bluuebunny/crossref_metadata_2025_split huggingface dataset:
Negative regulation of eosinophil production by TLR4 ligand (153.26)
Authors: Fulkerson Patricia , Schollaert Kaila , Rothenberg Marc 
Published: January 2023

Abstract
It has long been appreciated that a drop in eosinophils is associated with acute bacterial infection while, in contrast, blood levels of monocytes and neutrophils rise. The initial eosinopenic response is believed to be secondary to margination of circulating eosinophils to blood vessel walls, but the mechanism for prolonged eosinophil depletion with bacterial infection remains undefined. In a culture system developed to differentiate mature eosinophils from low density bone marrow progenitors, IL-5 stimulation of progenitors resulted in induced expression of six TLRs with highest expression of TLR2 and TLR4 noted throughout eosinophil development. Surface expression of TLR4 was confirmed on CD34+ progenitors by FACS. Stimulation of progenitors with LPS markedly inhibited IL-5-mediated eosinophil production. Further, LPS administration in vivo specifically reduced numbers of eosinophil progenitors in the bone marrow. Treatment of developing eosinophils with LPS induced the generation of pro-inflammatory mediators, including IL-6, IL-10, IFN-γ and TNF-α, but mechanistic studies demonstrated that suppression of eosinophil production by LPS was independent of these mediators. Taken together, these findings suggest a direct effect of LPS on eosinophil progenitors as an explanation for eosinopenia following bacterial infections and a potentially novel therapeutic strategy for depleting eosinophil progenitors and inhibiting peripheral eosinophilia in eosinophil-associated diseases.
ADAM10 overexpression dysregulates S2/S3 cleavage of Notch, shifting hematopoietic lineage commitment in favor of myelopoiesis (153.3)
Authors: Saleem Sheinei , Gibb David , Conrad Daniel 
Published: January 2023

Abstract
Although the mechanism by which A Disintegrin and Metalloproteinase 10 (ADAM10) regulates hematopoiesis remains unclear, recent reports indicate its essential role in performing S2 cleavage of Notch receptors. Thus to determine the role of ADAM10 activity in hematopoiesis, we generated ADAM10 overexpressing mice (A10Tg). Transgene expression alters hematopoiesis post-hematopoietic Lineage-Sca-1+c-kit+ (LSK) subset differentiation but prior to lineage commitment of progenitor populations, which are dramatically affected in A10Tg mice. This results in delayed T cell development, abrogated B cell development, and expansion of myeloid derived suppressor cells (MDSCs). Albeit being in a tumor-free host, A10Tg MDSCs are functionally and phenotypically analogous to tumor-derived MDSCs. Given ADAM10’s role in Notch signaling, we hypothesized that the observed hematopoietic alterations may be a consequence of perturbed Notch signaling. In fact, blockade of ADAM10 (S2) rescues B cell development and reduces myeloid cells in A10Tg LSKs. Additionally, inhibition of γ-secretase (S3) in wild type LSKs results in enhanced myelopoiesis, mimicking the phenotype of A10Tg mice. Collectively, these findings indicate that the differential cleavage of Notch into S2 and S3 products regulated by ADAM10 is critical to hematopoietic cell-fate determination. Additionally, A10Tg mice provide a novel model for the examination of MDSC-mediated immune suppression in a tumor free environment.
Role of IL-17 and Th17 cells in HSV-1 induced corneal immunopathology (154.10)
Authors: Suryawanshi Amol , Veiga-Parga Tamara , Rajasagi Naveen , Rouse Barry 
Published: January 2023

Abstract
Herpes Simplex Virus-1 (HSV) infection of the cornea leads to a blinding immuno-inflammatory lesion of the eye termed stromal keratitis (SK). Recently, IL-17 producing CD4 T cells (Th17) were shown to play an important role in many autoimmune as well as chronic inflammatory conditions, but the role of IL-17 and/or of Th17 cells in HSV mediated immunopathology is unclear. Here we show that, after HSV infection of the cornea, IL-17 is upregulated in a biphasic manner with an initial peak production around day 2 pi and a second peak at day 21 pi, a time point when clinical lesions are evident. Further studies demonstrated that innate cells particularly, γδ T cells, were the major producers of IL-17 early after HSV infection. However, during the clinical phase of SK, the predominant source of IL-17 was Th17 cells which infiltrated the cornea only after the entry of Th1 cells. The delayed influx of Th17 cells in the cornea was partly attributed to the local chemokine and cytokine milieu as well as the inhibitory effects of the cytokines produced by Th1 cells. Finally, HSV infection of the IL-17 receptor knockout mice, as well as IL-17 neutralization in WT mice showed diminished SK severity. This was attributed to the reduced level of pro-inflammatory cytokines and MMPs as well as reduced infiltration of neutrophils. In conclusion, our results show that IL-17 and Th-17 cells contribute to the pathogenesis of SK, the most common cause of infectious blindness in the western world.
Viral variant evades CD8+ T cell immunity following suboptimal antigen recognition and activation of responding effectors (154.26)
Authors: Shorter Shayla , Schnell Frederick , Edwards Lindsay , Evavold Brian 
Published: January 2023

