bluuebunny/crossref_metadata_2025_split huggingface dataset:
Establishment of nonmyeloablative bone marrow chimerism by double negative Treg cells through inducing T cell clonal deletion and suppressing NK cell function (126.1)
Authors: Zhang Zhu-Xu , Su Ye , Huang Xuyan , Wang Shuang , Min Wei-Ping , Yin Ziqin , Jevnikar Anthony 
Published: January 2023

Abstract
In bone marrow transplantation, T cells and NK cells play important role for graft rejection. In addition, graft-versus-host-disease and establishment of stable chimeric states without complete marrow ablation remain as major obstacles post bone marrow transplantation. In this study, we aimed to establish mixed chimerism in a non-irradiation condition. Our data indicate that adoptive transfer of donor-derived TCRαβ+CD3+CD4-CD8-NK1.1-(double negative, DN) Treg cells prior to C57BL/6 to BALB/c bone marrow transplantation, in combination with cyclophosphamide but not cyclosporine, FK506, or rapamycin, established stable mixed chimerism that led to acceptance of C57BL/6 skin allografts and rejection of 3rd party C3H skin grafts. Adoptive transfer of CD4+ and CD8+ T cells, but not DN-Treg cells, induced graft-versus-host diseases in this regimen. The recipient T cell alloreacitve responsiveness was reduced in the DN-Treg cell-treated group and T cell receptor Vβ2, Vβ7 and Vβ8 clonal deletions were observed in both CD4+ and CD8+ T cells. Furthermore, DN-Treg cell treatment suppressed NK cell-mediated donor bone marrow rejection in perforin-dependent manner. Taken together, our results suggest that adoptive transfer of DN-Treg cells can control both adoptive and innate immunity and promote a stable mixed chimerism and donor-specific tolerance in the non-irradiation regimen.
ICOS promotes T cell activation and function beyond phosphoinositide 3-kinase signaling (126.11)
Authors: Yu Xue-Zhong , Semple Kenrick , Leconte Julien , Suh Woong-Kyung , Li Jun 
Published: January 2023

Abstract
ICOS delivers an important costimulation to promote T-cell activation and function. Using a knock-in mouse strain, termed ICOS-YF, in which the cytoplasmic tail of ICOS cannot activate phosphoinositide 3-kinase (PI3K), we have shown that ICOS-PI3K signaling axis is critical for generation of follicular helper T cells. Although ICOS can potentiate TCR-mediated calcium flux in a PI3K-independent manner, its biological significance is unclear. To address this question, we studied the function of ICOS-YF T cells in comparison with ICOS wild-type (WT) and knock-out (KO) T cells in MHC-mismatched bone marrow transplantation (BMT) models. We found that CD4 T cells from ICOS-YF and KO mice were similarly impaired in their capacity to induce acute graft-versus-host disease (GVHD). In contrast, the pathogenic capacity of CD8 T cells from ICOS-YF mice was comparable to that of WT cells, whereas ICOS KO CD8 T cells were less pathogenic. In vitro, we observed that in both CD4 and CD8 T cells ICOS could potentiate TCR-mediated calcium flux in a PI3K-independent manner. These results suggest that although both CD4 and CD8 T cells depend on ICOS costimulation, the downstream signaling pathways they utilize are distinct: CD4 T cells depend on ICOS-PI3K signaling whereas CD8 T cells are more dependent on PI3K-independent pathways, probably calcium signaling. Taken together, our study reveals a complexity in ICOS signaling mechanisms in GVHD.
Recovery of thymic epithelial cells and immune reconstitution after cord blood transplantation depends on angiogenesis and neovascularization (126.12)
Authors: Politikos Ioannis , Kim Haesook , Cutler Corey , McDonough Sean , Antin Joseph , Ballen Karen , Ritz Jerome , Boussiotis Vassiliki 
Published: January 2023

