer-binding population by single cell TCR sequencing. We also reconstructed HIV-specific TCRs and expressed them in a reporter T cell line to measure TCR avidity to the tetramer.
We found that HIV controllers have: (1) a diverse TCR repertoire for the KF11 epitope presented by B*5701, in contrast, to non-B*5701 alleles presenting HIV epitopes that are heavily skewed; (2) high avidity HIV-specific TCRs for HIV epitopes presented by B*5701; and (3) TCR-expanded clones with a novel CD27dimCD45ROneg T cell phenotype. Importantly, lack of any three of these parameters resulted in the inability to control HIV, indicating that no single parameter is sufficient for natural control.
A rapid single-cell RNA-seq approach to identify neoantigen-specific T-cell receptors targeting tumor-specific mutations for use in gene-engineered T-cell immunotherapy
Authors: Lu Yong-Chen William , Zheng Zhili , Robbins Paul F , Tran Eric , Prickett Todd D , Gartner Jared J , Li Yong F , Ray Satyajit , Bliskovsky Valery , Fitzgerald Peter C , Rosenberg Steven A 
Published: January 2023

Abstract
Accumulated evidence suggested that effective cancer immunotherapies were associated with neoantigen-reactive T cells, and adoptive transfer of neoantigen-reactive tumor infiltrating lymphocytes (TILs) could result in tumor regressions. To improve the efficacy of adoptive T-cell therapy targeting tumor-specific mutations, we have proposed a new therapeutic approach, which involves the genetic modification of autologous T cells with neoantigen-specific TCRs and the transfer of these T cells back to cancer patients. However, the current techniques to isolate neoantigen-specific TCRs are labor-intensive, time-consuming and technically challenging, not suitable for clinical applications. To facilitate this process, a new approach was developed, which included the co-culture of TILs with tandem minigene (TMG)-pulsed antigen-presenting cells, single-cell RNA-seq analysis of T cells and the identification of paired TCR sequences associated with cells expressing high levels of IFN-γ and IL-2 mRNA. In this study, TIL fragment cultures were grown from tumors resected from three patients with metastatic gastrointestinal cancers, and then were screened against TMG libraries encoding tumor-specific mutations identified by whole-exome sequencing. Following this new approach, three TCRs were identified, synthesized and cloned into a retroviral vector. These TCRs were then transduced into donor T cells, and these transduced T cells were shown to specifically recognize the following neoantigens encoded by the TMGs: USP8 (R667H), KRAS (G12D) and NBAS (C144S). In conclusion, this approach provides a rapid procedure to isolate neoantigen-specific TCRs for basic and translational research, as well as clinical applications.