Abstract
Evasion of the CD8+ T cell response by mutating a dominant epitope is a common evasion strategy employed by many viruses. Citing the lack of a measurable antigen-specific CTL response, it is thought that this strategy allows for the virus to persist undetected by anti-viral effectors. Using an LCMV viral escape mutant, 35A, which contains a single amino acid mutation in the H-2Db restricted, gp33-43 CD8+ T cell epitope, we show that antigen specific P14 CD8+ T cells recognize the mutant antigen and mount an early response. Despite showing greatly reduced affinity for the presenting MHC class I molecule by RMAS assay, both in-vivo and in vitro data show that P14s undergo early proliferation in response to the mutated epitope but are sub-optimally stimulated as evidenced by lower levels of cell markers CD25 and CD28, increased apoptosis, and negligible IL-2 production. P14s responding to 35A also maintain a sustained level of SHP-1 phosphatase activity pointing to the initiation of a negative regulatory pathway that aborts full activation. In contrast to the viral escape mutation being ignored by antigen specific T cells, these observations show an active mechanism by which epitope mutation prevents the accumulation of a robust antigen specific CD8+ T cell response.
Early CD4 T cell help fails to rescue CD8 T cell-mediated control of neurotropic coronavirus infection (154.8)
Authors: Phares Timothy , Stohlman Stephen , Bergmann Cornelia 
Published: January 2023

Abstract
During murine neurotropic coronavirus infection CD8 T cells are critical anti-viral effectors, while CD4 T cells play a prominent role in the disease and demyelination associated with the infection. The failure of CD8 T cells to control infection in the absence of CD4 T cells implied that CD8 T cells require CD4 T help in this encephalomyelitis model. To explore whether CD4 T cell help was imprinted early during the priming and CD8 T cell expansion phase, CD4 T cells were not depleted until days 4 and 6 post-infection. Disease onset and progression was comparable between depleted and control groups, with CD4 depleted mice displaying slightly less severe clinical symptoms. Peripheral expansion of virus-specific CD8 T cells was also comparable to controls and recruitment of virus-specific CD8 T cells into the CNS was only slightly diminished. Nevertheless, control of infectious virus was impaired following CD4 T cell depletion and mice succumbed to infection. Failure to control virus correlated with reduced IFN-gamma in the CNS, as well as lower granzyme B and perforin mRNA levels in CD8 T cells. Impaired CD8 T cell activity coincided with the absence of CD4 dependent IL-21 expression in the CNS. These results indicate that the presence of CD4 T cells during priming does not rescue impaired CD8 T cell function within the CNS. The data rather suggest that CD4 T cell play a critical role in promoting CD8 T cell function and accumulation at the site of infection.
Efficient mucosal vaccination mediated by the neonatal Fc receptor (155.9)
Authors: Zhu Xiaoping , Ye Lilin , Zeng Rongyu , Bai Yu , Roopenian Derry 
Published: January 2023

Abstract
Almost all infectious diseases are initiated at mucosal surfaces, yet intramuscular or subcutaneous vaccination usually provides only minimal protection at sites of infection owing to suboptimal activation of the mucosal immune system. The neonatal Fc receptor (FcRn) mediates the transport of IgG across polarized epithelial cells lining mucosal surfaces. We mimicked this process by fusing a model antigen, herpes simplex virus type-2 (HSV-2) glycoprotein gD, to an IgG Fc fragment. Intranasal immunization, together with the adjuvant CpG, completely protected wild-type, but not FcRn knockout, mice after intravaginal challenge with virulent HSV-2 186. This immunization strategy induced efficient mucosal and systemic antibody, B- and T-cell immune responses, with stable protection for at least 6 months after vaccination in most of the immunized animals. The FcRn-IgG transcellular transport pathway may provide a general delivery route for subunit vaccines against many mucosal pathogens.
Adoptive transfer of multiple CTL clones limits antigen gene loss and tumor escape (156.22)
Authors: Kaluza Karen , Thompson Jill , Vile Richard 
Published: January 2023

