bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genomic and Proteomic Analysis of Hepatocellular Carcinogenesis in HBV Transgenic Mice (44.14)
Authors: Liu Yugang , Qu Zhonghua , Shi Yongyu , Liang Xiaohong , Sun Wensheng 
Published: January 2023

Abstract
Hepatitis B virus (HBV) transgenic mice invariably developed macroscopically evident tumors within the 20th month of life. Thus HBV transgenic mouse can be used as a natural model of hepatocellular carcinoma (HCC). We confirmed this phenomenon by observation of BALB/c mice harboring HBV complete genome (subtype ayw). To identify molecular changes involved in HCC, we performed both microarray profiling and proteomic analysis between HBV transgenic mice and their syngeneic controls at 6, 12, and 20-month-old. The total liver RNA was hybridized on an oligonucleotide chip containing 35,000 Mus musculus genes. The total liver protein was used for proteomic analysis. Out of all genes and proteins compared and screened, the number of differentially expressed transcripts and proteins was increased along with the growth of mice. We concluded a molecular portrait in different stages of HCC. These molecules were involved in important cellular pathways, including cell growth regulation, apoptosis, immune response, metabolisms, stress, cell adhesion, et al. The dynamics of molecular profiling during hepatocellular carcinogenesis would better elucidate the molecular mechanisms of HCC and help for the discovery of novel therapeutic targets.
Correspondence to Wensheng Sun: wsw@sdu.edu.cn
Supported by the National Nature Science Foundation of China, Grant No. 30440040, 30772031, and 30801036.
Modulation of CD4+ T cells by HCV+ hepatocytes (44.15)
Authors: Hall Caroline H.T. , Hahn Young S. 
Published: January 2023

Abstract
Hepatitis C virus (HCV) infection results in progression to chronic disease in approximately 80% of the infected population. Individuals who clear acute infection have a strong and sustained CD4 Th1 effector response, which is lacking in those who develop persistent infection. Within the liver, an additional immunosuppression is evidenced by a decrease in the percent of IFN-gamma producing CD4+ T cells relative to the peripheral blood. Viral replication occurs primarily within hepatocytes and there is consequently a high concentration of viral products in the liver. Our goal is to determine if HCV-infected hepatocytes have an immunosuppressive effect on CD4+ T cells. We have used a modified HCV cell culture system composed of a human hepatoma cell line, Huh 7.5, containing the full-length HCV genome or the non-structural portion of the genome only. The liver environment is modeled by a co-culture system that allows for direct contact between the hepatocytes and primary CD4+ T cells. We determined that expression of the full HCV genome within hepatocytes has a suppressive effect on CD4+ T cell IFN-gamma production. The CD4+ T cells can further be characterized by a decrease in the Th1 phenotype. These results suggest that local hepatocyte driven immunosuppression may contribute to the inefficient immune responses mounted by HCV infected patients. NIH DK066754 and U19 AI066328 funded this research.
Modulation of memory CD8 T cell function through the inhibitory receptors NKG2A and KLRG1 (44.21)
Authors: Cush Stephanie S 
Published: January 2023

Abstract
The ability of the immune system to establish and maintain functional CD8 T cell memory responses is critical for the control of many persistent viral infections. We used a murine model of g-herpesvirus infection, gHV68, to study the in vivo effects of a low load persistent viral infection on the function of memory CD8 T cells. The aim of this project is to determine how CD8 T cell functions are regulated in the presence of persistent herpesvirus antigens.

Our results show that the inhibitory receptors NKG2A and KLRG1 were consistently up regulated on CD8 T cells during long-term infection with gHV68. At 3 months after infection, more than 90% of the virus specific CD8 T cells expressed KLRG1 while 75% were positive for NKG2A. The expression pattern of NKG2A and KLRG1 during persistent gHV68 infection was fundamentally different than that of memory cells to non-persistent viruses. Our results indicate that the expression of these receptors on CD8 T cells does not impair their ability to produce cytokines or to degranulate. We are currently analyzing the effects of antibody blockade/depletion and of FACS sorting and adoptive transfer of memory populations expressing or not these receptors. Our results suggest that NKG2A and KLRG1 may have a critical role in controlling CD8 T cell responses during persistent herpesvirus infection.
IL-21 is a central component of CD4 T cell help required to purge a persistent viral infection (44.27)
Authors: Brooks David G , Elsaesser Heidi , Sauer Karsten 
Published: January 2023