Abstract
Umbilical cord blood (UCB) is increasingly used as an alternative source for allogeneic hematopoietic stem cell transplantation (HSCT). HSCT compromises thymopoiesis due to injury of thymic epithelial cells (TEC). Human UCB is enriched in endothelial precursors, which promote neovascularization and thymopoiesis in SCID mice transplanted with human thymus. We examined whether markers of angiogenesis, ANG1 and VEGF, might correlate with recovery of TEC, which support T cell development and expansion by producing IL-7 and SCF (a kit ligand). We analyzed 27 patients pre and 1, 2, 3, 6 and 12 months after UCBT. Early after UCBT, IL-7 and SCF levels inversely correlated with TREC, CD4, CD8 and Treg numbers suggesting that uptake of these cytokines by IL-7R and Kit on T cell progenitors induced their expansion. IL-7 and SCF were also inversely correlated with T cell differentiation into pathogen-specific effectors as determined by CMV-specific, IFN-γ ELISpot. Conversely, VEGF and ANG1 positively correlated with CD4, CD4CD25 and CD4CD45RA numbers. Interestingly, VEGF and ANG1 displayed inverse correlation with SCF and more so with IL-7 at early time points after UCBT, which converted to strong positive correlation (r=0.78, p=0.0004) at 12 months, when T cell recovery had occurred. Our results are consistent with a model of TEC recovery supported by VEGF and ANG1, leading to production and uptake of SCF and IL-7 by T cell progenitors, resulting in their differentiation and expansion.
Prior pancreatic islet transplantation mitigated the subsequent skin graft rejection against HY disparity in C57BL/6 with pre-existing immunological memory (126.15)
Authors: Kim Yong-Hee , Lee Sun-Ho , Park Chung-Gyu 
Published: January 2023

Abstract
Several successful immunomodulatory strategies in specific pathogen-free rodents have failed in outbred larger animals by pre-existing memories. In our previous report, we showed that the islet transplantation across the minor histocompatibility of HY (male antigen) could induce immunological tolerance to male antigen in female C57BL/6 mice. In this study, we tested whether islet-transplantation-induced immunological tolerance could be achieved against the barrier of pre-existing immunological memory. Splenocytes isolated from male C57BL/6 were injected into syngeneic female mouse intraperitoneally. The induction of memory was confirmed by accelerated male skin rejection from 27 days (naïve control mouse) to 19 days. The islets isolated from male C57BL/6 mice were transplanted into renal subcapsule of the sensitized mice. 37.5% of sensitized recipients rejected male islets, whereas 100% of naïve recipients accepted the grafts. Remaining 62.5% of sensitized recipients that accepted male islets did not accept subsequent male skin graft. Although male skin graft failed to be accepted in the male islet-accepted recipients, the intensity of inflammatory response was significantly lower than that shown in presensitized mice without male islet graft. In conclusion, the islet-transplantation-induced immunological tolerance could not circumvent the barrier of immunological memory, however, it could mitigate the intensity of inflammatory rejection process.
Regulatory-B cell and regulatory-T cell collaborate to promote transplantation tolerance (126.2)
Authors: Lee Kang Mi , Kim James , Yeh Heidi , Stott Ryan , Soohoo Julie , Deng Shaoping , Markmann James 
Published: January 2023

Abstract
Backgrounds: The role of regulatory-B cells (Bregs) in transplant tolerance remains unclear. We found that the anti-CD45RB/anti-TIM1 induced tolerance depends on both Bregs and regulatory-T cells (Tregs). This finding offers a unique opportunity to explore the interplay of Bregs and Tregs in a transplant setting. Methods: Islet allograft were prepared from BALB/c donors and transplanted into B6. Recipients were treated with anti-CD45RB and anti-TIM1 antibodies for tolerance induction. Naive B cells or B cells isolated from mice that have been transplanted and treated in combination antibody therapy transferred into µMT-KO and assessed Treg expression by FACS. Anti-CD25 antibody, PC61, was used for Treg depletion. Anti-IL-10 or anti-TGF-β was used to neutralizing cytokine activity. Results: Treg numbers significantly increase following adoptive transfer of B cells purified from recipients bearing long-term surviving grafts (LTS). Moreover, adoptive transfer of LTS B cells suppressed allograft rejection upon transfer to untreated B cell-deficient secondary host, but this effect was not observed in Treg-deleted recipients. The absence of IL-10 by using anti-IL10 antibody did not affect on induction of long-term islet allograft by anti-CD45RB/anti-TIM1, however anti-TGF-β treatment abrogated the graft survival. Conclusions: B cell regulation induced by anti-CD45RB/anti-TIM1 is associated with Treg generation. This effect may be mediated by Treg promoting cytokines such as TGF-β.
Persistence in graft of Foxp3,TGFβ and IL-10 is hallmark of successful retransplantation of grafts after primary transplantation under cover of CD200 expression (126.21)
Authors: Gorczynski Reg , Chen Zhiqi , Yu Kai 
Published: January 2023

