bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chlamydia muridarum recombinant MOMP nanovaccine induces cross-reactive antibodies against Chlamydia trachomatis human serovars
Authors: Sahu Rajnish , Sipowe Aguy Clemence Nguiakam , Tadjuidje Vanella , Geisler William M , Van Der Pol Barbara , Dennis Vida A 
Published: August 2023

Abstract
Chlamydia trachomatis (CT), caused by various serovars, remains the leading sexually transmitted bacterial infection worldwide. An efficient delivery system is critical to developing whole or subunit vaccines against CT, and some vaccine developmental efforts are targeting biodegradable nanoparticles-based vaccines with encapsulated antigens. We developed a nanovaccine employing the recombinant major outer membrane protein (rMOMP) of C. muridarum (Cm) encapsulated in poly(lactic acid-co-glycolic acid) 85/15 (PLGA-rMOMP). We previously compared the humoral responses of two priming routes, subcutaneous (SC) or intramuscular (IM-p), followed by two SC-boosts to evaluate rMOMP-specific serum antibodies [IgG2a, IgG2b and IgG1] produced by PLGA-rMOMP immunization of female BALB/c mice. Here we used an elementary body (EB)-specific ELISA to investigate the ability of systemic antibodies produced in immunized mice to recognize Cm and to cross-recognize human CT-serovars D, F and J. Serum from mice inoculated with PBS encapsulated PLGA served as negative control. We observed high recognition of Cm specific total IgG (8-fold) and isotypes IgG2a (8-fold), IgG2b (32-fold), IgG1 (8-fold) by SC in comparison to IM-p immunizations. We also observed IgG antibodies recognizing CT-serovars D, F and J from mice immunized via both routes. Evaluation of the Th1 (IgG2a, IgG2b)/Th2 (IgG1) antibody titer ratios revealed that immunization via the SC route induced predominantly Th1 antibodies recognizing both Cm and human CT-serovars. Our data show that mice immunized with PLGA-rMOMP produced high levels of systemic antibodies recognizing Cm, but more importantly, there was a robust serological cross-recognition of the human CT-serovars.
This research was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number NIH-R21AI111159, NIH-NIGMS-RISE (1R25GM106995-01) and the National Science Foundation (NSF)-CREST (HRD-1241701) and NSF-HBCU-RISE (HRD-1646729) grants.
Does intravenous vaccination with self-killing BCG lead to development of an immune response that protects against Mycobacterium tuberculosis?
Authors: Smith Alexander A , Rodgers Mark A , Bigbee Carolyn M , Schnappinger Dirk , Ehrt Sabine , Flynn JoAnne L 
Published: August 2023

Abstract
The use of intradermal BCG offers limited protection against tuberculosis. Intravenous (IV) administration of BCG has shown to produce robust protection against infection in a non-human primate model of tuberculosis. However, the duration of exposure and time required to elicit an efficacious, protective response is unknown and safety of IV administration of live BCG remains a concern. Here we investigated whether a self-limiting strain of BCG (killswitchBCG; ksBCG) could induce protective responses in macaques. ksBCG uses two transcriptionally repressible lysins; the inducer doxycycline is required for repression, with the absence of inducer leading to BCG death. Our first study demonstrated the robust in vivo killing of ksBCG once doxycycline is stopped IV ksBCG elicited a robust, multicellular immune response in airways and lung up to 8 weeks, similar to standard BCG. In the current study we are assessing protection conferred against Mycobacterium tuberculosis with IV ksBCG. Mauritian cynomolgus macaques (MCM) were vaccinated intravenously with ksBCG. 5 months post vaccination, animals were challenged with M. tuberculosis and later euthanized 12 weeks post infection. Bronchoalveolar lavages were routinely taken throughout the study, and alongside tissue samples, analyzed using spectral flow cytometry. The presence of multiple cell types (CD4, CD8, and gd T cells, B cells, NK cells), cytokines (IFNg, IL-2, IL-17, TNF) and cytotoxic molecules (granzymes B and K, perforin, granulysin), as well as M. tuberculosis bacterial burden will be assessed to determine the level of protection provided by IV ksBCG.
NIH R01 AI143788
Repetitive vaccination with intravenous BCG is safe and immunogenic in naïve and SIV-infected macaques
Authors: Lehman Chelsea C , King Hannah A , Pokkali Supriya , Bucsan Allison N , Seder Robert , Roederer Mario , Bolton Diane , Darrah Patricia A 
Published: August 2023

