bluuebunny/crossref_metadata_2025_split huggingface dataset:
Specific blockade of protein kinase C (PKC)-theta with a small molecule antagonist attenuates lung inflammation in a mouse model of established allergic airway disease (140.21)
Authors: DeClercq Charlene , Demers Delia , Bree Andrea , Fish Susan , Wei Shui-Quing , Brennan Aggie , Kasaian Marion , Chen Joan , Wang Daniel , Boschelli Diane , Chaudhary Divya , Williams Cara 
Published: January 2023

Abstract
PKCθ is primarily expressed in T cells and is recruited to the immunological synapse upon T cell receptor signaling. PKCθ deficient mice display a diminished pulmonary inflammatory response following allergen challenge. We investigated the allergic airway response in mice when PKCθ is specifically blocked with a small molecule inhibitor (compound A). Mice were sensitized with ovalbumin (OVA) in alum on days 0 and 14, then challenged with OVA on days 26, 27, and 28 (primary challenges). Four weeks later mice were aerosolized once with OVA (rechallenge). Airway inflammation was monitored after both primary challenges and rechallenge. Compound A was dosed 30 mg/kg intraperitoneally twice daily in 3 dosing regimens: for 4 days concurrent with and after the primary challenges only; for 3 days starting before the OVA rechallenge only; and both of the above regimens. Administration of compound A significantly inhibited the influx of eosinophils, lymphocytes and neutrophils into the airways. Ex vivo antigen induced Th2 cytokine production by splenocytes was also reduced in cells obtained from mice treated with Compound A in either the primary challenge phase, or both primary and rechallenge phases, but not the rechallenge phase alone. We conclude that blocking T cell activation with a PKCθ inhibitor may attenuate airway inflammation in established allergic disease and may be beneficial in treating chronic allergic asthma.
AIMP1 as a negative regulator of airway inflammation via suppression of Th2 responses (140.3)
Authors: Hong Hye Jin , Kim Myun Soo , Kim Eugene , Kim Tae Sung 
Published: January 2023

Abstract
AIMP1 (aminoacyl-tRNA synthetase-interacting multifunctional protein1)/p43 is found to related to Lupus-like autoimmune disease. Previously we demonstrated that AIMP1/p43 activated dendritic cells and macrophages, leading to the induction of Th1 responses. In this study, we investigated a role of AIMP1/p43 in the pathogenesis of lung airway inflammation by characterizing a phenotype of AIMP1 knockout mouse. Asthma-like pathological changes were observed in the lung and the sensitivity to methacholine was also severely increased in AIMP1 knockout mouse. Histologic studies showed that AIMP1 knockout mice had even thicker alveolar walls than the wild type mice. The expression level of gob-5, a gene related to mucus production, was highly enhanced in AIMP1 knockout mice. Additionally GATA-3, a transcription factor inducing Th2 development, was dramatically increased in the lung homogenates of AIMP1 knockout mice. Furthermore, in the AIMP1 knockout mice, the total IgE level was increased, along with the levels of IgG1 and IgG2a. The levels of IL-4 production were significantly higher in lymphocytes of AIMP1 knockout mice after OVA restimulation, compared with those in the wild type mice. Taken together, these results indicate that the Th2 responses were strongly increased in AIMP1 knockout mice and suggest that AIMP1 functions as a negative regulator of the Th2-response in asthma-like airway inflammation.
Divergent Roles for Airway Epithelial MMP7 and Retinoic Acid in Experimental Asthma (140.5)
Authors: Goswami Sangeeta , Angkasekwinai Pornpimon , Shan Ming , Greenlee Kendra J , Barranco Wade T , Polikepahad Sumanth , Seryshev Alexander , Sur Sanjiv , Woodruff Prescott , Dong Chen , Corry David B , Kheradmand Farrah 
Published: January 2023

