bluuebunny/crossref_metadata_2025_split huggingface dataset:
Suppression of IL-17 producing type 3 innate lymphoid cells and increased susceptibility to Klebsiella pneumoniae sepsis in neonatal mice exposed to extended empiric antibiotics
Authors: Mirpuri Julie , Babu Sarah Thomas , Niu Xinying , Kumar Dharmendra , savani Rashmin 
Published: January 2023

Abstract

Background/Objective
Over 75% of newborns admitted to the neonatal intensive care unit (NICU) receive antibiotics with no proven infection (empiric) at birth. Empiric antibiotic use in the NICU increases susceptibility to late onset sepsis, but the mechanisms are poorly understood.

Methods
Neonatal mice were exposed to broad-spectrum antibiotics through their dams from birth for either 3-(short exposure, SE) or 7-days (long exposure, LE). At 14 days, qRT-PCR was used for bacterial 16S rRNA (stool), antimicrobial proteins (AMPs) and cytokines (small intestine, SI). Blood, liver and spleen cultures was utilized to detect bacterial translocation. Histology of SI was performed for Paneth cells. Flow cytometric analysis of lamina propria (LP) cells determined the presence of type 3 innate lymphoid cells (ILC3), neutrophils, macrophages, T- and B-cells. Susceptibility to sepsis was determined by IP injection of K. pneumoniae in 12-day-old pups.

Results
Mice exposed to SE and LE showed significant expansion of Enterobacteriaceae (2 log difference). LE mice showed baseline translocation of Enterobacteriaceae into the liver and spleen and were more susceptible to sepsis. By flow cytometry, LE mice showed a persistent and significant 50% decrease in IL-17 producing ILC3 in the LP, with other cell types unaffected. Further, the ILC3 present were skewed to the IL-22 producing phenotype. AMPs and Paneth cells were also increased in the SI, likely mediated by the increase in IL-22.

Conclusions
Extended empiric antibiotic exposure resulted in persistent expansion of Enterobacteriaceae and increased susceptibility to neonatal sepsis, likely mediated by a microbiota-dependent decrease in IL-17-producing ILC3.
B7-H4 expression in donor T cells and host cells negatively regulates acute graftversus- host disease lethality
Authors: Saha Asim , Taylor Patricia A. , Lees Christopher J. , Panoskaltsis-Mortari Angela , Osborn Mark J. , Feser Colby J. , Thangavelu Govindarajan , Melchinger Wolfgang , Refaeli Yosef , Hill Geoffrey R. , Munn David H. , Serody Jonathan S. , Maillard Ivan , Kreymborg Katharina , van den Brink Marcel , Dong Chen , Huang Shuyu , Zang Xingxing , Allison James P , Zeiser Robert , Blazar Bruce R. 
Published: January 2023

Abstract
The B7 family members are critical in positive and negative regulation of immune responses by engaging various lymphocyte receptors. B7-H4 is a member of the B7 family that can negatively regulate T cell function. We investigated the role of host and donor B7-H4 in regulating acute graft-versus-host disease (GVHD). Allogeneic donor T cells infused into B7-H4−/− versus wild type (WT) recipients markedly accelerated GVHD-induced lethality in a C57BL/6 to BALB/c GVHD model. Chimera studies pointed toward B7-H4 expression on host hematopoietic cells as more critical than parenchymal cells in controlling GVHD. B7-H4−/− recipients had rapid mortality associated with increased donor T cell expansion, gut T cell homing and loss of intestinal epithelial integrity, increased Teffector function (proliferation, pro-inflammatory cytokines, cytolytic molecules) and reduced apoptosis. Higher metabolic demands of rapidly proliferating donor T cells in B7-H4−/− versus WT recipients required multiple metabolic pathways, increased extra-cellular acidification rates and oxygen consumption rates, and increased expression of fuel substrate transporters. Interestingly, during GVHD, B7-H4 expression was upregulated on allogeneic WT donor T cells. Consistent with these data, donor B7-H4−/− T cells given to WT recipients increased GVHD mortality and functioned similarly to WT T cells from allogeneic B7-H4−/− recipients. Graft-versus-leukemia responses were intact regardless as to whether B7-H4−/− mice were used as hosts or donors. Taken together, these data provide new insights into the negative regulatory processes that control GVHD and provide support for developing therapeutic strategies directed toward the B7-H4 pathway.
Alloreactive B cells acquire suppressive function in recipients tolerant to allografts
Authors: Khiew Stella , Yin Dengping , Jain Dharmendra , Wang Qiang , Alegre Maria-Luisa , Chong Anita S F 
Published: January 2023