Abstract
Tuberculosis (TB) is the leading cause of death in people with HIV (PWH) on or off antiretroviral therapy (ART); thus, new TB vaccine candidates should be evaluated in the setting of HIV. Mycobacterium bovis Bacillus Calmette-Guerin (BCG), the live attenuated bacterial TB vaccine given intradermally (ID) at birth, is contraindicated in PWH. We recently demonstrated that intravenous (IV) delivery of BCG elicits superior immunity and protection compared to ID delivery against Mycobacterium tuberculosis (Mtb) challenge in rhesus macaques. To assess IV BCG in an animal model of HIV infection, we immunized SIVmac251-infected macaques on suppresive ART (6m) and uninfected controls. Rhesus macaques received three doses of IV BCG, 8wks apart, and were monitored for the presence of BCG colony forming units (CFU) in peripheral blood and bronchoalveolar lavage (BAL) and Mtb-specific T cell responses in BAL. Repetitive IV BCG immunization was well-tolerated in SIV-infected and uninfected animals, with no difference in clinical parameters or BCG CFU in blood or BAL. T cell responses after IV BCG were similar, with both groups displaying a 10–50 fold increase in BAL T cell abundance after initial dose. Mtb-specific CD4 and CD8 responses peaked after the initial IV BCG dose and were maintained by subsequent immunizations for at least 6m. These data indicate that IV BCG is safe and immunogenic in SIV-infected, ART-suppressed macaques and that IV BCG boosting maintains T cell responses at the site of infection.
NIH Intramural Program
Characterizing the Plasma IgG Antibody Repertoire of Malaria-Naïve United Kingdom Adults Vaccinated with Novel Blood Stage Vaccine RH5.1/AS01B
Authors: Marchioni Jeffrey , Seeger Allison , Barrett Jordan R , McHugh Kirsty , Kain Jessica , MacGill Randall , Georgiou George , Draper Simon J , Lavinder Jason J , Ippolito Gregory C 
Published: August 2023

Abstract
Plasmodium falciparum, the most lethal species of human malaria, is susceptible to vaccine-induced antibodies, yet current efforts are challenged by limited efficacy, lack of durability, and parasite evasion. A promising blood-stage vaccine antigen, Reticulocyte Binding Protein Homologue 5 (RH5), has low polymorphism frequencies and uses a non-redundant red blood cell (RBC) invasion pathway. The Draper Lab (Oxford University) clinically tested RH5.1, an engineered variant of RH5, in AS01B adjuvant (GSK) designed to boost long-lasting, protective antibody titers.
RH5-specific B cell receptors (BCRs) from United Kingdom malaria–naïve adult volunteers vaccinated with RH5.1/AS01B were sequenced. Monoclonal antibodies (mAbs) were found to inhibit in vitro growth of RBC-invading parasite merozoites. Notably, the polyclonal plasma IgG of the volunteers exhibited high neutralizing potency, exceeding that of individual mAbs from the same donors, which raises the question: what is the disconnect between the BCR and circulating IgG repertoires? To examine this further, our lab completed BCR sequencing coupled with plasma IgG proteomics to characterize the plasma antibody repertoires of volunteers.
Plasma mAbs were identified that bind to known growth inhibitory epitopes, including those that block binding between RH5 and its RBC coreceptor, CD147, alongside a population of non-neutralizing, synergizing mAbs. Additionally, non-neutralizing mAbs targeting cryptic epitopes on the N- and C-terminus were found at consistently high levels. This dynamic between neutralizing and non-neutralizing plasma mAbs suggests a role for non-neutralizing mAbs, which should be further explored in future RH5 vaccine engineering.
Supported by grants from U.S. Agency for International Development (USAID) contract no. 7200AA20C00017 and the National Science Foundation Graduate Research Fellowship Program (NSF GRFP).
Evaluation of the time needed for BCG to be alive to protect against tuberculosis in intravenously BCG vaccinated SIV+ macaques
Authors: Jauro Solomon , Larson Erica C , Maiello Pauline , Rodgers Mark A , Ameel Cassaundra L , Gleim Janelle L , Sakal Jennifer , Kracinovsky Kara , Lin Philana Ling , Flynn JoAnne L , Scanga Charles A 
Published: August 2023

