bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of amyloid precursor-like protein 2 in Ewing’s sarcoma resistance to immunity (48.3)
Authors: Peters Haley , Wang Xiaojian , Nordgren Tara , Gilling Christine , Joshi Shantaram , Yan Ying , Solheim Joyce 
Published: January 2023

Abstract
Metastasized Ewing’s sarcoma, a childhood cancer of the bone, has a dismal 20-30% 5-year survival rate, despite multi-modal therapy. Ewing’s sarcoma cells have been shown to escape elimination by the immune system. We have found that several Ewing’s sarcoma cell lines express substantial levels of amyloid precursor-like protein 2 (APLP2). We sought to characterize cellular changes associated with Ewing’s sarcoma cells that avoid immune recognition, with emphasis on the function of APLP2 in this process. Ewing’s sarcoma cells that survived exposure to lymphokine-activated killer (LAK) cells exhibited higher levels of APLP2 expression, suggesting that elevated APLP2 in these cells may contribute to resistance to apoptotic cell death mediated by immune effectors. Consistent with this theory, we demonstrated that LAK-challenged cells have a reduced tendency to arrest in G2/M and to undergo apoptosis following exposure to gamma-irradiation. Transient transfection of a Ewing’s sarcoma cell line with APLP2 reduced irradiation-mediated G2/M arrest and apoptosis. IFN-gamma (a cytokine produced by LAK, NK and T cells) was found to up-regulate APLP2 production in a Ewing’s sarcoma cell line in vitro. This effect of IFN-gamma may contribute to the known ability of IFN-gamma to protect tumor cells from LAK-mediated lysis. Overall, our findings from these studies increase our understanding of mechanisms regulating immune recognition of Ewing’s sarcoma cells.
T cells expanded in presence of IL-15 exhibit elevated cell surface thiols, intracellular glutathione and increased long term survival (48.34)
Authors: Kaur Navtej , Naga Osama , Norell Hakan , Al-khami Amir , Chakraborty Nitya , Johnson Christina Voelkel , Mukherji Bijay , Mehrotra Shikhar 
Published: January 2023

Abstract
Persistence of effector cytotoxic T lymphocytes (CTLs) during an immunological response is critical for successfully controlling a viral infection or tumor growth. Various cytokines are known to play an important part in regulating the immune response. IL-2 and IL-15 possess similar properties, including the ability to induce T cell proliferation. The IL-2 family of cytokines that includes IL-2 and IL-15 are known to function as growth and survival factors for antigen-experienced T cells. Whereas long term IL-2 exposure has been shown to promote apoptosis and limit CD8+ memory T cell survival and proliferation, it is widely believed that IL-15 can inhibit apoptosis and maintains a memory CD8+ T-cell population. Our data show that human T cells and CTLs specific for melanoma reactive antigen Mart-127-35 and influenza matrix protein MP58-66 expanded in vitro in the presence of IL-15 had higher expression of CD56 and increased survival due to higher levels of anti-apoptotic proteins bcl-2, bcl-xl, cell surface thiols, and intracellular glutathione. Our study suggests that increased longevity of CTL cultured in the presence of IL-15 might depend on increased expression of antioxidant proteins and a subset of a highly expressed CD56hi population. Efforts to modulate the redox pathway may increase the T-cell survival without compromising cell functionality and confer therapeutic benefit to T-cell-based immunotherapy protocols.
IFNγ-inducible chemokines involved in lymphocyte trafficking and tumor regression are induced by immunotherapeutic Listeria monocytogenes (48.39)
Authors: Guirnalda Patrick , Wood Laurence , Pan Zhen-Kun , Crespo Joel , Paterson Yvonne 
Published: January 2023

