bluuebunny/crossref_metadata_2025_split huggingface dataset:
Subunit vaccine candidate with adjuvant confers protection against clinical Mycobacterium avium in wild type and immunocompromised mouse models
Authors: Larsen Sasha E , Reese Valerie A , Pecor TIffany , Brewer Guy , Berube Bryan , Njikan Samuel , Parish Tanya , Henao-Tamayo Marcela , Baldwin Susan L , Coler Rhea N. 
Published: January 2023

Abstract
Nontuberculous mycobacteria (NTM) are a group of 170 species of mycobacteria that are steadily increasing in incidence and prevalence. NTM cause a significant amount of morbidity and mortality and overtake Mycobacterium tuberculosis in prevalence in certain regions of the world, including the United States. M. avium, a clinically significant NTM, is known to cause a tuberculosis-like pulmonary disease that is often difficult to treat. Due to this quietly emerging health threat, we evaluated the ability of a subunit vaccine (ID91) with the Toll-like receptor agonist glucopyranosyl lipid adjuvant formulated in an oil-in-water stable nano-emulsion (GLA-SE) to confer protection in both the C57BL/6 (wild type) and Beige (immunocompromised) mouse models. We first established a challenge model with a low dose aerosol of the virulent clinical isolate M. avium 2–151 smt and monitored bacterial burden and immune responses to infection over time. Next, cohorts of mice immunized prophylactically with ID91+GLA-SE or bacillus Calmette–Guérin (BCG) were subsequently challenged with M. avium 2–151 smt. Not surprisingly, the immunocompromised Beige mice sustained a larger bacterial burden in the lungs and spleen compared to C57BL/6 mice regardless of treatment. However, both ID91+GLA-SE and BCG significantly reduced pulmonary bacterial burden in both models. Variable innate and adaptive pulmonary immune response kinetics in C57BL/6 and Beige mice in response to M. avium 2–151 smt infection were observed. This work demonstrates the first proof-of-concept study that a subunit vaccine candidate (ID91+GLA-SE) confers prophylactic protection against NTM and suggests further vaccine development may help reduce the burden of disease.
Antigen Presenting Cell Targetted, Adjuvant free Mucosal Vaccine Induces Protection against Pneumococcal Infection
Authors: Kumar Sudeep , Sunagar Raju , Gosselin Edmund J 
Published: January 2023

Abstract
Protein-based subunit vaccines require adjuvants to increase their immunogenicity. However, there is lack of FDA-approved mucosal adjuvant. We have previously shown that targeting antigens to antigen presenting cells increases their immunogenicity. Specifically, Streptococcus pneumoniae (Sp) protective antigen (PspA) fused to a single-chain bivalent-anti human Fc-gamma-receptor-I (hFcgRI) antibody, when introduced into mice, generates protection against Sp. To further enhance the immunogenicity of the anti-hFcgRI-PspA fusion protein, we have incorporated an additional hFcgRI-binding moiety, with the resultant trivalent-anti-hFcgRI inducing better protection compared to the bivalent anti-hFcgRI-based vaccine. This improved vaccine requires fewer immunizations to confer over 90% protection, significantly higher than that of bivalent-anti-hFcgRI-PspA (less than 60%). The trivalent- anti-hFcgRI-PspA also induces significantly higher levels of Sp-specific antibodies. In addition, we observed elevated levels of mucosal IgA responses in trivalent-anti-hFcgRI-PspA-immunized mice compared to that of bivalent anti-hFcgRI-PspA. Both trivalent-anti-hFcgRI-PspA and bivalent anti-hFcgRI-PspA enhanced antigen presentation in FcgRI-dependent manner. We have further observed that trivalent-anti-hFcgRI-PspA induces recruitment of innate immune cells in the nasopharyngeal lymphoid tissue upon intranasal immunization, which is a hallmark of adjuvant activity. Thus, our study suggests that anti-hFcgRI-PspA compensates for the lack of adjuvant by inducing adjuvant effects, and can be utilized as a mucosal vaccine platform.
The Development of Klebsiella Pneumoniae Vaccine Strategy via Eliciting the Serotype- Independent Immunity in the Lung
Authors: Iwanaga Naoki , Sandquist Ivy , Chen Kong , Norton Elizabeth B , Moreno Javier Rangel , Kolls Jay K 
Published: January 2023

Abstract

Objective
Multidrug-resistant of Klebsiella pneumoniae (K. pneumoniae) has emerged as a global threat. Vaccination for at high-risk populations could be a promising strategy to prevent and control the disease.