Abstract
Effective means of improving Bacille Calmette-Guérin (BCG) immunogenicity to protect against pulmonary tuberculosis (TB) is needed, especially among people living with HIV. Intravenous (IV) administration of BCG previously showed significant enhancement of immune responses and conferred ~75% protection against TB. Using our SIV/M. tuberculosis (Mtb) coinfection model (to mimic HIV/Mtb coinfection) in Mauritius cynomolgus macaques (MCM), we evaluated the length of time necessary for live BCG to elicit a protective immune responses by varying the timing of anti-BCG drug (isoniazid, rifampicin, and ethambutol: HRE) initiation. MCM were infected with SIVmac239 and randomly assigned into 4 different vaccination groups 16 weeks after SIV infection: intradermal BCG (BCG ID No HRE), BCG IV with HRE started 1 week post BCG (BCG IV 1wk HRE), BCG IV with HRE started 3 weeks post BCG (BCG IV 3wks HRE), and BCG IV without HRE treatment (BCG IV No HRE). The HRE treatment lasted for 8 weeks. Airways samples were collected by bronchoalveolar lavage (BAL), whereas lung tissue and lymph node were harvested at necropsy in a subset of BCG ID and BCG IV 3wks vaccinated MCM 20 weeks after BCG but before (Mtb) challenge. The remaining MCM were challenged with Mtb Erdman strain (~15 CFU). CD4 and CD8 T cell levels increased in BAL in all IV vaccinated MCM. CD4 T cells producing IFNg, IL-2, IL-17, and TNF peaked after BCG and were maintained until necropsy. There were increased tissue-resident T cells and cytokine production in lungs of IV vaccinated MCM compared to BCG ID. No signs of BCG dissemination were apparent in any IV vaccinated MCM. We are currently evaluating protection against infection and disease in the SIV+ MCM in each vaccination group.
Supported by grant NIH (R01 AI155345)
Antigen experience history directs distinct functional states of CD8+ CAR T cells during the anti-leukemia response
Authors: DeGolier Kole R , D’Antonio Marc , Danis Etienne , Yarnell Michael , Kohler Mark Eric , Kedl Ross , Scott-Browne James , Fry Terry 
Published: August 2023