Abstract
Using a transplantable HPV immortalized tumor model, TC.1, and a Listeria monocytogenes based immunotherapeutic that expresses HPV-E7, we have examined the requirements for T cell homing and infiltration following immunotherapy. Pro-inflammatory cytokines are produced in response to vaccination with Listeria based vaccines and underlie mechanisms of vaccine efficacy. Lymphocyte migration and infiltration into tumors and vaccine induced tumor regression are dependent upon a functional IFNγ receptor on the tumor cells. We hypothesized that vaccine induced and IFNγ dependent tumor regression relies on the presence of tumor derived, and IFNγ regulated chemokines. We noted the upregulation of CXCR3, CCR4 and CCR5 ligands at the mRNA and protein level in IFNγ and TNFα stimulated TC.1 tumor cells compared to unstimulated TC.1 cells. Vaccination with Listeria vaccines induced (i) the upregulation of tumor CCL4, CCL5 and CXCL9 and (ii) tumor infiltration of T cells bearing their cognate receptors. Tumor infiltrating lymphocytes consisted of both tumor antigen specific and Listeria antigen specific T cells. Chemokine receptor expression patterns varied among tumor infiltrating T cell subsets. IFNγ and TNFα stimulated CXCR3 ligand upregulation is a common feature among a variety of tumor cell lines and across a variety of tissues. Tumor cells are a source of effector T cell chemoattractants under vaccine induced pro-inflammatory conditions.
Expression of CXCL9/Mig in the tumor microenvironment is critical for recruitment of CD8 T cells and suppression of cutaneous tumor growth (48.4)
Authors: Gorbachev Anton , Petro Marianne 
Published: January 2023

Abstract
The role of T cell chemoattractants CXCL9/Mig and CXCL10/IP-10 in the suppression of tumor growth is not entirely clear. Our recent studies demonstrated the presence of tumor cells deficient in the expression of CXCL9/Mig in cutaneous murine fibrosarcomas. These Mig-deficient tumor variants were more tumorigenic than parental Mig-expressing tumor cells as indicated by accelerated growth kinetics of tumors induced by these tumor cells. Mig-deficient and Mig-expressing tumor variants were used as a tool to study the contribution of tumor-derived Mig to the intra-tumor recruitment of T cells. Using adoptive transfer of primed T cells into mice bearing either Mig-expressing or Mig-deficient tumors we demonstrated that CXCR3-expressing CD8 T cells were recruited into tumors producing Mig, but were marginal to undetectable in Mig-deficient tumors. The ability of Mig-deficient tumors to express IP-10 did not substitute for the expression of Mig to recruit CD8 T cells. Furthermore, knocking down expression of Mig (but not IP-10) by specific shRNAs resulted in increased growth of Mig-producing tumor variants in immunocompetent mice. Overall, the results suggested that Mig expression in the tumor microenvironment is non-redundant for the recruitment of CD8 T cells and for the suppression of tumor growth in the skin. The appearance of Mig-deficient tumor variants in growing tumors may indicate loss of Mig expression as a mechanism of tumor escape from host’s T cell-mediated immunity.
Increased Myeloid Derived Suppressor Cell recruitment and skewed TH2 response during influenza A virus infection in the absence of TLR7. (49.21)
Authors: Jeisy Scott Victoria , Bowzard J. , Davis William , Gangappa Shivaprakash , Katz Jacqueline , Sambhara Suryaprakash 
Published: January 2023

Abstract
Influenza A virus (IAV) is recognized by the innate immune system through RNA sensing pattern recognition receptors. One such receptor, Toll like receptor 7 (TLR7), and its downstream adaptor MyD88, have been shown to play an important role in the adaptive immune response to IAV infection. We investigated the specific contribution of TLR7 on the innate immune response to IAV. C57/B6 (WT) and TLR7-gene knock-out (TLR7-/-) mice were inoculated intranasally with a sub-lethal dose of Influenza A/PR/8/34 virus (PR8). Lungs were collected on days 3, 5, and 7post-infection to characterize cellular infiltrates and determine viral titers. TLR7-/- mice displayed increased morbidity following infection, despite having similar lung viral titers to WT mice. By day 7 post-infection, TLR7-/- mice had increased numbers of MDSC, neutrophil, and dendritic cell recruited to the lungs. Furthermore, on day 10 post-infection, the CD4+ T-cells of TLR7-/- mice expressed significantly higher levels of IL-4 compared with those of WT mice. To investigate functional differences between WT and TLR7-/- MDSCs, we purified MDSCs from day 7 infected mice and co-cultured them with OT-2 T-cells in the presence of ovalbumin peptide. On day 3, we observed increased numbers of IL-10 producing OT-2 T-cells when cultured with TLR7-/- MDSCs. Taken together, we conclude that TLR7 plays an important role in modulating the magnitude and quality of MDSCs and TH1/TH2 polarization during IAV infection.
Interferon gamma limits the magnitude of CD8 T cell responses in the lung during influenza infection (49.24)
Authors: Prabhu P Nayana , Ho Adrian , Kemeny David 
Published: January 2023