Methods
Mice were vaccinated via intranasal with a monovalent vaccine encoding Kp serotype 2 OMPx or a quadrivalent vaccine encoding OmpC, OmpW, Omplolb, and Omp36k, adjuvanted with heat-labile enterotoxin A1 domain (LTA1). Protective immunity was assessed by performing a pulmonary challenge with a heterologous mucoid serotype K1 strain of K. pneumoniae. Lung T-cell responses were evaluated mainly by FACS, ELISPOT, and LUMINEX. We also examined inflammatory cell changes by hematoxylin-eosin stain and immunofluorescence.

Results
Flow cytometry and ELISPOT analysis revealed that CD4+ T cells were significantly increased in the lungs of the monovalent or quadrivalent vaccinated mice and that a considerable fraction of them produced IL17A. And the supernatant of these cells also showed the significant production of IL-17A. Importantly, vaccinated mice showed a significant decrease in CFU in the lung and spleen, compared with mock-vaccinated mice. Finally, we observed that only lungs of intranasally vaccinated mice had lymphocytic structures around bronchi and vessels. Characterization of these structures by immunofluorescence confirmed the presence of inducible bronchus-associated lymphoid tissue (iBALT) – a structure that can not only support local induction of protective immunity but provide a long-term niche for vaccine-induced lymphocytes.

Conclusions
These data support the therapeutic value of intranasal vaccination strategies to prevent K. pneumoniae infections.
Protective role of TH17 cells in an FcγR targeting immunization strategy against the respiratory bacterial pathogen, Francisella tularensis
Authors: Bitsaktsis Constantine , McCauley James , Devenney Kyle 
Published: January 2023

Abstract
Francisella tularensis is a category A intracellular bacterial pathogen with no FDA approved vaccine. Understanding the immunological mechanisms involved in host defense can improve vaccine efficacy. Previously we have shown using an F. tularensis murine infection model that targets the inactivated bacterium (iFt) to Fcγ receptors on antigen presenting cells (APCs) using anti-LPS monoclonal antibodies (mAb-iFt) provides full protection against lethal pulmonary challenge with F. tularensis live vaccine strain (LVS). The protection provided by this mucosal vaccine strategy involves both innate and adaptive immune components, with a strong inflammatory response being critical for protection. This study focuses on elucidating the role of Th 17 cells, a T helper cell subpopulation involved in pathology and host defenses against bacterial pathogens. C57BL/6 mice were immunized with iFt and mAb-iFt on day 0 and 21 and then challenged i.n. with a lethal dose of LVS on day 35. Flow cytometry data revealed a significant population of Th17 cells (characterized as CD3+/CD4+/IL- 17a+/RORγ-T+ cells) in the lungs and spleen post-infection in the mAb-iFt immunized group when compared to PBS and iFt alone. Additionally, cytokine levels of IL-6 and TGF-β1 (two known synergistic Th17 polarizing cytokines) were elevated in the lungs and spleens of mAb-iFt immunized mice on days 2 and 5 post-infection when compared to iFt alone. Lastly, cytokine levels of IL-17a, an effector cytokine secreted predominantly by active Th17 cells, was shown to be significantly increased in mAb-iFt immunized mice when compared to iFt alone. Collectively, our data suggests that Th17 cells possess a protective role in the immune response to pulmonary F. tularensis infection.
Molecular Signatures of IL-10/IFN-γ Co-Producing CD4 T Cells During Acute Dengue Virus Infection
Authors: Tian Yuan , Seumois Gregory , de la Mata Sara Herrera , Goonawardhana Suraj , de Silva Aruna D , Premawansa Sunil , Premawansa Gayani , Wijewickrama Ananda , Grifoni Alba , Vijayanand Pandurangan , Peters Bjoern , Weiskopf Daniela , Sette Alessandro 
Published: January 2023

