bluuebunny/crossref_metadata_2025_split huggingface dataset:
MALT1 proteolytic activity is necessary for FcɛRI-mediated cytokine production in mast cells
Authors: Alfano Danielle N , Klei Linda R , Lucas Peter C , McAllister-Lucas Linda M 
Published: January 2023

Abstract
IgE mediated mast cell activation triggered by multivalent antigen crosslinking of FcɛRI receptors leads to the release of a wide range of inflammatory mediators. Recent studies revealed that B cell lymphoma 10 (Bcl10) and mucosa-associated lymphoid tissue 1 (MALT1) are required for IgE-induced nuclear factor κB (NF-κB)-induced proinflammatory cytokine production, while dispensible for mast cell degranulation and leukotriene synthesis. MALT1, a cytoplasmic signaling protein, possesses at least two functions: scaffolding activity and proteolytic activity. The contribution of MALT1 proteolytic activity to mast cell function has not yet been evaluated. We hypothesize that MALT1 proteolytic activity is necessary for FcɛRI-dependent proinflammatory cytokine production. In order to test our hypothesis, we compared the activation of bone marrow derived mast cells (BMMC) from wild type mice, MALT1 knockout mice (MALT1−/−), and mice harboring a mutant, proteolytically inactive MALT1 (MALT1PD/PD). We find MALT1 protease is active in wild-type BMMCs as evidenced by RelB cleavage, and is absent, as expected, in MALT1−/− and MALT1PD/PD BMMCs. Cytokine gene expression and production in response to IgE mediated FcɛRI crosslinking is equivalently abrogated in both MALT1−/− and MALT1PD/PD BMMCs as compared to wild type. These findings suggest that the contribution of MALT1 to IgE mediated responses can be largely attributed to its protease activity. The MALT1 protease has been shown to be drugable, and pharmaceutically targeting the MALT1 protease activity may represent a promising approach to abrogating late phase pathologic mast cell activation.
Is MHCII-restricted immunodominance determined by HLA-DM action?
Authors: Osan Jaspreet Kaur , Ferrante Andrea 
Published: January 2023

Abstract
Binding of antigenic peptides to class II MHC molecules (MHCII) and the activity of the “editing” molecule HLA-DM (DM) on the resulting peptide-MHCII complexes are mechanisms critical to immunodominance, yet not fully understood. Our previous results indicate that peptide binding to the human MHCII HLA-DR1 (DR1) is a flexible event in that different thermodynamic components compensate each other to the possible extent to achieve a stable complex, and the energetic mechanism responsible for binding is reflected in the structural lability of the resulting complex. Here we apply our thermodynamic-structural model of peptide binding to sequences derived from HA H1N1, for which a DR1-restricted classification in dominant, sub-dominant, weak and negative has been established. We measured peptide KD and t1/2, and correlated such measurements to the observed level of immunodominance. Our results show that immunodominant peptides have a binding affinity in the micromolar range in the presence and absence of DM, whereas negative and weak showed no binding, with one exception. Interestingly, DM effect on binding affinity is variable for these peptides, altering their affinity values on a sequence-by-sequence basis with no direct correlation to their level of immunodominance. Variations in complex kinetic stability in the absence of DM did not reflect variations in immunodominance, whereas all peptides showed a faster release in presence of DM. Taken together, our results suggest that there is no a unique peptide-binding or kinetic signature, which can be adopted as a proxy for immunodominance. Further investigation in the thermodynamic components of the binding process is required to corroborate or modify these initial conclusions.
Immunomodulatory potential of Weissella cibaria JW15 on dendritic cells..
Authors: Oh Eun young , Lee Sang-Myeong 
Published: January 2023