Abstract
We showed that overexpression of CD200, a molecule of the immunoglobulin supergene family, increases graft survival and suppresses inflammation/acquired immunity after binding to its receptors (CD200Rs). Skin/cardiac allograft survival is increased in transgenic mice over-expressing CD200 under control of a doxycycline-inducible promoter, (CD200tg), or following transplantation of CD200tg grafts to control mice, along with increased intra-graft expression of mRNAs implicated in Treg differentiation, including TGFβ, IL-10 and Foxp3. We have investigated whether C57BL/6 CD200tg grafts taken from control BALB/c mice at 14 days post transplantation can be successfully re-grafted into secondary control BALB/c mice, and whether such animals show perturbation of specific alloimmunity, and/or changes in graft gene expression in comparison to mice receiving control C57BL/6 grafts. Control grafts taken from mice at d14 and re-transplanted to control BALB/c mice were rejected rapidly, and with mice developing the expected CTL response in splenocytes detected by lysis of EL4 tumor targets in 4-hour 51Cr-release assays. In contrast, CD200tg grafts survived on secondary transfer to control BALB/c mice, with specific suppression of the cytotoxicitv response to EL4 targets. In these secondary recipients only, persistently high expression of Foxp3, IL-10 and TGFβ gene expression was seen. We conclude that expression of these latter genes is crucial for adoptive transfer of tolerance.
Role of MHC class II in Treg-mediated tolerance to allogeneic transplants (126.26)
Authors: LeGuern Christian , Martins Paulo , Germana Sharon 
Published: January 2023

Abstract
Transfer of donor MHC class II genes (MHCII) in the bone marrow of recipients of allografts induced tolerance spreading to all alloantigens. Tolerance was donor MHCII-specific, transferable only via host CD4 Tregs and selectively induced by MHCII gene therapy. Studies on the fate of transgenic MHCII molecules revealed that both the α and β chains provided peptides presented on the MHCII of bone marrow-derived antigen-presenting cells (APCs). Transgene-matched graft MHCII was critical for survival as donor MHCII-treated recipients promptly rejected MHCII-deficient transplants. Next, the role of thymic transgenic MHCII peptides on Treg development was studied in a model recapitulating MHCII peptide presentation: the IEα 52-68 peptide/IAb complexes recognized by the YA-e mAb and TEa transgenic TCR. With this model we showed that 1) the highly conserved IEα sequence was dominantly exposed on all dendritic cells located in the thymic medulla, the birthplace of Tregs, and 2) transfer of the IEα peptide into IEneg mice via bone marrow transplantation, converted host IEα-specific CD4 T cells into CD4+ CD25+ Foxp3+ Tregs. These data are consistent with a dominant role of MHCII peptides in shaping Treg TCR specificity and differentiation. They also propose a mechanism for MHCII-tolerance: MHCII-specific Tregs require activation by cognate MHCII peptide provided by MHCII-matched transplants prior to suppressing alloreactive T cells of different TCR specificity.
Rag1 and Rag2 Knockout rats to drive xenografts and oncology studies (126.30)
Authors: Xue Jeff , Keys Kelly , McCoy Aaron , Weinstein Edward 
Published: January 2023

Abstract
Rejection of grafted tissues by host immune systems is problematic during xenotransplantation. To help alleviate this and provide models amenable to the xenograft process, we created rats with genetic modifications of genes: Rag1 or Rag2. Rag1 and Rag2 play an important role in the rearrangement and recombination of genes of immunoglobulin and T-cell receptor molecules during the process of VDJ recombination. These rats were created via zinc finger nuclease (ZFN) targeted editing of Sprague-Dawley and F344 rat genomes. ZFN creates a double stranded break at the target locus. This lesion is repaired by the low fidelity repair mechanism non-homologous end-joining which commonly yields out of frame mutations in the gene, leading to non-functional proteins. Phenotypic characterization of these animals occurs when homozygosity is achieved. Rag1 and Rag2 protein ablation was demonstrated by western blot analysis. Flow cytometric analysis of peripheral blood cells in Rag1 KO animals at 4, 10 and 20 weeks demonstrated a significant decrease in mature T and B lymphocytes relative to age-matched wild type rats. Rag2 KO rats demonstrate a non-leaky depletion of mature T and B lymphocytes at 4 and 10 weeks. By employing the ZFN technique, we developed Rag1 and Rag2 KO rats with compromised immunology systems, which we believe will be helpful in the studies of xenografts, cancer, vaccine development, autoimmune, infectious and GVHD diseases.
Inhibition of the Heat Shock Protein receptor CD91 abrogate concomitant anti-tumor immunity (127.24)
Authors: Zhou Yu , Binder Robert 
Published: January 2023