Abstract
Dengue virus (DENV) is an emerging pathogen and a serious public health issue. It is estimated that ~390 million people are infected yearly. DENV infection is associated with a range of clinical manifestations, from asymptomatic to more severe presentations including dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). There is currently no specific therapy available for the treatment of dengue diseases other than supportive care. Thus, a better understanding of the host’s immune response against DENV, especially during the acute phase of infection, is of great public health interests. Although previous studies suggest that increased IL-10 level in the serum and plasma is associated with severe dengue disease, whether dengue-specific CD4 T cells produce IL-10 remains unknown. Here we show evidence of DENV-specific IL-10-producing CD4 T cells during acute DENV infection. Surprisingly, an IL-10+IFN-γ+ double positive (DP) CD4 T cell population is prominent in acute hospitalized DENV cases but disappear at the convalescent stage. These cells are not conventional regulatory T (Treg) cells as they lack Foxp3 expression. In contrast, RNA-sequencing and high-dimensional CyTOF analyses reveal that these cells partially exhibit previously identified signatures associated with cytotoxic CD4 T cells and type 1 regulatory T (Tr1) cells. Notably, we identified several novel signatures of DP cells including IL-19, IL-21, and IL-22, suggesting that these cells may have complex cytokine activities and functions. Overall, this study reveals unique phenotypes of DENV-specific IL-10/IFN-γ co-producing CD4 cells and provides insights into antigen-specific T cell responses during acute DENV infection.
Increased polyfunction in CD4 T cells from persons with acute vs. remote shingles
Authors: Jin Wenjie , Fang Michael , Vangipuram Ramya , Sayin Ismail , Golden Jackelyn , Cameron Cheryl , Cameron Mark , Tyring Stephen , Canaday David H. 
Published: January 2023

Abstract
Little is known about CD4 T cell function in Varicella Zoster Virus (VZV) infection. Here we sampled peripheral blood of individuals (n=31) with acute shingles as a result of reactivation of VZV and persons with a remote history of shingles (n=15). Mitogen and whole VZV were added to peripheral blood mononuclear cells (PBMC) for an in vitro stimulation assay. IFN-γ, IL-2, TNF-α, CD107a, T-bet, EOMES and KLRG1 were examined by flow cytometry. Mitogen- and whole VZV-stimulated total memory CD4 T cells showed significant differences in polyfunction (p&lt;0.0001) while the mitogen-activated groups were equal in acute vs history of shingles. There were significant differences between polyfunctional profiles in VZV-responding total and central memory CD4 T cells in acute vs remote shingles (p&lt;0.002). Acute patients’ VZV-specific total memory cells had higher frequencies of IFN-γ (p&lt;0.001) and IL-2 (p&lt;0.02) producing cells, perhaps due to differences in effector memory subsets. Acute patients' VZV-specific but not mitogen-responding CD4 T cells showed significantly higher levels of T-bet+ than donors with only a history of shingles (p&lt;=0.01). RNASeq analysis of ex vivo total memory CD4 T cells from these individuals showed differential regulation of T cell survival and differentiation pathways, including differential expression of IFN, TNF and mTOR signaling (p&lt;0.05 for genes/pathways). These gene signatures correlated with the frequency of IFN-γ and IL-2 producing cells responding to VZV above with linear regression analysis. In summary, CD4 T cells respond differently in individuals with acute vs. remote shingles, and recognize VZV-specific epitopes upon reactivation via T-bet-mediated pathways.
Supported by NIH and VA.
Human Vγ9Vδ2-T cells exhibit potent antiviral activity against EV71 infection
Authors: Tu Wenwei 
Published: January 2023