Abstract
Establishing a state of transplantation tolerance has been achieved in limited numbers of transplant recipients in the clinic. Clinical observations suggest that donor-specific antibodies can mediate allograft rejection, leading us to hypothesize that stable transplantation tolerance requires donor-specific B cell responses to also be suppressed. We hypothesize that this can be achieved through the induction of B cell-intrinsic tolerance. Using a well-established experimental model of transplantation tolerance induced to allogeneic B/c hearts with anti-CD154+donor spleen cell transfusion, we observed that donor-specific B cells are intrinsically tolerant. B cells from tolerant recipients did not produce anti-B/c IgG when adoptively transferred into naïve MD4 host and challenged with B/c splenocytes or heart grafts, and were blocked in their ability to differentiate into germinal center (GC) B cells. To test whether tolerant B cells could be rescued by ongoing GC responses, we adoptively transferred tolerant B cells into congenic hosts, followed by B/c spleen cell immunization. Tolerant B cells did not recover their ability to differentiate into antibody secreting cells, but instead they are able to suppress IgG production by naïve host B cells. Finally, we show that donor-specific tolerant B cells are not able to suppress third-party antibody responses nor exhibit linked suppression. Taken together, our data demonstrate that tolerant donor-specific B cells are profoundly altered compared to naïve B cells: they have significantly diminished ability to differentiate into antibody secreting cells, but instead acquired the ability to suppress donor-specific, but not third-party, antibody responses by naïve B cells.
C3 expression by lung transplants reduces alloimmune-mediated injury
Authors: Kulkarni Hrishikesh Satish , Liao Fuyi , Ma Lina , Wu Xiaobo , Kreisel Daniel , Atkinson John P , Gelman Andrew E. 
Published: January 2023

Abstract
Allograft injury is a major risk factor for death in lung transplant recipients. Complement protein C3 is primarily synthesized in the liver. However, a recent study by our group has demonstrated that C3 present in pulmonary epithelial cells can be augmented to facilitate their own ability to withstand acute environmental stress. Here, we hypothesized that pulmonary C3 expression reduces the severity of allograft rejection. To address this question, C3−/−left lungs [on a C57BL/6 (B6) background] were orthotopically transplanted into wildtype (WT) fully MHC mismatched (CBA/J) recipients. The right lung, native to the recipient, served as an internal control. CBA/J lungs transplanted into C3−/− recipients, C3−/− lungs transplanted into C3−/− recipients and CBA/J lungs transplanted into WT B6 recipients were also employed as controls. One week after transplantation, recipients were euthanized and evaluated for C3 levels and graft injury. C3 protein in C3−/− recipients of WT B6 and CBA/J lungs was not detected in the peripheral circulation but was expressed within graft tissues suggesting locally expressed C3 is sequestered within the lung. Signs of graft tissue injury were only observed in allogeneic recipients. However, acute rejection severity was significantly higher in C3−/− allografts when compared to WT allografts. Additionally, C3−/− allografts had a distinct pattern of acute alveolar damage and increased cellularity not evident in WT allografts. Taken collectively, our data indicate that expression of C3 by lung allografts attenuates alloimmunedependent responses that are associated with reduced survival in humans.
The prior exposure of CD4+ T cells to alloantigen blunts their secretion of the reparative growth factor amphiregulin
Authors: Zhang Xue , Mathews Lisa , Turnquist Heth R 
Published: January 2023

Abstract

Objective
Chronic rejection (CR) manifests after transplantation as function-limiting allograft fibrosis and vasculopathy. Insights into the mechanisms leading to CR are needed to develop effective therapies. The aggressive development of fibrosis during CR is similar to that observed in unproductive injury repair. Thus, we investigated how alloantigen (AlloAg) shapes novel mechanisms of immune-mediated tissue repair. Amphiregulin (Areg) is a growth factor secreted by innate and adaptive immune cells in response to IL-33 released during tissue injury. As IL-33 and Areg are implicated in muscle, epithelium, and nerve repair, we defined how AlloAg recognition by CD4+ T cells modulates IL-33-induced Areg secretion.

Method
C57BL/6 mice transgenic for a TCR recognizing a BALB/c-derived alloAg (Eα52–68) presented on I-Ab were administered IL-33 or Eα52–68 alone, or combined, 10 days before CD4+ T cell isolation. Areg secretion by isolated CD4+ during 4 days of culture with IL-33 or Eα52–68 was measured by ELISA. The signaling mediating Areg secretion was defined using inhibitors.

