bluuebunny/crossref_metadata_2025_split huggingface dataset:
Natural Killer cell function during foot-and-mouth disease virus infection (46.19)
Authors: Toka Felix N. , Nfon Charles K. , Endsley Janice J. , Ferman Geoffery S. , Estes D. Mark , Golde William T. 
Published: January 2023

Abstract
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. Immunological knowledge to assess more rapid acting vaccines against FMDV is presently limited. We examined the reactivity of swine and cattle NK cells following infection for their capability to express intracellular perforin, to kill a human tumor cell line target in vitro, and to secret IFN gamma. The cytotoxicity of NK cells from non-infected animals against the K562 cells is low with baseline levels at 5–15% in swine and 15–20% in cattle. Stimulation with rhIL-2 or rhIL-12 plus rhIL-15, increased the lytic activity against K562 cells. Infection with FMDV inhibited swine NK cell lytic activity but did not significantly increase IFN gamma secretion during the acute infection. Perforin expression increased but this did not correlate with the killing capability of the swine NK cells. Infection of cattle with FMDV initially activated the NK cells to increase target cell lysis. NK cell IFN gamma secretion and perforin expression were slightly elevated upon infection and coincided with the lytic activity in cattle. These results are a further indication of immune evasion by FMDV by inhibiting or limiting NK cell function. The potential to manipulate the innate immune response to block this evasion is discussed in the context of designing rapid acting vaccines for foot-and-mouth disease.
Increased socs-1 expression in dendritic cells may be responsible for reduced type 1 cytokine responses in HIV-1 transgenic rats (46.20)
Authors: YADAV ANJANA , REID WILLIAM 
Published: January 2023

Abstract
Dendritic cell (DC) function appears to be compromised in HIV infected individuals, although it is unclear whether the dysregulation is a result of direct infection of the DCs or an indirect effect of exposure to HIV proteins. Since effector/memory formation depends upon clonal expansion of naïve T cells following antigenic stimulation and since cytokines play a critical role in the initiation and regulation of immune responses by DCs, events that negatively affect antigen presentation, co-stimulation and cytokine production by DCs, also negatively affect T cell immune responses. Infection with HIV-1, results in cytokine dysregulation by DCs. Suppressor of cytokine signaling-1 (SOCS-1) is an inducible negative regulator of the JAK/STAT signal pathway; SOCS-1 is expressed in DCs and negatively regulates activation and type 1 regulatory cytokine production. We have earlier reported that HIV-1 transgenic rats have defects in type 1 cytokine production, type 1 cytokine responses and generation of effector/memory CD4 T cell subsets. Here we show that B cells from the HIV-1 Tg rats are also dysregulated in KLH specific primary and secondary Th1 isotype (IgG2a) responses. Additionally, bone marrow derived DCs (BMDCs) from HIV-1 Tg rats upon stimulation with LPS and/or rIFN-gamma, exhibit reduced immunostimulatory functions depicted by reduced production of IL-12+p40, increased expression of IL-10 and SOCS-1 and reduced STAT-1 phosphorylation relative to age matched controls. These outlined immune abnormalities together with the dysregulation in memory/effector development and Th1 responses suggests a link between SOCS-1 expression and IFN-γ-induction of IL-12 by BMDCs, resulting in impaired T helper type 1 responses reported in HIV-1 Tg rat. NIH-NIAID R01-A163171
4-1BB antibody significantly enhances SIV-specific CD8+ T-cell immunity and proliferation following naked DNA vaccination in cynomolgus macaques (47.10)
Authors: Hokey David A , Hirao Lauren A. , Calarota Sandra A. , Yan Jian , Boyer Jean D. , Dai Anlan , Yoon Hanna , Jure-Kunkel Maria N. , Balimane Praveen , Weiner David B. 
Published: January 2023

