bluuebunny/crossref_metadata_2025_split huggingface dataset:
TGF-beta1-induced expression of alphaE(CD103)beta7 integrin on Ag-specific tumor-infiltrating T lymphocytes promotes killing of E-cadherin-bearing tumor cells by triggering polarized cytotoxic granule exocytosis (50.22)
Authors: MAMI-CHOUAIB Fathia , JALIL Abdelali , VERGNON Isabelle , LE MAUX CHANSAC Béatrice , LAZAR Vladimir , BISMUTH Georges , CHOUAIB Salem , LEFLOC’H Audrey 
Published: January 2023

Abstract
Interaction of intercellular adhesion molecule (ICAM)-1 with lymphocyte function-associated antigen (LFA)-1 enhances target cell-T cell adhesion and thus promotes TCR-mediated killing. In the absence of ICAM-1, frequently lost during tumor cell dissemination, we demonstrate that the interaction of the epithelial cell marker E-cadherin with αE(CD103)/1 _7 integrin, often expressed by tumor-infiltrating T lymphocytes (TIL), plays a major role for an effective cancer cell lysis. Indeed, while tumor-specific CD103+ TIL are able to kill E-cadherin+/ICAM− autologous tumor cells, CD103− PBL-derived counterparts are inefficient. Furthermore, tumor cell killing by TIL is abrogated by anti-CD103 mAb and RNA interference targeting E-cadherin. Confocal microscopy analysis demonstrated that αEβ7 is recruited at the immunological synapse and that its interaction with E-cadherin is essential for cytolytic granule polarization and subsequent exocytosis. Interestingly, pre-incubation of PBL-derived T cells with anti-CD3 mAb and TGF-β1 induced CD103 expression resulting in a dramatic potentiation of anti-tumor effector function. Thus, CD8+/CD103+ tumor-reactive T cells infiltrating epithelial tumors most likely play a major role in anti-cancer CTL response.
Antigen priming in the presence of IL-12 selectively enhances the survival of CD62LhiCD8+ T cells and results in superior anti-tumor activity (50.23)
Authors: Diaz-Montero C. Marcela , EL-Naggar Sabry A. , Montero Alberto J. , Cole David J. , Salem Mohamed L. 
Published: January 2023

Abstract
It has been recently shown that early effector CD8+T cells are more efficient at killing tumors than late effector CD8+T cells. Since expression of CD62L is progressively lost as T cells differentiate, its expression on activated CD8+T cells has been associated with superior antitumor activity. In our study, we have shown that in vitro priming in the presence of IL-12 results in the accumulation of CD62LhiCD8+ T cells. The antiapoptotic effects of IL-12 were found to be selective for this population and was associated with downregulation of caspases 3 and 8, Bax, and Bad. We also found that IL-12 modulates the expression of CD62L. OT-1 cells primed with SIINFEKL with or without IL-12 were sorted into CD62Llo and CD62Lhi populations three days after priming and recultured with or without IL-12. Exposure of untreated CD62LloCD8+cells to IL-12 after sorting resulted in increased CD62L levels. CD62LloCD8+cells primed in the presence of IL-12 transiently upregulated CD62L expression. However, the levels decreased by day 7 unless cells were re-exposed to IL-12. The fact that effects were seen on sorted populations, suggests that IL-12 has a direct effect on the expression of CD62L. Finally, we found that CD8+CD62Lhi cells generated after priming with IL-12 had increased in vivo survival and enhanced recall capabilities. When compared with CD62Lhi cells primed with antigen alone, IL-12-treated- CD62Lhi cells showed enhanced anti-tumor capabilities. In conclusion, our data supports the findings that increased expression of CD62L on effector T cells results in enhanced anti-tumor activity, and suggests IL-12 treatment as an effective strategy for the generation and maintenance of such subpopulation.
NK Cell Mediated Tumor Cytotoxicity is enhanced by Mild Thermal Stress through Heat Shock Factor 1 (50.24)
Authors: Dayanc Baris Emre , Ostberg Julie R , Yuan Min , Repasky Elizabeth A. 
Published: January 2023

