bluuebunny/crossref_metadata_2025_split huggingface dataset:
CTLA-4 limits anti-CD20-mediated tumor regression
Authors: Ren Zhenhua , Peng Hua , Fu Yang-Xin 
Published: January 2023

Abstract
The inhibition of tumor growth by anti-CD20 antibody (Ab) treatment is mediated by antibody- and complement-dependent cytotoxicity in xenograft tumor models. Additionally, anti-CD20 therapy for B-cell lymphoma can result in intrinsic and extrinsic tumor resistance to further Ab treatment. However, adaptive immune response-related resistance has not been well studied in anti-CD20-mediated tumor control, and adaptive immunity has long been underestimated. However, using a syngeneic mouse B-cell lymphoma model, we observed that CD8+ T cells played an essential role for anti-CD20-mediated tumor regression. Mechanistically, anti-CD20 therapy promoted DC-mediated cross-presentation. Importantly, macrophages were also necessary for the increase in the tumor-specific CTL response after anti-CD20 treatment, via the production of type I IFN to activate DC function. Furthermore, adaptive resistance is gradually developed through the CTLA-4 pathway in Treg cells in larger lymphomas. Further blockade of CTLA-4 can synergize with anti-CD20 treatment in anti-tumor activities. Thus, this study reveals the essential role of adaptive immunity for anti-CD20-mediated tumor regression and opens a new avenue to combine anti-CD20 with immunotherapy to overcome the resistance of advanced B-cell lymphoma in the clinic.
Immunotherapy for melanoma using nonviral plasmid DNA based approach
Authors: Heller Richard , Lundberg Cathryn , Edelblute Chelsea , Arpag-Mcintosh Sezgi , Shi Guilan 
Published: January 2023

Abstract
Malignant melanoma is the deadliest form of skin cancer. New immunotherapy approaches including anti-CTLA4, anti-PD-1 and anti-PD1-L1 have improved this prognosis for several patients. While the results obtained with these therapies are encouraging, there is still a need to establish improved therapies. IL-12 is a potent cytokine mediating antitumor activity. However, it has only achieved modest antitumor effects in clinical trials, often accompanied by unacceptable adverse events. We evaluated if intratumoral delivery of a plasmid encoding IL-12 using gene electrotransfer (GET) could be used to induce an anti-tumor response. Effective protocols delivering plasmids with GET directly into tumors induced not only local immune response, but a systemic one as well. We have been evaluating the specific response following pIL-12 delivery as well as the potential to combine with checkpoint inhibitors to determine if the response could be further augmented. Using a B16F10 mouse melanoma model, we show that melanoma exhibited unique immune cell composition within the tumor microenvironment after intratumoral injection of pIL12 with GET. The total number of memory immune cells was markedly increased in pIL12 GET melanoma groups compared with control group. pIL12 GET displayed significant regulation of multiple immune cell types, including CD8+ cells, regulatory T cells and myeloid cells, which were induced to mount a CD8+ immune response. It was also observed that there was an increase in PD-1 expression following pIl-12 delivery. Taken together, these findings suggest a sequence of immune activity following pIL12 GET. Further, these results suggest that pretreatment or simultaneous treatment with IL-12 may augment anti-PD1 therapy.
The inhibitory NKR-P1B:Clr-b recognition axis facilitates detection of oncogenic transformation and cancer immunosurveillance
Authors: Tanaka Miho , Fine Jason H. , Aguilar Oscar A. , Kirkham Christina L. , Belcheva Antoaneta , Martin Alberto , Ketela Troy , Moffat Jason , Allan David S.J. , Carlyle James R. 
Published: January 2023