Abstract
A major challenge in the development of an effective vaccine for HIV is to stimulate potent cell-mediated immunity that includes the induction of long term memory. Ligation of the activation-induced co-stimulatory molecule, 4-1BB, stimulates CD8+ T-cell proliferation and the generation of memory in mice. We evaluated monoclonal antibody (mAb) agonists for 4-1BB to enhance DNA vaccines. Two studies were performed in cynomolgus macaques to evaluate the ability of these mAb to augment intramuscular DNA immunizations with either plasmid SIV-Gag or with codon-optimized consensus SIV-Gag, SIV-Env, and SIV-Pol plasmid DNA. Administration of anti-4-1BB mAb doubled the CD8+ T-cell killing ability as measured by granzyme B ELISpot assays. Furthermore, the proliferative response of SIV-specific CD8+ T-cells was increased by more than 5-fold after one immunization as measured by CFSE proliferation assays. Our data demonstrate the ability of 4-1BB stimulation to enhance cellular immunity following DNA vaccination in NHPs. These findings have clinical importance in that they represent a considerable improvement in the efficacy of naked intramuscular DNA vaccines in large animals and demonstrate the ability of 4-1BB agonists to direct CD8+ T-cell responses and memory.
This work is supported in part by NIH and NIAID funds.
Induction of human primary antibody responses in vitro by ovalbumin (OVA) or anthrax (rPA) on dendritic cells (DCs) and follicular dendritic cells (FDCs) in temporally controlled germinal center reactions (GCRs) (47.19)
Authors: Shikh Mohey Eldin M El , Sayed Rania M El , Wu Yongzhong , Szakal Andras K , Tew John G 
Published: January 2023

Abstract
Induction of potent primary human Ab responses entirely in vitro is challenging, although such in vitro responses could be helpful in assessing vaccines and therapeutics without jeopardizing human health. Given that DCs prime T cells and that FDCs promote B cell responses, we hypothesized that primary Ab responses could be induced in temporally controlled GCRs in vitro. To test this, monocyte derived-DCs were given OVA or rPA for processing, matured with LPS, and CD4+ T cells were added for priming. Ten days later, naïve B cells and OVA or rPA bearing FDCs were added and Ab production was subsequently assayed by ELISA. The CD4+ T cells clustered with DCs through the 10 days of priming and fused with FDC-B cell clusters when GCRs were induced. IgM anti-OVA levels were 79±15 ng/ml 7 days after adding B cells and declined to 2.4±0.5 ng/ml at day 14. In contrast, IgG anti-OVA rose from 9±1.7 at day 7 to 25±6.4 ng/ml at day 14 and the IgG1:IgG2 ratio was 1:3. Similarly, IgM anti-rPA levels were 10 ng/ml by just 5 days then IgM production switched to IgG. IgG anti-rPA rose from 3 ng/ml at day 5 to 9 ng/ml by day 12. In short, Ag specific primary human Ab responses, including Ig class switching from specific IgM to IgG, can be generated in GCRs entirely in vitro. Thus, potential vaccines can be rapidly and safely assessed using the human immune system.
(Supported and cleared by DARPA on 12/20/2006 - Approved for Public Release, Distribution Unlimited)
Incorporation of the TLR5-ligand flagellin in a recombinant Influenza A hemagglutinin vaccine provides enhanced immunogenicity and efficacy in vivo (47.2)
Authors: Huleatt James William , Tang Jie , Jacobs Andrea , Jock Dallas , Price Albert , McDonald William , Song Langzhou , Nakaar Valerian , Tussey Lynda , Powell T.J. 
Published: January 2023

Abstract
The linkage between innate and adaptive immunity is well established. The recognition of specific pathogen associated molecular patterns (PAMPs) by members of the Toll-like receptor (TLR) family plays a prominent role in the activation of adaptive immune responses. Previously, we have demonstrated that the linkage of specific PAMPs with defined antigens provides more potent and protective antigen-specific responses in the absence of adjuvants or complex formulations. Here we utilized this approach to develop a recombinant protein based influenza vaccine. The TLR5 ligand flagellin was linked to hemagglutinin (HA) to generate a single component vaccine. The recombinant E. coli expressed protein is recognized by influenza-convalescent antisera, indicating that the HA portion of the fusion protein is properly folded and presents conformational epitopes. In mice, the recombinant vaccine induces strong humoral and cellular responses that are quantitatively and qualitatively superior to those observed when the antigen is delivered with alum or co-delivered with flagellin unlinked. Analysis of serum antibody responses reveals strong reactivity with cells infected with influenza A and the ability to inhibit viral entry in vitro. Moreover, immunization with the fusion protein fully protects BALB/c mice from a lethal intranasal challenge with influenza A. These data demonstrate that linkage of the TLR5-ligand flagellin with HA provides significant promise as a non-egg based approach in the development of novel vaccines to combat seasonal and pandemic influenza.
ANTIBODY-MEDIATED TARGETING OF “INSIDE-OUT” ANIONIC PHOSPHOLIPIDS IN VIRAL DISEASE (47.21)
Authors: Soares Melina , Syed Sameer , Barbero Gustavo , Thorpe Philip E 
Published: January 2023