Abstract
Lactic acid bacteria (LAB) are well-known to exert various beneficial health effects. Especially, immunomodulatory activities of LAB have been highlighted by recent studies. Our previous findings demonstrated that Weissella cibaria JW15 possessed immunostimulatory activities through modulation of macrophages, T cells and NK cells. Here, we studied whether this LAB is able to modulate dendritic cells (DC) function since DC are the most effective antigen presenting cells, linking innate immunity with adaptive immunity. Bone marrow derived dendritic cells (BMDCs) from mice were prepared and stimulated with live JW15, heat-killed JW15 and EPS isolated from JW15, followed by evaluation of cytokine production, expression of maturation markers and activation of intracellular signaling pathways. Our results revealed that JW15 as well as EPS induced phenotypic maturation of DC as shown by increased surface expression of CD80, CD86, CD40 and MHC II. In addition, both of JW15 and EPS increased the production of various cytokines in a dose dependent manner including TNF-a and IL-12. We also found that JW15 activated NF-κB and MAPK signal pathways. Collectively, we propose that JW15 and EPS isolated from this bacteria is able to activate dendritic cells and possess a potential to be developed as functional materials to boost immune system. Further studies are needed to provide comprehensive information to support our conclusions and reveal molecular mechanisms by which JW15 modulate immune responses.
Identification of the physiological antigen presenting cell for glycolipid antigens
Authors: Arora Pooja , Porcelli Steven 
Published: January 2023

Abstract
A major subset of T cells that recognize complexes formed by the binding of glycolipids to CD1d are called invariant natural killer T (iNKT) cells. Unlike conventional T cells that need to proliferate and differentiate to become activated effector T cells, iNKT cells exist as an effector population and start responding rapidly after glycolipid recognition. The prototypical iNKT stimulatory antigen is synthetic α-galactosylceramide (αGC), initially designated as KRN7000. Administration of this antigen in vivo results in the production of cytokines associated with both T helper type 1 (Th1) or type 2 (Th2) responses. Several variants of this antigen having different fatty acyl- and phytosphinganine chains have been synthesized and polarize the response towards a predominance of either Th1 or Th2 cytokines, and are therefore described as Th1- or Th2-biasing analogues. It is not clear how the Th1- or Th2-bias in iNKT mediated response is induced, although selective presentation by distinct cell types has been proposed as a possible mechanism. By visualizing glycolipid antigen presentation directly with monoclonal antibodies specific for complexes of αGC bound to CD1d, we showed that the CD8αPosDEC-205Pos dendritic cells were the major APCs in the spleen for a range of αGC analogues, irrespective of their chemical structures and cytokine biasing activities. We found that DCs presenting Th1-biasing agonists upregulated ligands for stimulatory NK receptor molecules, while Th2-agonist presenting cells showed a marked upregulation of ligands for inhibitory receptors. Furthermore, modulation of accessory immune proteins was dependent on CD1d, suggesting direct cell-cell interactions between DC and iNKT cells.
A unique MHCII-binding register correlates with HLA-associated immunogenicity of the major K blood group antigen
Authors: Gunasekera Devi , Zimring James C , Pratt Kathleen P. 
Published: January 2023

Abstract
Kell is a glycoprotein expressed on red blood cells (RBCs). Its K and k isoforms contain either Met (K antigen) or Thr (k antigen) at position 193. Development of anti-K antibodies following K-mismatched blood transfusions can destroy RBCs, while cross-placental transfer of anti-K antibodies can destroy RBC precursors, resulting in Hemolytic Disease of the Newborn. HLA-DR11, DR13 and DR15 are over-represented among K-immunized patient populations. The immunogenicity of overlapping 15-mer Kell peptides with M193 or T193 at every possible position was investigated by Stephen et al (Blood. 2012;119(23):5563-5574). Surprisingly, peptide W179-M193, with the polymorphic M193T site at the peptide’s C-terminus, was the most effective at stimulating CD4 T cells from K-immunized women. The present study utilized MHCII binding prediction algorithms and quantitative peptide-MHCII binding assays to determine the binding registers, anchor residues and affinities of wild-type, truncated, and sequence-modified Kell peptides. Predictions were generated using IEDB and Propred algorithms. Competitive peptide-MHCII binding assays utilized 13 recombinant HLA-DRB1 monomers, Kell peptides and high-affinity reference peptides. Interestingly, the assays identified a unique binding register (W179-S187) restricted to HLA-DR11 and DR15, in addition to the polymorphic site, suggesting stronger MHCII avidity may explain the increased disease susceptibility associated with these alleles.
The role of transmembrane domains in the March-I-mediated downregulation of MHC-II and CD86.
Authors: Walseng Even , Bandola-Simon Joanna , Roche Paul 
Published: January 2023