Abstract
Chimeric antigen receptor (CAR) T cells are highly effective in treating hematologic cancers, but patients can relapse due to downregulation of the CAR-targeted antigen, leading to the hypothesis that CAR T cells require high density of target antigen to be effective. Non-engineered CD8+ memory T cells generate more rapid and robust effector T cell responses upon secondary infection, relative to their naïve counterparts, and can display higher antigen sensitivity. Our objective was to characterize the role of prior CD8+ T cell antigen experience (AgEx) in dictating functional CAR-mediated anti-tumor responses. To study the effect of T cell AgEx history on CD8+ CAR T cell (CAR8) function in vivo, we used a vaccine model to control prior T cell AgEx history, and a syngeneic adoptive transfer model with murine anti-CD19 CAR8 targeting murine leukemia engineered to express a low or high level of CD19. We analyzed CAR8 expansion and leukemia burden using flow cytometry and performed ATACseq and RNAseq at timepoints pre- and post-CAR transduction and after reinfusion of CAR8 into leukemia-bearing mice to ask whether CAR8 function was determined by the epigenetic and/or transcriptomic state of the starting cell population. We found that starting cell state dictated in vivo CAR8 effector functions, expansion, and differential chromatin accessibility in regions containing binding motifs for AP-1, Runx, Tcf and other transcription factor (TF) families. Motif chromatin accessibility and gene expression of many TFs showed distinct patterns temporally. We conclude that CAR8 maintain functional traits epigenetically imprinted in the starting cell and predict these analyses could inform methods to transcriptionally modulate CAR T cell function.
Sensitizing solid tumors to respond to Cytokine-induced apoptosis and CAR T-cell therapy
Authors: Guttormson Matthew C , Ting Adrian 
Published: August 2023

Abstract
Chimeric antigen receptor (CAR) T-cells are T-cells engineered with synthetic receptors that directs their specificity and function toward an antigen of choice. This application has been highly successful in treating a variety of blood-borne cancers such as B-cell lymphomas. CAR T-cells against solid tumors have been less efficacious, potentially due to the resistance of tumor cells to T cell-mediated cytotoxicity. Current paradigm suggests that tumor cells are killed by perforin/granzyme-secreting CD8 cytotoxic T cells (CTLs). However, activated CD8 T cells also produce TNF and IFNg, two cytokines that have been reported to have anti-tumor function, but it is not clear whether these cytokines can directly kill tumor targets. It has been reported that defects in kinases in the TNFR1 signaling pathway can sensitize cells to TNF-induced death. Therefore, we hypothesize that by inactivating these kinases, we can sensitize solid tumors to killing by CD8 CAR T-cells producing TNF and IFNg. We intend to conduct proof-of-concept studies to show that sensitizing tumor cells to cytokine-mediated cytotoxicity will enhance the effectiveness of CAR T-cell therapy against solid tumors.
Institutional Funding
Predicting the therapeutic efficacy of TLR stimulated macrophages for cancer treatment
Authors: Kurt Robert A , Newman Thalia , Liew Chun Wai 
Published: August 2023

Abstract
Macrophages play a variety of roles in immunity including both pro- and anti-inflammatory functions, which can be controlled by toll-like receptor (TLR) stimulation. Previously, we generated a computational model of interconnected TLR signaling cascades and validated the model using experimental data from the THP-1 monocyte cell line. This project aimed to adapt the computational model to provide insights into the activation of bone marrow-derived macrophages (BMDM) to enable the accurate prediction of BMDM therapeutic efficacy following TLR stimulation. Initially, we quantitated expression of genes encoding TLR signaling proteins in BMDM and THP-1 cells and found distinct differences between the cell types. For instance, IRAK2 was expressed in BMDM at a level over 150 times the expression level in THP-1 cells, while other genes, such as TLR4, were expressed at relatively similar levels. Next, these data were used to adapt the computational model to BMDM. In order to determine whether the model predicted the optimal way to activate the BMDM, the cells were treated with lipopolysaccharide and flagellin and IL-1b production was quantified by ELISA. The data revealed that the model did not predict the optimal way to achieve pro-inflammatory cytokine expression in the BMDM. We are currently altering the model to improve its ability to predict pro-inflammatory cytokine expression following TLR stimulation. In addition, studies are underway exploring the therapeutic efficacy of the TLR-stimulated BMDM using a murine mammary carcinoma model. The ultimate goal of this project is to produce a computational model capable of predicting how BMDM respond to TLR stimulation and to use the model to predict the anti-tumor efficacy of the BMDM.
Triple checkpoint blockade, but not anti-PD1 alone, enhances the efficacy of engineered adoptive T cell therapy in advanced ovarian cancer
Authors: Anderson Kristin G , Su Yapeng , Burnett Madison , Bates Breanna , Suarez Magdalia Rodgers , Voillet Valentin , Gottardo Raphael , Greenberg Philip D 
Published: August 2023