Abstract
Natural killer (NK) cells express receptors specific for MHC class I (MHC-I) molecules involved in “missing-self” recognition of cancer and virus-infected cells. Here, we elucidate the role of MHC-I-independent NKR-P1B:Clr-b interactions in the detection of oncogenic transformation by NK cells. Ras oncogene overexpression was found to promote a real-time loss of Clr-b on mouse fibroblasts and leukemia cells, mediated in part via the Raf/MEK/ERK and PI3K pathways. Ras-driven Clr-b downregulation occurred at the level of the Clrb (Clec2d) promoter, nascent Clr-b transcripts, and cell surface Clr-b protein, in turn promoting missing-self recognition via the NKR-P1B inhibitory receptor. Both Ras- and c-Myc-mediated Clr-b loss selectively augmented cytotoxicity of oncogene-transformed leukemia cells by NKR-P1B+ NK cells in vitro and enhanced rejection by WT mice in vivo. Interestingly, genetic ablation of either one (Clr-b+/−) or two Clr-b alleles (Clr-b−/−) enhanced survival of Eμ-cMyc transgenic mice in a primary lymphoma model, despite preferential rejection of Clr-b−/− hematopoietic cells previously observed following adoptive transfer into naïve wild-type mice in vivo. Collectively, these findings suggest that the inhibitory NKR-P1B:Clr-b axis plays a beneficial role in the innate detection of oncogenic transformation via NK cell-mediated cancer immune surveillance, in addition to a pathological role in the immune escape of primary lymphoma cells in Eμ-cMyc mice in vivo. These results provide a model for the human NKR-P1A: LLT1 system in cancer immunosurveillance in lymphoma patients, and suggest it may represent a target for immune checkpoint therapy.
Thymoquinone enhanced the tumoricidal activity of NK Cells against Lung Cancer
Authors: Singh Santosh Kumar , Mishra Manoj K. , Lillard James W. , Singh Rajesh 
Published: January 2023

Abstract
Among different immune cell populations, NK cells are an important component in unleashing the therapeutic activity of immune cells. Therefore, in order to enhance the tumoricidal activity of NK cells, identification of new small-molecule natural products is prominent. In this study, we used a natural compound Thymoquinone (TQ), a constituent of Nigella sativa to enhance the tumoricidal activity of NK cell. We expanded human NK cell (NK-92MI) population derived from human peripheral blood mononuclear cells (PBMCs). Next, expanded NK cells were co-cultured with A549, which is a non-small cell lung cancer cell lung cancer (NSCLC) cell, with or without TQ and after 24h of co-cultured, harvested supernatants were analyzed for IFN-g secretions by ELISA method. This study was further validated using flow cytometry; immunofluorescence, qRT-PCR and western blot techniques. Our result showed TQ increased IFN-γ secretion by NK cells and enhanced NK cell-mediated killing of NSCLC cells. The up-regulation of pro-apoptotic (BAX, BID) and down-regulation of anti-apoptotic (BCL-2, MCL-1) makers confirmed that TQ enhanced the NK cell tumoricidal activity, which is a simple, economic and reliable method to create a new strategy to improve the treatment of NSCLC and has the potential to create a modern class of treatments for advanced lung cancer.
Role of liver-resident innate lymphocytes in tumor progress
Authors: Song Hao , Peng Hui , Sun Rui , Tian Zhigang 
Published: January 2023

Abstract
Over the past decade, the incidence of hepatocellular carcinoma (HCC) is increased dramatically around the world, especially in Asia. There are numerous reasons aggravating the development of HCC. As a cytotoxic cell, the NK cell is vital for controlling cancer development. However, NK cells are heterogeneous with distinct subsets, among which CD49a+NK cells, namely tissue-resident NK cells (trNK), has been barely researched in tumor models. Given the enrichment of trNK cells within the liver, our study aims to investigate the specific role of trNK cell in HCC.
In our study, we used a mouse model of colorectal cancer with liver metastasis, through intrasplenic injection of MC-38 tumor cells. In this model, liver CD49a+ NK cells as well as iNKT cells decreased with the tumor development, whereas DX5+ conventional NK (cNK) cells increased. Furthermore, liver cNK cells upregulated the expression of TIGIT, which is not apparently changed on liver trNK cells. These findings suggested that cNK and trNK cells might play opposite roles in liver tumor growth. Consistent with our hypothesis, we found that liver tumor was greatly restricted in mice devoid of iNKT cells, further confirming that iNKT cells exert a pro-tumorigenic role. With respect to trNK cells, it remains to be further validated whether they really facilitate tumor development. Therefore, based on our findings, we hypothesized that liver-resident innate lymphocytes, including iNKT and trNK cells, can promote the immune escape of tumor cells. However, the underlying mechanism in this progress still needs to be investigated deeply and thoroughly.
Reduced germinal center formation is not sufficient to promote cross-reactive protection against influenza.
Authors: Johnson Jenny Lynn , Keating Rachael , Thomas Paul G. , McGargill Maureen A. 
Published: January 2023