Abstract
Therapies to cure persistent viral infections have largely been unsuccessful because it is unclear what is required of the immune system to purge virus once it proceeds past acute infection. T cell immunity is crucial to eliminate viral infection. However, CD4 and CD8 T cell functions are rapidly aborted during persistent infection, preventing viral clearance. To date the majority of work examining T cell function during viral persistence has focused on CD8 T cells. Yet, many aspects of immune exhaustion are associated with functions performed by CD4 T helper cells (which are also rapidly exhausted after persistent infection), suggesting that loss of CD4 T cell function is a key mechanism permitting viral persistence. CD4 T cell help is required throughout persistence to sustain CD8 T cell responses and resolve persistent viral infection, suggesting that CD4 T cells maintain some yet undefined, but essential, helper functions. We demonstrate that IL-21 as an essential component of CD4 T cell help during viral persistence. In the absence of IL-21 signaling, despite elevated CD4 T cell responses, help to CD8 T cells is not provided. CD4 T cells utilize IL-21 to prevent CD8 T cell deletion, sustain immunity and resolve persistent infection. Thus, we identify a novel mechanism of CD4 T cell help during viral persistence and propose that CD4 help is not lost during viral persistence, but instead is altered from promoting memory development toward the production of factors that ultimately sustain effector responses to control viral replication.
Cross-protective humoral and cellular immune responses to A/H3N2 drifted influenza viruses following immunization with Fluzone® with a synthetic TLR4 agonist formulation (45.11)
Authors: Shaverdian Narek , Baldwin Susan L. , Reed Steven J. , Raman Vanitha , Goto Yasu , Mompoint Farah , Vedvick Thomas S. , Clegg Christopher H. , Coler Rhea N. , Reed Steven G. 
Published: January 2023

Abstract
Influenza has a worldwide impact with significant human mortality and economic burden. Although the composition of influenza vaccines is reviewed annually to ensure protection against prevalent influenza strains, antigenically drifted strains emerge after the vaccine composition is decided. Vaccine mismatch reduces vaccine efficacy and produces significant epidemiological consequences. Current licensed vaccines poorly stimulate Th1 anti-influenza cellular responses and provide weak protection against drifted influenza strains not included in the vaccine. We investigated the ability of EM005, a synthetic TLR4 agonist formulated in an oil-in-water emulsion, to adjuvant the Fluzone influenza vaccine. EM005 enhanced the potency of the Fluzone vaccine and broadened its serological protection against drifted A/H3N2 influenza variants compared to vaccination with Fluzone alone or Fluzone formulated with emulsion. Equally important, the EM005 adjuvanted vaccine induced a potent Th1 response cross-reactive to all the A/H3N2 strains evaluated. These results suggest that rational adjuvants can enhance both the magnitude and quality of protective humoral and cellular responses of influenza vaccines.
This work was supported in part by Grant #42387 from the Bill & Melinda Gates Foundation
Thalidomide and emergence of Mycobacterium leprae (ML)-specific responses in erythema nodosum leprosum (45.20)
Authors: Pereira Geraldo M B , Esquenazi Danuza de Almeida , Robottom Ferreira Anna Beatriz , Moraes Milton Ozório , Carvalho Daniel Serra , Nery José Augusto da Costa , Alvim Iris Maria Peixoto , Spencer John Stewart , Sampaio Elizabeth Pereira , Pessolani Maria Cristina Vidal , Sarno Euzenir Nunes 
Published: January 2023

Abstract
Acute inflammatory lesions are seen in lepromatous leprosy (LL) during reactions, such as ENL. Necrotic ENL as the initial manifestation of LL was diagnosed at the Souza Araujo Outpatient Unit of FIOCRUZ in Rio de Janeiro (female patient, 26 yrs old). Biopsies and peripheral blood mononuclear cells (PBL) were obtained on days 0, 7 and 30 of treatment with thalidomide (negative pregnancy test). At days 7 and 30 progressive healing of the lesions was observed. The quantification of specific mRNAS for IL-6, TNF-α, ninjurin and IL-10 by qPCR detected an initial high level of IL-6, TNF-α and ninjurin mRNAs, and after thalidomide treatment a decrease of the first 3 mRNAs in the lesion, but IL-10 message increase. ML-specific synthetic peptides induced progressively lower levels of IL-6 in the culture supernatants, but high-level IFN-γ, IL-13 and TNF-α in PBL responses after the healing of the lesions (ML 1419c protein; multiplex assay). CD4+, CD8+, CD45-RO+ and γδ T cells were present in the initial biopsy, but were nearly absent in the last one. Activated (CD69+) CD4, CD8 and γδ T cells were observed in short-term ML-stimulated cultures. Taken together these observations suggest that thalidomide may contribute to the emergence of a ML-specific T cell response with a potential role in the healing process of ENL.
Toll-like receptor 2: An important regulator of the Mycobacterium tuberculosis granuloma (45.23)
Authors: McBride Amanda B , Salgame Padmini 
Published: January 2023