Abstract
The anionic phospholipid phosphatidylserine (PS) is found exclusively in the inner leaflet of the plasma membrane of resting mammalian cells. We hypothesized that certain events that occur during virus replication (eg cell activation or membrane rearrangement) would trigger the exposure of anionic phospholipids on the outer surface of virus- infected cells and subsequently on the enveloped viruses that bud out of these virus- infected cells. We further hypothesized that these exposed anionic phospholipids would serve as targets for anti-viral therapy. We demonstrate here that anionic phospholipids become exposed on the enveloped Pichinde Virus (a model virus for Lassa Fever virus, a potential bioterrorism agent) and on Pichinde virus-infected cells. To detect anionic phospholipids, we used a chimeric monoclonal antibody, bavituximab, that binds anionic phospholipids in a B2-glycoprotein I dependent manner. We show that bavituximab treatment is able to cure overt disease in guinea pigs lethally infected with Pichinde virus. Bavituximab treatment reduced the amounts of virus in multiple tissues and caused direct clearance of virus from the blood. Direct clearance of free virus and antibody-dependent cellular cytotoxicity of virus-infected cells appear to be the major mechanisms that contribute to the anti-viral effect of bavituximab. Bavituximab-treated survivors were immune to reinfection. Furthermore, the murine version of bavituximab, 3G4, shows therapeutic efficacy in a lethal murine model for human cytomegalovirus. Our study demonstrates the promise of anionic phospholipids as targets for new broad-spectrum anti-viral drugs.
Creating Retroviral Vector Encoding Two Homologous Chimeric Immune Receptors with Degenerated Codons (47.28)
Authors: Yang Wen , Lu Li , Ma Qiangzhong , Junghans Richard P 
Published: January 2023

Abstract
T cells genetically modified to express chimeric immune receptors (CIR) have been shown to be promising for immunotherapy. T cell receptor (TCR)-based CIR can be created in that the extracellular domains (Ec) of alpha (a) and beta (b) chains are fused to signaling cytoplasmic domain (Cy) of CD3 zeta (z) (TCRa-z and TCRb-z). To reduce homologous recombination associated with high homology between two nucleotide sequences in the same vector, we applied a novel approach by substituting the nucleotide sequence of one of the two homologous genes with degenerated genetic codes without change in amino acid sequence. In this study, we created a mutated DNA sequence encoding the Cy of z with degenerated codons (muz). A retroviral vector encoding a pair of TCR based CIRs (TCRa-z and TCRb-muz; tcsv-p36z) was created, with specificity via the pair of Ec of TCR a/b to a JC virus peptide p36/HLA-A2. 293T cells were transduced with retrovirus from PG13 cells with the tcsv-p36z. The transduced cells that expressed substantial TCR Vb of the CIR were able to bind p36/A2 tetramer, indicating the expression of both a and b chains in the single cell level. Western blot confirmed that both a and b chains were cellularly expressed intact. This approach, which we employed in creating retroviral vector encoding tc-CIRs with high homology, can be universally applied to creating viral vectors encoding two or multiple homologous molecules for gene therapy.
Adjuvant properties of trout C5a in subunit protein and DNA vaccines (47.31)
Authors: Zhang Yong-An , LaPatra Scott , Li Jun , Sunyer Oriol 
Published: January 2023

Abstract
The activation of the complement system results in the generation of C3a, C4a and C5a anaphylotoxins. C5a is the most potent and pro-inflammatory of all three anaphylatoxins. In mammals, C5a has been shown to influence the generation ofantigen-specific antibody responses, thereby acting as a molecular adjuvant. We have previously reported that rainbow trout C5a has potent chemotactic properties. In addition, we have also characterized a trout C5a receptor that is expressed at high levels both in granulocytes and B cells. In this study, we tested the possible role of trout C5a as a molecular adjuvant. To this end, we generated recombinant infectious hematopoietic necrosis virus glycoprotein (G), and a G-C5a fusion protein in E.coli. We showed that in contrast to G alone, G-C5a displayed a potent chemoattractant activity, indicating that the C5a portion of the fusion protein was functional. G-C5a was injected in incomplete freunds adjuvant (IFA) to one group of trout, while two other groups of trout were inoculated with the same dose of recombinant G in incomplete or complete freunds adjuvant (CFA) respectively. At six week postinjection, the serum antibody titers of the group injected with recombinant G-C5a in IFA were significantly higher than those of the group injected with G protein in IFA. Thus, these results suggest for the first time a role of C5a as molecular adjuvant in teleost fish. Experiments in progress will evaluate the effect of a boost injection on enhancing secondary and memory immune responses in the same fish. In addition, the presence of neutralizing antibodies against IHNV in the immunized fish will be reported.
Are Broadly Neutralizing Antibodies against HIV-1 Unusual? (47.32)
Authors: Scott Jamie Kathleen , Wang X. , Wu S. , Lepik C. , Singh H. , Richman D. D. , Utz P. J. , Breden F. 
Published: January 2023