Abstract
Less than 50% of ovarian cancer patients survive five years after diagnosis. This rate has changed little in the last 30 years, highlighting the need for novel therapies. A promising strategy employs T cells engineered to target tumor-expressed proteins. Mesothelin (Msln) is overexpressed in ovarian cancers, contributes to the malignant and invasive phenotype and has limited expression in healthy cells, making it a candidate immunotherapy target.
We used the immune-competent ID8VEGF mouse model of advanced ovarian cancer to show that T cells engineered to express a Msln-specific T cell receptor (TCRMsln) can preferentially accumulate within established tumors, delay tumor growth and significantly prolong mouse survival. However, elements in the tumor microenvironment limited engineered T cell function. Tumor-bearing mice were therefore treated with TCRMsln T cells plus PD1, Tim-3 and/or Lag-3 checkpoint-blocking antibodies, alone or in combination. Triple checkpoint blockade, but not single- or double-agent treatments, dramatically increased effector cytokine production by intratumoral TCRMsln T cells. Single cell RNA-sequencing revealed gene expression changes in intratumoral TCRMsln T cells and myeloid cells consistent with immune activation and inflammation. Moreover, combining adoptive immunotherapy with triple checkpoint blockade prolonged survival in treated mice, relative to TCRMsln with or without single- or double-agent checkpoint blockade.
These results suggest that disrupting multiple immune-inhibitory pathways simultaneously, which can be more safely pursued in a cell intrinsic form through genetic engineering, may be necessary to improve outcomes for ovarian cancer patients.
XCL1 facilitates the interaction with CD8+ T-cells and XCR1+ DCs within the Cervical Lymph Nodes during WNV encephalitis
Authors: De La Torre Kristen , Durrant Douglas M 
Published: August 2023

Abstract
West Nile Virus (WNV), a neurotropic flavivirus, causes encephalitis, especially in elderly and immunocompromised individuals. Previous studies have demonstrated that CNS-localized dendritic cells (DCs) protect against WNV neuroinvasive disease by establishing antiviral T-cell responses in the CNS. In this study, we investigated the role of the XCL1-XCR1 interaction since its primary function is to facilitate DC-mediated CD8+ T cell immunity. XCL1, also known as lymphotactin, is expressed by various immune cells, including activated CD8+ T cells, and binds to XCR1, a chemokine receptor expressed predominately by a subset of DCs. Following subcutaneous infection with WNV, XCL1 expression increased within the CNS on day 6 post-infection after local viral replication began. In contrast, the expression of its receptor, XCR1, increased by day 9 post-infection in wild-type (WT) mice. We found that WNV-infected XCR1-deficient (XCR1−/−) mice demonstrated increased symptomatic disease and mortality in conjunction with the increased viral burden correlating with decreased CD8+ T-cell recruitment and activation at day 12 post-infection in the CNS compared with their WT counterparts. We found that XCR1+ DCs were present in the cervical lymph nodes at days 6 and 9 post-infection and in the CNS at day 12 post-infection. Taken together, these results suggest that the XCL1-XCR1 interaction promotes viral clearance and protection against WNV neuroinvasive disease.
NIH-SC3GM130472
Studies in humans and mice reveal a critical role for CCR2 in trafficking of pDCs during infection with SARS-CoV-2
Authors: Zhang Howard H , Lu Xinping , Garner Davis , Parween Farhat , Singh Satya , Majumdar Shamik , Weaver Joseph , Stein Sydney , Lesho Patricia , Damcott Coleen , Lutfi Forat , Petrick Kendra , Rock Peter , Chertow Daniel , Kelsall Brian L , Murphy Philip M , Dahiya Saurabh , Hardy Nancy , Farber Joshua 
Published: August 2023

