bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD40-targeted S1 subunit vaccine protects against MERS-CoV and S1-associated pulmonary immunopathology in transgenic human DPP4 mouse model
Authors: Hashem Anwar M , Algaissi Abdullah , Agrawal Anurodh , Al-amri Sawsan , Almasoud Abdulrahman , Alharbi Naif , Peng Bi-Hung , Li Xuguang , Tseng Chien-Te 
Published: January 2023

Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus that emerged in 2012. While infection control measures have played a major role in limiting human/camel-to-human transmissions, development of safe and effective human or camel vaccines is warranted. Here, we extended and optimized our previous rAd5-based vaccine platform characterized by in vivo amplified and CD40-mediated specific responses to generate MERS-CoV S1 subunit-based vaccine. We generated rAd5 constructs expressing CD40-targeted S1 fusion protein (rAd5-S1/F/CD40L), untargeted S1 (rAd5-S1), and GFP (rAd5-GFP), and evaluated their efficacy and safety in human DPP4 transgenic (hDPP4 Tg+) mice. Immunization of hDPP4 Tg+ mice showed that a single dose of rAd5-S1/F/CD40L elicited robust and significant specific IgG and neutralizing antibody responses as those induced in mice immunized with two-doses of rAd5-S1. After MERS-CoV challenge, both vaccines conferred complete protection against morbidity and mortality, as evidenced by significantly undetectable/reduced pulmonary viral loads as compared to control group. However, rAd5-S1 but not rAd5- S1/F/CD40L immunized mice exhibited marked pulmonary perivascular hemorrhage post MERS-CoV challenge despite the observed protection. Collectively, these data indicate that incorporating CD40L into this Ad5-based MERS-CoV S1 subunit vaccine as targeting molecule and molecular adjuvant can not only enhance immunogenicity and efficacy but also prevents induction of inadvertent pulmonary pathology in immunized and challenged mice, thereby offering a promising strategy to enhance the safety and potency of antigen-specific immune responses.
Recombinant live attenuated influenza virus expressing G protein fragment in a chimeric hemagglutinin molecule induces G-specific antibodies and confers protection against respiratory syncytial virus
Authors: Jung Yu-Jin , Lee Yu-Na , Kim Ki-Hye , Lee Youri , Lee Jongsang , Kim Cheol , Kang Sang-Moo 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is one of the most important pathogens associated with significant morbidity and mortality in infants and the elderly. Live attenuated influenza vaccine (LAIV) is a licensed platform for vaccination in humans and known to induce broader immune responses locally and systemically. RSV G attachment proteins mediate virus binding to the target cells and contain a conserved central domain with neutralizing epitopes. Here, we generated recombinant LAIV based on the attenuated A/PR8/34 virus backbone, expressing a RSV G conserved domain in a chimeric hemagglutinin (HA) fusion molecule (HA-G). The attenuated phenotypes of chimeric HA-G LAIV were evident by restricted replication in the upper respiratory (nose) but not in the lower respiratory track lung tissues of the mice. Chimeric HA-G LAIV grew better at low temperature 33□. Prime and boost intranasal vaccination of mice with HA-G LAIV induced G-protein specific IgG and IgG2a (T helper type 1) isotype antibodies. Immunization of mice with chimeric HA-G LAIV showed significant increases in G-protein specific IgG and antibody-secreting cell responses in lung, bronchioalveolar fluid, bone marrow, and spleens after challenge. Vaccine-enhanced disease typically caused by inactivated-RSV vaccination was not observed in chimeric HA-G LAIV as analyzed by lung histopathology and infiltrating innate and adaptive immune cells after RSV challenge. Immunity of chimeric HA-G LAIV against influenza virus was not compromised. These results in this study suggest a novel approach of developing RSV vaccine candidates using LAIV, potentially dual vaccines conferring protection against both pathogens.
A Computationally Optimized Broadly Reactive Antigen (COBRA) Elicits Broadly Neutralizing Antibodies Against a Conserved Influenza Virus Hemagglutinin B-Cell Epitope
Authors: Sautto Giuseppe Andrea , Kirchenbaum Greg A. , Ecker Jeffrey William , Abreu Rodrigo Bessa , Pierce Spencer Reynolds , Ross Ted M. 
Published: January 2023

