bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fli-1 transcription factor affects inflammatory cell infiltration into the kidneys independent of autoantibodies (P4150)
Authors: Sato Shuzo , Williams Sarah , Zhang Xian 
Published: January 2023

Abstract
Background and Purpose: Expression levels of Fli-1, a member of the Ets family of transcription factors, are a mitigating factor in the development of nephritis in lupus model mouse. In this study, we examined the effects of reduced expression of Fli-1 on inflammatory cell infiltration into the kidneys in vivo. Methods: We generated MRL/lpr transgenic mice expressing green fluorescent protein (GFP), and isolated spleen cells from these mice. One million cells were injected into 9 Fli-1 heterozygous (+/-) and 13 wild-type (WT) MRL/lpr mice at the age of 4 to 8 months. Mice were sacrificed 18 hours after injection, kidneys were removed and frozen sections were made. Inflammatory cells with GFP were counted by immunofluorescence microscopy. The expression of chemokines in the kidney tissue was analyzed by real-time PCR. Results: The GFP positive cells were significantly higher in kidneys from WT MRL/lpr mice compared to Fli-1+/- MRL/lpr mice despite similar autoantibody titers found. We identified the majority of these cells as T cells, B cells and macrophages. Fli-1+/- MRL/lpr mice had less proteinuria and the expression level of chemokines in the kidneys had lower trends after GFP cells injection. Conclusion: Our data indicate that Fli-1 expression impacts lupus nephritis development by modulating expression of inflammatory chemokines in the kidney and infiltration of inflammatory cells independent of autoantibodies in MRL/lpr mice.
Transition from acute to persistent infection: Antigen persistence and cytotoxic T cells response in infectious bursal disease virus infected SPF chickens (P6129)
Authors: Rauf Abdul , Vlasova Anastasia , Murgia Maria , Jung Kwonil , Saif Yehia 
Published: January 2023

Abstract
Described by Cosgrove about a half century ago, infectious bursal disease (IBD) is as an acute, highly contagious viral infection of young chickens, which last for 7 to 10 days and leads to immunosuppression. Here, we are reporting for the first time that in IBD virus (IBDV) -infected, specific pathogen free (SPF) chickens bursa, the virus antigen, macrophages, CD4+, CD8+ cells, Fas, Fas ligand (FasL), and caspase-3 were consistently detected by immunohistochemistry up to 56 days post inoculation (DPI). Two-week-old SPF chickens were inoculated intraocularly with standard challenge strain (STC) using a 104embryo infectious dose. Histopathology of bursal tissues revealed severe lymphoid follicular depletion at DPI 5-28. A distinctive pattern of bursal follicles in the form of small and large follicles was detected from DPI 35 to 56. At 42 and 56 DPI a characteristic clustered pattern of IBDV antigen, macrophages, CD4+, CD8+ cells, Fas, FasL, and caspase-3 were detected in or around the small bursal follicles as compared to large bursal follicles. This is the first report on the persistence of IBDV antigen causing a latent infection, in bursal tissues of SPF chicken. Detection of persistent IBDV infection is highly significant in the elucidation of virus pathogenesis, immunology and epidemiology and it raises many questions that need to be answered in future studies.
N-acetylcysteine modulation of JAK2 and JAK3 results in decreased STAT3 activation in human B lymphocytes (P1473)
Authors: Bonnaure Guillaume , Néron Sonia 
Published: January 2023

Abstract
Activation of STAT3 is crucial for human B lymphocyte differentiation and inhibition of its phosphorylation in CD40-activated B lymphocytes results in impairment of in vitro generation of immunoglobulin-secreting cells. Conversely, redox homeostasis plays a role in B lymphocyte maturation. Thus, the presence of antioxidants can potentially modulate their outcome. We showed that N-acetylcysteine (NAC), an antioxidant molecule, can inhibit STAT3 phosphorylation, which is paralleled with strong inhibition of immunoglobulins secretion. Western analysis reveals that NAC-treatment of CD40-activated human B lymphocytes, in the presence of IL-2, IL-4 and IL-10, causes a reduced phosphorylation of JAK2 and JAK3, which are important activators of STAT3. Furthermore, this inhibition appears to be dose-dependant. Our results indicate that other antioxidant molecule such as α-tocopherol and trolox do not affect these two kinases phosphorylation. Other kinases or their inhibitors associated to STAT3 phosphorylation do not seem affected by the presence of antioxidant in the media. In conclusion, these observations reveal that STAT3 phosphorylation down regulation in NAC conditions is due to its association with JAK2 and JAK3.
Metabolic strategies used by long-lived bone marrow plasma cells (P1474)
Authors: Minges Wols Heather , Witte Pamela 
Published: January 2023

