bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combined B cell specific deletion of the deubiquitinating enzymes CYLD and A20 leads to a dramatic accumulation of CD5+ B cells (TUM5P.936)
Authors: Hoevelmeyer Nadine , Hahn Matthias , Waisman Ari 
Published: January 2023

Abstract
Mutations in NFκB signaling molecules and in negative regulators of NFκB, such as CYLD and A20, have been identified in B cell lymphomas. These deubiquitinating (DUB) enzymes remove activating lysine-63 linked ubiquitin chains from numerous NFκB signaling molecules, including oncogene Bcl-3, thus controlling their activity. We found that mice lacking CYLD specifically in B cells develop with age a marked population of CD5+ B cells in the blood and the body, a population that resembles cells found in CLL patients. Importantly, we found that doubleBKO mice, that lack both A20 and CYLD in B cells, develop CD5+ B cells in an enhanced fashion, already at two months of age, with more than 95% of all B cells expressing CD5 before one year of age. Apart of CD5 expression, these B cells also increasingly express CLL markers, such as ZAP70 and Bcl-2. Starting with 14 month of age, these mouse mutants start to die most probably from CLL pathology. Thus, we could assign a separate, yet essential role for each of these two DUB enzymes in the control of B cell function, and have shown that loss of both together resulted in the initiation of a previously not identified chain of events probably leading to lymphoma/leukemia. In addition, we were able to identify a splice variant of CYLD that we previously found in mice, also in human CLL patients, thus indicating that this molecule, we termed sCYLD, may have a tumor-associated role not only in mice but also in human.
Targeting gender specific mechanisms to enhance anti-tumor immune responses against hepatocellular carcinoma (TUM5P.939)
Authors: Barrios Kelly , Vidrine Amy , Watkins Stephanie 
Published: January 2023

Abstract
In cancer, gender disparity has been observed in disease aggressiveness, incidence and prognosis without a clear understanding of the role the immune system plays into this sexual dimorphism. Hepatocellular carcinoma (HCC) occurs much more frequently in men than in women, however mechanisms associated with antitumor immunity in each gender is unclear. We introduce the use of a novel and clinically relevant mouse model of HCC to extricate the role of hormone signaling on immune cell function and the impact on immunotherapy. This model mirrors human disease since it’s driven by the overexpression of the oncogenes c-met and β−catenin. We used hydrodynamic delivery of these oncogenes to induce mutations and HCC. Male and female mice had comparable rates of tumor development; however in the absence of Foxo3, female mice were protected from HCC development. This protection was dependent upon immune cells lacking Foxo3 expression. Our data suggest that in females, silencing of Foxo3 permits the activation and migration of effector cells to the liver and that Foxo3 can control DC function by altering hormone receptor expression. Our results show that immune response and inhibition of tumor growth is differentially regulated by gender. A better understanding of the mechanisms regulating immune mediated tumor protection in females, will aid in devising ways to improve antitumor immune responses in males, and identifying proper targets to design or enhance treatment for all HCC patients
Examination of the role of dendritic cells, macrophages and B cells in the function of vaccine adjuvants. (VAC6P.944)
Authors: Mosaheb Munir , Reiser Michael , Toussi Deana , Liu Xiuping , Masari Paola , Wetzler Lee 
Published: January 2023

Abstract
Vaccines are vital in the fight against infectious diseases. Many vaccines are successful because of adjuvants added to their formulations or in the case of live attenuated vaccine because the adjuvant is inherent, however they have been used with little understanding of their mechanisms. PorB, a TLR2 agonist, is the major outer membrane protein of Neisseria meningitidis with potent adjuvant activity. In this study we investigated the role of individual antigen-presenting cell (APC) types in the adjuvant activity of PorB with comparisons to other vaccine adjuvants. WT mice and MyD88 APC floxed mice were immunized three times two weeks apart with ovalbumin (OVA) and either PorB, Alum, MF-59, MPL or CpG. We confirmed MyD88 deletion by qPCR on the purified cells and by measuring cytokines after stimulation with TLR ligand. Analysis of serum indicated that MyD88 in B cells, macrophages and dendritic cells is important for the production of PorB/OVA induced anti-OVA IgG. Alum, a MyD88 independent adjuvant, was able to induce strong anti-OVA IgG in mice that lacked MyD88 in any of these APCs. Interestingly the adjuvant activity of MF-59, which may be MyD88 dependent, were not affected by deletion of MyD88 in the different APC cell types, suggesting that if MyD88 is involved in its adjuvant activity, it must be due to stimulation of other cell types. These insights will aid in vaccine development by allowing more intelligent and judicious use of these and other vaccine adjuvants.
Biological activity of dialyzable leukocyte extracts in a mouse cutaneous herpes model. (VAC6P.946)
Authors: Pérez-Tapia Sonia , Salinas-Jazmín Nohemí , Estrada-Parra Sergio , Limón-Flores Alberto , Vázquez-Leyva Said , Becerril-García Miguel , Medina-Rivero Emilio , Pavon Lenin , Velasco-Velázquez Marco 
Published: January 2023