Abstract
The development of broadly protective influenza vaccines will represent the main countermeasure to overcome influenza virus spread and improve the coverage offered by the current standard of care.
A computationally optimized broadly reactive influenza virus hemagglutinin (HA) antigen (COBRA), named P1, elicits a broadly reactive antibody (Ab) response against multiple H1N1 strains.
In order to understand the mechanism of P1-conferred breadth of response, we first characterized the breadth of the Ab response at the B-cell level. Specifically, the reactivity of secreted Abs from mice immunized with P1 or seasonal/pandemic vaccine strains was assessed against a panel of H1N1 recombinant HA. Interestingly, while Abs from seasonal/pandemic immunized mice showed a predominant homologous response, those from P1- immunized animals exhibited a broader recognition.
In order to dissect the antibody response, a panel of P1-specific B-cell hybridomas was generated. Subsequently, the corresponding purified monoclonal Abs (mAbs) were assessed for the breadth of HA binding and hemagglutination inhibition (HAI) activity against different H1N1 viruses. Collectively, P1-specific mAbs exhibited a broad spectrum of binding and functional activities, spanning from strain-specific to broadly reactive and neutralizing mAbs, while seasonal/pandemic-specific mAbs displayed a narrower spectrum of activity. Epitope mapping studies revealed that P1-elicited broadly reactive and neutralizing Abs recognize unique conserved epitopes within the HA receptor binding site. Collectively, these studies shed light on the mechanism of breadth conferred by P1 and leverage its optimization for the development of a more effective influenza vaccine.
Computer-Optimized Immunization Strategy to Boost Highly Cross-Reactive Antibodies to RSV
Authors: Anderson Christopher S , Mariani Thomas J. 
Published: January 2023

Abstract
Respiratory Syncytial Virus (RSV) is a tremendous burden on public health. RSV infects humans within two years of life and causes continual infections throughout their lifetime. RSV is estimated to cause approximately 30 million episodes of acute lower respiratory infection resulting in over 100,000 deaths annually (in children ages 0 – 5 alone). Despite this burden, no vaccine is currently available for RSV. RSV has been classified into two subtypes (A and B) based on genomic and antigenic properties. Evolutionary rates are similar to other RNA viruses with estimates of 1.83–1.95 x 10−3 nucleotide substitutions/site/year. The high mutation rates of RSV produce an evolutionary landscape that has led to genetic and antigenic variation across and within subtypes. The attachment glycoprotein (G-protein) is must abundant on the surface of the virus and contains an almost completely conserved region, the central conserved domain (CCD). We developed a computational model of the humoral immune system that simulates B cell responses to G-protein antigen. We use a sequence-based approach to approximate antigenic differences across (&gt;1000) RSV strains. We then use computer simulations to identify an immunization strategy that can boost antibody levels to the CCD region. Last, we demonstrate that the immunization strategy results in a highly cross-reactive antibody response.
A knock-in mouse to test vaccines for the broadly neutralizing influenza response
Authors: Li Wenjuan , Ota Takayuki , Nemazee David 
Published: January 2023

Abstract
Influenza virus is a major public health burden controlled by vaccination. However, influenza viruses develop mutants at a high frequency. The yearly vaccine elicits a specific response that fails to protect against most influenza strains, and its efficacy and production is risky every flu season. It would be ideal to develop a universal vaccine conferring life-long, broad protection. So far, the academic field has isolated several broadly neutralizing antibodies that target conserved regions of viral proteins from donors. However, how to elicit these antibodies in human through designed vaccines remains challenging and inefficient. Herein we generated humanized BCR mice to facilitate testing of strategies to induce broadly neutralizing antibodies in humans. CR9114 is one of the most broadly neutralizing antibodies known with activity against group 1 and group 2 influenza A and also influenza B virus. Most contacts are with the H-chain only, which uses VH1-69. We knocked in a promoter-germline VDJ element to the physiological Igh locus, allowing analysis of the response of B cells capable in principle of broad neutralization and the testing of vaccine candidates.
Intravenous Bacille Calmette-Guerin provides protection across a dose spectrum in a Rhesus macaque model of tuberculosis
Authors: Zeppa Joseph John , Darrah Patricia A , Pokkali Supriya , Hackney Joshua A , Maiello Pauline A , Scanga Charles A. , Laddy Dominick J , Lin Philiana Ling , Seder Robert A , Roederer Mario , Flynn JoAnne L 
Published: January 2023