Abstract
Maintaining effective humoral immunity is an important aspect of healthy aging and the goal of most vaccines. Evidence from multiple laboratories agrees that long-lived humoral immunity derives from long-lived plasma cells (LLPC) that reside in the bone marrow. Because serum antibodies have a relatively short half-life and must be constantly produced, an urgent need is a clearer understanding of how LLPC maintain secretion of protective antibodies for years or a lifetime. PC secrete an estimated 1000 molecules of antibodies/sec, and some investigators make a compelling argument that the only recourse for a PC is a short lifespan, limited by oxidative and proteosomal stress. Our work begins to address how these important cells survive the extreme metabolic demands of constant antibodies secretion that is essential to lasting, protective immunity. We identified PC using the fluorescent probe ER Tracker - as highly-secretory cells, PC have large amounts of rough ER. In parallel, we stained cells with a reduced MitoTracker probe that fluoresces upon oxidation in respiring mitochondria. Our data show that ER Tracker-hi cells were found in both MitoTracker-hi and -lo populations, suggesting PC may use different metabolic strategies: aerobic respiration and glycolysis. Further studies will assess what governs each metabolic strategy, providing a deeper understanding of the cellular metabolic phenotype of long-lived plasma cells.
Analysis of the differentiation of single antigen-specific B cells early in the primary response (P1477)
Authors: Taylor Justin , Pape Kathryn , Jenkins Marc 
Published: January 2023

Abstract
Following activation with antigen, naïve B cells undergo a robust proliferative expansion and differentiation that results in the production of numerous distinct cell types with different roles in protective immunity. While some of the transcriptional regulators and extrinsic factors directing differentiation are known, these mechanisms cannot account for the heterogeneity of the response. In this study we tracked the response of single naïve B cells and found that individual cells do not differentiate into all of the cell types found within a polyclonal response. Thus, the heterogeneity found within the polyclonal response is in part the result of different naïve B cells differentiating differently. Ongoing work is focused on determining the contributions that B cell receptor affinity and stochastic competition play in differentiation.
The translesion DNA synthesis polymerase Rev1 recruits uracil DNA glycosylase to switch regions and enhances dU glycosylation for immunoglobulin class switch DNA recombination (P1478)
Authors: White Clayton , Zan Hong , Thomas Lisa , Mai Thach , Li Guideng , Xu Zhenming , Zhang Jinsong , Casali Paolo 
Published: January 2023

Abstract
Immunoglobulin (Ig) class switch DNA recombination (CSR) plays a critical role in the maturation of the immune response by diversifying the biological effector functions of antibodies. CSR is initiated by activation-induced cytidine deaminase (AID)-mediated deoxycytosine (dC) deamination, yielding deoxyuridine (dU), and dU glycosylation by uracil DNA glycosylase (Ung) in Ig switch (S) region DNA. Translesion DNA synthesis (TLS) polymerases, such as Rev1, play important roles in somatic hypermutation (SHM), but their role in CSR has not been defined. We found that Rev1 directly interacts with Ung to recruit it to S region DNA and enhance dU glycosylation for CSR. Rev1 targets S regions undergoing CSR in an AID-dependent and Ung-independent fashion. Ung recruitment to S regions, DNA-dU glycosylation and CSR are reduced in Rev1-/- B cells. Together with a mutation spectrum in S region DNA that is similar to that of Ung-/- B cells, our findings suggest that Rev1 operates in the same pathway as Ung during CSR, as emphasized by further decreased CSR in Rev1-/- Msh2-/- B cells. Rescue of CSR by a catalytically inactive Rev1 mutant in Rev1-/- B cells shows that the role of Rev1 in CSR is mediated by Rev1’s scaffolding function, not its enzymatic function. Thus, we describe a novel and important scaffolding function for Rev1 in CSR, which together with its known enzymatic function in SHM highlights a central role of this TLS polymerase in the maturation of the antibody response.
Increased autoimmunity in men with Klinefelter’s syndrome: Sex chromosome dosage effect (P3309)
Authors: Okorogu Rita , Surampudi Prasanth , Valera Isela , Halder Ramesh , Okereke Chetachi , Wang Christina , Swerdloff Ronald , Singh Ram 
Published: January 2023