Abstract
Tubercle granulomas are highly organized immune structures that protect the host against Mycobacterium tuberculosis (Mtb) infection. The organization of immune cells around infected macrophages serves to sequester Mtb and enhance macrophage microbicidal responses. In addition, the granuloma localizes inflammation, thereby preventing damage to surrounding lung tissue. Although there is controversy regarding the involvement of Toll-like receptor 2 (TLR2) in host defense against Mtb, our findings indicate that TLR2 may be playing a key role in regulating these two opposing functions of the Mtb granuloma. Comparative studies between wild type and TLR2 gene-deficient mice demonstrated that the absence of TLR2 did not affect the magnitude of the Th1 effector response generated following aerosol infection with Mtb or the induction of recall Th1 memory immunity in response to Mtb challenge. However, the consequence of TLR2 absence was manifested at the level of the granuloma. Analysis of lungs from chronically-infected TLR−/− mice revealed increased bacterial burden and loss of granuloma integrity in comparison with infected WT mice. The infected lungs of TLR2−/− mice also exhibited enhanced inflammation and reduced Foxp3+ regulatory T cells. Together, these findings provide insights into how TLR2 might regulate the tubercle granuloma.
B7.1 and B7.2 are crucial for primary response to Mycobacterium tuberculosis infection but redundant for the induction of memory response (45.24)
Authors: Bhatt Kamlesh , Rafi Wasiulla , Salgame Padmini 
Published: January 2023

Abstract
Costimulation plays a crucial role in priming of naïve T cell to an effector cell type. In a recent study, we have shown that lack of B7.1 and B7.2 (DKO) causes the host to generate a dampened Th1 effector response, disabling it to contain the bacterial burden during chronic phase of infection. Since the development of T cell memory is connected to the effector T cell response, we sought to investigate how a defective primary effector response will affect the induction of the memory. Memory mice were generated by treating infected wildtype (WT) and DKO mice with antibiotics and resting them after the treatment. These memory mice along with age matched naïve controls were then challenged with low dose infection with M. tuberculosis. A significant decrease in bacterial burden was seen in both WT and DKO memory mice at 4 and 6 weeks post infection when compared to naive WT and DKO mice respectively. This decreased bacterial burden was accompanied with a faster Th1 effector response in both WT and DKO memory lungs as indicated by increased number of IFNγ producing cells in the lungs. These results indicate that despite its inability to generate a defective primary response the DKO host is able to mount a memory response against M.tuberculosis infection. Further studies are in progress to study the mechanism of memory induction in the absence of a successful primary response in the DKO mice.
This study was supported by NIH Grant AI 49778, Potts Memorial Foundation Award and UMDNJ Foundation Award.
Parenchymal expression of CD40 exacerbates liver injury in viral hepatitis (45.25)
Authors: Gupta Shalini , Yan Jiabin , Sun yixiao , Chan Tehsheng , Sun Jiaren 
Published: January 2023

Abstract
T cells contribute to immune-mediated liver injuries in viral hepatitis. Although high levels of costimulatory molecules, e.g. CD40 and CD86, are found in activated Kupffer cells and hepatocytes in patients, the precise role of hepatic CD40 expression in antigen presentation, T cell activation and liver injury in viral hepatitis is not very clear. In this study, we generated conditional transgenic mice having the liver-specific expression of CD40, and induced transgene expression by i.v. injection with 2×109 pfu recombinant replication-deficient adenovirus with Cre. The transgenic mice and their non-transgenic littermates were sacrificed, and the serum and tissue samples were collected at 3, 6, 11, 18 and 27 days post infection. In control mice, the serum alanine aminotransferase (ALT) levels were slightly elevated at days 6 and 18, and normal at other time points. Although transgenic mice had relatively normal serum ALT levels at day 3, they displayed a significant elevation of ALT levels to an average of 501 and 366 U/L on days 6 and 11, respectively, followed by decreased levels thereafter. Pathological examinations revealed a relatively mild lymphocyte infiltration in isolated portal areas in control animals at day 6. In contrast, there was significant lymphocyte infiltration in multiple portal areas, as well as in the liver parenchyma, that was accompanied by various degrees of hepatic apoptosis (Councilmen's bodies) and necrosis in transgenic mice at days 6 and 11. These results suggest that CD40 expressed on hepatocytes play a critical role in virus-induced hepatitis.
Impaired Anti-viral Immunity Following Syngeneic Bone Marrow Transplant (45.7)
Authors: Coomes Stephanie M. , Wilke Carol A. , Moore Thomas A. , Moore Bethany B. 
Published: January 2023