Abstract
The innate immune response of airway epithelial cells to aeroallergen likely initiates the development of T cell responses that are central to allergic inflammation. Although proteolytically active allergens induce the expression of interleukin (IL)-25/IL-17E, we show that epithelial matrix metalloproteinase 7 (MMP7) is expressed in human asthma, and is required for maximal activity of this cytokine in promoting T helper type 2 cell differentiation. Allergen-challenged MMP7-/- mice showed reduced airway hyperreactivity, allergic inflammatory cytokine, and increased expression of retinal dehydrogenase (RALDH)-1. Inhibition of RALDH-1 restored the asthma phenotype in MMP7-/- mice and inhibited lung T regulatory cell responses while exogenous administration of retinoic acid attenuated the asthma phenotype. Thus, MMP7 coordinates allergic lung inflammation by activating IL-25 while simultaneously inhibiting retinoid-dependent T regulatory cell development.
Disparate immune responses to Mi-HAg occur in vitro and in vivo (141.1)
Authors: Xu Hong , Bozulic Larry D , Zhu Ziqiang , Huang Yiming , Hussain Lala R , Ildstad Suzanne T 
Published: January 2023

Abstract
Historically, insights into the mechanism of alloresponses to minor histocompatibility antigen (Mi-HAg) disparities have focused on T cell activity and used in vitro assays. It was believed that prior in vivo immunization was required for T cell responses to Mi-HAg to occur in vitro. Here, we investigated the generation of adaptive immune responses to Mi-HiAg using an in vivo model. We found that AKR (H-2k) mice rejected skin grafts from MHC-matched, Mi-HAg-disparate B10.BR (H-2k) mice and generated significantly high levels of anti-donor Ab, suggesting that skin graft rejection was associated with the initiation of strong adaptive humoral responses. As B cell activation requires signaling from activated T cells, in vitro and in vivo MLR assays were performed to examine the immune response to Mi-HAg. AKR cells exhibited no reactivity to Mi-HAg disparate stimulators in in vitro MLR assays. In contrast, T cells from AKR mice exhibited strong reactivity in in vivo MLR assays to Mi-HAg in B10.BR mice. The reason for this disparity in response to Mi-HAg in vitro vs. in vivo MLR is not clear. The in vitro culture environment may lack some factors that were present in vivo and that are critical for generation of T cell immune responses to Mi-HAg. T cells were indeed activated in response to Mi-HAg, as CD44highCD62L-/low effector/memory T cells were generated.
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A novel method to assess the level of immunosupression in transplant patients using Amnis ImageStream technology (141.24)
Authors: Pankewycz Oleh , Minderman Hans , Wallace Paul , Feng Lin , Laftavi Mark R. 
Published: January 2023

Abstract
Contemporary immunosuppressive protocols rely on drug dosing guidelines gleaned from large clinical trials, but the effect of this therapy in a transplant recipient remains difficult to predict. To assess global immunosuppression in individual patients, we measured the degree to which NFkB translocation in peripheral T cells was impaired by conventional immunosuppressive therapy. NFkB is a ubiquitously expressed transcription factor normally sequestered in the cytoplasm. Upon activation, NFkB inhibitors are degraded, releasing NFkB to translocate to the nucleus. The abundance of NFkB in the nucleus vs. the cytosol reflects the activation state of a particular cell type. Peripheral blood cells were isolated from 17 subjects, stimulated in culture for 24 hrs with PMA and ionomycin and stained for T cell surface markers and the major intracellular isoform of NFkB, RelA or p65. Using Amnis Imagestream technology, we compared nuclear NFkB in resting and activated CD3, CD4 and CD8 T cell subsets. Compared to normal controls (n=8), transplant recipients on tacrolimus, MPA and prednisone showed reduced translocation of NFkB to the nucleus. Patients with stable allograft function (n=5) had a 36-75% reduction in nuclear NFkB translocation in all 3 T cell subsets. In contrast, patients with acute rejection (n=2) had an increase in ability to translocate NFkB in CD3 and CD8 cells and only a 9% suppression in CD4 cells. Patients with viral infections (n=2) had severely compromised NFkB translocation with 100% suppression in CD3 and CD4 subsets and 92% suppression in CD8 T cells. NFkB inhibition strongly correlates with overall immunosuppression and clinical outcomes. The novel Amnis Imagestream assay may be a useful guide to individualizing immune therapy or directing minimization strategies. (unfunded)
T cells reconstituting the periphery in thymoglobulin treated kidney transplant recipients are preferentially donor-senescent when compared to basiliximab treated patients (141.25)
Authors: Keslar Karen , Cherkassky Leonid , Valujskikh Anna , Shoskes Daniel , Fairchild Robert , Poggio Emilio 
Published: January 2023