Abstract
Autoimmune diseases are more common in women than in men. Studies in animal models suggest a role of sex chromosome genotype in modulating autoimmune susceptibility. Here, we begin to translate these murine findings onto humans. Our hypothesis is that X chromosome dosage, rather than the female sex itself, imparts susceptibility to autoimmune diseases. Our results show that levels of IgG anti-chromatin and anti-nucleosome autoantibodies were significantly higher in men with sex chromosome aneuploidy (47,XXY) compared to 46,XY men. XXY men, however, did not have non-specific B cell hyper-reactivity, as the levels of anti-thyroid peroxidase antibody that is associated with autoimmune thyroid disease were similar between the two groups. A preliminary analysis of completed questionnaire revealed an increase in autoimmune and inflammatory conditions when compared with known population prevalence of these diseases. Analysis of immune cell phenotype and responses conducted thus far showed a significant reduction in the frequency of natural killer T cells in 47,XXY men as compared to 46,XY men. These data suggest a role of X chromosome dosage in inducing autoimmunity in humans. Since animals and humans with autoimmune diseases have reduced numbers and/or functions of natural killer T cells, our observation of reduced natural killer T cells in 47,XXY men may be related to increased autoimmunity in these men.
Dual signals of TCR and Notch promote antigen-specific T cell development from pluripotent stem cells (P4370)
Authors: Lei Fengyang , Song Jianxun 
Published: January 2023

Abstract
The pivot in conducting a successful adoptive T-cell immunotherapy for cancer is the obtaining of a large number of naïve phenotyped, stem-cell like tumor antigen specific T cells. Previously we have reported that both conventional and antigen-specific T cells could be generated from induced pluripotent stem (iPS) cells by different methods. Here, we observed that dual signals of TCR and Notch played a significant role in antigen-specific T lineage differentiation from pluripotent stem cells. We used a murine MHC-I restricted, tyrosinase-related protein 2 (TRP2) specific TCR and Notch ligands Delta-like (DLL) 1 and 4 as a model system for inducing iPS cells to differentiate into melanoma-specific CD8+ T cells. Murine iPS cells were retrovirally transduced with TRP2-specific TCR (iPS-TRP2), and then co-cultured on OP9 bone marrow stromal cells overexpressing both DLL1 and DLL4 ligands (OP9-DL1/DL4) in the presence of Flt3L and IL-7. This approach has a better effect in terms of directing an in vitro iPS-TRP2 differentiation towards TRP2-specific CTLs. In addition, adoptive transfer of in vitro developed iPS-TRP2-derived precursor T cells into C57BL/6J mice bearing xenografted B16 melanoma significantly increased anti-tumor immunosurveillance and protected animal from tumor challenge. These data suggest that signals from TCR and Notch can cooperatively promote antigen-specific T cell development from pluripotent stem cells.
Quantitative single-cell functional characterization of CD19-specific chimeric antigen receptor T cells (P4344)
Authors: Varadarajan Navin , Liadi Ivan , Romain Gabrielle , Singh Harjeet , Cooper Laurence 
Published: January 2023

Abstract
Adoptive cell therapy (ACT) utilizing chimeric antigen receptor (CAR) T cells rendered specific for CD19 have demonstrated significant anti-tumor effects in patients with CD19+ chronic lymphocytic leukemia (CLL). In spite of the clinical promise of ACT in achieving complete responses, their efficacy remains unpredictable and new approaches are needed to a priori define the therapeutic potential of T-cell based therapies. In our current work, we characterize the in vitro functionality of CD19-specific (CD19RCD28) CAR+ T cells propagated using artificial antigen presenting cells, by employing a novel methodology single-cell nanowell screening (SNS).We show that CAR+ T cells exert specific cytotoxicity against NALM6 cells (31 ± 8 %) when co-incubated at a 1:1 ratio in nanowell containers. Furthermore, single CAR+ T cells were capable of engaging and killing multiple targets; 17 ± 8% of T cells killed two target cells within the 6 hour window of observation. Hierarchical clustering indicated that interferon-gamma (IFNγ) secretion is not correlated to cytotoxicity of T cells. CAR+ T cells that secreted IFNγ upon target ligation did not undergo activation-induced cell death (AICD) whereas T cells that engaged in repeated killing showed an increased propensity to undergo AICD (p = 0.04). In summary, our SNS based methodology allows the deep functional characterization of clinical grade CAR+ T cells and can be used to determine in vitro functions of CAR+ T cells.
Poly(dG) conjugated protein immunization targets dendritic cells and induces T cell immunity in vivo. (P4528)
Authors: Toussaint Helene , Erdos Geza , Larregina Adriana , Carey Cara , Falo Louis 
Published: January 2023