Abstract
Enterovirus 71 (EV71) is a major pathogen of hand, foot and mouth disease (HFMD). It is highly contagious and usually causes mild disease. However, occasionlly HFMD caused by EV71 would develpoed into severe neurological complications or even deaths in young children. There is no effective anti-viral therapy against EV71 infection. Human Vγ9Vδ2-T cells, as the first line of human immune defense, exhibit antiviral activities against different viruses. However, whether human Vγ9Vδ2-T cells have antiviral activity against EV71 remains unkown. In this study, we used EV71-infected primary human monocyte-derived macrophages (MDMs) to examine the anti-EV71 activity of Vγ9Vδ2-T cells. We found that phosphoantigen pamidronate-expanded Vγ9Vδ2-T cells could kill EV71-infected MDMs, and thus reduce the viral load. The cytotoxicity of Vγ9Vδ2-T cells against EV71-infected MDMs required cell-cell contact and mediated by Fas/FasL pathway. Vγ9Vδ2-T cells also displayed potent capacity to produce IFN-γ and thus inhibit EV71 viral replication. CCR5 pathway was involved in the migration of Vγ9Vδ2 T cells towards EV71-infected cells. These results demonstrated that Vγ9Vδ2-T cells have both cytotoxic and non-cytolytic antiviral activities target EV71. Our data suggests a novel therapeutic approach against EV71 by boosting Vγ9Vδ2 T cell immunity.
This work is supported in part by Health and Medical Research Fund, Hong Kong SAR government (13120972) and the General Research Fund (17115015 and 17126317), Research Grants Council of the Hong Kong SAR, China.
Single-Cell Transcriptomics Identifies TOX as a Key Transcriptional Regulator of Stem- Like CD8 T Cells in Chronic Infection
Authors: Wu Tuoqi , Yao Chen , Sun Hongwei , Lacey Neal , Gattinoni Luca , McGavern Dorian B , O'Shea John J , Schwartzberg Pamela L 
Published: January 2023

Abstract
Stem-like CD8 T cells maintain long-term antiviral immunity during chronic infection and share regulatory pathways with memory precursor cells generated after acute infection. However, it is unclear whether stem-like CD8 T cells require distinct transcriptional and epigenetic regulation for their longevity and adaptation to the immunosuppressive environment of chronic infection. By comparing single-cell transcriptomes and epigenetic profiles of CD8 T cells responding to acute and chronic viral infections, we found that stem-like CD8 T cells became distinct from memory precursors prior to the termination of clonal expansion. We discovered a co-expression gene module containing Toxthat exhibited higher transcriptional activity associated with more active histone marks in stem-like T cells than memory precursors. Moreover, TOX promoted persistence of antiviral CD8 T cells and was required for stem-like CD8 differentiation. Thus, stem-like CD8 T cells require distinct transcriptional and epigenetic programs for their differentiation and persistence during chronic viral infection.
iNKT cell stimulation by glycolipid ligands modified from α-galactosylceramide results in differential interleukin-2 secretion profiles
Authors: Schafer Carolina E. , Santibáñez Álvaro , Gutiérrez-González Matías , Toro Jessica , Marcelain Katherine , Howell Amy R. , Porcelli Steven A. , Carreño Leandro J. 
Published: January 2023