Abstract
Natural killer (NK) cells directly kill tumor cells and regulate downstream adaptive immune responses. Previous studies in our lab suggest that mild thermal stress (39.5 C) significantly enhances the cytotoxicity of human peripheral blood NK cells compared to normothermic conditions (37 C) against human colon tumor cells. This enhancement was not seen when syngeneic human PBMC were used as targets, nor when the IL-2 dependent human NK cell lines NKL or NK92MI, or IL-2 activated primary NK cells were used as effectors. Thus, we hypothesized that resting NK cell cytotoxic activity is regulated by mild thermal stress. Blocking of the activating receptor NKG2D and its stress-induced ligand MICA with blocking antibodies suggested the involvement of these molecules in the thermal regulation of NK cytotoxicity. We found that inhibitory receptor KIR2DL1 levels on NK cells were not changed with thermal stress. Increased clustering of NKG2D on NK cells was observed with thermal stress, suggesting the potential for enhanced efficiency of activating signals. However prevention of lipid raft clustering with methyl B-cyclodextrin on NK cells shows no significant affect on thermally enhanced cytotoxicity. On target cells, MICA expression appeared to be upregulated by mild thermal stress in a manner that is dependent on the transcription factor HSF-1. Overall, these data suggest that thermal therapy might be clinically utilized to help enhance NK mediated tumor cell killing.
Supported by NIH P01 CA94045, R01 CA71599, Komen Foundation DISS0402487 and Fulbright Program.
OX40-ligand inhibits function of IL-10-producing intratumoral CD4+CD25+ regulatory T cells in human follicular B-cell lymphoma (50.25)
Authors: Voo Kui Shin , Bover Laura del Carmen , Neelapu Sattva S , Kwak Larry W , Liu Yong-Jun 
Published: January 2023

Abstract
Recent studies showed that infiltrating T regulatory cells inhibit effector T-cell function in B-cell non-Hodgkin lymphomas. Therefore, it was suggested that strategies that inactivate or remove regulatory T-cell populations should be done in conjunction with established clinical protocols in B-cell lymphomas bearing patients. In the present study, we purified CD4+CD25−, CD4+CD25highICOS+ and CD4+CD25highICOS- cell subsets by flow cytometry from a follicular lymphoma patient. We examined their intracellular FoxP3 expression and determined their IL-10 secretion in cell supernatants under different conditions. The cell subsets were stimulated with anti-CD3 and anti-ICOS antibodies in the presence of IL-2 on parental L cells or OX40L-L cells. Cytokine production in supernatants was measured by ELISA. Both CD4+CD25highICOS+ and CD4+CD25highICOS- cells showed FoxP3+ expression by FACS analysis, while it was negative in CD4+CD25−. After 3 or 6 days of culture, only ICOS+ cells produced large amounts of IL-10 (470 and 3,250 pg/ml respectively) and OX40L dramatically inhibited this production (87 and 94% of inhibition). Immunohistochemistry staining demonstrated simultaneous expression of FoxP3 and ICOS on T-cells infiltrating B-cell lymphoma tissues. These results suggest that OX40L could be potentially used as a therapeutic agent to modulate T regulatory cells function in B-cell lymphoma patients.
Depletion of CD4+CCR4+ cells enhances T cell-mediated responses in vitro and vaccine-induced anti-tumor immune responses in mice. (50.27)
Authors: Biragyn Arya , Olkhanud Purevdorj B , Sumitomo Kenya , Baatar Dolgor 
Published: January 2023

Abstract
CCR4 is a chemokine receptor which is preferentially expressed on T helper 2 (Th2) CD4+ cells and CD25+CD4+ T regulatory cells (Tregs). Tregs and Th2-type cytokines (IL-10 and IL-4) were implicated in suppression of anti-tumor immune responses. We have hypothesized that depletion of CCR4+CD4+ cells, particularly Tregs and Th2 cells, may improve anti-tumor T cell immune responses. To test this, we generated a recombinant protein consisting of TARC (CCL17, ligand for CCR4) fused with 38kD truncated form of Pseudomonas exotoxin (PE38), designated TARC-PE38. TARC-PE38 efficiently and specifically killed CCR4+ cells, including sorted human peripheral blood CCR4+CD4+ and CCR4+ Tregs. We demonstrate that, while CCR4+non-Tregs mostly suppress Th1-type polarization without affecting T cell proliferation, CCR4+Tregs primarily regulate T cell proliferation via contact-mediated suppressive pathways. As a result, depletion of CCR4+ cells enhance proliferation of T cells and promote the Th1-type polarized responses. Moreover, the depletion also significantly enhanced tumor antigen-specific T cell responses in vitro. Importantly, in vivo depletion of CCR4+ cells augmented tumor - specific CD8+ CTL responses improving survival of tumor bearing mice, when combined with cancer DNA vaccines. Taken together, our data suggest that poor immune responses to tumors can be reversed by transiently depleting suppressive CCR4+ cells.
This work was supported by the Intramural Research Program of the National Institute on Aging, NIH.
Aberrantly expressed neutrophil elastase (ELA2) in the nucleus and cytoplasm of acute lymphocytic leukemia (ALL) cells cleaves cyclin E (CCNE) into low-molecular-weight forms (LMWFs) yielding novel HLA-A2 restricted determinants (50.28)
Authors: Ishiyama Ken , Kondo Yukio , Wieder Eric , Lu Sijie , Molldrem Jeffrey 
Published: January 2023