Abstract
Induction with anti-thymocyte globulin (ATG) results in better 1 yr renal transplant outcomes than basiliximab in immunologically sensitized transplant patients. This observation contrasts with the potential homeostatic proliferation of surviving donor-reactive T cells in the post-ATG lymphopenic environment. Donor-specific reactivity was measured longitudinally in 8 (n=4 for basiliximab, n=4 for ATG) kidney transplant recipients who completed a 6 mo follow-up. Peripheral blood cells at pretransplant and monthly posttransplant were tested for donor-reactivity by IFN-γ ELISPOT assays. Patients induced with ATG or basiliximab had similar decreases in their absolute lymphocyte counts (and total T cells) at 1 wk post-tx. While these counts returned to near baseline by 3 mo in basiliximab treated patients, counts in ATG-treated patients were 61% of baseline at 6 mo. Mean post-tx donor-specific alloreactive T cell frequencies at 6 mo follow-up were similar to pretransplant levels for basiliximab patients but were &lt;20% of pretransplant levels for ATG-treated patients. Furthermore, peripheral T cells in ATG-treated patients are not preferentially reconstituted with RO+ memory cells as might be expected in an environment promoting homeostatic proliferation. These data suggest that reconstitution of the periphery in ATG-treated patients is not preferentially donor-reactive.
Host-derived plasmacytoid dendritic cells prolong cardiac allograft survival and regulate alloreactive T cell responses via B7 homolog-1 (141.28)
Authors: Matta Benjamin M , Sumpter Tina L , Colvin Bridget L , Thomson Angus W 
Published: January 2023

Abstract
This study evaluated the contribution of the inhibitory molecule B7 homolog-1 (B7-H1) to the regulation of alloimmune responses by pDC. mPDCA-1+ splenic pDC from wild type (WT) or B7-H1 knockout (KO) mice were pulsed with donor antigen (Ag). Cell phenotype was analyzed for Ag-presenting and co-regulatory molecules and T cell allostimulatory capacity was assessed in mixed leukocyte reaction (MLR). WT or KO pDC pulsed with donor Ag were injected into syngeneic mice and CD4+ T cells were purified 7 days later for secondary MLR. T cell proliferation was assessed by 3H-thymidine incorporation and cytokines quantified by ELISA. BALB/c hearts were transplanted into untreated C57BL/6 controls or into mice receiving syngeneic pDC pulsed with donor Ag 7 days before grafting. pDC expressed low levels of CD40, CD80, CD86, MHC I/II, and B7-H1 and showed poor CD4+ T cell stimulatory capacity compared to conventional myeloid DC. pDC from KO mice exhibited greater T cell stimulatory capacity compared to WT controls. T cells from mice given immature, donor Ag-pulsed KO pDC showed greater proliferation and secreted more IFN-γ in secondary MLR compared to WT controls. Recipient-derived, immature WT pDC pulsed with donor Ag prolong allograft survival. These results reveal that B7-H1 on pDC contributes to regulation of alloreactive T cell responses. This work was funded by the transplantation immunology training grant NIAID T32 AI 074490.
Prolonged Exposure of Myeloid Dendritic Cells To Rapamycin enhances LPS-induced IL-12p70 production, but does not promote Th1 responses (141.29)
Authors: Turnquist Heth Roderick , Macedo Camila , Cardinal Jon S. , Geller David A , Metes Diana , Thomson Angus W. 
Published: January 2023

