bluuebunny/crossref_metadata_2025_split huggingface dataset:
Elimination of Intracranial Gliomas via Immune Rejection of Subcutaneous Allogeneic Glioma Cell Lysate (48.40)
Authors: Stathopoulos Apostolos , Milbouw G. , Chen T. 
Published: January 2023

Abstract
Allogeneic glioma cell lines, coupled with allogeneic and syngeneic tumor cell lysates, could be used to induce immune mediated rejection of syngeneic glioma cell lines in two strains of rat. Fisher344 and Sprague Dawley (SD) rats were implanted subcutaneously with 9L and C6 glioma cell lines. After tumor formation, rats were injected subcutaneously with a combination of allogeneic tumor cell line and combined allogeneic and syngeneic tumor lysate. As a further study, treated rats, who had decreased tumor size from the above treatment, were tested for immune memory. Same experiments were also performed in the intracranial model. Rats treated with the same methode were able to significantly reduce (Fisher344), or completely reject (SD rats), their tumor. “Cured” rats re-injected with five times the original number of malignant cells and remained tumor free. Analysis of these tumors shows numbers of CD4, CD8, B-lymphocytes, and dendritic cells. In the intracranial model, rats had significantly less intracranial spread of tumor than the control group. Our in vivo experiments demonstrate that established subcutaneous or intracranial gliomas may be treated with a combination of allogeneic cells and tumor lysates (allogeneic and syngeneic tumor cells) to reduce its size or eliminate it, and establish lasting memory.
Allogeneic melanoma vaccine expressing alphaGal epitopes induce anti-tumor immunity to autologous melanoma antigens without signs of toxicity (48.7)
Authors: Rossi Gabriela R , Mautino Mario , Lejukole Henry , Koenigsfeld Marie , Vahanian Nicholas , Link Charles J 
Published: January 2023

Abstract
The hyperacute rejection of a xenotransplant is a potent mechanism of tissue destruction mediated by complement-fixing natural Ab that recognize alphaGal (aGal) epitopes. These epitopes are not present on human cells. Humans develop high titers of naturally acquired Ab against these epitopes. We hypothesized that this mechanism could be used to generate a potent anti-cancer vaccine. In this study we tested this hypothesis by using an allogeneic whole cell cancer vaccine to treat murine melanoma. We used the aGal KO mice (aGT H-2bb) and the aGal negative melanoma cell lines B16 and S91M3. We genetically engineered S91M3 (H-2dd) cells to produce the aGal(+)S91M3 cells. Mice with pre-existing subcutaneous and pulmonary B16 (H-2bb) tumors were efficiently treated with aGal(+)S91M3 melanoma vaccines. Control S91M3 vaccines had no impact on tumor growth. Vaccination with aGal(+)S91M3 induced cytotoxic CD8+ T cells recognizing the “autologous” aGal(-)B16 tumors measured by IFN-gamma release and by adoptive transfer experiments. The presence of allo-antigens did not dominate the induction of immunity to “cryptic” tumor antigens tested in the pre-existing melanoma metastasis model. The effective anti-tumor immunity was not associated with autoimmunity or other signs of toxicity in long-term toxicology studies for breast, lung and melanoma allogeneic vaccines. This study establishes for the first time the safety and efficacy data of allogeneic aGal(+) whole cell vaccines and constituted the basis for the initiation of human clinical trials to treat human cancers currently underway.
Lipid Accumulation and Dendritic Cell Function in Cancer (49.10)
Authors: Herber Donna L. , Kroeger Jodi L. , Makris Melissa , Gabrilovich Dmitry I. 
Published: January 2023