Abstract
Patients receiving autologous hematopoietic stem cell transplantation (HSCT) or bone marrow transplantation (BMT) as therapy for various malignancies or autoimmune diseases have an increased risk for infectious complications post-transplant, especially in the lung. We have used syngeneic BMT in mice and the murine gammaherpesvirus, MHV-68, to study the efficacy of adaptive immune responses post-BMT. Using this model, we have shown that mice 5 weeks post-transplant have fully reconstituted their hematopoeitic lineages in both the lung and periphery. However, the BMT mice have a reduced ability to clear lytic virus from the lung, despite increased expression of interferon gamma, interleukin-2, and TNF-alpha in the lungs of BMT mice. Recruitment of immune cells to the lungs after MHV-68 infection is similar in BMT mice and untransplanted controls, suggesting functional defects. While bone-marrow derived dendritic cells from transplanted mice are able to stimulate allogeneic T cell proliferation in mixed leukocyte reactions (MLRs), CD4+ T cells from BMT mice show reduced proliferative responses in MLRs. These results suggest defects in CD4+ T cell responses post-BMT may contribute to enhanced susceptibility to viral infections. Additionally, levels of TGF beta are significantly increased in the lungs of BMT mice before infection, and numbers of CD4+Foxp3+ regulatory CD4+ T cells are increased in the BMT lung (but not spleen) both before and at day 7 after viral infection. We speculate that TGF beta-induced expansion of Tregs in the lung limits anti-viral host defense post-bone marrow transplantation.
Reduced c-myc expression levels limit follicular mature B cell cycling in response to TLR signals (46.6)
Authors: Bandaranayake Ashok D , Meyer-Bahlburg Almut , Andrews Sarah F , Rawlings David J 
Published: January 2023

Abstract
The splenic B cell compartment is comprised of two major, functionally distinct, mature B cell subsets, i.e. follicular mature (FM) and marginal zone (MZ) B cells. Whereas MZ B cells exhibit a robust proliferative response following stimulation with the TLR4 ligand, LPS, FM B cells display markedly delayed and reduced levels of proliferation to the identical stimulus. The current study was designed to identify a potential mechanism(s) accounting for this differential responsiveness. In contrast to the delay in cell cycle entry, FM and MZ B cells exhibited nearly identical LPS-driven alterations in the expression level of cell surface activation markers. Further, both the NFκB and mTOR signaling cascades were similarly activated by LPS stimulation in FM vs. MZ B cells, while inducible activation of ERK and AKT were nearly absent in both subsets. MZ B cells, however, exhibited higher basal levels of pAKT and pS6 consistent with a pre-activated status. Importantly, both basal and LPS activation-induced c-myc expression was markedly reduced in FM vs. MZ B cells; and enforced c-myc expression fully restored the defective proliferative response in FM B cells. These data support a model wherein TLR responses in FM B cells are tightly regulated by limiting c-myc levels thereby providing an important checkpoint to control non-specific FM B cell activation in the absence of cognate antigen.
Distinct regulatory roles for Janus Tyrosine Kinase 3 (Jak3) in generation of T-helper 17 (TH17) and inducible regulatory T cells (Tregs) (47.10)
Authors: Khattar Mithun , Chen Wenhao , Stepkowski Stanislaw 
Published: January 2023