Abstract
We reported that CCNE-specific CTL elicited from healthy donors (HDs) kill ALL and CML cells, which aberrantly express CCNE. Further, 5 LMWFs of CCNE1 are present only in malignant cells and constitutively active to promote cell division. In addition, we showed ELA2-specific CTL also kill AML and CML, and ELA2 is normally expressed only in myeloid cells. To determine whether ELA2 might be aberrantly expressed in ALL and whether ELA2 cleaves CCNE to LMWF’s, we first found CCNE of LMWF to be overexpressed by western blot of cell lysates from ALL, but not from ALL in remission or HD B-cells. ELA2 added to ALL lysates increased LMWF’s and was inhibited by the ELA2 inhibitor elafin. Subcellular fractionation and co-immunoprecipitation showed ELA2 to be aberrantly associated with CCNE in nucleus, cytoplasm, and membrane-bound organelles in ALL, but not normal bone marrow cells. Because determinants recognized by CCNE-CTL are also found in the LMWFs and lysis of leukemia correlates with target protein overexpression, we studied the effect of induced CCNE expression on target susceptibility to cytolysis. While normal PBMC were not killed, PHA or α-CD3/28 stimulated PBMC were killed by CCNE-CTL, although non-specifically. We conclude that in ALL, aberrant expression of the myeloid protein ELA2 results in mistrafficking of ELA2 to cytoplasm and nucleus, thus mediating CCNE cleavage to LMWF’s and increasing susceptibility to CCNE-CTL killing. This shows novel tumor antigen interaction that may cooperatively reverse CTL tolerance.
TNF-related apoptosis inducing ligand (TRAIL) enhances TCR-mediated renal cancer recognition by a novel CD4+ T cell clone (50.29)
Authors: Wang Qiong , Hanada Ken-ichi , Yang James C 
Published: January 2023

Abstract
A human CD4+ T cell clone (HC/2G-1) was established by stimulating peripheral blood T cells with dendritic cells pre-incubated with the autologous apoptotic renal tumor line in the presence of IFN-α. It recognizes multiple MHC-mismatched RCCs by IFN-γ release (10 of 11 lines) and lysis (9 of 10 lines), but does not recognize multiple EBV-B cells or fibroblasts. It shows little or no recognition of a panel of melanomas, breast cancer and non-small cell lung cancers. Phenotypically, HC/2G-1 is CD3+CD4+TCRα/β+, but CD161−CD16−NKG2D−. HC/2G-1 does not utilize Vα or Vβ TCRs described for classical NKT cells, but rather Vα14, Vβ2.1. Tumor recognition was TCR α/β mediated as demonstrated by conferring reactivity by transducing third party T-cells with the cloned TCR from HC/2G-1. Tumor recognition by clone HC/2G-1 was not blocked by antibodies to HLA-class I or class II, but was significantly reduced by both anti-TCRα/β and anti-TRAIL Ab. HC/2G-1 expresses low level of TRAIL, and can be upregulated with type I IFNs. Although most RCC tumor lines were resistant to TRAIL induced apoptosis, treatment of soluble TRAIL trimer enhances tumor recognition such as IFN-γ production by HC/2G-1 as well as by HC/2G-1 TCR-transduced T cells. Furthermore, tumor recognition by HC/2G-1 TCR-transduced T cells was blocked by anti-TRAIL Ab. This suggests that TRAIL may be inducing the antigen, or a restriction element on tumor cells. Attempts to understand the molecular mechanism of tumor recognition by HC/2G-1 are underway.
Host-derived CCL5 is critical for mammary tumor growth and metastasis (50.30)
Authors: Liu Jianguo , Ma Xiaojing 
Published: January 2023