Abstract
Immune cell dysfunction in cancer is an area of intense study, with particular focus on dendritic cell (DC) abnormalities. The mechanism of these abnormalities remains unclear. We examined the potential role of lipid metabolism in defective DC function in cancer. Different mouse tumor models (CT26, MC38, or EL4) were used for in vivo studies. Tumors grew for up to 3w and then spleens were harvested for DC analysis. Phenotypic analysis by flow cytometry included CD11c, CD11b, MHCII, CD40, CD80, and CD86. Lipid levels were determined using Bodipy 493/503. For the analysis of DC function, cells with high and normal lipid content were sorted and used in allogenic mixed leukocyte reaction. For in vitro studies, DCs were harvested from spleen and cultured overnight with a variety of stimulants (LPS, tumor supernatant, GM-CSF, or IL-4). The control level of lipids in DCs was established in naïve tumor-free mice. 30–40% of freshly isolated splenic DCs from tumor-bearing mice had increased levels of intracellular lipid compared to naïve mice, and this accumulation was time dependent. Phenotypically, DCs with increased amounts of lipid had increased levels of MHCII and co-stimulatory molecules. However, despite that increase, DCs with high lipid content had substantially reduced ability to stimulate T cells in allogenic mixed leukocyte reaction than DCs with normal lipid amount. These results indicate a role for lipid accumulation in dendritic cell dysfunction. The data also indicate that the tumor microenvironment is necessary to induce the lipid accumulation and dysfunction.
Role of reactive oxygen species in immune suppression in cancer (49.12)
Authors: Corzo Cesar Alexander , Nagaraj Srinivas , Kusmartsev Sergei , Gabrilovich Dmitry 
Published: January 2023

Abstract
The inability of the host immune system to develop and maintain an effective anti-tumor immune response is one of the main mechanisms of tumor progression. Recent findings suggested that hyper-production of reactive oxygen species (ROS) by myeloid cells in cancer may be one of the major factors responsible for T-cell non-responsiveness. However, the mechanism of ROS accumulation as well as their precise role in immune defects in cancer remains unclear. Previous reports have also stated that myeloid-derived suppressive cells (MDSC) contain increased levels of active STAT3 and we set out to address the relationship, if any, between STAT3 and the observed high ROS production. Our data indicate that MDSC isolated from tumor-bearing mice have a significant increase in the expression of two critical components of NADPH oxidase: gp91phox and p47phox than their wild type counterparts. The NADPH oxidase has long been considered a strong source of ROS in myeloid cells and we suggested that STAT3 may be partially responsible for this up-regulation. Our results show that STAT3 does indeed binds to the promoter region of the p47 gene. Furthermore, in vitro inhibition of STAT3 decreased the expression of both gp91phox and p47phox. In addition, the presence of an active NADPH complex was critically important for MDSC-mediated immune suppression. Tumor-bearing mice with defective NADPH oxidase had significantly lower presence of MDSC than their wild-type counterparts. MDSC from gp91phox KO mice had significantly lower suppressive activity against antigen-specific T cells than MDSC isolated from wild-type tumor-bearing mice. Arginase activity and nitric oxide production, factors implicated to mediate MDSC-mediated immune suppression, were also decreased in gp91phox KO mice. These findings may help to clarify the role of ROS in cancer and to develop new effective cancer treatment.
The effects of vitamin C on the interaction between COX-2 expression and IGF-I axis in the human melanoma cell line, SK-MEL-2 (49.27)
Authors: Lee Seung Koo , Kang Jae Seung , Kim Jee Eun , Jung Da Jung , Hahm Eunsil , Bae Seyeon , Hwang Young Il , Shin Dong Hoon , Lee Wang Jae 
Published: January 2023