Abstract
Invariant natural killer T (iNKT) cells are a specialized group of unconventional T cells that recognize glycolipids presented by the surface protein CD1d, expressed by most antigen-presenting cells. This results in the rapid secretion of a wide array of cytokines without the need for prior antigen immunization, thus making iNKT cells an attractive target for immunotherapy. Marine sponge sphingolipid α-galactosylceramide (α-GalCer) is a potent agonist for iNKT cell activation, characterized for inducing both Th1- and Th2-like cytokine secretion profiles. There is an increasing interest for the development of Th1- and Th2-polarizing ligands for more focused immunotherapeutic applications. Our aim is to evaluate the iNKT cell activation ability of C6” modified α-GalCer derivatives. We generated and isolated clones from iNKT cell hybridomas derived from a partially humanized mouse, consisting of a knock-in of the human CD1d gene. We selected by flow cytometry iNKT cell populations reactive to α-GalCer presented in the context of the human CD1d molecule. Additionally, we evaluated activation of iNKT cells by C6” modified α-GalCer derivatives and obtained a diverse interleukin-2 response depending on the structural modifications of the ligands and the individuality of the iNKT cell clones. In several cases, this response was more potent than that induced by stimulation with α-GalCer. Our results highlight differences between mouse and human lipid antigen presentation, as well as differences in the magnitude of the cytokine response amongst clonal populations. Our ongoing work aims to elucidate the mechanism for this observation by performing Next Generation Sequencing of the TCR chains of the responding clones.
MHC class I and II genes dependent hypersensitivity to nevirapine and efavirenz in Indian HIV-1 patients
Authors: Dalal Bhavik Ashok , Shankarkumar Aruna , Pazare Amar 
Published: January 2023

Abstract

OBJECTIVE
The present study demonstrates and investigates the involvement of differences in human leukocyte antigens (HLA) class I and class II genes in the risk of hypersensitive reaction to nevirapine and efavirenz among Indian HIV- 1 infected patients.

METHODS
Thirty-three HIV-positive patients treated with efavirenz and nevirapine were studied. The HLA class I and II molecular typing in both Non-tolerant and tolerant patients were performed using the sequence-specific primers (SSP), the Olerup HLA-A-B-C and DQ-DR SSP Combi Tray. Odds ratios (OR) with 95 % confidence intervals (CI) were calculated to assess patients’ increased risk of developing hypersensitivity in comparison with Tolerant.

RESULTS
Thirteen patients (39.39%), developed a hypersensitive reaction thus requiring the drug to be discontinued. AnHLA-B*40 and HLA-C*15 allelesrevealed a significant association in 8/13 (61.53%) p = 0.028, OR= 9.6; 95% CI=1.04–222.49 and 7/13 (53.84%)p = 0.05, odds ratio (OR)= 8.75; 95% CI= 0.92–205.44 of hypersensitivity reaction in HIV patients compared with only 4% in Tolerant group, respectively. MHC class I genes, HLA-A*24 allele and MHC class II genes, DRB1*10:01:01:01 allele showed significant association with skin rash in 5/13 (38.46%) and 4/13 (30.76%) respectively (p&lt;0.05). One patient developed potentially life-threatening condition i.e. Steven Johnson Syndrome.

CONCLUSION
A strong significant associations were seen between the MHC genes alleles (HLA-B*40 and HLA-C*15) and cART drug (nevirapine and efavirenz) induced skin rash in the Indian population. It can be utilized to avoid a subset of hypersensitivity reaction in HIV infected patients in Indian population.
Identification of novel subsets amongst dendritic cells and macrophages generated from the culture of bone marrow
Authors: In Hyunju , Na Hye Young , Ryu Seul Hye , Sohn Moah , Choi Wanho , Shin Hyun soo , Park Ji Soo , Park Chae Gyu 
Published: January 2023