Abstract
Human dialyzable leukocyte extracts (DLE) are heterogeneous mixtures of low-molecular-weight peptides. DLE improve the clinical responses in infections, allergies, cancer, and immunodeficiencies. TransferonTM is a human DLE registered as a drug by Mexican health authorities and commercialized nationally. In order to establish a model that could be used routinely to assess the activity of TransferonTM, we standardized and validated a mouse model of HSV-1 cutaneous infection. In such model, we evaluated the biological activity of 27 TransferonTM batches that complied with other quality standards. All batches improved survival of HSV-1-infected mice; average survival increased from 20.9% in control mice to 59.6% in TransferonTM-treated mice. Biological activity of TransferonTM correlated with increased serum levels of IFN-γ and reduced IL-6 and TNF-α concentrations at 7-9 days post-infection (high mice mortality period). Our results show that: a) the mouse cutaneous herpes model allows the evaluation of the biological activity of TransferonTM; b) the assay can be used as a routine tests for batch release; c) TransferonTM is produced with high homogeneity between batches; and d) the protective effect of TransferonTM in this model correlates with differential changes in cytokines related to innate or adaptive immunity, suggesting that immunomodulation is associated to the reported therapeutic effects on herpes infections. Supported by FTU/IB/012/010/PRO.
Adoptive B cell immunotherapy accelerates control of chronic viral infection (VAC6P.947)
Authors: Moseman E. , Heydari Sara , Zinselmeyer Bernd , McGavern Dorian 
Published: January 2023

Abstract
Many viruses can establish persistence and cause disease in humans. Studies have established a role for T cells in resolving persistent viral infections; however, the importance of a humoral response is less clear. In this study, we sought novel insights into the importance of humoral immunity in resolving a persistent infection by utilizing the lymphocytic choriomeningitis virus (LCMV) clone 13 model. Infection with aggressive LCMV variants results in a chronic viral infection during which antiviral T cells pass through a state of functional exhaustion, but the infection is nevertheless controlled over time. We postulated that humoral immunity was required for LCMV control and could be harnessed as an immunotherapy. We observed dramatic increases in splenic plasma cells as early as one week following LCMV infection, which coincided with the emergence of non-neutralizing antibodies. B cell depletion starting at the peak of T cell exhaustion (day 15) resulted in a failure to control virus. Moreover, adoptive transfer of LCMV immune B cells into mice with an active infection dramatically accelerated viral clearance. We found using two-photon microscopy that therapeutic B cells home to the infected recipient spleen where donor antibody secreting cells proliferate to amplify antiviral humoral responses. These data demonstrate that humoral immunity can contribute to the resolution of a persistent viral infection and that B cells can be harnessed for adoptive immunotherapy.
Type A-derived genetically modified live attenuated Francisella tularensis is immunogenic and efficacious in rabbits (VAC7P.965)
Authors: Reed Douglas , Caroline Amy , Stinson Elizabeth , Cole Kelly , Santiago Araceli , Mann Barbara , Barry Eileen 
Published: January 2023

Abstract
Francisella tularensis is a gram negative non-motile coccobacillus that is the causative agent of tularemia. There is concern that F. tularensis could be used as a biological weapon and development of vaccines is a high priority. Type A strains are highly virulent but the current live vaccine strain (LVS) is based on a type B strain and is only partially protective against aerosol challenge with SCHU S4, a type A strain. A series of live attenuated vaccine candidates were derived from the SCHU S4 strain containing specific deletion mutations in target genes. After initial studies in mice, these attenuated strains were evaluated in rabbits. Rabbits were vaccinated by scarification which induced a mild fever and eschar formation at the inoculation site but no other clinical signs of disease. Serum IgG responses could be detected in all vaccinated rabbits as early as day 7 and continued to increase through day 21 before declining slightly on day 28. CD4+ T cells but not CD8+ T cells proliferated when cultured in vitro with heat-killed SCHU S4. Three of the vaccine strains conferred partial protection against death (ranging from 27-40%) and prolonged time to death in animals that did succumb. Vaccination with LVS prolonged survival by 1-2 days but did not protect against death. Type A-derived vaccines are immunogenic and provide better protection than LVS in the rabbit model against a robust aerosol challenge. These candidates warrant further study and evaluation.
ManLAM-aptamer as an immunoadjuvant improves immune-protective effects of BCG (VAC7P.968)
Authors: Zhang Xiao-Lian , Sun Xiao-ming , Pan Qin 
Published: January 2023

