bluuebunny/crossref_metadata_2025_split huggingface dataset:
Minced muscle grafts reduce the proinflammatory immune response and partially restore musculoskeletal healing
Authors: Wager Chrissy M Leopold , Goldman Stephen M , Pollot Beth E , Greising Sarah M , Wenke Joseph C , Corona Benjamin T 
Published: January 2023

Abstract
The immune response plays a key role in healing of bone and muscle injuries. An unrestrained proinflammatory response generally results in impaired healing. Extremity fracture is often complicated by traumatic volumetric muscle loss (VML), which can lead to delayed or non-union of the fracture (i.e., impaired healing). In a rat model, we recently showed that the VML injury of the tibialis anterior muscle prevented healing of the adjacent tibia fracture and induced a robust proinflammatory response in the musculature early post-injury that was not observed with the fracture alone. In the current studies, we show that a fracture treated with rhBMP-2, a highly osteoinductive protein, was impaired by the VML injury and corresponded with a heightened proinflammatory response. We utilized an autologous minced muscle graft (MMG) to repair the VML which, interestingly, reduced the proinflammatory response. Gene transcript analysis of the muscle injury indicates that MMG repair results in decreased gene expression of proinflammatory mediators including ITGAM, IL18, and CCL12, compared to non-repaired muscle. In the bone defect, MMG repair also induced a modest proinflammatory response with lower expression of IFNG and TNF than the non-repaired group. The restrained proinflammatory response corresponded with restored healing of the bone fracture and, remarkably, partial restoration of muscle function which does not normally occur clinically. Altogether, data suggest that MMG repair controls the proinflammatory response in a manner conducive to improved healing responses in both muscle and bone. These data reveal that immunomodulation to dampen proinflammatory responses could provide an effective therapy for musculoskeletal trauma.
Deletion of CCL2 in mesenchymal stem cells suppresses experimental aortic aneurysms
Authors: Xu Baohui , Deng Hongping , Li Gang , Fujimura Naoki , Furusho Yuko , Dalman Ronald L 
Published: January 2023

Abstract
Abdominal aortic aneurysm (AAA) is a macrophage-mediated chronic inflammatory disease. Aortic mural macrophages arise from bone marrow-derived circulating monocytes, recruited to the aneurysmal aorta by CCL2/CCR2 signaling. Given the importance of mesenchymal stem cell (MSC)-derived CCL2 in bone marrow monocyte mobilization, we hypothesized that MSC-derived CCL2 contributes to AAA pathogenesis. AAAs were created in 10–12 weeks old, male MSC-specific CCL2 knockout (Nestin+/CCL2 Flox+/+) and wild type (CCL2 Flox+/+) mice via intra-aortic elastase infusion. AAA incidence and progression were assessed via serial in vivo ultrasound aortic diameter measurements and histopathological analysis at sacrifice. MSC cell-specific CCL2 ablation significantly reduced circulating Ly-6Chigh inflammatory monocytes at baseline as well as 3 hours following lipopolysaccharide injection. Following elastase infusion, wild type mice demonstrated progressive, time-dependent aortic enlargement beginning on day 3. In MSC-specific CCL2 knockout mice, elastase-induced aortic enlargement was substantially suppressed, and medial elastin lamellae and smooth muscle cell cellularity were preserved compared to that demonstrated in wild type AAA mice. Mural macrophages, T and B cell infiltration were all also significantly attenuated in knockout, as compared to wild type, mice. Additionally, aortic neoangiogenesis, as defined by mural CD31+ capillary vessel density, was significantly reduced in knockout mice. Taken together, this evidence suggests that MSC-specific CCL2 deletion suppresses progression of experimental AAAs, in conjunction with attenuation of aortic mural inflammation and angiogenesis.
Correction of mitochondrial and peroxisomal defects by lovastatin and AICAR in EAE.
Authors: Singh Inderjit , Saxena Nishant , Singh Avtar K , Won Jeseong 
Published: January 2023