Abstract
Our laboratory has demonstrated that intravenous (IV) vaccination with Bacille Calmette-Guerin (BCG; 5x107CFU) provides remarkably robust protection against low-dose Mycobacterium tuberculosis (Mtb) infection in Rhesus macaques (9/10 animals protected [&lt;50 thoracic Mtb CFU]; 100,000-fold reduction compared to intradermal [ID] BCG). IV BCG also resulted in a 100-fold increase in mycobacterium-specific T cells in the bronchoalveolar lavage (BAL) of animals compared to other routes of administration (ID or aerosol). These results led us to hypothesize that significant protection could be observed at lower IV BCG doses. We therefore vaccinated macaques with decreasing doses of BCG (5x107, 1.6x107, 5x106, 1.6x106, 5x105and 1.6 x105CFU) followed by six months of immune monitoring and a three-month low-dose Mtb challenge. As expected, a mycobacterium-specific T cell dose-dependent response was observed in both the BAL (FACS) and PBMCs (IFNgELIspot) prior to infection, as well as in the lung at time of necropsy (FACS). Remarkably, 17/25 animals had no thoracic Mtb CFU across all groups, and at least one animal from each group was protected (30–100% protection). Mycobacterium-specific CD4 T cell numbers in the BAL appear to be a correlate of protection. These results demonstrate that even at low doses IV BCG can be efficacious in preventing tuberculosis with a potential role for T cell-dependent immunity.
SOCS1 antagonist-expressing recombinant BCG enhances anti-tuberculosis protection in a mouse model
Authors: Yasutomi Yasuhiro 
Published: January 2023

Abstract
Suppressor of cytokine signaling 1 (SOCS1) plays a key role in the negative regulation of JAK/STAT signaling, which is involved in innate immunity and subsequent adaptive immunity. Bacillus Calmette-Guerin (BCG) induces up-regulation of SOCS1 expression in host cells, which may lead to the suppression of immune responses by BCG via inhibition of the JAK/STAT signaling pathway. This might cause reduction in the protective effect of a BCG vaccine. In the present study, we assessed the immune responses and protective efficacy of a recombinant BCG vaccine expressing an SOCS1 antagonist molecule (rBCG-SOCS1DN). C57BL/6 mice were immunized with rBCG-SOCS1DN or parental BCG Tokyo vaccine strain harboring an empty plasmid vector (rBCG-pSO). rBCG-SOCS1DN enhanced induction bone marrow-derived dendritic cell activation and T cells activation compared to rBCG-pSO. The amount of IFN-g, TNF-a and IL-6 produced by splenocytes of rBCG-SOCS1DN-immunized mice were larger than those produced by splenocytes of rBCG-pSO-immunized mice. Moreover, the rBCG-SOCS1DN-immunized mice showed a substantial reduction in the number of colony-forming units (CFU) of Mycobacterium tuberculosis (Mtb) in the lungs and spleens in comparison with that in control BCG-immunized mice when the immunized mice were infected with a highly pathogenic Mtb strain by inhalation. These findings provide evidence for the possibility of rBCG-SOCS1DN being an effective Mtb vaccine with a novel concept of rBCG as a tool for immunomodulation in host cells.
Immunity of a Klebsiella Pneumoniae Vaccine Independent of Antigen Specific Antibody
Authors: Iwanaga Naoki , Sandquist Ivy , Chen Kong , Norton Elizabeth B , Moreno Javier Rangel , Kolls Jay K 
Published: January 2023

Abstract

Objective
We demonstrated the serotype independent immunity of Klebsiella pneumoniae (K. pneumoniae) vaccine. We would like to declare next if the noble effect is dependent on the antibody specific manner.

Methods
Mice were vaccinated via intranasal with Kp serotype 2 OMPx adjuvanted with heat-labile enterotoxin A1 domain (LTA1) and boosted at 3 weeks after the initial immunization. Protective immunity was compared among the C57Bl/6 mice, STAT3 fl/fl×CD4 Cre mice, and Il21r −/− mice by infecting with K1 strain of K. pneumoniae. Lung T-cell responses were evaluated by FACS and ELISPOT as well as K1 strain specific antibodies were assessed by ELISA.

Results
CD4+ T-cells of the vaccinated lungs in the CD4+ STAT3 deficient mice were significantly decreased compared to the WT mice and the Il21r −/− mice, and IL17A response by ELISPOT were significantly reduced as well. Meanwhile, K1 strain specific IgG in the serum and IgA in the lung homogenates of vaccinated WT mice were significantly increased in vaccinated mice and these responses were reduced in vaccinated CD4+ STAT3 deficient mice and Il21r −/− mice. Furthermore, vaccine induced protection was compromised in vaccinated CD4+ STAT3 deficient mice whereas the protection as well as antigen elicited lung IL-17 responses were maintained in Il21r −/− mice.