Abstract
Influenza infection continues to be a major health threat with over 9 million cases annually in the United States alone. Vaccination is the best method of preventing infection; however, current vaccines are only 10–60% effective due to a high mutation rate of the virus and difficulty predicting which strains will circulate. Furthermore, current vaccines will have little to no impact on a novel pandemic virus. A universal influenza vaccine would generate an immune response to conserved portions of the virus, allowing for protection against multiple influenza subtypes. We showed that a low dose of rapamycin, given during influenza vaccination, altered the antibody response such that mice were protected against secondary infection with multiple strains of influenza. Rapamycin reduced formation of germinal centers (GC) resulting in a decrease in antibodies specific for the variable region of the vaccine HA. Thus, we hypothesized that rapamycin enhances heterosubtypic protection by reducing generation of high affinity antibodies specific for the variable region of HA, which allows antibodies specific for the conserved regions to be more prevalent. To test this, we investigated whether the reduction in GCs was sufficient to induce cross-reactive antibodies. We depleted GCs during vaccination by removing follicular helper T cells or by transiently blocking CD40L. The reduction of GC reduced the amount of class-switched, influenza-specific antibody, similar to rapamycin-treated mice. However, this was not sufficient to enhance protection following challenge with a virus of a different subtype. These data suggest that while rapamycin reduces germinal center formation, this alone is not sufficient to promote cross-reactive antibodies.
Protective efficacy against live Chlamydia muridarum intravaginal challenge in BALB/c mice immunized with MOMP encapsulated PLGA 85/15 nanoparticles
Authors: Sahu Rajnish , Verma Richa , Dixit Saurabh , Duncan Skyla A. , Singh Shree R. , Dennis Vida A. 
Published: January 2023

Abstract
Chlamydia trachomatis (Ct) remains the leading sexually transmitted bacterial infection worldwide but yet no vaccine to prevent its spread. Attempts to develop vaccines using whole or subunits have failed due to inefficient delivery systems. We have recently developed a Chlamydia nanovaccine by encapsulating its recombinant major outer membrane protein (rMOMP) in poly (lactic acid-co-glycolic acid) (PLGA) 85/15 nanoparticles (PLGA-rMOMP). In the current study, we assessed the systemic and mucosal antibody responses and the protective efficacy of the PLGA-rMOMP nanovaccine against a live C. muridarum (Cm) challenged infection in immunized mice. Female BALB/c mice were immunized subcutaneously three times at two weeks interval with 50 μg each of either PLGA-rMOMP or bare rMOMP and sacrificed two weeks after the last immunization to measure rMOMP-specific systemic and mucosal antibodies. Serum and cervico-vaginal washes were collected to measure IgG including subtypes IgG2a, IgG2b (Th1) and IgG1 (Th2) responses by ELISA. Immunized mice were challenged intravaginally with 105 inclusions forming units (IFUs) of Cm, swabs were collected for three weeks and cultured to quantify bacterial shedding by inoculating McCoy cells. PLGA-rMOMP immunized mice showed enhanced systemic and mucosal IgG antibodies response followed by IgG1 and IgG2b, as compared to bare rMOMP. Moreover, the PLGA-rMOMP immunization induced complete (60% of mice) and partial (40% of mice). Our results show that PLGA-rMOMP stimulates systemic and mucosal antibodies and provides protective efficacy against a Chlamydia infection in mice.
Aberrant innate immune activation facilitates EcoHIV evasion from vaccine-induced protection
Authors: Liu Li , Peng Jie , Tang Hangying , Kwok Hauyee , Nishiura Kenji , Lin Qingqing , Fang Jun , Chen Zhiwei 
Published: January 2023