Abstract
Tuberculosis (TB) remains an important worldwide health threat. However vaccination with Mycobacterium bovis bacille Calmette-Guerin (BCG), the only available licensed vaccine of TB, provides limited protection against adult pulmonary tuberculosis. Mannosylated lipoarabinomannan (ManLAM) is a kind of glycolipid on the cell wall surface of BCG and has been implicated with remarkable immunosuppressive effect on the host immune cells. In the present study, we used Systematic Evolution of Ligands by EXponential enrichment (SELEX) to generate a single-stranded DNA aptamer that specifically bound to ManLAM of BCG with high affinity. We found that vaccination with BCG combined with BM2 recruited dendritic (DC) cells into peripheral lymph nodes and decreased the numbers of regulatory B cells and regulatory T cells. It could also significantly enhanced the Th1-type cellular response and induced conversion of IgG2a isoforms in mice. We demonstrated that vaccination with BCG plus BM2 significantly reduced the progression of Mycobacterial tuberculosis (M. tb) H37Rv infection, bacterial loads in lungs and spleens of mice via aerosol infection with M. tb H37Rv, and provided much better protective immune response against M. tb than vaccination with BCG alone. Our data provide a new strategy for TB control and the aptamer BM2 holds great promise for developing a new potential immunoadjuvant of BCG.
Increased T and B cell responses in M. bovis BCG vaccinated pigs by co-immunization with pDNA encoding a prototype tuberculosis antigen (VAC7P.973)
Authors: Huygen Kris , Bruffaerts Nicolas , Vandermeulen Gaëlle , Romano Marta , Préat Véronique , Stockhofe-Zurwieden Norbert 
Published: January 2023

Abstract
The only vaccine currently available to prevent tuberculosis, M. bovis BCG, has limited efficacy. BCG is a poor inducer of CD8+ T cells, which are particularly important for the control of latent tuberculosis and protection against reactivation TB. The induction of strong CD8+ T cell responses is a hallmark of DNA vaccines. We postulated that combining the existing BCG vaccine with plasmid DNA encoding immunodominant antigens of M. tuberculosis, could improve the vaccine by generating stronger CD8+ (and CD4+) T cell responses. As a prototype tuberculosis antigen, the mycolyl-transferase Ag85A was selected. Three groups of six pigs each, were vaccinated intradermally with BCG alone, BCG combined with empty vector or BCG combined with codon-optimized V1j.ns-tPA-Ag85A vector. The BCG-pAg85A DNA combination induced strongly increased and sustained humoral and Th1 type cellular responses against Ag85A and against the purified protein derivative of tuberculin. Increased IFN-γ producing CD8 T cells were detected by intracellular cytokine staining and an Ag85A T cell epitope with predicted MHC class I motif, was promiscuously recognized by 5/6 pigs vaccinated with the combination. In summary, our study provides the first proof of concept in a large mammalian species, for a new TB vaccine, based on a BCG-pDNA combination.
Oral immunization with Escherichia coli-expressed enterovirus 71 VP1 and chitosan adjuvant protect mice against lethal challenge (VAC7P.984)
Authors: Li Aqian , Zhang FuShun , Zhang Shuo , Hao Chunsheng , Zhang QuanFu , Wu Wei , Liu Lin , Liu Chuan , Liang Mifang , Li Xiuling , Li Dexin 
Published: January 2023