Abstract
In response to a viral challenge, antigen specific CD8 T cells proliferate and gain effector functions while simultaneously acquiring the capacity to secrete cytokines such as interferon gamma (IFN-γ). IFN-γ is thought to play a role in the homeostatic control of antigen specific CD8 T cell numbers although the mechanism by which this occurs is still unclear. Infection of mice deficient for IFN-γ (IFN-γ-/-) with PR/8 influenza was associated with increased antigen-specific CD8 T cell counts in the lung and the BAL fluid when compared to wildtype (WT) C57BL/6 mice. Using in vitro experiments, we show that there is no difference in the rates of proliferation of the WT and IFN-γ-/- CD8 T cells. However upon stimulation, we observed increased cell death in WT CD8 T cells compared to their IFN-γ-/- counterparts. This effect could be reversed upon addition of IFN-γ to the IFN-γ-/- CD8 T cells in a dose-dependent manner. We also observed decreased cell death in the WT CD8 T cells after blocking IFN-γ signaling. These experiments suggest that increased antigen specific CD8 T cell counts in IFN-γ-/- mice is not the result of increased proliferative capacity but is instead due to the absence of IFN-γ-induced CD8 T cell death. Our results suggest that IFN-γ plays a role in promoting cell death that limits the CD8 T cell response during an influenza viral infection.
Increased IL-17 production in post-septic mice exacerbates respiratory syncytial virus induced immunopathology of the lung (49.25)
Authors: Mukherjee Sumanta , Lukacs Nicholas , Kunkel Steven , Carson IV William 
Published: January 2023

Abstract
Survivors of severe sepsis exhibit increased morbidity and mortality in response to secondary infections. Although bacterial secondary infections have been widely studied, there remains a paucity of data with viral infections post sepsis. Here, we show in an experimental mouse model of severe sepsis (cecal ligation and puncture, CLP) followed by respiratory syncytial virus (RSV) infection, exacerbated immunopathology in the lungs of CLP mice compared to RSV infected sham surgery mice. The immunopathology associated with the viral infection in CLP mice was in part due to increased mucus production in the lungs and impaired ability of septic mice to clear the virus. These findings correlated with increased IL-17 production in the lungs of RSV-infected CLP mice. RSV-infected CLP mice had increased levels of Th2 cytokines and reduced IFNγ in the lungs and lymph nodes compared to RSV infected sham mice. In addition, CD4 T cells from CLP mice ex vivo increased IL-17 production irrespective of the skewing conditions. Further, in vivo neutralization of IL-17 prior to RSV infection led to a significant reduction in virus induced mucus production and Th2 cytokines. Taken together, these data provide evidence that post septic CD4+T cells are primed toward IL-17 production, which may contribute to the immunopathology of a secondary viral infection.
Local inflammatory chemokines support the chronic activation of naïve influenza-specific CD8 T cells during the recovery from influenza infection (49.4)
Authors: Wu Tao , Suarez-Ramirez Jenny , Lee Young-Tae , Bouchard Keith , Cauley Linda 
Published: January 2023

Abstract
We previously found processed peptide antigens in the draining lymph nodes of the respiratory tract two months after primary influenza infection. These antigens were presented to naïve (but not central memory) CD8 T cells by CD11b+ DC. Many partially-activated F5 cells remained clustered in the paracortex of the MLN during late antigen presentation. Further studies show that the MLN remains enlarged &gt;30dpi and contains larger numbers of CD11b+ DCs than resting LNs but the DC are not phenotypically mature. Consistently, RT-qPCR analysis shows very small quantities of cytokines in the lungs and DLNs during antigen persistence, which may contribute to the immature phenotype of the DCs and incomplete activation of naïve F5 cells. During this time mRNA levels for several inflammatory chemokines remain elevated in the lungs and DLNs including CCL8 and CXCL9. Consistent with this, naïve F5 cells lacking CXCR3 and CCR2 chemokine receptors show impaired proliferative responses when transferred to influenza infected mice 30dpi, as compared to wild type cells. These data indicate that a mild inflammatory response is induced during antigen persistence and supports chronic activation of naïve CD8 T cells through local chemokine expression.
Dynamic capping of CD25 occurs at point of contact prior to delivery of IL-2 from dendritic cell to Treg hybridoma. (50.12)
Authors: Field Elizabeth , Kulhankova Katarina , Rouse Todd , Nasr Mohamed 
Published: January 2023