Abstract
Overexpression of cyclooxygenase-2 (COX-2) frequently occurs in several malignancies and the type I insulin-like growth factor receptor (IGF-IR) is important for growth, transformation, proliferation and protection from apoptosis. However the interaction between COX-2 expression and IGF-I axis in human melanoma remains obscure. Although inhibition of COX-2 could reduce tumor volumes in various murine models, selective COX-2 inhibitors could increase cardiovascular risk. Therefore, we studied the effects of vitamin C, which is known as the potent inhibitor of proliferation of human melanoma cell line, SK-MEL-2. We confirmed that the optimal concentration of vitamin C for the inhibition of proliferation without inducing apoptosis is 1.0 mM. ELISA for IGF-I and IGF-II conducted after treatment with NS-398 and vitamin C revealed that the production of IGF-II was decreased by COX-2 inhibition. RT-PCR for IGF-IR after COX-2 inhibition using NS-398 or COX-2 siRNA showed the inhibition of COX-2 expression led to dramatic down-regulation of IGF-IR expression which is restored by adding exogenous PGE2, which is produced by the enzymatic action of COX-2. Next we observed that both COX-2 and IGF-IR expression were significantly decreased after treatment of 1.0 mM vitamin C. Interestingly exogenously treated IGF-II induced IGF-IR expression and exogenous PGE2 treatment restored IGF-IR expression which is down-regulated by vitamin C. These findings suggest that vitamin C could modulate IGF-II production and then down-regulate the expression of IGF-IR by the inhibition of COX-2 expression, followed by suppression of the proliferation of human melanoma cell line, SK-MEL-2.
Analysis of lymphocyte subpopulations stimulated in vitro with human dialyzable leukocyte extracts (DLE) (49.33)
Authors: GRANADA MARISOL DE LA FUENTE , ESTRADA-PARRA IRIS CITLALLI , CHACON-SALINAS ROMMEL , ESTRADA-PARRA SERGIO ANTONIO , PEREZ-TAPIA SONIA MAYRA 
Published: January 2023

Abstract
Dialyzable leukocyte extracts (DLE) are obtained from freezed-thawed buffy coats and subsequent dialysis ( &lt;12 KDa). They can transfer cellular immune response from an immune donor to a naïve recipient. DLE have been used as immunomodulators for the last 50 years with good results. Until now little research has been done regarding their mechanism of action. The aim of this work was to analyze, using flow cytometry, if in vitro stimulation of PBMC with a range of DLE concentrations (50 microg, 50 ng, 50 pg, 50fg and 50ag) induced proliferation or activation of different lymphocyte subpopulations. Peripheral blood mononuclear cells (PBMC) were purified using a density gradient and viability was assessed using trypan blue exclusion stain. CFSE stain was performed to measure proliferation of CD3+/CD4+ (Th), CD3+/CD8+ (Tc), CD19+ (B) and CD56+ (NK) cells after 24, 48 and 72h. CD69 expression was measured on the same cell subpopulations after 3, 6, 24 and 48 h. Controls included PMA+ionomicin and non stimulated cells. Results showed that DLE increased CD69 expression only in the Th (CD3+/CD4+) population (50 microg, 50 ng, 50 pg). This subpopulation also proliferate more in comparison with Tc, B or NK cells. Next experiments will analyze if these cells that activated and proliferate upon DLE stimulation express other markes, such as CD25 and Foxp3. Our results may explain in part the immunomodulatory effects that these extracts have.
This work was supported in part by SIP/IPN. 1Are fellows of COFAA, EDI and SNI.
Ligation of CD40 expressed constitutively on breast cancer cell line induces increased TGF-β production (49.8)
Authors: Jung Da Jung , Kang Jae Seung , Kim Jee Eun , Hahm Eunsil , Lee Seung Koo , Bae Seyeon , Hwang Young-Il , Shin Dong Hoon , Lee Wang Jae 
Published: January 2023