Abstract
In multiple sclerosis, the CNS disease progression continues despite treatment with FDA approved immunomodulatory drugs. Recent studies report that loss of mitochondria and peroxisomes contributes to the CNS disease of MS. In EAE, the loss of myelin proteins paralleled the loss of peroxisomes (catalase and PMP70) and loss of mitochondria (complex I and citrate synthase) as well as levels of ATP. Using EAE animal model, we investigated the efficacy of immune modulatory drugs (lovastatin or AMPK activator AICAR) for protections of mitochondria, peroxisomes, and myelin in relation to the clnical disease. While lovastatin or AICAR attenuated the EAE disease, but individually these drugs provided only partial protection of mitochondria and peroxisomes. However, combination therapy of lovastatin and AICAR was able to protect mitochondria, peroxisomes, and myelin as well as greater efficacy against clinical disease of EAE as compared to individual drug treatments. Lovastatin-mediated inhibition of RhoA induced the expression of transcription factors, PPARα, PPARγ, and catalase and thus biogenesis of peroxisomes and AICAR-mediated activation of AMPK induced expression of transcription factors, PGC-1α, SIRT1, and MnSOD and thus biogenesis of mitochondria in EAE spinal cords. In summary, these studies document that oral medication of combination of immunomodulatory drugs (lovastatin and AICAR) protect against the loss of mitochondria and peroxisomes and energy homeostasis in EAE resulting in greater efficacy in EAE supporting a potential therapeutic approach for progressive MS. Supported in-part by VA:BX002829 and NIH:NS037766.
Breadth of function and homing capacity of peripheral CD4+ Th responses to intradermal (ID) immunization with enterotoxigenic E. coli fimbrial tip adhesin CfaE plus adjuvant LTR192G
Authors: Trop Stefanie , Laird Renee M , Gutierrez Ramiro L. , Riddle Mark S. , Porter Chad K , Maciel Milton 
Published: January 2023

Abstract
ETEC are a leading cause of traveler’s diarrhea and a vaccine development priority for the US military. A subunit vaccine comprised of ETEC fimbrial tip adhesin, CfaE, adjuvanted with single-mutant heat-labile toxin, LTR192G, was given ID 3 times, 21 days apart, and evaluated for immunogenicity and efficacy against moderate to severe diarrhea (MSD) following CFA/I+ ETEC oral challenge. The homing phenotype, activation status, and cytokine production of circulating CD4+ T cells following vaccination were assessed by flow cytometry to understand the response generated by mucosal adjuvant, LT, introduced via the skin, and the potential mechanisms involved in protection from an enteric pathogen.
Following in vitro stimulation with CfaE or LT 15mers, PBMCs of most vaccinated subjects expressed IFNγ or IL-17, within multiple homing subsets. Specifically, activated CfaE-specific α4β7+IFNγ+ cells were detected in 69% of vaccinated subjects, suggesting potential for gut homing, while 58% subjects generated CD62L+IFNγ+ responses. LT-specific IFNγ was observed in CD62L+ and CLA+ cells (52% and 50% subjects, respectively). Both CfaE- and LT-specific IL-17 were identified in CCR6+ (65%, 66% subjects, respectively) and CD62L+ cells (65%, 58%).
Subjects without MSD following experimental challenge had significantly higher CfaE-specific activated (CD154+) α4β7+ cells (p=0.043, t-test), as well as IFNγ (p = 0.036) and IL-17 (p = 0.008) responses to vaccination. LT-specific activation of CLA+ cells (p = 0.035) and IL-17 (p = 0.032) levels were also higher in subjects without MSD. Overall, ID vaccination with CfaE and LTR192G elicited broadly homing and functional Th cell responses that may play a role in protection at the mucosal level.
Immune response and early transcriptional changes after primary and booster vaccination against Hepatitis B in young and old adults
Authors: Weinberger Birgit , Haks Marielle C , Katzgraber Franz , Ottenhoff Tom HM , Grubeck-Loebenstein Beatrix 
Published: January 2023

Abstract
Many currently used vaccines are less immunogenic and effective in the elderly compared to younger adults due to age-related changes of the immune system. Most vaccines utilized in the elderly contain antigens, with which the target population had previous contact. Therefore, most studies investigating vaccine-induced immune responses in the elderly do not analyze responses to neo-antigen but rather booster responses. It can be hypothesized that age-related differences in the immune response are distinct for primary and recall responses. We therefore aimed to investigate primary and recall responses using the same antigen in young and older adults and chose Hepatitis B vaccine as a model antigen. Young (20–40 years) and elderly (&gt;60 years) healthy volunteers received either a primary series (no prior vaccination) or a single booster shot (documented primary vaccination more than 10 years ago) of the registered vaccine Twinrix. Expression of immunity-related genes was measured before and 1 day after vaccination and antibody titers were determined at several time points with a long-term follow-up 6 months after the last vaccination. Antibody responses were lower and seemed to be delayed in the elderly compared to young adults. Non-responders after the 3-dose primary series were more frequent in the elderly group. Antibody titers after booster vaccination increased in all participants. Associations between early expression-profiles on day 1 and antibody responses were found.
Antigenic competition affects the magnitude and breadth of CD8 T cell immunity following immunization with a nanoparticle neoantigen cancer vaccine
Authors: Blobel Nicolas J , Ramirez-Valdez Andrei , Ishizuka Andrew S , Lynn Geoffrey M , Seder Robert A 
Published: January 2023