Abstract
Although GWAS have implicated chemokine receptors in affecting disease severity, the role of chemokines in COVID-19 pathogenesis has not been defined. To bridge this gap, we surveyed expression of 14 chemokine receptors on blood leukocytes from 19 hospitalized COVID-19 patients vs. healthy controls. Among many differences between patients and controls, we found that CCR2, which we have shown mediates lymphocyte transendothelial migration, was down on patient MAIT cells and pDCs, whose numbers in blood are depressed in COVID-19. If a chemokine receptor mediates extravasation, lower levels of the receptor on blood cells could result from efficient, and thereby preferential migration of leukocytes expressing higher levels of the receptor into infected tissue. In support of this mechanism, we found enrichment of CCR2high cells among MAIT cells migrating to the CCR2 ligand, CCL2, in vitro. To test if CCR2 mediates leukocyte extravasation in COVID-19 we infected transgenic K18-hACE2 Ccr2 reporter mice that were Ccr2+/RFP (Ccr2 sufficient) or Ccr2RFP/RFP (Ccr2 deficient) with SARS-CoV-2 intranasally and analyzed pDCs. Blood pDCs were RFP+ and their numbers were decreased in Ccr2RFP/RFP mice at baseline. Over seven days after infection with SARS-CoV-2 (LD50), pDCs increased and then decreased in the blood, while increasingly progressively in lungs in both Ccr2+/RFP and Ccr2RFP/RFP mice. However, absolute numbers of lung pDCs and ratios of lung/blood pDCs were markedly diminished, as was survival, in Ccr2RFP/RFP mice. We propose that increased mortality in Ccr2-deficient mice may be due in part to inadequate numbers of lung pDCs, leading to local deficiencies in type 1 and III interferons, which are critical for defense against SARS-CoV-2.
Diabetic wound keratinocytes induce macrophage JMJD3-mediated Nlrp3 expression via IL1R signaling
Authors: Wolf Sonya , Audu Christopher , Moon Jadie Y , Joshi Amrita D , Melvin William J , Barrett Emily C , Mangum Kevin D , Gallagher Katherine A 
Published: August 2023

Abstract
Type 2 diabetic (T2D) wound Macrophages (Mϕs) display a delayed chronic inflammatory state that prevents the resolution of wound inflammation. Increased NLRP3 inflammasome gene expression and activation in Mϕs contributes to impaired wound healing in T2D and is induced by diabetic wound environment factors, yet the molecular mechanisms regulating this are unclear. Our lab has shown that keratinocytes regulate Mϕ phenotype during wound healing by producing proinflammatory cytokines. Thus, this project explores the molecular mechanisms by which keratinocytes drive increased Nlrp3 expression in diabetic wound Mϕs. Here, we found that Nlrp3 expression is increased in diabetic wound Mϕs late post-injury, and this can be induced by stimulation of Mϕs with diabetic wound keratinocyte supernatant. Further, we found that stimulation of Mϕs with diabetic wound keratinocyte supernatant decreased H3K27me3 deposition at the Nlrp3 promoter. Using genetically engineered mice (Jmjd3 fl/fl lyz2cre), we demonstrate that JMJD3, a demethylase specific for H3K27, controls Mϕ-mediated Nlrp3 expression and inflammasome activation during wound healing. Importantly, we show that diabetic wound keratinocytes induce Jmjd3 expression in Mϕs via IL1R/MyD88 upstream signaling. Interestingly, we found that IL1α is increased in human and murine wound diabetic keratinocytes compared to non-diabetic controls and stimulation of Mϕs with IL1α increases Nlrp3 expression via a JMJD3-mediated mechanism. Thus, our data suggest a role for keratinocyte IL1α in driving increased Nlrp3 expression in diabetic wound Mϕs via JMJD3. Continued investigation into the keratinocyte-Mϕ axis is key to developing novel treatments for non-healing T2D wounds.
P30 DK089503, 1K99DK133828-01, 5K12 GM111725