Conclusions
STAT3 activation in CD4+ T-cells regulated by IL-21R signaling are required for the K. pneumoniae specific antibodies, but these antibody responses were dispensable for the serotype independent immunity LTA1 adjuvanted K. pneumoniae vaccine. However, CD4 STAT3 was essential for Th17 immunity and therapeutic efficacy of the vaccine.
Phospholipase A2 group IID is indispensable for coronavirus-specific humoral immune memory in middle aged mice
Authors: Zheng Jian , Perlman Stanley 
Published: January 2023

Abstract
Phospholipases hydrolyze phospholipids to release long-chain fatty acids that are further metabolized sequentially by cyclooxygenases and synthases to yield various lipid mediators. Phospholipase A2 group IID (PLA2G2D) is a member of phospholipases family and shows age-dependent increases in the lungs. In our previous study, we found that infection of mice lacking PLA2G2D expression (Pla2g2d(−/−) mice) converted a lethal severe acute respiratory syndrome-coronavirus (SARS-CoV) infection to a nonlethal one (&gt;80% survival), subsequent to enhanced respiratory dendritic cells (DCs) migration to the draining lymph nodes, augmented antivirus T cell responses, and diminished lung damage. However, the long-term effects of PLA2G2D on host immune system against coronavirus remained unknow. In this study, we compared coronavirus-specific cellular and humoral immune memory induced by immunization or infection in middle-aged Pla2g2d−/− mice and wild type mice. Surprisingly, virus-derived replicon (VRP)-immunized or sublethal dose-infected Pla2g2d−/− mice failed to generate virus-specific neutralizing antibodies, which was accompanied by a lack of antibody-producing plasma cells and reduced follicular T cells (Tfh) compared to wild type mice. We hypothesize that PLA2G2D deficiency in myeloid cells, likely DCs, play a critical role in the formation of CD4 Tfh cells and humoral immune memory against respiratory coronavirus infection.
Characterization of HLA-DP and DQ restricted dengue virus-specific CD4+T cell responses reveals lower magnitude responses associated with a differential pattern of immunodominance as compared to DR restricted responses
Authors: Weiskopf Daniela , Grifoni Alba , Angelo Michael , Moore Eugene , Sidney John , Phillips Elizabeth J , Mallal Simon Alexander , de Silva Aruna D , Peters Bjoern , Sette Alessandro 
Published: January 2023

Abstract

Background
Assessment of adaptive immunity is an important element to understand immunopathology and to define the correlates of protection against dengue virus (DENV). While previous studies characterized responses restricted by the HLA DRB1 locus, the responses associated with the DP and DQ loci have not been characterized to date. Accordingly, here we mapped HLA DP and DQ restricted DENV-specific CD4 T cell epitopes in PBMCs from individuals previously exposed to DENV from the general population of the Sri Lanka.

Methods
We studied the DP (*0101, *0201, *0401, *0402) and DQ (*0501, *0602, *0301, *0302, *0201) allelic variants which provide a greater than 80% population coverage worldwide. CD4+ T cells purified by magnetic bead negative selection were cultured for two weeks with autologous APC and pools of HLA –specific predicted binders and tested for reactivity in a standard IFNγ ELISPOT assay.

Results
Magnitude of responses for both the DP and DQ loci were significantly lower than previously observed for the DRB1 locus. This was correlated with lower membrane expression of HLA- DP and DQ molecules on the PBMCs as compared to HLA DR. While responses against Capsid were dominant for the various DR and DP alleles this protein was hardly recognized by DQ alleles. Responses against NS3 were dominant in the case of DR and DQ but absent in the case of DP, and response to NS1 were prominent in the case of the DP alleles, but negligible in the case of the DR and DQ alleles tested.

Conclusion
The HLA DP and DQ loci mediate DENV-CD4 specific immune responses of lower magnitude as compared to HLA DRB1. The responses mediated by the DP and DQ loci are also associated with distinct and characteristic patterns of immunodominance.
EVALUATION TH17 AND TREG CELLS DURING CHRONIC CHIKUNGUNYA INFECTION
Authors: Guerra-Gomes Isabel Cristina , Leite Jefferson , Santana Alynne , Macêdo Bruna , Fonseca Rephany , Keesen Tatjana , Carregaro Vanessa , Silva João 
Published: January 2023