Results
CD4+ T cell exposure to IL-33 prior to alloAg secreted Areg upon de novo AlloAg exposure or secondary IL-33 stimulation. Exposure to AlloAg before IL-33, however, limited the capacity of alloreactive T cells to secrete Areg in response to either IL-33 or alloAg. Using the P38 inhibitor SB203580 and NF-κB inhibitors MG132 and TCPA-1 we established that both NF-κB and p38 are critical CD4+ T secretion of Areg.

Conclusion
Our studies suggest that recognition of AlloAg disrupts a reparative response mediated by CD4+ T cells. Limiting the reparative capacity of non-self T cells may aid pathogen clearance, but support development of CR in after transplant.
CD4+ T Cells are both necessary and sufficient to induce acute graft vs. host disease in lymphopenic recipients
Authors: McDaniel Brianyell T , Enriquez Josue , Furr Kathryn , Grisham Matthew 
Published: January 2023

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is a potentially life-saving treatment for refractory/relapsing hematological malignancies, blood disorders or autoimmune diseases. However, approximately 40–50% of patients undergoing allogeneic HSCT will develop a multi-organ, inflammatory disorder called acute graft vs. host disease (aGVHD). Experimental and clinical studies suggest that intestinal injury due to toxic, pre-transplant conditioning protocols (e.g. lethal irradiation and/or chemotherapy) may play a major role in the development of aGVHD. However, recent studies from our laboratory as well as others, suggest that this may not be the case.

Objective
We wished to quantify and compare the onset and severity of aGVHD induced by the adoptive transfer of allogeneic T cells into untreated lymphopenic mice.

Methods
Four million allogeneic or syngeneic CD4+CD62L+CD25− T cells were transferred (i.p.) into NK cell-depleted RAG1−/− mice or RAG2−/−IL2rγ−/− double knock-out (DKO) mice and assessed daily for signs of aGVHD.

Results
We found that adoptive transfer of allogeneic but not syngeneic T cells into NK cell-depleted RAG1−/− or DKO mice induced many of the clinical and histological features of aGVHD including weight loss, inflammatory cytokine production and tissue inflammation. In addition, adoptive transfer of allogeneic T cells into each recipient induced severe anemia as well as dramatic reductions in bone marrow and spleen cellularity.

Conclusions
We conclude that naïve allogeneic CD4+T cells are both necessary and sufficient to induce aGVHD in lymphopenic recipients in the absence of toxic, pre-transfer conditioning.
RNA origami nanostructures for potent and safe anti-cancer immunotherapy
Authors: Matiski Lawrence , Qi Xiaodong , Liu Xiaowei , del Villar Ryan Rodriguez , Zhang Fei , Yan Hao , Chang Yung 
Published: January 2023

Abstract
Nucleic acids released from microorganisms or damaged cells are perceived by the host cells as danger signals. They can be recognized by various types of pattern recognition receptors (PRRs), leading to activation of the innate immunity. Recently, we have identified a novel self-assembled RNA-origami nanostructure that shows high structural stability, feasibility of manufacture in large quantities, and demonstrates potent innate stimulatory activity. We further test its activity in murine tumor models both in vitro and in vivo. Although this RNA-origami nanostructure exerts no direct cytotoxicity on the tumor cells, it induces significant delay or regression of tumor growth in tumor-bearing mice with minimal toxicity. The mice showing tumor regression developed systemic long-term immunity and became resistant to further challenges of tumor cells. This anti-tumor immunity is dependent on both the innate and adaptive immune system. Thus, RNA-origami nanostructures represent a new line of platforms for rational design and construction of effective, safe and affordable immunotherapeutics.
A novel vaccination platform for epicutaneous allergen-specific immunotherapy based on beta-glucan neoglycoconjugates
Authors: Korotchenko Evgeniia , Machado Yoan , Strandt Helen , Joubert Isabella , Schiessl Viktoria , Bauer Renate , Sarajlic Muamera , Neuper Theresa , Wildner Sabrina , Horejs-Hoeck Utta , Gadermaier Gabriele , Scheiblhofer Sandra , Thalhamer Josef , Weiss Richard 
Published: January 2023