Abstract
MHC class II molecules on APCs bind to and presents peptides to antigen-specific CD4 T cells. DCs are the most potent subset of APC and can activate naïve T cells. Immature DCs possess relatively low expression of the costimulatory molecule CD86 and have a rapid rate of turnover of MHC-II. This is due in part to ubiquitin-mediated degradation of MHC-II and CD86 by the transmembrane (TM) ubiquitin ligase March-I in immature DCs. Curiously, March-I activity is suppressed by CD83, a TM protein that is upregulated by DC activation. Despite the well-documented role of March-I in specifically regulating MHC-II and CD86 expression, little is known about how these substrates interact with March-I and how CD83 is able to suppress March-I function. In this study we have examined the importance of TM domain interactions in March-I function by replacing TM domain sequences in March-I or CD83 with that of ICAM-1. A March-I TM1 mutant had reduced ability to affect MHC-II expression and had no effect on CD86 expression. Binding studies demonstrated that that March-I TM1 mutant lost all ability to bind to CD86. Whereas wild-type CD83 was able to block March-I downregulation of MHC-II and CD86, replacement of the CD83 TM domain with that of ICAM-1 completely abrogated CD83 suppression of March-I function. Binding studies confirmed that the CD83 TM mutant had lost all ability to bind to March-I. Our data therefore suggest that March-I binds to MHC-II and CD86 by TM domain interactions and that CD83 blocks March-I function by competitively inhibiting the binding of March-I to MHC-II and CD86.
Changes in MHC-II/peptide binding affinity following protein glycation, as observed in Diabetes
Authors: Santambrogio Laura , Clement Cristina , Negroni Pia , Stern Lawrence 
Published: January 2023

Abstract
Increased oxidized ribose, glucose, glyoxal and methylglyoxal concentrations are observed in several metabolic diseases including diabetes. These highly reactive moieties bind to the cellular proteome to form advanced glycation end-products (AGEs), which can alter the structure and function of targeted proteins. To address whether the AGE-mediated modification of immunologically relevant proteins could have functional consequences we incubated soluble recombinant HLA- DR1 (HLA-DRA*01:01/DRB1*01:01) in presence of glucose at concentration commonly observed in diabetic subjects (15–20 mM). Analysis of the AGE adducts was achieved by using bottom-up proteomics and tandem mass spectrometry analysis. Major observed modifications were glyoxal-derived hydroimiadazolone, argpyrimidine, dihydroxy and methylglyoxal adducts on arginine residues, pyralline and glyceraldehyde-derived pyridinium compound adducts of lysine and carboxy ethylation and methylation of both arginine and lysine residues. These AGE-related modifications were comparable to what observed on MHCII, immunoprecipitated from the PBMC of Diabetic patients. AGE PTMs mapped to both the binding groove and HLA-DM binding domains. AGE-modified and unmodified HLA-DR1 were then assessed for their binding affinity of several relevant peptides in a fluorescence polarization assay in the presence or absence of HLA-DM. Changes in binding affinity for the selected peptides was observed in AGE-HLA-DR1 as compared to the unmodified HLA-DR1. Altogether our data indicate that protein PTM, as commonly observed in conditions of metabolic and oxidative stress can change HLA-DR1/peptide binding affinity thus compromising MHC-II restricted immune responses.
PB1 as a potential target for increasing the breadth of T-cell mediated immunity to Influenza A
Authors: Uddback Ida Elin Maria , Steffensen Maria A. , Pedersen Sara Ram , Nazerai Loulieta , Thomsen Allan Randrup , Christensen Jan Pravsgaard 
Published: January 2023