Abstract
Our aim was to evaluate the role of Th17 and its balance with Treg cells in defining CHIK chronification pathogenesis, as well as its association with clinical scores. We hypothesized that given that Chikungunya (CHIK) is caused by an arthritogenic alphavirus and possess several similarities with rheumatoid arthritis (RA), both characterized by Th17 cells playing a pathogenic role, in association with limited studies on the role of T cells regarding this CHIK chronification process. To reach this aim, without suitable animal models of CHIKV to properly assess the development of disease clinical signs, we developed a model of CHIK in DBA1/J mice associated with collagen-induced arthritis (CIA). Virus titration was obtained through One Step qPCR. Flow cytometry analyses were performed through cells isolation of draining lymph nodes from CHIKV previously infected (n=6), CIA (n=6) and CHIKV+CIA (n=7). As for humans, peripheral blood samples were obtained, controls (n=6), acute patients (n=6) and chronic patients (n=6). It was observed that our model was able to address clinical scores, such as develop joint swelling. Results showed that association with CHIKV infection increased clinical scores, number of paws involved, and the incidence of arthritis. Analyses of transcripts demonstrated an increased IL-17a and FoxP3 mRNA, however anti-inflammatory cytokines, such as IL-10, was diminished in animals infected with CHIKV. In humans, Treg frequencies presented diminished Treg number and percentage in patients in the chronic and acute stage compared with control individuals. Therefore, our results suggest an important role of T cell response in CHIK infection chronification.
Cytotoxic CD4 T cells control host immunity during viral persistence
Authors: Thiault Nicolas , Husain Mushtaq , Chen Angeline , Verstichel Greet , Larange Alexandre , Patil Veena Shivagouda , Vijayanand Pandurangan , Kronenberg Mitchell , Cheroutre Hilde 
Published: January 2023

Abstract
The CD4 T helper cell lineage is initially established and controlled in the thymus by the transcription factor, ThPOK. Sustained expression of ThPOK by mature T cells in the periphery maintains the CD4 T helper commitment and allows for plasticity and differentiation to different helper fates (Th1, Th2, Th17...) in response to stimulation with cognate antigens. Our group demonstrated before that in the intestine, repeated stimulation of mature CD4 T cells results in the loss of ThPOK expression followed by the reprogramming of the T helper cells to CD4 cytotoxic T lymphocytes (CTL), expressing notably CD8 T cells attributes (such as Runx3, granzyme B).
Continuous activation is also characteristic of chronic viral infections. We therefore investigated if viral infections lead to the generation of reprogrammed CD4 CTL and moreover if these reprogrammed CD4 CTL play essential roles for the organism’s protection. Using the well-defined lymphocytic choriomeningitis virus (LCMV) acute (strain LCMV-Amstrong) versus chronic (LCMV-Clone 13) infection model, we observed that solely chronic infection enables activated CD4 T helper cells to lose ThPOK and convert to CTL with potent killer activity. Moreover, viral-induced CD4 CTL precursors produce various cytokines, notably Il-10, which is required for the induction of the reprogramming process. Finally, we engineered two unique mouse model systems, in which either all the CD4 T cells are forced to reprogram to CTL (gain of function) or else, prevented from reprogramming to CTL (loss of function). Using these models, we show that the reprogrammed CD4 CTL generated under chronic infection conditions, play critical roles in protective immunity.
Investigating contributions of the accessory molecule HLA-DO to development of autoimmune diseases
Authors: Welsh Robin A , Song Nianbin , Foss Catherine , Boronina Tatiana , Cole Robert , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
Used biochemical and biophysical studies, our laboratory has previously shown that HLA-DO (DO) enhances binding of DM-insensitive peptides, and reduces binding of DM-sensitive peptides to HLA-DR1 molecules. We hypothesized that these properties of DO might be important for ensuring proper presentation of immunodominant epitopes in the thymic medulla, a main DO expressing area; thereby ensuring proper negative selection. To test this hypothesis, we investigated the development of experimental autoimmune encephalomyelitis (EAE) in mice that were H2-O wild type (DO-WT), or knock-out (DO-KO). With this model, we found that DO-KO mice displayed faster disease onset in comparison to DO-WT mice. Accelerated disease onset correlated with increased recovery of MOG specific CD4 cells at all stages of disease in DO-KO mice. More recently, these observations were confirmed with a second autoimmune disease model, collagen induced arthritis (CIA), in transgenic HLA-DR1 expressing mouse. Using in vivo Near Infrared Fluorescence (NIRF) imaging, we found that DR1+DO-KO mice had increased levels of active collagen degradation relative to DR1+DO-WT mice. Diseased DR1+DO-KO mice were also found to have a greater accumulation of CD4 T cells in the affected areas. These observations suggest that DR1+DO-KO mice have larger pools of collagen specific CD4 T cells. Indeed, DR1+DO-KO mice immunized with the immunodominant collagen epitope showed an increased percentage of collagen specific CD4 T cells as compared to DR1+DO-WT mice. As a whole, data from both autoimmune models suggest that loss of DO function leads to an increased peripheral pool of autoreactive CD4 T cells, likely due to faulty negative selection.