Abstract
Dendritic cells (DCs) play pivotal roles in various immune responses by sampling and presenting antigens to naïve T cells and inducing the subsequent T-cell activation or tolerance. In vitro culture systems have been widely used to study DC biology. The culture of bone marrow (BM) with the aids of a hematopoietic cytokine, Fms-like tyrosine kinase 3 ligand (FLT3L), yields plasmacytoid DCs (pDCs) and classical DCs (cDCs), whereas the culture of BM in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) produces BM-derived DCs and macrophages considered to possess inflammatory phenotypes. In the present study, various antibodies including DC-specific monoclonal antibodies were utilized to carefully describe the subsets of cells present in the cultures of BM with GM-CSF or FLT3L. We identified new subsets of DCs and macrophages: 3 different subsets of DCs and macrophages were characterized from the CD11c+ MHCIIhi cells in the BM culture with Flt3L by flow cytometric analyses. The newly discovered subsets of DCs and macrophages present in the BM culture require further investigation to distinguish and elucidate their phenotypes and functions in details.
Characterizing the MHC-II immunopeptidome of HIV using a cell-free antigen processing system and peptide:MHC-specific antibodies
Authors: Sengupta Srona , Douglass Jacqueline , Boronina Tatiana , Wu Fengting , Moskovljevic Milica , Reed Madison , Yu Jeanna , Tabdili Yasmine , Siliciano Robert , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
The breadth and specificity of CD4+T cell responses in HIV-1 infection are critical determinants of viral load and an individual’s ability to control infection. Most studies of CD4+T cell responses against HIV have relied on indirect evidence from peptide-pulsing analyses. Direct studies of antigen presentation have been hampered by the technical difficulties inherent in isolating peptide:MHC (p:MHC) complexes from HIV-infected cells. This has made it challenging to understand the HIV immunopeptidome, particularly for MHC-II epitopes. Using a cell-free antigen processing system, we have generated a near complete map of potential DR1-restricted HIV-1 epitopes. To confirm that epitopes identified using this system are actually presented by HIV-infected cells, we generated novel reagents termed single-chain diabodies (scDbs), or bispecific antibodies, which contain one Fab fragment against a p:MHC and another against CD3. These antibodies enable the use of CD8+ or CD4+T cells as readouts for antigen presentation. In proof of concept experiments, scDbs directed against immunodominant HIV MHC-I epitopes enhance cytokine production by CD8+ T cells, confirming presentation of these peptides. Preliminary experiments showed similar results with scDbs targeting MHC-II Gag epitopes on dendritic cells overexpressing the relevant peptides. We are now profiling the use of scDbs to detect antigen presentation in HIV-infected macrophages and CD4+T cells, the two target cells of infection. By establishing an immunopeptide map and tools to detect HIV peptide presentation, we can begin to address the kinetics, pathways, and cell types contributing to HIV antigen presentation on MHC-II, which will inform improved vaccine therapies.
Biodegradable Nanoparticles Containing Antigen and Vitamin D3 Efficiently Induce Antigen-specific Immune Suppression
Authors: Lee Chong-Kil , Jeong Seong-Un , Jung Ho-Hyun , Kim Sang-Hyun , Kim Kyungjae 
Published: January 2023

Abstract
Biodegradable poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles (NPs) encapsulated with antigens can efficiently deliver the antigens to antigen-presenting cells such as dendritic cells (DCs). Active form of vitamin D, 1α,25-dihydroxyvitamin D3 (VD3), is known to have various immunosuppressive effects. Encapsulation of VD3 within NPs would enhance the delivery of VD3 to APCs, while preventing the potential systemic side effects of VD3. In this study, we examined the effects of PLGA NPs encapsulated with ovalbumin (OVA) and VD3. PLGA NPs containing OVA and VD3 (NP[OVA+VD3]) were prepared by W1/O/W2 double emulsion solvent evaporation method. Bone marrow-derived DCs treated with NP[OVA+VD3] were decreased in the expression of co-stimulatory molecules, and produced higher levels of immunosuppressive cytokines, while producing lower levels of pro-inflammatory cytokines, compared with DCs treated with NP[OVA]. Addition of NP[OVA+VD3] to allogeneic MLR reduced the proliferation of allogenic T cells. In addition, NP[OVA+VD3]-treated DCs efficiently generated CD4+CD25+Foxp3+Treg cells from CD4+CD25− T cells. Furthermore, mice injected i.v. with NP[OVA+VD3] were suppressed in inducing OVA-specific cytotoxic T cells. These results show that phagocytic delivery of antigen and VD3 to APCs using biodegradable NPs efficiently suppresses antigen-specific immune responses.