Abstract
CD40 has been shown to play a critical role in normal B-cell development and function. Interestingly, CD40 has also demonstrated to be expressed at high levels on some human cancers, including the breast cancer. However, the function of CD40 expressed on tumor cells is still not fully understood. In this study, therefore, the function of CD40 on tumor cells in the aspect of the production of Transforming Growth Factor-beta (TGF-β) was investigated after CD40 stimulation. First, high expression of CD40 on the human breast cancer cell line, MDA-MB231 was observed. The tumor cells were incubated for 24 hr in the presence of agonistic anti-CD40 antibodies or sCD40 ligands, the productions of TGF-β were dramatically increased in each cases. Human T cells purified from peripheral blood were stimulated with PHA and ionomycin. And then cells were cultured with MDA-MB231 for 24 hr. As a result, significantly increased production of TGF-β was observed. Taken together, TGF-β production by breast cancer cells increased via the ligation of CD40 on their surface with CD40L on the surface of T cells. Thus, considering that TGF-β serves dual roles not only in promoting tumor progression and metastasis, but also in immune suppression, these results give an important clue for understanding the mechanism of the immune suppression by tumor cells.
Differential antitumor activity between IFN-lambda and IL-21 (50.18)
Authors: Sato Atsuko , Ohtsuki Mamitaro , Kobayashi Eiji , Murakami Takashi 
Published: January 2023

Abstract
Interferon (IFN)-lambdas are the latest members of the class II cytokine family and exhibit antiviral activity. Murine B16 melanoma and Colon26 cancer cells were transduced with mouse IFN-lambda to determine whether IFN-lambda possesses antitumor activity. Over-expression of IFN-lambda induced Fas, promoted significantly caspase-3/7 activity, and increased p21Waf1/Cip1 and dephosphorylated Rb (Ser780) in B16 cells in vitro. IFN-lambda expression in tumor cell lines markedly inhibited subcutaneous and metastatic tumor formation in vivo. Moreover, an antibody-mediated immune cell depletion assay showed that natural killer (NK) cells were critical to IFN-lambda-mediated tumor growth inhibition. Hydrodynamic injection of IFN-lambda cDNA successfully targeted liver metastatic foci of Colon26 cells, and moderately decreased the mortality of mice with tumors. IFN-lambda over-expression in the liver increased NK cells (2.4 fold) and enhanced their tumor killing activity. However, the antitumor activity of IFN-lambda was weaker than that of IL-21, which also promotes NK-cell proliferation and stimulation. Thus, although IFN-lambda induced both tumor apoptosis and NK cell-mediated immunological tumor destruction, the antitumor effect of IFN-lambda was smaller than that of IL-21.
In vivo analysis of early events leading to T helper cell differentiation (51.15)
Authors: Rozo Cristina T , Liu Qian , Gause William 
Published: January 2023

Abstract
CD4+ T cells are able to respond to various pathogens by differentiating into highly polarized T helper effector (Th) cells that regulate distinct immune responses. Bacteria and viruses can elicit a Th1 response and most helminth parasites can induce a Th2 response. Although, dendritic cells (DCs) are the most potent activators of naïve T cells in vitro, it remains unclear which cell population is most important under in vivo conditions to cause the polarization into effector groups. Naïve CD4+ T cells reside in T cell areas of secondary lymphoid organs and DC’s are the predominant MHC-II expressing cells in the T cell areas. To examine in vivo what antigen presenting cells (APCs) and accessory cells are associated with early stages of the Th2 response C57BL/6 mice received a subcutaneous ear injection in the context of an infectious helminth infection or Th1 inducing agent either in the presence or absence of Ealpha peptide. Local draining lymph nodes were harvested and flow cytometry in conjunction with immunohistochemical techniques were used to visualize antigen presentation using Y-Ae antibody directed towards MHC II complexed with I-E alpha chain and to identify the cell populations participating in the development of the Th2 versus the Th1 immune response. We observed that under Th2 conditions, there was an increase in DX5+ basophils and CD11c+ APCs as early as four hours after immunization.
Neutrophils play critical role for the type 2 protective responses against nematode parasite N. brasiliensis (51.5)
Authors: Liu Zhugong , Pesce John T. , Hamed Hossein , Alem Farhang , Lin Hongxia , Chaussabel Damien , Liu Qian , Urban Joseph F. , Gause William C. 
Published: January 2023