Abstract
Coupling of long chain carbohydrates to allergens can mask IgE epitopes and thus reduce the allergenicity of the neoglycoconjugates. The resulting molecules can target and activate dendritic cells via C-type lectin receptors. Neoglycoconjugates therefore have a high potential for epicutaneous immunotherapy (EPIT) by delivering a hypoallergenic vaccine to a tissue rich in dendritic cells. In the current study we test the therapeutic efficiency of the beta-glucan laminarin conjugated to ovalbumin (LamOVA) in a mouse model of allergic asthma.
Sensitized Balb/C mice were treated epicutaneously via micropores created by fractional laser ablation. Humoral and cellular responses were evaluated by ELISA, RBL assay and BAT. Lung function was analyzed by whole body plethysmography.
Laminarin remained biologically active after coupling to ovalbumin as indicated by binding to Dectin-1 receptor. IgE epitopes were successfully masked according to in vitro IgE binding aasay and skin reaction scores in therapeutic setup.LamOVA was more potent in stimulating humoral immune responses in comparison to OVA and induced the highest IgG1 and IgG2a production. Epicutaneous treatment with OVA and LamOVA reduced lung inflammation and improved lung function of sensitized mice more efficiently compared to s.c. injection with OVA/alum.
Epicutaneous immunotherapy can alleviate lung inflammation and reduce airway hyperresponsiveness in sensitized mice more efficiently than s.c. injection with alum. Coupling of allergen to laminarin reduces local side effects and increases the immunogenicity, resulting in a safer therapy.
Natural Adjuvants for in situ Vaccination Lymphoma Immunotherapy
Authors: Aleynick Mark , Peng Paul , Upadhyay Ranjan , Hammerich Linda , Roudko Vladimir , Yellin Michael , Brody Joshua 
Published: January 2023

Abstract

BACKGROUND
In situ vaccination (ISV) against lymphoma can be achieved with Flt3-ligand treatment to recruit dendritic cells (DC), radiotherapy to load DC with tumor antigens, and intratumoral injection of a pattern-recognition- receptor agonist (PRRa) to activate antigen-loaded DC. While clinical trials of ISV using synthetic PRRa have yielded remissions, the optimal approach to activate DC is unknown. We hypothesize that ‘natural’ PRRa, such as the attenuated pathogens in common prophylactic vaccines, could target multiple PRR, leading to more robust activation of DC as compared to synthetic PRRa.

METHODS
20 FDA-approved vaccines (BCG, MMR, etc.) were screened in vitro, for their effect on DC phenotype and function. DC tumor antigen cross-presentation was assessed using CRISPR gene-edited β2m−/−GFP-lymphoma cells and a GFP-specific CD8 T cell system. Mechanisms of immune activation were interrogated using MyD88, TRIF, MAVS, and IFNAR knockout mice, as well as a library of CRISPR-edited PRR-null Raw264 macrophages. Potent vaccines functioning through distinct mechanisms were evaluated in vivo in an ISV using the A20 murine lymphoma model.

RESULTS
Several vaccines induced robust DC activation, cross-presentation and increases in T cell activation, proliferation, and tumor killing. Some vaccines were more effective than synthetic PRRa in activating DCs to induce a T cell response. Distinct PRR and signaling pathways were engaged by different vaccines. In vivo, vaccine combination therapy induced tumor regression in a majority of animals.

CONCLUSIONS
Prophylactic vaccines, used alone or in combination, are effective DC activators and can be repurposed for use in ISV, with immediate translation into the clinic.
Optimization of human T cell activation and expansion protocols improves efficiency of genetic modification and overall cell yield
Authors: Yu Jessie , Chen Priscilla , Watson Ashley , Truong Danielle , Antonchuk Jennifer , Kokaji Andy I , Woodside Steven M. , Eaves Allen C. , Thomas Terry E. 
Published: January 2023

Abstract
Cancer immunotherapy using CAR T-cells is a rapidly progressing field and manufacturing these cells is a complex process that requires multiple optimization steps. We have developed reagents for the isolation, activation and expansion of human T cells that will be available for clinical cell therapy manufacturing. Soluble ImmunoCult™ Human T Cell Activators induce T cell activation via cross-linking CD3 and co-stimulatory molecules on the surface of cells. Activated T cells then can be genetically modified and subsequently expanded in ImmunoCult™-XF, a serum- and xeno-free T cell expansion medium. Here, we present several optimization strategies with ImmunoCult™ products in order to obtain high transfection efficiency and maximum cell yield. By evaluating activation dynamics of T cells and determining the optimal transfection time points, the transfection efficiency can be substantially improved in both CRISPR/Cas9- and lentiviral-mediated gene-modification methods. Our study also suggests that maintaining T cells at lower cell density after the third day following activation greatly improves cell viability and cumulative cell growth, resulting in an &gt;1000-fold expansion of total human T cells with &gt;85% viability over 10–12 days of culture. Expanded T cells co-express CD45RO+CD62L+ with low expression of PD-1. As an example, we applied the workflow described here to generate TCRαβ KO T cells from healthy donors with up to 90% knockout efficiency. The purity of TCRαβ KO T cells can be further increased with the use of an EasySep™ Human TCRαβ depletion kit. Taken together, the processes outlined in this study can be easily and rapidly implemented to improve T cell manufacturing efficacy.
IMPACT OF GUT MICROBIOTA ON VACCINE RESPONSE IN HIV EXPOSED INFANTS
Authors: Nyangahu Donald David , Plumlee Courtney , Wendoh Jerome , Urdahl Kevin B , Jaspan Heather 
Published: January 2023