Abstract
To date, AIDS vaccines cannot induce broadly(b)-neutralizing (Nt) antibodies (Abs) against HIV-1. It has been suggested that HIV’s Nt epitopes mimic “self” epitopes, and that bNt Abs are rare because they develop from autoreactive B cell clones that are normally deleted or made anergic by self antigens (Agns); such Abs are thought to have long CDR-H3 loops (H3s). Thus, self-tolerance must be broken to accumulate self-reactive naïve B cells whose Abs recognize Nt, HIV-1 epitopes. It has been reported that most of the bNt monoclonal (M)Abs have very long H3s, and that most react with self molecules (e.g., cardiolipin, CL). Alternatively, bNt Abs may arise out of “normal” Ab responses, with HIV Agns selecting bNt Abs with long H3s, and/or long H3s may develop via processes driven by persistent Agn.
We have begun to test these models by several approaches. Using autoAgn ELISAs and microarrays, bNt MAbs were shown not to be unusually self-reactive, compared to genuine autoreactive MAbs or autoimmune sera; also, bNt sera did not react significantly with CL. Finally, sequence analysis of ~700 human MAbs indicated that the bNt MAbs are no different than non-Nt MAbs against HIV-1 or other chronic viruses, or autoimmune anti-protein MAbs; but they are very different from autoimmune MAbs against non-protein autoAgns. Taken together, our results support the conclusion that HIV-bNt Abs are not unusual in their reactivity or structures.
IL-21 augments Monocyte Derived Dendritic Cell (mDC) maturation and upregulates PD-L1 gene expression (47.37)
Authors: Desai Seema , Liu Huanliang , Pahwa Savita 
Published: January 2023

Abstract
IL-21 and IL-4 bind overlapping epitopes on the common γ-chain, and IL-21 is reported to inhibit DC maturation and function in a murine model. To investigate the role of IL-21 on different stages of DC, monocyte derived immature dendritic cells (iDC) were generated in GMCSF and IL-4 in the presence and absence of IL-21, 20ng/mL for 5 days, termed as IL-21 iDC and control (c) iDC. IL-21iDC had significantly higher CD83 expression than ciDC on Day 5 (n=5, P&lt;0.004) with equivalent CD86 and HLADR expression. IL21iDC had 30–50 times greater PD-L1 mRNA expression compared to ciDC as quantified by real-time PCR, but with similar expression of SOCS-1 mRNA at three doses of IL21 (20,50,100ng/ml). Day 5 iDC were further matured by the addition of maturation cocktail (TNF-α, IL1β and PGE2) for 2 days. Day7 cDC and IL-21DC expressed high PD-L1 and did not differ in their expression of phenotypic markers. Autologous T cell response to flu antigen presenting cell (APC) function of IL-21DC was 2.6 fold greater (IFN-γ Elispot response) than of cDC, whereas allogeneic T cells had a similar MLR proliferative response to IL21DC as to cDC. We conclude that IL-21 enhances iDC differentiation with mild augmentation of mDC APC function. DC maturation under optimal culture conditions is unaffected by IL-21.
Source of Research Support: Institutional Support
Antibody-mediated protection against lethal respiratory tularemia (47.5)
Authors: Kirimanjeswara Girish Soorappa , Golden Jacqueline M , Metzger Dennis W 
Published: January 2023