Abstract
We study mechanisms of tumor escape from adoptive therapy in the mouse melanoma model B16ova. In this model, transfer of OT-I T cells can significantly prolong the survival of mice. However, intense OT-I pressure promotes tumor cell genomic instability, and most mice eventually develop recurrent ova gene-loss tumors. We hypothesized that simultaneous transfer of two CTL clones would limit tumor recurrence by preventing the selection of single gene-loss cells. To accomplish this, we co-transferred OT-I T cells and Pmel T cells. Co-transfer three days following tumor challenge prevented tumor growth, while neither clone alone could. And in mice with established tumors, co-transfer provided significantly better therapy than either clone alone. This improvement was due to both fewer tumors recurring and the delayed appearance of those that did recur. Additionally, we found that recurrent tumors from mice treated with co-transfer continued to express both target antigens, and the mice maintained both T cell populations. We therefore hypothesized that it should be possible to exploit those cells to further prolong the survival of mice. We used cyclophosphamide to stimulate reactivation of the immune system in mice with recurrent tumors and found that this supplemental therapy significantly prolonged the lives of only those mice which had previously received co-transfer. Overall, our results reveal several benefits of targeting multiple tumor antigens.
Selective expansion of NK cells from PBMCs for cancer immunotherapy (156.28)
Authors: Lim Seon Ah , Kim Tae-Jin , Kim Kwanghee , Lee Kyung-Mi 
Published: January 2023

Abstract
NK cells have the ability to kill transformed or tumor cells without any prior stimulation. Despite this ability, the function of NK cells in tumor patients is greatly suppressed due to multiple factors. Therefore, efforts have been focused on isolating and expanding NK cells from tumor patients and re-infusing them after in vitro activation. These processes require multiple procedures to isolate and purify NK cells from the patients' PBMCs and high cost cytokines. To devise a simple and low-cost method to expand and activate NK cells with high purity, we have screened numerous feeder cell lines. Among these cell lines, we discovered KL-1 cell line, selectively enriched NK cells from whole PBMCs. After 15 days of culture, the number of CD56+ CD3- NK cells increased approximately 130-fold with the NK cell purity of over 90%. Expanded NK cells mounted superior cytotoxicity against NK-sensitive K562 cells as well as KL-1 cells. These results indicated that KL-1 feeder cell line could be exploited to generate NK cells for adoptive transfer with this cost-effective yet highly functional NK expansion method.
SA-4-1BBL as an adjuvant component of tumor vaccines achieves better therapeutic efficacy than an agonistic 4-1BB antibody by limiting the frequency of regulatory T cells (156.8)
Authors: Sharma Rajesh , Srivastava Abhishek , Yolcu Esma , Shirwan Haval 
Published: January 2023

Abstract
Costimulation through 4-1BB receptor is important for the generation of CD8+ T cell responses critical for the control of tumors. As such, this pathway has been targeted for tumor immunotherapy using agonistic 4-1BB Abs. However, the use of Abs is associated with severe toxicity. We recently developed a nontoxic form of soluble 4-1BBL, SA-4-1BBL, and demonstrated that it delivers quantitatively and qualitatively improved signals to T cells than an agonistic 4-1BB Ab (3H3). We herein demonstrate that SA-4-1BBL as the adjuvant component of HPV E7 and survivin TAA-based vaccines shows better therapeutic efficacy than the 3H3 Ab in E7 expressing TC-1 cervical and survivin over expressing 3LL lung cancer mouse models. The better therapeutic efficacy of SA-4-1BBL was associated with its ability to restrict the frequency of CD4+CD25+Foxp3 Treg cells. In marked contrast, mice vaccinated with 3H3 Ab had dramatically increased numbers of Treg cells. Importantly, in vivo depletion of Treg cells using an Ab to CD25 significantly enhanced the efficacy of 3H3, but not SA-4-1BBL, based vaccine. Taken together, these data demonstrate that SA-4-1BBL serves as a better alternative to agonistic Abs to 4-1BB receptor for the development of cancer vaccines due to its safety and ability to diminish the incidence of regulatory T cells.
Essential role of NLRP3 and AIM2 inflammasomes in IL-1β production induced by the extracellular pathogen, enterohemorrhagic Escherichia coli (157.1)
Authors: Kailasan Vanaja Sivapriya , Rathinam Vijay , Kalantari Parisa , Fitzgerald Katherine , Leong John 
Published: January 2023