Abstract
Induction of T cell immunity against neoantigens of cancers is a promising personalized vaccine approach against tumors. We developed a novel vaccine platform to enhance the breadth and magnitude of neoantigen-specific CD8 T cell responses by coupling neoantigen peptides to a polymer containing a Toll-Like Receptor-7/8 agonist that self-assembles into nanoparticles (termed SPP-7/8a). As the SPP-7/8 vaccine platform can be formulated to immunize against multiple neoantigen peptides, a critical issue is how the delivery influences the magnitude and breadth of CD8 T cell responses. Thus, C57BL/6 mice were injected subcutaneously against a single neoantigen at either a single site, or divided across four sites. CD8 T cell responses were ~3% in the mice immunized at multiple sites compared to ~1% at a single injection site. To assess breadth, mice were immunized with four antigens, with either a single antigen at each of four injection sites or a combination of all four antigens across four injection sites. In mice that received all four antigens in each site, CD8 T cell responses against two of the antigens were undetectable. By contrast, in animals that received 1 antigen per site, responses were detected against all four antigens, and the total magnitude was higher than animals receiving 4 antigens per site. Collectively, these data suggest that despite increasing the CD8 T cell response of a single antigen via multi-site injection, vaccination even with as few as 4 antigens can interfere with the induction of neoantigen-specific CD8 T cells. Current work is focused on altering the delivery of the SPP-7/8a immunization approach to optimize both magnitude and breadth of CD8 T cell immunity that will be critical for tumor treatment and protection.
Heat shock protein gp96-Ig vaccine induces antigen specific CD8 T cell response in the placenta
Authors: Strbo Natasa , Romero Laura , Garcia Dennise 
Published: January 2023

Abstract
The placenta acts as a barrier against infections, due to multiple unique structural, cellular, and immune properties. The detrimental effects of congenital viruses, including Zika virus (ZIKV), on pregnancy and fetal outcomes occur in part because of impaired placental function and profound pathological changes have been observed infected placentas. Our hypothesis is that induction of appropriate vaccine-induced immune responses in the placenta are key to successful prevention of congenital viruses’ infections in developing fetus. The main goal of our study is to developed novel heat shock protein (HSP) based vaccine approach that will induce antigen-specific responses in the placenta and will lead to the prevention of the pathogen transmission to the fetus.
HSP gp96 is a biological adjuvant that activates antigen-presenting cells and simultaneously delivers antigen specific peptides to MHC I for CD8 T cell activation. B6 females were mated with B6 mice and gestation day (GD) 0.5 was determined by the presence of vaginal plug. At GD 5.5, 0.5 million of OT1-gfp cells were injected in the tail vein. Two days after, 2.5 million 3T3-gp96-OVA-Ig or 3T3-gp96-Ig cells were injected subcutaneously (SC). Five days later, pregnant uteri were collected and placental and decidual single cells suspension was obtained. Frequency of OT1-gfp cells was determent by flow cytometry.
We found that secreted gp96 vaccine delivered by SC route induces very high frequency of antigen specific CD8 T cells in the placenta (15%) and decidua (10%) without compromising placental or fetal viability.
Our findings are highly supportive of development of novel gp96-Ig vaccine as unique systemic and placenta-homing, antigen-specific CD8 CTL vaccine strategy.
IgH isotype-specific B cell receptor dosage regulates B cell fate
Authors: Wesemann Duane R , Granato Alessandra , Tong Pei , Zuo Teng , Chaudhary Neha , Han Seung Seok 
Published: January 2023