Abstract
Rapamycin (RAPA) is a tolerance- and Treg-sparing immunosuppressant. Prolonged exposure of myeloid dendritic cells (mDC) to RAPA confers resistance to phenotypic and functional maturation following exposure to inflammatory stimuli. Infusion of RAPA-conditioned mDC (RAPA-DC) promotes transplant survival and inhibits GVHD. However, RAPA is also reported to have pro-inflammatory properties, mediated by increased DC IL-12. Our aim was to define how DC cytokine production and CD4+ T helper (Th) cell polarization is modified by prolonged RAPA exposure.
Methods: Bone-marrow derived murine mDC were generated in 7 day (d) cultures, with RAPA added d2-d7. Following purification, RAPA-DC or control mDC (CTR-DC) remained unstimulated or were exposed to LPS. Co-stimulatory molecule expression and intracellular IL-12p40 was determined by flow cytometry. Th polarization was assessed using these DC to stimulate alloreactive CD4+ T cells, followed by analysis of T cell proliferation, apoptosis, and intracellular cytokines. Western blot was used to assess total and phosphorylated signaling molecules.
Results: RAPA-DC displayed reduced costimulatory molecules and allostimulatory capacity before and after exposure to LPS. IL-12p40 and p70 were reduced in unstimulated RAPA-DC compared to CTR-DC. LPS induced a significant increase in IL-12p70 production by RAPA-DC, due to failed inhibition of glycogen synthase kinase-3. However, RAPA-DC stimulated with LPS remained poor allostimulators that induced apoptosis in alloreactive CD4+ T cells and failed to promote Th1 responses.
Conclusions: Prolonged exposure of mDC to RAPA, although promoting LPS-induced IL-12p70 production, inhibits T cell stimulatory capacity and Th1 polarization.
Treg versus TH17 lymphocyte lineages are cross-regulated by LIF versus IL-6 (141.34)
Authors: Metcalfe Su M , Gao Wenda , Thompson Lorraine , Zhou Qiang , Putheti Prabhakar , Fahmy Tarek , Strom Terry 
Published: January 2023

Abstract
In an in vivo/ex vivo murine heart allograft model of allo-tolerance versus allo-rejection, that retains the complexity of the in vivo environment, novel molecular links to Foxp3 and tolerance have been revealed. These include leukaemia inhibitory factor (LIF), a member of the IL-6 cytokine family. However, IL-6 itself was linked to allo-rejection. This raises several questions: (i) does LIF play an active role in immune tolerance? and (ii) does the differential between LIF and IL-6 occur within the T cell itself? Using CD4+Foxp3+ Treg (nTreg) from Foxp3-GFP knockin mice we tested the effect of exogenous LIF, versus exogenous IL-6, on the respective lineage-specific transcription factors, Foxp3 (Treg) and RORγt (TH17). LIF supported Foxp3 transcription but not RORγt: in contrast IL-6 profoundly repressed Foxp3 and induced RORγt. For the LIF-specific receptor subunit, gp190, the response to LIF versus IL-6 was the polar opposite: gp190 transcripts were increased by LIF but strongly inhibited by IL-6, a result that indicated a potential mechanism for cross-regulation, wherein IL-6 would reduce LIF signalling by repressing synthesis of the LIF receptor. Overall, we suggest LIF is a nodal regulator of the Treg lineage, acting to oppose IL-6, the nodal regulator of the TH17 lineage. Our discoveries identify novel drug targets and we provide proof of concept using targeted LIF-nanotherapy to enhance donor-specific Treg cells in vivo. Funding BHF, NIHR, NIH, JDRF.
Manipulating The Liver to Induce Systemic Tolerance to Heart Allografts Through CD4+CD25+ Regulatory T cells and NKT Cells (141.39)
Authors: Li Wei , Carper Katie , Wang Weigang , Meng Zihui , Reyes Jorge D , Perkins James D 
Published: January 2023