Abstract
Nippostrongylus brasiliensis (Nb) infection induces a potent Th2 response; however, little is known about the innate host-pathogen interactions that lead to the protective Th2 immunity. To identify these early events, microarray analysis was performed on draining skin lymph nodes shortly after Nb inoculation. Expression of several chemokines, including CCL2 and CXCR3 ligands, were found increased. We observed a transient and marked increase in Gr-1+ cells in the draining lymph node after Nb inoculation, which was CCL2-dependent. The Gr-1bright population was identified as neutrophils expressing TGFβ and TNFα. We further demonstrated that neutrophils were required for host protection against Nb infection and for the optimal Th2 development. After neutrophil depletion, Nb infection resulted in early IFN-γ expression, high mortality and delayed worm expulsion. Decreased Th2 cytokine expression and elevated serum IgG2a were also observed. In contrast, infection with Nb larvae treated with antibiotics cocktail to eliminate associated bacteria induced a protective Th2 response in neutrophil-depleted hosts. These studies suggest that the primary function of neutrophils in this model was to eliminate bacteria associated with parasite infection. Neutrophils thus play an essential role in depleting Th1-inducing bacteria, thereby promoting optimal development of the host protective Th2 response.
Perforin-2 Expression in Fibroblasts is Upregulated during dsRNA Stimulation. (52.12)
Authors: Ramos Jahir , Shiratsuchi Motoaki , Deyev Vadim V , Podack Eckhard R 
Published: January 2023

Abstract
Perforin-2 (P2), also referred to as Mpeg1, Mpg1, and MPS1 in the literature, was originally reported as a macrophage specific molecule. Previous reports demonstrated that P2 mRNA was exclusively detected in macrophages and macrophage-like cells. This is the first report that P2 is upregulated in murine fibroblasts during dsRNA or interferon exposure.
Cells were transfected via lipofection with Poly I:C, an analog of viral double stranded RNA. In a separate trial the culture supernatant of Poly I:C- transfected cells was harvested and added to a separate cell culture. Since Poly I:C induces the secretion of interferon, recombinant interferons were used individually and in concert to stimulate a boost in P2 transcription. We also performed promoter assays using luciferase constructs containing various lengths of P2 promoter sequences.
Real time RT_PCR analysis revealed that upon treatment with Poly I:C, P2 mRNA level was increased dramatically. Similar effects were observed following treatment with culture supernatant from Poly I:C-transfected cells. Interferon-alpha, beta, and gamma all induced P2 mRNA upregulation and we found the effect to be synergistic. Our luciferase promoter assay demonstrated that Poly I:C strongly activated an ectopic promoter in exon 1.
Our results indicate that dsRNA stimulation followed by interferon induction increased P2 transcription from an ectopic promoter. We have identified a putative anti-viral responsive gene in murine fibroblasts encoding the membrane anchored P2 protein.
Catalytically inactive cyclooxygenase 2 (COX-2) and absence of PGE2 biosynthesis in murine peritoneal macrophages following in vivo phagocytosis of heat-killed Mycobacterium bovis BCG (52.17)
Authors: Shibata Yoshimi , Shinohara Tsutomu , Tsuji Shoutaro , Henriksen Ruth Ann , Nishiyama Akihito , Myrvik Quentin N , Yamashita Makiko 
Published: January 2023