Abstract
Enterovirus 71 (EV71) is identified as the etiologic agent of hand-foot-and- mouth disease (HFMD) in the Asia-Pacific region while there are currently no vaccines or effective antiviral drugs available. VP1, one of the major immunogenic capsid proteins of EV71, has shown considerable promise for preventing EV71 infection by intramuscular inoculation in mice. In this study, we found that oral gavage vaccination (O.V.) of ICR mice with recombinant VP1 (rVP1) and chitosan adjuvant induced VP1-specific sIgA antibodies in intestine, feces, vagina, respiratory tract and virus-specific neutralization IgG antibodies. The splenocytes collected from the rVP1-immunized mice exhibited significant cell proliferation and induced high levels of Th1 immune responses cytokine IFN-γ and Th2 immune responses cytokine IL-4 after stimulation. Moreover, the rVP1 immunized mother mice conferred protection (survival rate up to 30%) to neonatal mice against the lethal (1000 plaque form unit, PFU) EV71 challenge. All these data indicate that oral immunization with rVP1 is safe and effective in inducing broad-spectrum immune responses and being a promising subunit vaccine candidate for prevention of EV71 infection. Corresponding author: Prof. Dr. Dexin Li Email: lidx@chinacdc.cn
An optimized, synthetic DNA vaccine encoding the toxin A and toxin B receptor binding domains of Clostridium difficile induces protective antibody responses in vivo (VAC7P.958)
Authors: Baliban Scott , Baliban Scott , Shammassian Berje , Michael Amanda , Zetner Isaac , Latimer Brian , Cocklin Simon , Welles Seth , Bouillaut Laurent , Sonenshein Abraham , Kahn Amir , Sardesai Niranjan , Jacobson Jeffrey , Weiner David , Kutzler Michele 
Published: January 2023

Abstract
Clostridium difficile-associated disease (CDAD) constitutes a large majority of nosocomial diarrhea cases within industrialized nations and is mediated by the effects of two secreted toxins, toxin A (TcdA) and toxin B (TcdB). Key toxin-neutralizing epitopes have been discovered within the carboxy-terminal receptor-binding domain (RBD) of the toxins, which has generated interest in developing the RBD as a vaccine target. While numerous platforms have been studied, very few data describe the potential of DNA vaccination against CDAD. Therefore, we created highly optimized plasmids encoding the RBD from TcdA and TcdB and immunized C57BL/6 mice and rhesus macaques intramuscularly followed by in vivo electroporation. In these animal models, vaccination induced significant levels of anti-RBD antibodies within the serum and feces that could neutralize toxins in an in vitro cytotoxicity assay. Moreover, mice that were either immunized with plasmids or given non-human primate immune sera were protected from a lethal intraperitoneal challenge of purified C. difficile toxins. Finally, immunized mice were significantly protected when challenged with strains of C. difficile spores that were homologous (VPI 10463; n=10/10) and heterologous (UK1; n=4/8) to our vaccine antigens. These data demonstrate the robust immunogenicity and efficacy of a TcdA/B RBD-based DNA vaccine in preclinical models of acute toxin-associated disease.
HIV broadly neutralizing antibodies and immunological tolerance (VAC7P.988)
Authors: Yang Guang , Liu Mengfei , Wiehe Kevin , Nicely Nathan , Haynes Barton , Mascola John , Bjorkman Pamela , Kelsoe Garnett 
Published: January 2023

Abstract
It is generally thought that many human antibodies (Ab) that neutralize multiple clades of HIV-1 (bnAbs) are self-reactive. We previously identified kynureninase as the primary self-antigen recognized by 2F5, suggesting that generation of Ab to the 2F5 epitope of HIV-1 may be proscribed by immune tolerance, a notion supported by impaired B-cell development in mice expressing the 2F5 VH and VL regions. However, there is a lack of quantitative and systemic assessment of polyreactivity among HIV bnAbs. In addition, it is unknown whether other classes of HIV-1 bnAbs and non-neutralizing antibodies are also influenced by immunological tolerance. Here, we used protein microarrays to assess bnAb binding to over 9,600 human proteins and compared bnAb binding profiles to HIV non-neutralizing antibodies. We found that bnAbs as a class are more autoreactive and more polyreactive than non-neutralizers. Interestingly, 4 of 7 CD4bs bnAbs screened from 2 distinct lineages (VRC01, VRC02, CH103 and CH106) bind human protein ubiquitin ligase E3A (UBE3A). We confirmed this crossreactivity in ELISA, and demonstrated that UBE3A competitively inhibits gp120 binding to VRC01. Our results demonstrate that HIV-1 bnAbs are significantly more polyreactive and self-reactive than non-neutralizers, which may subject them to immunological tolerance control in vivo. Our identification of UBE3A as a self-antigen of bnAbs provides a mechanism for the rarity and delayed development of certain CD4bs bnAbs.
Evaluation of human cell mediated immune responses to VZV antigens (VAC7P.993)
Authors: Cox Kara , Prokop Michael , Dubey Sheri , Vora Kalpit 
Published: January 2023