Abstract
Immunoglobulin heavy chain (IgH) isotypes (e.g. IgM, IgG and IgE) are generated as secreted/soluble antibodies (sIg) or as membrane-bound (mIg) B cell receptors (BCR) through alternative RNA splicing. IgH isotype dictates soluble antibody function, but how mIg isotype influences B cell behavior is unclear. We examined IgH isotype-specific BCR function by engineering polyclonal Ighγ1/γ1 and Ighɛ/ɛ mice, which initially produce IgG1 or IgE from their respective native genomic configurations. Transcripts producing IgM, IgG1, and IgE are produced in an alternative splice form bias hierarchy, where mIgμ&gt;mIgγ1&gt;mIgɛ—influencing respective BCR densities and fitness for populating the periphery—a pattern replicated in activated and memory B cells from wild-type mice. Restrained B cell development from Ighγ1/γ1 and Ighɛ/ɛ mice was proportional to sIg/mIg ratios, and was rescued by enforced expression of the respective mIgs. In addition, enhanced BCR signaling by way of PTEN deletion in IgE B cells generated functional IgE memory responses from IgE memory B cells themselves, which are normally absent under normal conditions, indicating that weak signal strength normally restrains IgE responses. Thus, IgH isotype-specific mIg dosage regulates B cell fate by influencing quantitative baseline BCR signal strength.
Sex-specific differences in the CD8+ T cell response to infection is linked to differences in IL-12 sensitivity
Authors: Mon Kristel Joy Yee , Goldsmith Elizabeth , Wang Jocelyn , Watson Neva B , Smith Norah , Rudd Brian D 
Published: January 2023

Abstract
It is well known that males and females respond differently to intracellular pathogens. In general, females mount more robust immune responses than males, which decreases susceptibility to infection but comes at the cost of increasing auto-immunopathology. However, the underlying basis for these sex-related differences is poorly understood. In this study, we investigated cell-intrinsic differences between male and female CD8+ T cells. First, we compared the ability of male and female CD8+ T cells to proliferate following in vitro stimulation. Interestingly, we found that female CD8+ T cells proliferate sooner and faster than their male counterparts. Next, we asked whether cell-intrinsic differences between male and female CD8+ T cells contribute to changes in the CD8+ T cell response to infection in vivo. We co-transferred equal numbers of male and female donor CD8+ T cells from TCR transgenic mice into the same host and compared their ability to respond to Listeria monocytogenes. The data indicated that female CD8+ T cells preferentially become short-lived effectors (SLECs), while male donor CD8+ T cells give rise to more memory precursor cells (MPECs), even when placed in the same environment. To gain insight into why female CD8+ T cells are inherently more responsive to infection, we performed a series of in vitro and in vivo experiments, whereby either antigen (signal 1) or IL-12 (signal 3) was titrated. These studies showed that male and female CD8+ T cells respond similarly to low amounts of cognate antigen, but female CD8+ T cells appear to be more sensitive to IL-12. Collectively, our data demonstrates that female CD8+ T cells are biased towards forming SLECs after infection because of their enhanced capacity to respond to IL-12.
Oxidative phosphorylation is progressively programmed during plasma cell differentiation
Authors: Price Madeline J , Barwick Benjamin G , Scharer Christopher D , Boss Jeremy M 
Published: January 2023

Abstract
The differentiation of immune cells requires a transition in metabolic programming to achieve effector function. Unlike T cells, the metabolic changes and epigenetic regulation of this process during the differentiation of antibody secreting plasma cells is not well characterized. Using in vivo and ex vivo LPS driven models of B cell differentiation, the metabolic changes and epigenetic regulation of metabolism was investigated. Naïve B cells (B220+GL7−CD138−) are metabolically quiescent, and upon activation through TLR4, B cells rapidly shift their metabolic program to use oxidative phosphorylation for their energetic demands. Activated B cells (B220+GL7+CD138−) use oxidative phosphorylation, but maintain a large spare respiratory capacity. In contrast, terminally differentiated, post-mitotic plasma cells (B220−GL7−CD138+) use oxidative phosphorylation at their maximal ability, with no spare respiratory capacity, suggesting unique metabolic changes are programmed upon differentiation. Next, an adoptive transfer model system that utilizes CFSE labeled B cells was used to distinguish molecular events during the proliferative/activation versus the differentiation phase. Analysis of epigenetic and transcriptome data in the two phases identified a subset of metabolic genes that underwent differentiation specific programming. Additionally, the role of key transcription factors was examined in terms of their control of the metabolic reprogramming steps. These results provide novel insights into how the step-wise changes in metabolism are programmed as quiescent B cells differentiate into antibody-secreting plasma cells.
The intracellular C5 system is critical to DAMP sensing and cellular responses in human monocytes
Authors: Niyonzima Nathalie , Freeley Simon , Arbore Giuseppina , Le Friec Gaelle , Lappegård Knut Tore , Mollnes Tom Eirik , Kemper Claudia , Espevik Terje 
Published: January 2023