Abstract
The liver has been demonstrated in many models to favor the induction of peripheral tolerance. The mechanisms mediating this phenomenon remain undefined. In this study, we delivered the C3H spleen cells (SCs) into the B6 liver through the portal vein (p.v.) in contrast to tail vein (i.v.) injection to characterize the liver leukocytes by flow cytometry and their function by MLR, ELISPOT, and immunohistochemistry assays. Heterotopic heart transplantation was performed from C3H donors to B6 recipients at day 7 post-treatment. The TCRαβ +NK1.1+ cells (NKT) were significantly increased in the liver, and CD4+CD25+Foxp3+ regulatory T cells (Treg) were markedly increased in the both liver and spleen after p.v. but not i.v. injection of C3H SCs. The proliferative activity of T cells from the p.v. treated mice was significantly suppressed and IL-2 was decreased, IL-4 and IL-10 were increased in vitro, suggesting that the role of the Treg may contribute to these responses. Further, significantly increased IL-4, IL-10, and IFN-γ production was detected from liver nonparenchymal cells under αGalCer stimulation, suggesting the role of liver NKT cells. Heart allograft survival was significantly prolonged in the C3H SCs p.v. treated recipients than in the i.v. treated recipients. Therefore, the liver can serve as a site of systemic tolerance induction. Both NKT cells and CD4+CD25+Foxp3+ Treg appear to be involved in the regulation of peripheral immune responses after antigen immunization of the liver.
In Vivo Introduction of an Extrachromosomal Plasmid Expressing Long-term Human Alpha-1-antitrypsin protects Islet Allografts (141.48)
Authors: Moser Hadas , Shahaf Galit , Bellacen Keren , Mizrahi Mark , Lewis Eli C 
Published: January 2023

Abstract
Alpha-1-antitrypsin (AAT) exerts anti-inflammatory and tolerogenic activities during islet allograft transplantation in diabetes mouse models. Although a serine protease inhibitor, evidence suggests that AAT possesses activities that are independent of protease inhibition. The previously reported plasmid, pEF-hAAT, contains the genomic sequence of human AAT (hAAT) and sustains circulating levels after single hydrodynamic tail-vein injection (HDI) in mice. To asses whether expression of hAAT by pEF-hAAT protects islets from acute rejection, pEF-hAAT (100 μg, n = 6) or PBS (n = 4, control) were hydrodynamically introduced to mice. Liver expression and circulating hAAT levels were determined. Supernatant of transfected Hepa1c cells exhibited anti-elastase activity. Sixteen days post HDI mice were injected streptozotocin (225 mg/kg) and grafted with 450 allogenic islets. Circulating hAAT levels (1-350 μg/ml) persisted for over 30 days. Normoglycemia was achieved and islets were accepted in 6 out of 6 recipients that expressed hAAT. All 4 control animals exhibited acute islet allograft rejection by day 12. We conclude that plasmid-derived long-term expression of as low as 1 μg/ml hAAT exhibits islet-allograft protection. Using site-directed mutagenesis of pEF-hAAT we will examine whether activities that are independent of protease inhibition partake in the protection of islet allografts.
COMPENSATION FOR PKC-theta DEFICIENCY IN ALLOIMMUNE RESPONSES (141.6)
Authors: Yin Deng Ping , Ma Lian Li , Gao Xiudan , Williams Phillip , Chari Ravi 
Published: January 2023