Abstract
Enterohemorrhagic Escherichia coli (EHEC) induces elevated levels of IL-1β in infected patients. However, the molecular mechanisms involved in the activation of IL-1β by EHEC are unknown. Inflammasomes are multiprotein complexes in the cytosol that process proIL-1β to bioactive IL-1β. To identify the inflammasomes involved in IL-1β induction by EHEC, we infected macrophages derived from wild type and mice deficient in inflammasome components with EHEC and analyzed the processing and release of mature IL-1β. EHEC-induced IL-1β production was dependent on caspase-1 and the adaptor ASC. Notably, both NLRP3, a cytosolic sensor for microbial and endogenous products, and AIM2, a cytosolic sensor for DNA, were absolutely essential for EHEC-induced IL-1β production. Though EHEC is a non-cytosolic pathogen, bacterial products such as DNA were observed in the cytosol of infected cells, providing a potential means by which cytosolic inflammasomes might be activated. This also indicated that phagosomal degradation of bacteria is a prerequisite for inflammasome activation by EHEC. Supporting this hypothesis, blocking lysosomal acidification and type I interferon signaling that are essential for bacterial lysis in phagosome, led to a marked decrease in EHEC-induced IL-1β secretion. Thus, our results demonstrate that EHEC potently activates multiple inflammasomes as a result of release of bacterial products from phagosomes into the cytosol.
IL-22 enhances Calgranulin A expression by human macrophages to inhibit virulent Mycobacterium tuberculosis growth (158.21)
Authors: Dhiman Rohan , Paidipally Padmaja , Barnes Peter , Tvinnereim Amy , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
Previously, we found that IL-22 produced by NK cells inhibits intracellular mycobacterial growth in human macrophages (Mϕ) by enhancing phagolysosomal fusion. In the current study, we determined the mechanism/s by which IL-22 inhibits virulent M. tuberculosis H37Rv (M. tb) growth in Mϕ. First we asked whether IL-22 restricts M. tb growth in phagosomes that leads to enhanced phagolysosomal fusion. W7, a phagolysosomal fusion inhibitor abrogated IL-22 dependent M. tb growth in Mϕ (11.38 ± 3.3 x 106 vs 2.95 ± 1.7 x 106 CFU per well, p=0.05) suggesting IL-22 inhibits M. tb growth after phagolysosomal fusion. In microarray analysis we found that mRNA expression for 41 genes was &gt;1-fold higher in rIL-22 cultured infected Mϕ compared to infected Mϕ. Of these genes, we selected Indolamine 2,3-dioxegenase (IDO1) and Calgranulin A for further study. rIL-22 enhanced Calgranulin A expression in M. tb infected Mϕ by three fold (3.3 ± 0.83 vs 1.0 arbitrary units, p=0.04) compared to infected Mϕ as measured by real time PCR and confirmed by confocal microscopy. In contrast, IDO1 expression of infected Mϕ was not effected by rIL-22. Calgranulin A siRNA abrogated rIL-22 dependent growth inhibition of M. tb in macrophages (18.4 ± 1.8 x 106 vs 7.97 ± 0.8 x 106 CFU per well, p=0.0005). Studies are underway to determine the effect of Calgranulin A siRNA on phagolysosomal fusion using early endosomal, late endosomal and lysosomal markers.
Autophagy protects against infection and inflammation in a murine model of tuberculosis (158.9)
Authors: Castillo Eliseo , Dekonenko Alex , Delgado-Vargas Monica , Arko-Mensah John , Lyons Rick , Virgin Herbert , Deretic Vojo 
Published: January 2023