Abstract
Most tumors are capable of generating immune response in the bearing host, a phenomenon described as concomitant immunity. Since tumors lack the typical (foreign) PAMPs that provide signal 2 for T cell priming, the mechanism of how concomitant immunity is established remains a mystery. We provide evidence here that, heat shock proteins (HSPs), when released into the tumor microenvironment, is the immunogen necessary for priming concomitant tumor specific immunity. These responses require CD91- the endocytic and signaling receptor for the immunogenic HSPs. Our study abrogates the interaction of tumor-derived HSP with CD91 in vivo by over-expression of a natural inhibitor of CD91. The data shows that: first, tumors expressing CD91 inhibitor grew with significantly faster kinetics than the non-inhibitor expressing counterparts in wild type mice, although this difference was non-existent in immunocompromised mice and the inherent proliferative rates of tumors with or without inhibitor were identical; second, inhibitor-expressing tumors, when used as an immunogen in tumor prophylaxis, were significantly less efficient in priming immune responses. We also examined the competition for CD91 between the inhibitor and tumor-derived HSP in vivo. In summary, our study demonstrates that the HSP-CD91 pathway is critical for establishment of concomitant immunity. We propose a novel mechanism of immune evasion for tumors expressing competing ligands for immune receptors such as CD91.
Immunogenetic Hypothesis for Cancer Survival (127.26)
Authors: Williams R.Michael 
Published: January 2023

Abstract
Immunosurveillance reflects an ongoing war between the immune system and the specific cancer. If the honeymoon of no detectable cancer lasts a lifetime, we say there was a cure by primary therapy. Adjuvant therapy alters the outcome statistically. Adjuvant trials are balanced for known relevant patient and tumor characteristics but NOT for the genetics of the host or the tumor. Small absolute differences seen in adjuvant trials likely reflect an imbalance in these important characteristics, and adjuvant drugs or radiation alter the immune response in addition to killing some tumor cells. In short, survival probably depends as much on the immune system as it does on the cancer treatment. We proved that survival with a histocompatible tumor was genetic and immunological. Single AA differences in H-2 can change survival. Adaptive immune responses are antigen or epitope specific and almost always involve class I and class II MHC genes. T-cells are involved and the immune response is regulated so that decreasing T-suppressor cells (Tregs) can result in increased survival (ipilumumab in melanoma). Immunization, even after the cancer and its antigens are in abundant supply, can prolong survival (Spiruleucel-T in prostate cancer). We propose that the clinical oncology community investigate the Immunogenetic Hypothesis for Cancer Survival, and predict that HLA genes and T-cell immunoregulation effects will participate importantly in how long a cancer patient will survive.
Evaluation of the serum level of soluble E -cadherin and cell surface expression of α5β1 integrin in patients with acute leukemia whether lymphocytic or myeloid ,in order to extrapolate their prognostic value (127.3)
Authors: Abdelrahman Zakia , Abo shamaa Lobna , Sadek Nadia , El sayed Amina , Mekway Nada 
Published: January 2023

Abstract
Background:E-cadherin is expressed on CD34+ stem cell,where it likely contributes to intracellular interactions during hematopoiesis. Also integrin such as α5β1 integrin represent an essential component of the bone marrow microenvironment that regulates cell survival .In leukemia, immature blasts are not retained within the marrow, suggesting a breakdown of adhesive mechanisms and cell migration. Objective: this study was done to evaluate serum soluble E -cadherin and cell surface expression of α5β1 integrin on PBMCs in patients with acute leukemia whether lymphocytic or myeloid Twenty patients (11 AML and 9 ALL)and ten age and sex matched normal individuals were studied. soluble E -cadherin were assayed using ELISA and cell surface expression of α5β1 integrin on PBMCs by indirect immunoflourescence . Results: Soluble E-cadherin was significantly lower in all leukemic and AML patients than control ( p=.0108,p=.0049 respectively).However no significant difference between AML and ALL. α5β1 integrin cell surface expression was significantly higher in both AML,ALL groups than control(p=.0001,0.0038 respectively). E-cadherin has negative significant correlation (r= -.573, p= .008) with bone marrow blast percentage after the induction of chemotherapy. Conclusion this suggests that a dual double way interaction between increased expression of α5β1 integrin and loss or decreased expression of E-cadherin that governs the process of tumour progression and metastasis.
Characterization of a novel mouse model for human B-cell lymphomas (127.31)
Authors: Bommireddy Ramireddy , Besselsen David , Rimsza Lisa 
Published: January 2023