Abstract
CCL5 is a chemokine produced by a variety of human cells under stress. It plays an important role in inflammation by recruiting T cells, macrophages and eosinophils to inflammatory sites. It has been shown in animal studies that tumor-derived CCL5 is able to help tumors to form new blood vessels (angiogenesis), thus fostering their growth. Tumor-derived CCL5 can also inhibit the T cell response and enhance the growth of breast cancer. Human studies have shown high incidence of CCL5 expression in tissue sections of breast carcinomas. Furthermore, in stage II breast cancer patients, the expression of CCL5 increased significantly the risk for disease progression. However, the mechanisms of CCL5 promote the tumor growth and metastasis is still unknown.
Using a syngeneic 4T1 mammary tumor model that closely resembles human breast cancer in primary tumor growth characteristics and metastatic properties, we have observed that, compared to control wild type mice of identical background, mice that are genetically deficient in CCL5 (CCL5-ko) are highly resistant to 4T1 tumor growth and to tumor-induced death. In addition, the remaining tumors in CCL5-ko mice developed little lung metastasis after 60 days. The CCL5-ko mice that regressed the tumor promptly and strongly rejected rechallenge with 4T1 tumor, indicating establishment of protective immunity. Furthermore, mAb-mediated neutralization of CCL5 in vivo prevented 4T1 tumor growth and lung metastasis. Our data also indicate that 4T1 tumor cells spontaneously produce high levels of CCL5 driven by highly active NF-kB and other yet to be characterized transcription factors. In conclusion, CCL5 plays a critical role in the growth and metastasis of mammary tumors, and that CCL5 represents a desirable and accessible target in breast cancer therapy.
Substance P modulation of anti-tumor responses: enhanced tumor resistance by adoptive transfer of SP-treated T cells and increased NK cell activity (50.5)
Authors: Manske Jill , Brand Lucas , Kastelic Amanda , Westin Angelene 
Published: January 2023

Abstract
Substance P (SP) is a neuropeptide that has been shown to have immunoregulatory properties including effects on mediators involved in anti-tumor immunity. In previous studies we have shown that treatment of mice with SP provides protection against tumor growth. This protection requires both T cells and NK cells, and adoptive transfer of cells from SP-treated animals can transfer tumor protection. These studies suggest a model in which SP treatment prior to tumor challenge primes immune mediators to prevent or delay tumor establishment. In this study we examined the role of NK cells and T cells in tumor protection. The cytotoxic activity of NK cells against Yac-1 targets was examined by labeling cells with annexin-V and measuring apoptotic cell death by flow cytometry. NK cells from mice treated with SP had significantly higher cytotoxic activity compared to NK cells from controls. To examine whether SP-primed T cells could increase tumor resistance, purified T cells from SP-treated donor mice were adoptively transferred into animals prior to tumor challenge. Mice adoptively transferred with T cells from SP-treated donors developed significantly fewer tumors than did controls. These results suggest that SP mediates anti-tumor immunity in part by increasing NK cytotoxic activity. Since SP-primed T cells can transfer tumor protection, these cells may play a role in this increase in NK activity.
Targeting Activation Induced Cytidine Deaminase Promotes Rejection of Large Established Tumors by Cytotoxic T Lymphocytes in a Mouse Myeloma Model (50.8)
Authors: Bai Xue-Feng , Liu Jin-Qing , Joshi Pramod S. , Liu Chang-Gong , Wu Lai-Chu , Liu Yang 
Published: January 2023

Abstract
Activation induced cytidine deaminase (AID) is an essential enzyme that controls the generation of antibody diversity in B cells and is considered a general gene mutator. Using transgenic T cells specific to P1A (P1CTL) to treat mice with large established plasmacytoma J558 tumors, we frequently observed mutations in P1A antigenic epitope, which lead to cancer cell evasion of destruction by P1CTL. To determine whether AID plays a role in CTL-mediated rejection of plasmacytoma, we generated AID-silenced plasmacytoma J558 cells and controls and tested whether large established AID-silenced J558 tumors could be rejected by P1CTL. Complete rejection of large established tumors by P1CTL was rare (0–10%) in mice with AID+ J558 tumors while the rejection rates were 30–90% in mice with AID-silenced J558 tumors. AID knock down did not reduce mutation rate of P1A gene, but rather caused more genetic instability and altered gene expression profile. Notably, we observed strong up-regulation of OX-2 (CD200) gene and down-regulation of CaMKII-δ gene in AID-silenced J558 cells, whose roles may be related to enhanced immune rejection of AID-silenced J558 tumors. Our results suggest that simultaneously targeting AID in CTL-targeted therapy of myeloma or other AID+ tumors may be of therapeutic value.
Agent based model of the TH1/TH2 differentiation decision (51.3)
Authors: Lawrence Michael Brandeau , Schmidt Brian J. , Chavali Arvind K. , Glaw Lydia 
Published: January 2023