Abstract
CD4+CD25+ regulatory cells (Tregs) depend on external IL-2 for expansion, homeostasis and function, but the process of IL-2 delivery to Treg is poorly understood. We reported that IL-2 producing dendritic cells (DCs) control Treg function in vitro and that IL-2 delivery to Treg requires cell:cell contact and CD25. To further explore DC delivery of IL-2 we demonstrated that IL-2 producing dendritic cells similarly controlled suppressor function of CD4+CD25+ Treg hybridoma cells RD6, then studied the fate of DC-derived material and CD25 using live cell imaging and confocal microscopy of dye-labeled or IL-2mCherry transfected DCs and dye-labeled or CD25-AcGFP transfected RD6. Conjugate formation between DCs and RD6 was independent of IL-2 and CD25. However, the uptake of DC-derived material into RD6 cells required both IL-2 sufficient DCs and CD25. RD6 internalized DC-derived material from dye-labeled WT DCs and IL-2mCherry-expressing IL-2 KO DCs but not mCherry-expressing IL-2 KO DCs. Anti-CD25 blocked internalization of DC-derived material. RD6 exhibited dynamic capping of CD25 to the point of contact with IL-2 sufficient DCs, but showed no redistribution of CD25 after contact with IL-2 deficient DCs. CD25 capping preceded the release of IL-2 from the DC. The data indicate that IL-2 delivery from DC to RD6 involves membrane remodeling with spatial redistribution of CD25 to the DC contact site, raising the possibility for actin-cytoskeletol control of IL-2 delivery in Tregs.
Lymphoma resulting from β-catenin stabilization are sensitive to mTOR inhibition: a unit on cancer and the immune system (51.8)
Authors: Lo Oanh , Gounari Fotini , Dose Marei 
Published: January 2023

Abstract
The Wnt/β-Catenin pathway is an important cell-signaling pathway that affects the development of T-cells. If certain signals are not working properly, lymphomas may arise and drugs can be used as treatment. β-Catenin stabilization in mice results in thymic lymphomas which suggest that there are parallels to human leukemias and T-cell lymphomas. Recently, the immunosuppressive drug rapamycin has been under investigation for its potential in cancer therapy. This lab experience inspired me to create a unit that can be used in any type of high school biology, anatomy, or health class in understanding the role of the immune system in controlling cancer. First, students will learn about the anatomy of the immune system. They will gain an understanding of how cancers arise and the role of the immune system and cancer. Students will also participate in a laboratory activity that allows them to differentiate between normal and cancer cells by viewing slides under a microscope in a blood-based cancer diagnosis lab activity. The unit will conclude with an on-line research component allowing students to investigate how cancer cells are treated with immunosuppressive drugs.
CpG-induced inflammatory DCs promote strong Th1/CTL responses to particulate antigens (52.11)
Authors: De Koker Stefaan , De Geest Bruno G. , Willart Monique , Remon Jean Paul , Lambrecht Bart , Grooten Johan 
Published: January 2023

Abstract
Increasing safety issues are now driving the vaccine field to develop vaccines composed of recombinant antigens. Most protein antigens are however poorly immunogenic and require the addition of immunepotentiators to evoke immunity. Moreover, compared to particulate antigens, uptake and presentation of soluble antigens by DCs is far inferior. By encapsulating antigens into polyelectrolyte-based particles modified with TLR ligands, we aim to create entirely synthetic but non-pathological mimics of micro-organisms. Encapsulating OVA in these particles strongly increased antigen presentation by DCs via MHCI and MHCII. Although antigen encapsulation in these particles was sufficient to elicit immunity, the strength of the induced Th1 and CTL response was dramatically increased by functionalizing the particles with the TLR9 ligand CpG, yielding protective immunity against B16-OVA melanoma. Importantly, the particulate formulation was also far superior to a mixture of soluble antigen and CpG. Injection of CpG resulted in a strong recruitment of monocyte-derived inflammatory DCs in the draining lymph nodes, which were totally absent in CCR2-/- mice. Although immune responses to soluble antigen and CpG remained unaffected in CCR2-/- mice, the strong Th1/CTL response observed in wild type mice to particulate antigen/CpG was almost totally abrogated in the CCR2-/- mice, pinpointing to important differences in how T-cell responses are initiated to particulate and soluble antigens.
Investigation of TLR ligand-generated inflammatory signals for the induction of high magnitude memory CD8+ T lymphocyte responses (52.14)
Authors: Nelson Michelle , Johnson Alison , Milligan Gregg 
Published: January 2023