Abstract
The efficacy of current vaccines results primarily from the induction of protective antibodies. However, we still lack efficacious vaccines against many infectious agents that, like cancer, require durable and protective T cell immunity. Efficient and selective delivery of antigen and adjuvant to antigen presenting cells in vivo remains a critical challenge to vaccine design. Therefore, we have designed an antigen and adjuvant delivery platform based on simple and direct coupling of Poly(dG) oligo deoxynucleotides to protein antigens. In initial studies we found that immunization of naïve animals by direct injection of conjugates of Poly(dG) and Ovalbumin, Poly(dG)-OVA induces efficient priming of potent antigen specific CTL responses, differentiation of naïve CD4 T cells into Th1 cells, induction of specific memory T cell responses and generation of antigen-specific antibody responses. Applying this strategy to HIV, we immunized mice with conjugated Poly(dG)-SIV-Gag and demonstrated potent induction of Gag-specific CTL responses in vivo. Further, in human models we show that intradermal injection of Poly(dG)-antigen conjugates result in efficient uptake by human skin migratory DCs, preferentially dermal DCs, that also acquire potent T cell stimulatory functions. Our data suggests that the Poly(dG)-protein conjugate immunization platform can both selectively target antigens to skin APC and function as an APC-directed adjuvant to induce broad based humoral and T cell immunity.
Disrupted T cell central memory following vaccination in patients with sickle cell disease (P4536)
Authors: Adami Alexander , Szczepanek Steven , Lewis Deirdre , Farrington Nayre , Bracken Sonali , Natarajan Prabitha , Secor, Jr. Eric , Jellison Evan , Thrall Roger , Andemariam Biree 
Published: January 2023

Abstract
Increasing evidence suggests that a systemic state of disrupted immunity and inflammation accompanies sickle cell disease (SCD) despite the erythrocytic nature of the SCD mutation. Frequent infection plagues patients, who require aggressive vaccination from an early age. However, recent experimental and clinical evidence suggest diminished vaccine efficacy in SCD. We hypothesize that patients with SCD exhibit disrupted T cell memory in response to vaccination. We collected blood from nine healthy adult controls and eleven SCD patients before and 4-6 weeks after routine vaccination against influenza virus (IRB #11-026-1). Peripheral blood mononuclear cells were isolated and analyzed by flow cytometry. As previously reported, relative percentages of CD8+ central memory (CM) T cells significantly contracted following vaccination in control subjects (from 48.2 to 39.3, p &lt; 0.01). Conversely, relative percentages of CD8+ CM T cells did not change in SCD patients (44.3 to 44.4, p &gt; 0.05). Further, CD4+ CM T cells were significantly higher in patients with SCD versus controls before vaccination (58.7 vs. 44.7, p &lt; 0.02) and trended higher following vaccination (56.8 vs. 40.7, p = 0.08). These data indicate that patients with SCD exhibit both disrupted CD8+ CM kinetics and altered CD4+ CM levels in the context of vaccination. Our findings suggest that at least a portion of the poor clinical response to vaccination in SCD may arise from disrupted T cell memory.
Metabolic engineering of Salmonella vaccine bacteria to boost human γδ T cell immunity (P4538)
Authors: Workalemahu Grefachew , Wang Hong , Puan Kia-Joo , Kuzuyama Tomohisa , Jones Bradley , Morita Craig 
Published: January 2023

Abstract
Human Vγ2Vδ2 T cells monitor isoprenoid metabolism by recognizing foreign (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), an intermediate in the 2-C -methyl-D-erythritol-4-phosphate (MEP) pathway used by most Eubacteria and Apicomplexan parasites, and self isopentenyl pyrophosphate (IPP), an intermediate in the mevalonate pathway used by humans. Whereas microbial infections elicit prolonged expansion of memory Vγ2Vδ2 T cells, immunization with synthetic prenyl pyrophosphates or aminobisphosphonates elicit short-term expansion with rapid anergy and deletion. We hypothesized that a live attenuated bacterial vaccine metabolically engineered to overproduce HMBPP would elicit Vγ2Vδ2 T cell immunity by mimicking natural infections. Here we report on an aroA- Salmonella enterica serovar Typhimurium SL7207 strain metabolically engineered by deleting the gene for lytB (the downstream enzyme from HMBPP) and complementing with genes encoding mevalonate pathway enzymes. lytB- SL7207 had extremely high HMBPP levels, infected human cells like the wild type bacteria, and stimulated large ex vivo expansions of Vγ2Vδ2 T cells in all 6 human donors tested. Moreover, vaccination of rhesus monkeys with live lytB- SL7207 expanded Vγ2Vδ2 T cells without significant side effects. Thus, these studies provide proof-of-principle for the use of metabolic engineering to derive Salmonella vaccines that boost human γδ T cell immunity.
IL-27Rα is required for shaping the magnitude and repertoire of T cells responses to subunit immunization (P4500)
Authors: Pennock Nathan , Kedl Ross 
Published: January 2023