Abstract
Zostavax© is a safe and effective vaccine against Herpes Zoster. The precise immune correlate of protection and the corresponding VZV antigens responsible for protective immunity are not currently defined. A few key pieces of data suggest that cell mediated immune responses are more important than humoral responses in the immune control over viral re-activation from latent infection. Primarily, the majority of Herpes Zoster outbreaks occur in the aging population in which antibody titers against VZV are maintained but cell mediated immune responses wane. Additionally, individuals who are immunocompromised as a result of disease or drug treatment have difficulty controlling the virus. A group of ten viral proteins of Varicella were chosen for evaluating human cell mediated immune responses. Donor samples were recruited in the following cohorts: i; Herpes Zoster outbreak within the last 5 years, ii; Subjects age 35+ with no history of Shingles or Zostavax© vaccination, and iii subjects pre and post Zostavax© vaccination. Blood samples from these donors were stimulated with the set of VZV antigens and a panel of cytokine responses from the plasma was measured. Here we show a ranking of VZV viral protein immunogenicity from these cohorts, as well as the cytokine profiles for each. These studies will help to define the viral proteins involved in generating protective cell mediated immunity correlated with protection.
TLR3 preconditioning enhances the therapeutic efficacy of umbilical cord mesenchymal stem cells in DSS-induced colitis (VAC8P.995)
Authors: Carrion Flavio , Fuenzalida Patricia , Fernandez Catalina , Ibañez Cristina , Vega Ana Maria , Fuentealba Rodrigo , Kurte Monica 
Published: January 2023

Abstract
Background and Aim: Mesenchymal stem cells (MSCs) have attracted much attention since exert anti-inflammatory and immunomodulatory effects. Recently, diverse studies have been showed that the stimulation of specific toll-like receptors affects the immunomodulatory properties of MSCs. The aim of this work was to evaluate the effects of TLR preconditioning on therapeutic efficacy of umbilical cord MSCs (UC-MSCs) in a murine model of acute colitis. Methods: C57BL/6 mice were exposed to 2,5 % dextran sulfate sodium (DSS) in drinking water. At days 1 and 3 were injected intraperitoneally with 1 x 106 of UC-MSCs or TLR3 and TLR4-preconditioning UC-MSCs. We evaluated Clinical signs of disease, body weights daily, plasma cytokines levels, colon length and histological changes. Results: The infusion of TLR3-preconditioned UC-MSCs significantly ameliorated the clinical and histopathologic severity of DSS-induced colitis compared with wild type UC-MSCs or TLR4-preconditioned UC-MSCs. In contrast, infusion of TLR4-preconditioned UC-MSCs significantly increased the clinical signs of disease, colon shortening and histological disease index. Conclusions: These results suggest that the immunomodulatory properties of UC-MSCs can be differentially modulated by TLR agonists. Also, these data showed that the short in-vitro TLR3-preconditioning enhances the therapeutic efficacy of UC-MSCs, which can be very useful for treatment or amelioration of proinflammatory diseases.
Developing bacteriophage Qβ as versatile carrier for anti-carbohydrate cancer vaccine: influence of antigen design on antibody responses (VAC8P.998)
Authors: Yin Zhaojun , Gildersleeve Jeffrey , Finn M. g. , Huang Xuefei 
Published: January 2023

Abstract
Vaccines that aim to stimulate the immune system to eradicate tumor cells are very attractive method for cancer therapy. One such strategy, active vaccination against tumor associated carbohydrate antigens (TACAs), which are overexpressed on malignant cells and strongly correlated with tumor progression and metastasis, has shown high potential in clinical studies. However, two major challenges need be circumvented to develop effective TACA anticancer vaccines. First, TACAs are self-antigens and tolerated by the immune system; Second, TACAs are weak T cell independent antigens and cannot generate long lasting immune responses when administered alone. We are interested in exploration of bacteriophage Qβ as versatile platforms for the development of carbohydrate based cancer vaccine. The highly repetitive array of subunit in Qβ permits the organized display of antigens on the surface of the particle. The Qβ also contains natural T-helper epitopes and pathogen associated molecular patterns, which can engage innate immunity and activate the immune system. Herein, we present the vaccination studies with Tn antigen conjugated to bacteriophage Qβ. The effect of antigen density, nature of linker, and immunization routes on the quality and diversity of antibody responses was analyzed. With the optimal construct, significant preventive and therapeutic effects were achieved on highly aggressive TA3Ha tumor model.