Abstract
The conclusion that peritoneal macrophages (MØ) isolated from mice given heat-killed BCG (HK-BCG) intraperitoneally do not release PGE2 ex vivo has been established for over 25 years. However, when peritoneal MØ from untreated mice are treated in vitro with HK-BCG, MØ express COX-2, a rate-limiting enzyme for PGE2 biosynthesis, with increased PGE2 release. The present study of C57BL/6 mouse peritoneal MØ treated in vitro or in vivo with HK-BCG was undertaken to determine whether COX-2 is expressed and whether PGE2 release is dependent on its catalytic activity. The results indicate that MØ treated with HK-BCG in vivo expressed COX-2 that was catalytically inactive and localized subcellularly in the cytoplasm but not associated with the nuclear envelope (NE). In contrast, MØ treated in vitro expressed catalytically active COX-2 that was localized in the NE and diffusely in the cytoplasm. In addition, COX-1, constitutively localized in the NE and ER in untreated M , was dissociated from the NE in response to in vivo but not in vitro HK-BCG treatments. These results indicate that COX-1 and COX-2 dissociated from the NE are catalytically inactive, which accounts for the lack of PGE2 production by local MØ activated in vivo with HK-BCG. Our studies suggest that following phagocytosis of intracellular bacteria, local MØ express catalytically inactive COX-2, which may enhance M -mediated innate and Th1 acquired immune responses against intracellular infections sensitive to endogenous PGE2. (NIH RO1 HL71711, DOD DAMD 17-03-1-0004)
Priming of γδ T cells (52.4)
Authors: Hedges Jodi Fern , Buckner Diana , Rask Kerri , Kerns Hannah , Jackiw Larissa , Trunkle Theresa , Pascual David , Dobrinin Erin , Radke Miranda , Holderness Jeff , Jutila Mark 
Published: January 2023

Abstract
We have begun to elucidate the role of γδ T cells in innate immunity. Purified human and bovine γδ T cells display a subtle activation, akin to priming, in response to a wide variety of pathogen associated molecular patterns (PAMPs) such as lipopolysaccharide and peptidoglycan. A primed γδ T cell displays an enhanced response to downstream secondary signals compared to resting cells, but is not overtly activated. The signature response of a primed γδ T cell include increases in Mip1α, GM-CSF, and IL-2Rα gene and protein expression, that renders them more sensitive to IL-2. Priming is distinct from overt activation by known TCR agonists in that upregulation of IFNγ and TNFα, the signature of an activated γδ T cell, are rarely detected. Priming has also been detected in vivo in γδ, but not αβ, T cells derived from the gut mucosa of Salmonella infected calves. In an effort to manipulate γδ T cells to provide an innate protective response, we have identified compounds and herbal products that mimic the priming activation of PAMPs on γδ T cells. Treatment of bovine calves with one of the plant extracts protects against Salmonella-induced enterocolitis. γδ T cell priming may be a critical step in innate immune protection and clearance, especially for pathogens that invade mucosal surfaces, where the majority of γδ T cells are found.
Immune opsonins modulate BLyS/BAFF release in a receptor-specific fashion (53.6)
Authors: Li Xinrui , Su Kaihong , Szalai Alexander J , Zhou Tong , Kimberly Robert P , Edberg Jeffrey C 
Published: January 2023

Abstract
TNF ligand superfamily member 13B (B lymphocyte stimulator (BLyS), B cell activating factor (BAFF)) promotes primary B cell proliferation and immunoglobulin production. It exists in both membrane-bound and soluble forms. Because the soluble form is thought to be the primary biologically active form, we investigated factors that might regulate its cleavage and processing.
In both myeloid cell lines and primary human monocytes, the shedding of membrane BLyS can be regulated by CRP and IgG. Within 10 minutes, both of these opsonins trigger significant shedding of BLyS (up to 52.2±3.0%, p=0.003) by engaging FcγRs. In U937 cells, a primary role of FcγRI is demonstrated both by direct receptor cross-linking (FcγRI 45.69±2.05% verses FcγRIIa 2.11±1.33%) and by the lack of enhanced cleavage when FcγRI and FcγRIIa are co-crosslinked. The shed BLyS is biologically functional since it promotes B cell survival. The generation of a B cell proliferation and survival factor by Fcγ receptor engagement, especially FcγRI, which can also enhance antigen uptake and presentation mediated by ligands of both innate and adaptive immune systems, provides a unique opportunity to facilitate antibody production. These functions suggest that Fcγ receptors may provide a link between immune complex mediated autoimmune diseases and elevations in circulating BLyS in autoimmune disease patients.
Supported by NIH R01AR042476 and R01AR033062.