Abstract
Current influenza vaccines are inducing antibody-mediated immunity toward the surface proteins hemagglutinin and neuraminidase. High variability of these proteins results in costly annual evaluation and production of new vaccines, not providing protection against heterosubtypic or emerging pandemic strains. Consequently, there is a need for a new universal influenza vaccine strategy. Unlike the surface proteins, the internal proteins of influenza are highly conserved. By vaccinating systemically and locally with nucleoprotein encoded in an adenovirus (AdNP) we could induce a protective CD8 T-cell response, that lasted up to at least 8 month post vaccination. To increase the breadth of our vaccine by combining construct, we studied the vaccine potential of another influenza protein. We showed that an adenovirus encoding PB1 elicits high numbers of antigen-specific CD8 T cells in mice. However, in vivo killing of peptide loaded cells was lower in PB1-vaccinated mice than in NP-vaccinated mice. This correlated with a poorer protection against challenge. To explain this, T cells from vaccinated mice were co-incubated with naïve splenocytes loaded with decreasing concentrations of peptide. If T cells were immediately added to the co-cultures, similar dose/response curves regarding IFNγ production was observed. However, if the T cells were added 6 hours after loading, cells from PB1-vaccinated mice required a much higher initial concentration of peptide to be activated compared with NP specific cells, indicating a lower stability of the MHC-PB1peptide complex. These results underscore that not only are T-cell quality and quantity crucial to consider in future vaccine design, but also MHC-peptide interactions.
Live attenuated Leishmania donovani Centrin knock out parasites generate non-inferior protective immune response in aged mice against Visceral Leishmaniasis
Authors: Bhattacharya Parna , Dey Ranadhir , Dagur Pradeep K , Joshi Amritanshu B , Ismail Nevien , Gannavaram Sreenivas , Debrabant Alain , Akue Adovi D , KuKuruga Mark , Selvapandiyan Angamuthu , McCoy John P , Nakhasi Hira L 
Published: January 2023

Abstract
Visceral Leishmaniasis (VL) causes significant mortality. Age appears to be critical in determining the clinical outcome of VL and there is no effective vaccine available against VL for any age group. Previously, we showed that gene deleted live attenuated L. donovani parasites (LdCen−/−) induced a strong protective immune response in young mice. In this study we analyzed LdCen−/− mediated modulation of immune response in aged mice (18 months) and compared to young (2 months) mice. Analysis of innate response in dendritic cells (DCs) from both young and aged mice upon infection with LdCen−/− parasites, showed significant enhancement of innate effector responses, which consequently augmented CD4+ Th1 cell effector function compared to LdWT infected DCs in vitro. Similarly, parasitized splenic DCs from LdCen−/− infected young and aged mice revealed induction of proinflammatory cytokines and down regulation of anti-inflammatory cytokine genes compared to LdWT infected mice. Furthermore, immunization with LdCen−/− induced higher IgG2a antibodies, lymphoproliferative response and stimulated splenocytes for heightened leishmanicidal activity in young and aged mice. Additionally, upon virulent L. donovani challenge, LdCen−/− immunized mice from both age groups displayed multifunctional Th1-type CD4 and cytotoxic CD8 T cells leading to significantly reduced parasite burden in the spleen and liver compared to naïve mice. Although, there was no difference in the LdCen−/− induced innate response in DCs between aged and young, the adaptive response in terms of T cell and B cell activation in aged was reduced, which correlated with LdCen−/− induced significant but diminished protective response in aged compared to young mice against VL.
Development of a Multivalent Subunit Vaccine against Respiratory Tularemia
Authors: Mansour Ahd A , Banik Sukalyani , Malik Meenakshi , McCormick Alison A , Bakshi Chandra Shekhar 
Published: January 2023