Abstract
The development of inducible Foxp3+ regulatory T (iTreg) cells is reciprocally related to that of pro-inflammatory T helper (Th) 17 cells. Both processes depend on transforming growth factor (TGF)-β. Recent studies indicated that interleukin (IL)-2, a Janus kinase 3 (Jak3)-dependent cytokine, is required for the development of iTreg cells, while exhibiting an inhibitory effect on Th17 differentiation. This raises the question about distinct roles of Jak3 in iTreg vs. Th17 differentiation. To address this question, we cultured naïve CD4+ T cells with TGF-β alone for iTreg cell polarization or TGF-β with IL-6 for Th17 cell polarization without or with selective Jak3 inhibitor, NC1153. We found that a dose-dependent inhibition of Jak3 reduced the TGF-β-mediated iTreg differentiation. In contrast, similar NC1153 concentrations under Th17 polarizing condition increased the percentage of Th17 cells. When we examined the proportion of regulatory T (Treg) cells within the CD4+ T-cell population from Jak3-dificient mice, we found that these mice lack Treg cells. Thus, inhibition of Jak3 has differential effects on iTreg/Th17 balance with preferential generation of pro-inflammatory Th17 cells.
This work was supported by grants from NIH (grant number HL 69723)
MicroRNA Profiles of TH Cells (47.11)
Authors: Batts LaKeisha , Coarfa Cristian , Creighton Chad , Gunarante Preethi , Corry David 
Published: January 2023

Abstract
Naïve CD4+ T cells differentiate into two classes of helper effector (TH) cells during exposure to antigen within peripheral lymphoid tissues. TH1 cells are generated in the presence of IL-12 and IFN-γ whereas TH2 cells result from the influence of IL-4 and IL-6. These cells orchestrate adaptive immune responses by regulating: T cell effector differentiation, macrophage activation, and antibody isotype switching. The mechanisms that control CD4+ T cell differentiation are not yet fully defined. These short transcripts regulate gene expression and are capable of defining and altering cell fate. The importance of miRNA function in T-helper cell differentiation and function has yet to be determined. Using Next Generation Sequencing, we have begun to dissect the smallRNAome of TH cells before and after differentiation. We found enhanced induction of miRNAs in TH1 cells (64% vs. 55% in naïve) relative to TH2 cells (45% vs. 55% in naïve) suggesting a role for miRNAs in TH cell differentiation. miRNAs mmu-let-7c and mmu-mir-181a, which have previously been shown to function in TH cell development and activation were increased in expression in TH1 cells. Expression of two additional miRNAs, mmu-mir-199a and mmu-mir-101a, were also enhanced in both TH1 and TH2 cells and based on their putative immune targets STAT6 and IL-4Rα, we propose that they critically regulate TH1 and TH2 cell differentiation. We further discovered novel miRNAs that putatively target GATA3 and several genes related to cell survival and apoptosis. Understanding how differentially expressed miRNAs function in TH cell differentiation will pave the way for novel and highly specific therapies for TH cell-mediated diseases.
Increased number of systemic T regulatory cells but not Th1 or Th2 cells in tobacco smokers (47.14)
Authors: Malmhall Carina Anna , Andersson Anders , Jackute Jurgita , Houltz Birgitta , Lötvall Jan , Lindén Anders , Bossios Apostolos 
Published: January 2023

Abstract
Tobacco smokers with severe chronic obstructive pulmonary disease (COPD) display a local accumulation of T helper cells and it has recently been proposed that smokers have an increased number of local and systemic T regulatory (Treg) cells as well. However, it is not known whether smokers have an altered number systemic T helper (Th) cells matching that of Treg cells. The current transectional study addressed this issue in human subjects.
Blood samples were obtained from 6 chronic smokers with COPD (GOLD stage II-III), 5 chronic smokers with normal lung function and 6 healthy never-smokers. Treatment with cortisone, long acting beta-2-agonists, anticholinergics and NSAIDs, respectively, was not allowed 4 weeks prior to sampling. T cells (CD3+ and CD4+ respectively), Tregs (CD4+FOXP3+), Th1 (CD4+T-bet+) and Th2 (CD4+GATA-3+) cells were determined using immunocytochemistry.
We found no pronounced or statistically significant difference in the number of T cells or Th1 cells, when comparing the three groups of subjects. Systemic Th2 cells were not detectable in either group. However, we confirmed that the mean [SEM] number of Tregs (CD4+Foxp3+) was higher in chronic smokers with and without COPD (3.30 [0.63] x104/ml and 2.72 [0.49] x104/ml, respectively) than in never-smokers (1.36 [0.19] x104/ml) (p = 0.02).
Our results suggest that chronic smoking has no pronounced effect on the number of systemic Th1 or Th2 cells and that chronic smoking rather than COPD per se accounts for the increased number of systemic Tregs.
Funded by the Heart-Lung Foundation of Sweden