Abstract
Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, infects approximately one-third of the world’s population. Recently, autophagy, an intracellular process that sequesters portions of the cytoplasm, including damaged or surplus organelles into autophagosomes and degrades them upon fusion with lysosomes, was found to be crucial in macrophages for controlling MTB ex vivo. Additionally, the autophagy pathway and proteins have been shown to be critical in immunity and inflammation. Despite the known roles of autophagy in killing intracellular bacteria and regulating the immune system it is unknown how autophagy controls MTB in vivo. Using Atg5f/f LyzM-Cre mice, we investigated the in vivo role of autophagy in macrophages in controlling MTB infection and shaping the immune response. Compared to wild-type mice, Atg5f/f LyzM-Cre mice displayed greater weight loss, lung pathology and death in a dose-dependent manner. An increase in neutrophils and macrophages was found in the lung of infected Atg5f/f LyzM-Cre mice whereas the wild type mice had an increase in CD4 and CD8 T cell infiltrates. In this context, luminex assay also revealed an increase in IL-1α, GM-CSF, IL-17 and CXCL1 in the lung lavage of the infected Atg5f/f LyzM-Cre mice when compared to the wild type mice. Taken together, these results show the importance of the autophagy pathway in macrophages in vivo in controlling MTB infection, and indicate a broader role in shaping the immune response.
Essential pro-survival role for caspase-8 during T-cell activation and embryonic development through inhibition of RIPK3-dependent necrosis (159.9)
Authors: Dillon Christopher , Oberst Andrew , Weinlich Ricardo , Martinez Jennifer , McCormick Laura , Fitzgerald Pat , Pop Cristina , Hakem Razq , Salvesen Guy , Green Douglas 
Published: January 2023

Abstract
Caspase-8 is required for initiation of death receptor-mediated apoptosis, but counterintuitively is also required for normal T-cell proliferation and embryonic development. Recent work demonstrated that upon activation, caspase-8-deficient T cells fail to proliferate due to high rates of necroptosis, an alternative form of programmed cell death. To investigate these seemingly contradictory functions, we bred caspase-8 knockouts, which are embryonic lethal at E10.5, with necroptosis deficient RIPK3-/- animals. Unlike Casp8-/- animals, Casp8-/- RIPK3-/- double knockout (DKO) animals were generated at expected Mendelian frequencies and appeared grossly normal. DKO thymocytes responded normally to stimuli inducing intrinsic cell death, but resisted death induced by ligation of CD95. DKO animals, but not their normal littermates, were resistant to liver damage and death induced by injection of agonistic anti-Fas Ab Jo2. In contrast to casp8-/- T cells, DKO T cells proliferated normally in response to TCR ligation and displayed normal expansion and subsequent peripheral deletion in vivo when challenged with SEB. Interestingly, DKO animals display lymphoaccumulation of an aberrant B220+CD3+ population similar to that seen in gld or lpr mice. The survival of Casp8-/- RIPK3-/- mice and the normal proliferation of T cells from these animals point to an essential role for caspase-8 in limiting RIP3-dependent necrosis both during development and immune cell activation.
Chronic IFN-gamma expression alters the migration pattern of peripheral T cells (160.7)
Authors: Lin Fanching , Saleh Bahara , Hodge Deborah , Young Howard 
Published: January 2023

Abstract
We report here on preliminary data from an IFN-gamma(IFN-γ)ARE-deleted Balb/c mouse model developed in our laboratory.This mouse has a 162 nt deletion in the 3’untranslated region of the IFN-γ mRNA to eliminate an AU-rich sequence. The deletion has a positive impact on IFN-γ mRNA accumulation by dramatically increasing its half-life. In the absence of stimulatory factors,these mice express low level of IFN-γ constitutively. IFN-γ ARE-deletion results in a severe phenotype on Balb/c mice. The knockout (KO) and heterozygous (Het) mice are smaller in size than the littermate controls, have a rough coat and a low survival rate after weaning. Additionally, these mice have an enlarged thymus with an increased percentage of mature naïve T cells. The mature thymocytes from Het animals are able to leave the thymus and enter peripheral lymphoid tissues. However, the results from chimera experiments indicate that IFN-γ may impact the thymic environment, thus resulting in abnormal T cell development. In addition, we found the migration pattern of peripheral T cells from Het mice differs from that observed with cells from control animals, further suggesting an abnormality in the Het T cell physiology. Current experiments focus on analyzing changes in the thymic environment and T cell development in newborn mice. In addition, we are analyzing chemokine receptor expression to determine if changes in these receptors may be a mitigating factor in the T cell migration patterns.