Abstract
Francisella tularensis (Ft); the causative agent of a fatal human disease tularemia is classified as a Category A Select Agent. No licensed vaccine is available for prevention of tularemia in the U.S.A. In this study, we used a novel Tobacco Mosaic Virus (TMV) based delivery platform for development of a fully protective multi-antigen subunit tularemia vaccine. Previously we have published that a trivalent TMV-conjugate vaccine confers 50% protection in immunized mice against respiratory Ft LVS challenge. In this study, we refined TMV-conjugate vaccine formulation to improve the level of protection in immunized C57BL/6 mice against respiratory tularemia.
We developed a tetravalent vaccine by conjugating OmpA, DnaK, Tul4 and SucB proteins of Francisella to TMV. CpG adjuvant was also included in the vaccine formulation. C57BL/6 mice were immunized intranasally (i.n.) on days 0, 14 and 28 and challenged with 10LD100 of Ft LVS on day 49 post-primary immunization (PPI). Mice were monitored for survival, weight loss, antibody and recall responses, and duration of immune protection.
100% of immunized mice were protected against a 10LD100 i.n. challenge dose of Ft LVS. Mice vaccinated with TMV-tetravalent vaccine showed high levels of TH1 antibodies than those with trivalent vaccine formulation. The challenged mice exhibited significantly reduced bacterial burden in lungs, liver and spleen. Strong ex-vivo recall responses were observed in immunized mice as late as day 84 PPI and 80% of mice survived when challenged 163 days PPI. Collectively, this study demonstrates that tetravalent vaccine formulation provides complete protection, induces strong protective and memory immune responses against respiratory challenge with F. tularensis.
Vaccine self-assembling immune matrix is a non-viral delivery platform that improves overall vaccine performance while maintaining safety and stability
Authors: Shollenberger Lisa M , Harn Donald A 
Published: January 2023

Abstract
Vaccination remains the most effective public health tool to prevent infectious disease. Many vaccines are marginally effective, and are less so when administered to immune-compromised populations. To enhance vaccine immunogenicity, we exploited the biphasic property of the (RADA)4 synthetic oligopeptide to create a new vaccine delivery method, VacSIM® (vaccine self-assembling immune matrix). Vaccine components are mixed with the VacSIM® solution for injection, after which the (RADA)4 peptides self-assemble into hydrated nanofiber gel-matrices, forming a vaccine depot with the vaccine antigens in the aqueous phase, allowing for slow-release of vaccine components over time. Thus, we have a non-viral, inert, biodegradable delivery system, not requiring formulation that can deliver various types of vaccines. We hypothesize that slow-release of vaccine components provides antigen persistence, driving enhanced vaccine-specific responses that are improved in both quantity and quality. We have tested VacSIM® with live bacterial vectors, inactivated virus and multiple recombinant protein vaccines. VacSIM® delivery of different protein immunogens, including Hepatitis B and HIV Envelope, is superior to delivery in licensed and unlicensed adjuvants and requires only a single injection. VacSIM® improves existing vaccines by reducing the number of injections needed, improving vaccine performance, especially in difficult-to-protect populations like the elderly and immune compromised, and reducing unwelcome side effects. VacSIM® can be stockpiled and stored without a cold chain, which is especially beneficial in resource-limited settings where vaccine access and compliance are issues.
Maternal Pertussis Vaccination with a Mono-component Pertussis Toxoid Vaccine is Sufficient to Protect Newborns
Authors: Kapil Parul , Zimmerman Lindsey I , Wagner Leslie D , Papin James F , Merkel Tod J 
Published: January 2023

Abstract
Bordetella pertussis is a highly infectious human pathogen causing serious illness in infants, children and adults. The first pertussis vaccination is administered at two months of age, therefore, infants under two months of age are most vulnerable to infection. Rising incidence of pertussis resulted in the 2012 recommendation by the Advisory Committee on Immunization Practices (ACIP) to immunize all pregnant women to protect infants from birth until their first vaccination. Using a baboon model, we demonstrated that maternal vaccination with the three or four component acellular pertussis vaccines protected newborn baboons from the disease but not from the colonization. We hypothesized this protection was due to the transfer of maternal anti-PT antibodies to the infant. Therefore we evaluated the protection conferred by maternal vaccination with a pertussis toxoid (PTx) only vaccine. Infant baboons born to mothers vaccinated with the PTx-only vaccine were free from disease and had low WBC counts following exposure to B. pertussis at five weeks of age. In contrast, infants born to unvaccinated mothers experienced severe disease with high WBC counts. Our results demonstrate that vaccination of baboons during pregnancy with a PTx-only vaccine is sufficient to protect newborn baboons from disease following exposure to pertussis. Because pertussis toxin is a secreted protein that can be purified from bacterial supernatants, a single-component vaccine comprised of pertussis toxoid is very likely the most cost-effective acellular pertussis vaccine possible. The development and use of mono-component PTx vaccines may enable the extension of maternal vaccine programs to low and middle income countries.