Abstract
Adaptive immune responses tend to polarize to predominantly TH1 or TH2-directed, and this decision is critical for the clinical outcome of many diseases. For example, Leishmania species that elicit a TH2 response, such as donavani, can lead to visceral leishmaniasis, which is fatal if not treated. In contrast, the body is able to combat leishmaniasis and recover from species that tend to elicit a TH1 response, such as L. major. The type of response the immune system mounts is governed by a multifactorial process involving the nature of the antigen and co-stimulatory molecules expressed on antigen presenting cells (APCs). We developed an agent-based computational model using knowledge of immunological interactions recently elucidated by in vivo two-photon microscopy studies to investigate whether APCs could be engineered to elicit a TH1 response after a TH2 response was previously established by a pathogen. The model supports the stochastic repertoire scanning model and further suggests that this mechanism may help T-cells coordinate their response by sampling the cytokine cues left by other differentiating T-cells. Additional cell types, such as natural killer and memory cells, may be important for establishing the dynamics of the response and the ratio of TH1 to TH2 cells. Our model additionally suggests that engineered APCs have the potential to be a viable treatment method for diseases such as visceral leishmaniasis.
Supported by NIH R01 HL54614.
A soluble Dectin-1 Fc fusion protein enhances innate immunity against invasive pulmonary aspergillosis (51.8)
Authors: Mattila Polly E , Metz Allison E , Repaka Rekha R , Steele Chad 
Published: January 2023

Abstract
The Dectin-1 beta-glucan receptor is essential for alveolar macrophage (AM)-mediated defense against Aspergillus fumigatus. Increased incidence of invasive pulmonary aspergillosis (IPA) over the last several decades has necessitated the development of novel vaccines and immunotherapies to augment host defense against A. fumigatus in susceptible individuals. In our studies, the extracellular carbohydrate binding domain of Dectin-1 was fused with murine IgG1 Fc (sDect-Fc) by PCR and cloned into a mammalian expression vector. sDect-Fc was also cloned into a non-replicating adenovirus for in vivo expression. The viability index (VI) in AM cultures containing sDect-Fc was 80% lower than untreated AM cultures, indicating sDect-Fc significantly augments AM-mediated killing of swollen conidia. sDect-Fc also minimized the AM inflammatory response to A. fumigatus. IV administration of AdsDect-Fc, but not AdLuc, to naive mice resulted in sDect-Fc protein in the lungs. In preliminary studies, mice receiving AdLuc had 66% mortality within 16 days after A. fumigatus challenge whereas mice that received AdsDect-Fc had no mortality within a 21 day period. These results support the use of sDect-Fc as an immunotherapeutic strategy to augment host defense against IPA. Grant HL080317 from the NIH and grant CI-21330-N from the ALA.
Cyclophosphamide effects on avian macrophages in vitro (52.10)
Authors: Kolawole Abimbola Olayinka , McCorkle Fred Miller 
Published: January 2023

Abstract
Cyclophosphamide is an alkylating agent used in the treatment of a variety of cancers. It has been shown to cause immunodepression by reducing T cell, B cell and antibody production, but little is known about the effects of the drug on macrophages. Avian macrophages are good models for human macrophages because they have the same functions in maintaining immunity. This study examined the cytotoxic effect of cyclophosphamide on the activated avian macrophage cell line MQ-NCSU, as well as the effects of the drug on attachment to a plastic substrate, phagocytosis, nitrite production and RNA synthesis for macrophage chemoattractant protein-1 (MCP-1) and nitrous oxide synthetase-1 (NOS-1). Avian macrophages were exposed to doses ranging from 101 to 10−7 mg/mL for a period of 30 min. Cyclophosphamide was not cytotoxic to the avian macrophages and did not affect the ability to phagocytize sheep red blood cells and E. coli. Macrophage attachment was reduced by 21% after treatment at concentrations of at least 10−4 mg/mL. At 10 mg/mL, cyclophosphamide reduced nitrite production by 67% and was associated with a qualitative reduction in the amount of nos-1 RNA extracted from macrophages. The total cellular RNA extracted from and the amount of mcp-1 RNA in macrophages was not affected. This study shows that the clinical doses of cyclophosphamide used for cancer patients are not detrimental to avian macrophage functions.
Acknowledgements: Dr. Jennifer Schisa, Graduate student research and creative endeavors grant, Central Michigan University.