Abstract
It has been previously shown that B cells are necessary and LPS-specific antibodies are sufficient for vaccine-induced immunity to Francisella tularensis, suggesting that antibodies play a major role in protection against tularemia. However, due to the intracellular nature of the bacterium, it is widely believed that T cells, but not antibodies, are critical for protection. We assessed the relative contribution of serum antibodies in protection against respiratory tularemia by investigating the ability of serum antibodies to clear bacteria from the lungs of intranasally infected mice and prevent systemic infection. Passive administration of immune serum collected from immunized mice was found to confer protection to naïve mice against lethal F. tularensis (LVS) challenge. Adoptively transferred antibodies rapidly cleared bacteria from the lungs and prevented spread of bacteria to liver and spleen. This protection was dependent on FcγRs and phagocytes such as macrophages and neutrophils, but not complement. Interestingly, phagocytosis and killing of opsonized bacteria was dependent on the presence of IFN-γ. Consistent with this finding, in vivo antibody-mediated protection required IFN-γ and T cells. Together, these data suggest that a close and critical interaction of humoral and cellular immune responses is necessary for providing sterilizing immunity to F. tularensis. Furthermore, serum antibodies conferred protection against lethal respiratory tularemia when given 24 to 48 hr post-exposure and thus this study provides evidence for the use of antibodies for both prophylactic and therapeutic purposes.
(This work was supported by NIH Grant PO1 AI056320 (DWM) and UO1 AI057158 (GSK))
Memory T cells Require Costimulation and Dendritic Cell Migration to Purge an Established Persistent Viral Infection (47.7)
Authors: McGavern Dorian B. , Lauterbach Henning , Truong Phi 
Published: January 2023

Abstract
Adoptive immunotherapy relies on transfer of exogenously derived pathogen-specific (memory) T cells and is a highly successful approach to purge an established persistent viral infection. For example, administration of adoptive immunotherapy to mice persistently infected with lymphocytic choriomeningitis virus (LCMV) (carrier mice) results in systemic eradication of the virus, including noncytopathic clearance of persistently infected CNS neurons. By visualizing GFP-tagged LCMV-specific memory CD8+ T cells on brain reconstructions, it was revealed that cytotoxic lymphocytes (CTL) distributed throughout the CNS within 8 days after immunotherapy. Flow cytometric and microscopic analyses revealed that conventional dendritic cells arrived in the CNS concurrent with the memory CTL and induced anti-viral cytokine production (e.g., TNF-alpha). When adoptive immunotherapy was performed in carrier mice depleted of DCs or with memory T cells deficient in TNF-alpha, the adoptively administered memory T cells failed to purge the persistent infection. We further demonstrated by employing CTLA-4-Fc blockade and CCR2 deficient recipients that memory CTL rely on costimulation and DC migration, respectively, for optimal secondary expansion and efficient viral clearance. These studies underscore the importance of DC mobility and memory CTL-DC interactions in the clearance of a persistent viral infection.
Codelivery of GM-CSF by hydrogel overcomes non-responsiveness to hepatitis B vaccine through recruitment and activation of dendritic cells (47.9)
Authors: Tao Mi-Hua , Chou Ho-Yuan , Wu Pin-Yi 
Published: January 2023

Abstract
Nonresponsiveness to hepatitis B vaccine, composed mainly the major hepatitis B surface antigen (HBsAg), has been described in approximately 5–10% of healthy immunocompetent subjects. The underlying cellular and molecular events for the nonresponsiveness are not completely understood, but are likely associated with impaired T helper cell responses to HBsAg in subjects with particular HLA-DR alleles. In this study, we describe a novel formulation of HBsAg vaccine containing GM-CSF in a thermosensitive biodegradable copolymer (hydrogel) system, acting as a sustained-release matrix for the loaded proteins. In responder mouse strains, hydrogel HBsAg/GM-CSF vaccine elicited high titers of anti-HBs antibody and strong T-cell proliferative responses. Importantly, hydrogel HBsAg/GM-CSF vaccine also elicited strong anti-HBs antibody and T-cell immune responses in B10.M mice, a mouse strain with H-2 gene-linked nonresponsive phenotypes. The local and sustained release of GM-CSF is required for its immunostimulatory function in nonresponder mice, because administration of hydrogel/HBsAg and hydrogel/GM-CSF in separate locations or a free mixture of HBsAg and GM-CSF failed to induce detectable humoral and cellular immune responses. Analysis of draining lymph nodes of hydrogel HBsAg/GM-CSF-treated mice revealed an elevated number of CD11c+ dendritic cells with enhanced expression of CD80 and CD86. These results suggest that the local adjuvant activity of GM-CSF helps nonresponder mice generate T helper cells, which subsequently provide help for B cells to produce anti-HBs antibodies, by promoting DC migration to and activation in the draining lymph node. This approach may be applicable to design of vaccines for a variety of other diseases.