Abstract
Maternal HIV infection has been shown to alter the composition and diversity of gut microbiota in their infants. We hypothesized that inheritance of an altered microbiota in HIV-exposed uninfected (HEU) infants is associated with poor vaccine responses. Here, using a germ free mouse model, we investigate the causative relationship between stool microbiota in HEU infants and BCG vaccine response.
BCG vaccine Th1 responses were assessed using a whole blood assay and an intracellular cytokine flow cytometry panel. Stool samples from time of BCG vaccination were collected, and those from high or low BCG responder infants transferred by oral gavage to germ free pups at day 5 after delivery. Five days after gavage, pups were sacrificed to analyze inherent immunity or vaccinated with 10^6 CFUs of BCG. Vaccinated animals were sacrificed 28 days post vaccination. BCG specific T cells were quantified using TB10.4 (class I) tetramers by flow cytometry.
At d10 post-delivery, pups gavaged with high responder stool (Phr) had significantly lower spleen cell count compared to those gavaged with low responder stool (Plr). Furthermore, Phr pups had significantly higher proportions but not numbers of effector memory CD4 T cells (CD4+CD44hiCD62Llo) relative to Plr. Total number of B cells and neutrophils were significantly reduced in Phr vs Plr. When vaccinated with BCG, Phr displayed significantly higher number of TB10.4-specific CD8 T cells compared to Plr.
Our data shows a direct role of the microbiota in inherent immunity in infants. Furthermore, the microbiota in low responder HEU infants caused reduced number of vaccine specific CD8 T cells. Together, we show for the first time that intestinal microbiota in human infants at birth impacts vaccine response.
Replicating Ad-SIV recombinant vaccines elicit mucosal humoral immunity in rhesus macaques at both rectal and vaginal sites with potential protective efficacy
Authors: Hogge Christopher James , Hait Sabrina Helmold , Enyindah-Asonye Gospel , Mushtaq Zuena , Hoang Tanya , Robert-Guroff Marjorie 
Published: January 2023

Abstract
Humoral immunity, especially at mucosal sites, is important for preventing HIV acquisition. Replicating adenovirus (Ad)-SIV priming/gp120 boosting regimens have elicited mucosal immunity and protection against intrarectal SIV challenge in Rhesus macaques. Here we investigated mucosal responses in female macaques primed intranasally/orally, then intratracheally with Ad5hr-SIV(Env/gag/nef), boosted twice intramuscularly with SIV gp120, and challenged vaginally with repeated low-dose SIVmac251. Vaginal washes and rectal swabs and biopsies were obtained 3-weeks post-immunizations. gp120-specific antibodies were assayed by ELISA and memory B-cells by flow cytometry. Gp120-specific rectal memory B-cells increased post-Ad priming but waned post-boosting, whereas rectal and vaginal antibodies increased post-boosting. Compared to pre levels, rectal gp120-specific IgA (p = 0.011) and IgG (p&lt; 0.0001) were elevated post-2nd boost; vaginal IgA/IgG antibodies differed post-1st boost (both p&lt;0.0001) and IgG levels &gt; IgA post-2nd boost (p=0.001). Rectal memory B cells post- 1st and 2nd Ads positively correlated with gp120-specific rectal IgA (p = 0.015, p = 0.0013, respectively) post-1st Ad, indicating vaccine targeting to mucosal effector sites. Significantly delayed SIV acquisition was not seen, but vaginal IgG post-1st Ad correlated with number of challenges (p=0.029). Vaccinated animals had lower acute viremia than controls (p = 0.054), consistent with higher vaginal gp120-specific IgG 2 weeks post-infection (p &lt; 0.0001). Our data show that upper respiratory tract immunization with replicating Ad vectors elicits strong mucosal immunity with potential protective efficacy against rectal/genital transmission.