Abstract
Cholesterol crystal (CC)-induced inflammasome activation is a key contributor to atherosclerosis. Phagocytosis of CC causes lysosomal damage, assembly of the NLRP3 inflammasome, activation of caspase-1 and release of mature IL-1β. We have demonstrated that complement activation by CC in serum leads to C5a-C5aR1 driven CR3 up-regulation, increased uptake and IL1B gene expression in monocytes. Intracellular complement activation now emerges as critical regulator of cellular responses for induction of Th1 cells. In this study we investigated if CC-induced NLRP3 activation and IL-1β release in monocytes involves intracellular C5 activation. We observed that resting monocytes contained stores of C5 and C5a that increased upon CC uptake. Reduction of intracellular C5 by siRNA treatment abrogated IL-1β secretion to the same extent as NLRP3 inhibition. We confirmed the importance of the intracellular C5a-C5aR1 axis using monocytes from a serum C5-deficient patient, which produced intracellular C5 and detectable C5a and mounted equally strong IL-1β response when compared to healthy donor monocytes. Monocytes expressed also active Factors B and D and contained C3 and C3 activation fragments suggesting that an intracellular C5 convertase could mediate intracellular C5 activation. This notion is supported by the finding that knockdown of Factor B and usage of a cell-permeable convertase inhibitor both reduced CC-induced IL-1β. These results indicate that intracellular C3/C5 convertase-driven C5 activation is required for DAMP sensing, such as for CC, and suggest a critical role of intracellular complement in sterile inflammation and related disease states – and, importantly, that this novel axis can be pharmacologically targeted.
Glioblastoma usurps host extracellular adenosine to circumvent host anti-tumor Responses
Authors: Bynoe Margaret S 
Published: January 2023

Abstract
Glioblastoma (GB) is the most aggressive, fatal, and least successfully treated of all solid brain tumors due to its high capacity to proliferate and extreme chemoresistance. Treatment for GB patients is managed with multimodal therapies including surgical resection (if possible), radiotherapy and chemotherapy with temozolomide (one of the few drugs shown to have some effect). Despite this, patient survival ranges between 6–18 months. Because GB is highly infiltrative, it often invades normal brain tissue, sometimes preventing surgical resection, which inevitably leads to recurrence. Adenosine is an immune suppressive signaling molecule that is generated by the enzymatic action of CD73. I tested the hypothesis that GB usurps host extracellular adenosine to circumvent host anti-tumor mechanisms by down regulating macrophage/microglial cell immune responses and promote M2 instead of M1 macrophage/microglial phenotype. To test this, I isolated macrophage/microglial cells from the tumor microenvironment of mice lacking or expressing CD73. Compared to WT mice, the frequency of M1 verses M2 phenotypes showed that eighty percent of the macrophage/microglial cells present in the tumor microenvironment of CD73−/− mice exhibited an M1 phenotype, while WT mice had about 30 percent M1 phenotype.
Exosomal MEK1 delays tumor growth by promoting anti-tumor immunity
Authors: Searles Stephen , Lyden David C , Bui Jack D 
Published: January 2023

Abstract
Exosomes mediate intercellular communication and regulation in many processes, including cancer-immune cell interactions. Exosomes are small (~100 nm) membrane vesicles that arise from the late endosomal pathway and contain bioactive molecules that are transferred to and affect the function of target cells. Tumor-derived exosomes are important regulators of anti-tumor immunity and have been shown to exert stimulatory and inhibitory effects on immune cells, depending on the experimental setting. Using a proteomic approach, we found that exosomes derived from tumors that undergo immune rejection (“regressors”) contain different proteins than exosomes from progressively growing tumors (“progressors”). Specifically, the signaling molecule MAP2K1 (“MEK1”) is highly enriched in regressor compared to progressor exosomes, leading us to hypothesize that exosomal MEK1 stimulates anti-tumor immunity and inhibits tumor progression. We found that forced expression of MEK1 in progressor exosomes delays tumor growth and increases anti-tumor immune cell infiltration, and that MEK1-hi exosomes are sufficient to delay tumor growth, but MEK1-low exosomes are not. Moreover, we found that MEK1 protein can be physically transferred from tumor cells to macrophages via exosomes, and that MEK1-hi exosomes induce pro-inflammatory cytokine production in macrophages, but MEK1-low exosomes do not. We propose that tumor-derived MEK1 is transferred to macrophages via exosomes, which enhances their anti-tumoricidal effector function and delays tumor growth. This cell extrinsic, anti-tumor activity of MEK1 contrasts with the known oncogenic properties of MEK1 and could limit the efficacy of MEK1 inhibitors currently in clinical trials.