Abstract
The risk of developing lymphoma is markedly increased after solid organ transplantation. Up to 20% of people who receive a solid organ transplant develop a lymphoma. Calcineurin (CN) A beta knockout (Cnab-/-) mice do not reject allogeneic tumors suggesting an immunosuppressive state, is also a unique model for reverse immunosurveillance. Currently, there is no mouse model for low grade lymphomas based on immune dysregulation. Since we observed a spontaneous activation of T cells, and B-cell lymphomas in CNAβ deficient mice, we hypothesize that CN is involved in T-cell regulation, the lack of regulation causes smoldering inflammation which in turn activates B cells and results in lymphomagenesis. The lymphomas in Cnab-/- mice are positive for B cell surface marker (CD21) but negative for T cell (TCRβ) markers and they are clonal in origin and exhibit malignant (destroying the normal architecture of adjoining area) phenotype. In addition, we do see spontaneous activation of T cells in Cnab-/- mice early in life and continue to accumulate, and B cells later when they are old with mild inflammatory lesions in the gut and liver in Helicobacter-free (clean) colony. Since inflammation is considered to be one of the hallmarks of cancer and also the root cause of cancer, this is a novel mouse model for understanding a slow growing, difficult to cure form of lymphomas.
Tumor-targeted T cells secreting TLR5 ligand effectively treat melanoma-bearing mice (127.34)
Authors: Geng Degui , Srivastava Ratika , Li Chaoyang , Davila Eduardo 
Published: January 2023

Abstract
T-cell-based therapies can kill tumor cells expressing immunodominant tumor antigens (TAg). However, this process allows the escape and metastases of tumor cells expressing low levels of TAg or expressing subdominant TAgs. Our preliminary data and published reports indicate that stimulating TLR2 or TLR5 directly on tumor-reactive T-cells reduces the activation threshold to poorly immunogenic TAgs. Genetically engineering T-cells expressing the MART-1 TCR and producing the TLR5 ligand flagellin (Fg) will generate potent and long-lived antitumor activity by costimulating T-cells and antigen-presenting cells at the tumor site. Human T-cells engineered to express the MART-1 TCR-Fg showed significantly greater proliferation, survival, and cytolytic activity than MART-1 T-cells. TLR5-stimulated T-cells responded to 10-to-50 times lower levels of TCR stimulation. TLR5 stimulation also augmented the production of various cytokines including IL-2, GM-CSF, IFN and TNF. Unexpectedly, we found that TLR5 stimulation on melanoma tumors induced the production of cytokines and chemokines which further promoted the recruitment of CD4 and CD8 T-cells, and NK cells. We are currently examining the therapeutic efficacy of using TCR-Fg T cells to treat NOD-scid IL2Rgammanull (NSG) mice bearing melanoma tumors.
Thymosin beta 4 is involved in interleukin 18 enhanced interferon gamma secretion in human NK cell line, NK-92MI (127.8)
Authors: Lee Ha-Reum , Cho Daeho 
Published: January 2023

Abstract
Thymosin beta 4 (Tβ4), which is the major G-actin sequestering protein, has been shown to have ubiquitous distribution and multiple biological activities. However, what regulates Tβ4 expression and what its function is on natural killer (NK) cells are still unknown. In this study, recombinant human IL-18 (rhIL-18) enhanced the endogenous level of Tβ4 through p38 MAPK and JNK signaling pathway in the human NK cell line, NK-92MI. IFN-γ production is stimulated by over-expression of endogeneous Tβ4. Furthermore, pretreatment with an inhibitor for Tβ4 decreased IL-18-enhanced IFN-γ secretion, and transfection with Tβ4-specific short hairpin RNA resulted in reduction of IFN-γ production in primary NK cells as well as in the human NK cell line. This suggests that Tβ4 is regulated by IL-18 and is involved in IL-18-enhanced IFN-γ secretion in NK cells.