Abstract
We have recently shown that PKC?-/- T cell-reconstituted Rag2-/- mice failed to reject fully MHC mismatched (H-2d) cardiac allografts. Sub-optimal blockade of costimulation, insufficient to prolong survival of cardiac allografts in wild-type (WT) mice, induced donor specific tolerance in PKC?-/- mice. The current results showed that cardiac allografts were rejected in PKC?-/- T cell-reconstituted Rag2-/- mice by an adoptive transfer of PKC?-/- B cells. Cardiac allografts were accepted in PKC? and B cell double knockout mice (PKC?-/-/µ MT) and PKC? and CD28 double knockout (PKC?-/-CD28-/-) mice, whereas acute rejection occurred in either CD28 or B cell single knockout mice. Bioluminescence imaging and flow cytometry analyses showed reduced NF-(B activation in the cardiac allografts, decreased frequency of Th1, Th17, and Treg subsets in the spleen cells, and diminished the percentage of IE-reactive V-beta11 subset in the peripheral blood lympocytes in PKC?-/-, and PKC?-/-CD28-/- mice. A single TLR engagement with CpG ODNs prevented tolerance induction in both PKC?-/-/µ MT and PKC?-/-CD28-/- mice, associated with the elevation of NF-(B activation and specific T cell subsets. We conclude that B cells have the ability to compensate for the deficiency of PKC?, which is CD28 dependent. CpG ODNs restore allograft rejection bypassing PKC?, CD28, and B cell signaling pathways.
Ag-specific humoral immune responses: effects of dialyzable spleen extracts from immunized mice (33.12)
Authors: Valderrabano-Ortiz Estela , Perez-Tapia Sonia Mayra , Calderon-Amador Juana , Estrada-Parra Sergio , Flores-Romo Leopoldo , Estrada-Garcia Iris 
Published: January 2023

Abstract
Murine dialyzable spleen extracts (DSE) are similar to Transfer Factors in that they can transfer cell mediated immunity (CMI) from an immunocompetent organism into a naïve one in an antigen-specific manner. Lymphoid organs from rats treated with transfer factor have shown histological changes and increased lymphoblasts and plasmocytes. In this work we investigated if treating mice with DSE obtained from OVA immunized mice (DSEova) could modify the humoral immune response against OVA in naïve mice. Methods. DSEova preparation: splenic cell suspensions from OVA immunized BALB/c mice were disrupted by freeze-thaw cycles, extracts were then dialyzed with a membrane pore of 10 kDa. BALB/c mice were treated with 20μg of DSEova alone. Serum samples were obtained between days 0-36, and IgM and IgG anti-OVA were assayed by ELISA. Mice were sacrified on days 9, 15 and 23 post-immunization, and germinal centers (GC) in lymph nodes were identified by immunohistochemistry using PNA and THY-1. Results. Compared to control saline-treated animals, DSEova treated mice produced IgM and IgG anti-OVA. Compared to saline-treated mice, DSEova-mice showed increased numbers of GC, but of smaller size than OVA-treated mice. Conclusion. In this in-vivo model, compared to control-mice, DSEova would appear to stimulate the Ag-specific antibody responses and the appearance of GC.
CD4+ T cells during priming help naive CD8 +T cell activation (33.19)
Authors: Chen Shu-Ching , Liao Chi-Chia , Lai Yo-Ping 
Published: January 2023

Abstract
Upon encountering their antigens, naïve T cells are activated and driven to clonal expansion and differentiation into armed effector cells. According to current understanding, induction of an optimal CD4+ T-cell response requires both antigen-specific and costimulatory signals. While, stimulating naïve CD8+ T cells with specific antigens and costimulatory signal is not sufficient to induce optimal clonal expansion and effector functions. In this study, an in vitro approach was utilized to dissect the cellular and molecular requirement for CD8+ T-cell priming and differentiation. Naïve CD62LhiCD44lo CD8+ T cells were sorted and stimulated by anti-CD3 plus anti-CD28 antibodies (anti-CD3/CD28). Firstly, mitomycin C-treated splenocytes did not aid CD8+ T-cell activation. Secondly, supernatant from activated CD8+ T cells-depleted splenocytes may contain certain mediator(s) that effectively helps CD8+ T-cell activation. Furthermore, we demonstrated that the certain mediator was provided by activated CD4+ T cells and acted at priming to promote clonal expansion of naïve CD8+ T cells. These new findings advance our understanding of the regulation of CD8+ T cell activation and differentiation.