Abstract
Most currently used vaccines elicit antibody and CD4+ T cells, but protection against some pathogens also require a CD8+ T cell response. CD8+ T cells can be readily elicited using live attenuated vaccines or non-viable vaccine antigens in combination with adjuvants. However, little is known about how adjuvants affect the elicited CD8+ T cell response. We examined the ability of different adjuvant stimuli (TLR ligands CpG, MPL and Poly(I:C)) to elicit critical signals in bone marrow-derived dendritic cells (BMDC) necessary for appropriate development of antigen-specific CD8+ memory T cells. Mice vaccinated with MPL- or CpG-stimulated, peptide-pulsed BMDC exhibited increased CTL activity compared to those given Poly(I:C)-treated BMDC. Greater numbers of antigen-specific memory cells were detected in the spleen and iliac LN of the MPL-stimulated BMDC group. While minimal variation was observed in the expression of chemokine receptors on the surface of CD8+ T cells among the various immunization groups, there were differences observed in cytokine production. Less TNF-α was generated by CD8+ T cells from CpG-stimulated BMDC vaccinated mice. In summary, we determined that various TLR ligands differentially affected the magnitude, homing, and effector functions of memory CD8+ T lymphocytes. The results of these studies suggest that specific TLR ligand adjuvants may affect the quality of vaccine-elicited CD8+ T cells which may, in turn, impact vaccine-mediated protection.
Distinct populations of CD4+ helper T cells generate CD4 and CD8 memory responses to infection (52.23)
Authors: Weber K. Scott , Li Qi-Jing , Persuad Steve , Campbell Jeff , Davis Mark , Allen Paul 
Published: January 2023

Abstract
Helper T cells are critical for protective immunity, CD8 T cell memory, and CD4 recall responses required for effective vaccines. Despite the importance of CD4 T cells in immune responses, studies examining T cell responses to Listeria monocytogenes have focused primarily on CD8 T cells. We have developed two CD4 T cell transgenic mouse lines specific for an immunodominant L. monocytogenes epitope, providing a novel means of examining CD4 T cell activation and memory formation. These mice, called LLO118 and LLO56, differ by only 14 amino acids in their TCR sequences and have identical responses to peptide and L. monocytogenes infection in vitro. Comparison of the in vivo CD4 T cell responses to L. monocytogenes infection revealed LLO118 T cells proliferate much more strongly in a primary infection and cause a more robust CD8 response to secondary infection than LLO56. Surprisingly, we found that LLO56 has a superior CD4 recall response to secondary infection. CD5 expression and TCR downregulation correlate with the discordant primary and secondary responses. CD5 is higher in LLO56 prior to infection whereas LLO118 TCR levels downregulate more after the primary infection. Thus, a strong initial proliferation results in a better CD8 recall response, but a decreased CD4 recall response due to downregulated TCR levels. These findings imply that there are two populations of helper T cells: one that works best in primary responses and another in the secondary response.
CD8+ T cells adversely effect their own memory potential via the regulation of the IL-12/IL-15Rα axis (52.32)
Authors: Kohlhapp Frederick , Zloza Andrew , McCracken James , O'Sullivan Jeremy , Jagoda Michael , Lacek Andrew , Guevara-Patiño José 
Published: January 2023

Abstract
Current paradigms establish that CD8+ T cell memory potential is adversely affected by the degree of inflammation during an effector response. However, the regulation of this balance between inflammation and memory potential remains poorly understood. Here we demonstrate that CD8+ T cells, themselves, limit their own memory potential through direct regulation of inflammatory cytokine production by antigen presenting cells (APCs) during vaccination. By depleting CD8+ T cells during vaccination, we show augmented formation of memory precursor effector cells (MPECs), which results in the enhanced quantity (antigen-specific CD8+ T cell number) and quality (cytolytic function) of the resulting memory responses. Furthermore, we show that CD8+ T cells promote inflammation and shift the balance towards terminal effector CD8+ T cell responses by increasing IL-12 production and decreasing IL-15Rα expression by APCs in an IFN-γ-dependent manner. These results identify an unrecognized role for CD8+ T cells in the regulation of their own effector differentiation fate and a previously uncharacterized relationship between the balance of inflammation (IFN-γ, IL-12) and memory formation (IL-15Rα).