Abstract
A goal of cellular immune vaccines is the generation of large numbers of antigen specific T cells in response to subunit immunization. A broad spectrum of cytokine and cell surface costimulatory molecules are known to shape the magnitude and repertoire of T cells responding to vaccination. We show here the majority of TLR agonist-based vaccine adjuvants unexpectedly depend on IL-27 for eliciting CD4 and CD8 T cells responses. This is in sharp contrast to infectious challenge which generates T cell responses that are IL-27 independent. Mixed bone marrow chimera experiments demonstrate that IL-27 dependency is T cell intrinsic, requiring T cell expression of IL-27Rα. Further, we show that IL-27 dependency is not only necessary for the maximal expression of vaccine elicited T cell responses, but also critical for representation of high affinity T cells in the responding T cell pool. This observation complements the biology from other reports demonstrating dependence of low affinity T cells on the TNFR family member CD27. Collectively our data highlight the unexpected central importance of IL-27 in the generation of robust, high affinity cellular immune response to subunit immunization.
Anti-viral vaccines license T cell responses by suppressing granzyme B levels in human plasmacytoid dendritic cells. (P4510)
Authors: Fabricius Dorit , Nussbaum Bendikt , Busch Danie , Panitz Verena , Mandel Birgit , Vollmer Angelika , Kaltenmeier Christof , Lunov Oleg , Tron Kyrylo , Nienhaus Ulrich , Debatin Klaus-Michael , Jahrsdörfer Bernd 
Published: January 2023

Abstract
Human plasmacytoid dendritic cells (pDC) are important modulators of adaptive T cell responses during viral infections. Recently, we found that human pDC produce the serine protease granzyme B (GrB), thereby regulating T cell proliferation in a GrB-dependent manner. Here, we demonstrate that intrinsic GrB production by pDC is significantly inhibited in vitro and in vivo by clinically used vaccines against viral infections such as tick-borne encephalitis (TBEV). We show that pDC GrB levels inversely correlate with the proliferative response of co-incubated T cells and that GrB suppression by a specific antibody or a GrB substrate inhibitor results in enhanced T cell proliferation, suggesting a predominant role of GrB in pDC-dependent T cell licensing. Functionally, we demonstrate that GrBhigh, but not GrBlow pDC transfer GrB to T cells, and may degrade the zeta-chain of the T cell receptor in a GrB-dependent fashion, thereby providing a possible explanation for the observed T cell suppression by GrB-expressing pDC. Modulation of pDC-derived GrB activity represents a previously unknown mechanism by which both anti-viral and vaccine-induced T cell responses may be regulated in vivo. Our results provide novel insights into pDC biology during vaccinations and may contribute to an improvement of prophylactic and therapeutic vaccines.
Exposure to low doses of chemically attenuated blood stage malaria parasites rapidly activates CD4+ T cells and induces strain and species transcending immunity (P4477)
Authors: Good Michael , Reiman Jennifer , McPhun Virginia , Rodriguez Bibiana , Ito Koichi , Raja Amber , De Sai , Engwerda Christian , Lee Moses , Stanisic Danielle 
Published: January 2023

Abstract
There has been a re-appraisal of the role of cell-mediated immunity in protection against malaria. We have developed a vaccine strategy using chemical attenuation of infected red cells to induce protective cellular immunity. Our focus has been on attenuation using a class of chemicals typified by centanamycin, which binds in AT-rich regions of the parasite genome. Chemically attenuated parasites (CAPs) do not grow in immunodeficient mice. We observe that low doses of Plasmodium chabaudi ring stage CAPs induce profound protection against different strains and species of Plasmodium. CAPs prepared using other species of Plasmodium are also effective. CAPs are rapidly removed from the peripheral blood following vaccination and are taken up by dendritic cells and macrophages in the spleen and liver. Vaccination does not induce detectable antibodies but protection is T-cell dependent. Following vaccination there is a rapid increase within the blood of activated CD4+ T cells (defined as CD49d+ CD11a+). Transfused ovalbumin-specific transgenic OTII cells are not activated following vaccination. This novel approach is now being applied to P. falciparum where we plan to vaccinate humans with low doses of P. falciparum CAP, determine their immune responses and then challenge vaccinees with viable parasite-infected human red cells to determine vaccine efficacy.