Abstract
Aberrant innate immune response during acute phase of infection causes persistent viral infection through suppression of T-cell function. Understanding the impact of this aberrant innate immune activation on vaccine-conferred protection is critical regarding development of effective vaccine against chronic disease. Here, using EcoHIV mice model and a novel DC-targeting DNA vaccine that elicited high frequencies of GAG-specific, broadly reactive, and polyfunctional cytotoxic CD8+ T cells, we show that aberrant innate immune activation at early stage of infection rendered EcoHIV resistant to vaccine protection by suppressing T cells proliferation and function. We found that despite high frequencies of p24-specific IFN-γ+CD8+ T cells elicited by vaccination, EcoHIV have fallen short of effective recall of antiviral T cell immunity induced by prophylactic vaccination in vivo. Provirus DNA was readily detected in PBMC and spleen in all of the infected mice without detectable resistant mutations. Compared to non-immunized mice, provirus copies in immunized mice were decreased only at 1 and 2 weeks post viral inoculation without further decrease over the following weeks. Further investigation revealed that EcoHIV infected multiple DC subsets, and CD11b+ myeloid derived cells within 7 dpi, and induced quick expansion of eosinophils and monocytic cells in spleen, which peaked at 14 dpi and remained high at 21 dpi. The expansion of monocytic cells was associated with down-regulation of CD80 and CD86, up-regulation of PD-L1/L2 on multiple dendritic cell subsets, PD-1+ and Tim+ CD8 T cells, compromised capacity of DCs in stimulating CD4 and CD8 T cells proliferation, and suppressed viral-specific CD8 T cells proliferation in vitro.
CD8+ T cell-mediated inhibition of HIV-1.
Authors: Xu Yinyan , Abad Maria , Warren Joanna A. , Clutton Genevieve , Goonetilleke Nilu 
Published: January 2023

Abstract
CD8+ T cells contribute to the control of viral infections, including HIV-1. Previous studies have reported that CD8+-mediated virus inhibition is predictive of subsequent HIV plasma viremia and CD4+T cell decline in HIV-infected, antiretroviral naïve individuals; higher virus inhibition is associated with a better clinical outcomes.
Ex vivo CD8+ T cell-mediated inhibition of HIV replication is measured after co-culture of CD8+ T cells with HIV-1 super-infected, autologous CD4+ T cells, at different effector to target ratios. We adapted and standardized the CD8+ T cell virus inhibition assay (CD8-VIA) for use as an endpoint assay in therapeutic testing of HIV-1 T cell vaccines.
CD8+ T cells from HIV infected, durably (&gt;2 years) antiretroviral suppressed participants (n=11) were co-cultured with JR-CSF infected autologous CD4+ T cells at E:T ratios ranging from 1/2:1 to 1/160:1, generating CD8+ T cell virus inhibition curves for each participant. IC50 values varied by 100-fold across participants suggesting that CD8+ T cells harbor a broad range of virus inhibitory capacity even when viremia in participants is durably suppressed.
Ongoing studies are comparing IC50 values to other measurements (magnitude, specificity, avidity, proliferation and production of cytokines and lytic molecules) of HIV-specific T cell immunity to understand the critical determinants of CD8+ T cell inhibitory capacity.
A viral homologue of IPS-1 Interacts with TRAF3 leading to a diminished antiviral state.
Authors: He Buyuan , Sanchez Julio C. , Sanchez David Jesse 
Published: January 2023

Abstract
Kaposi’s Sarcoma (KS) is an angioproliferative disease with symptoms including skin lesions that appear at different sites of the body caused by Kaposi’s Sarcoma Associated Herpesvirus (KSHV). The KSHV genome encodes genes pirated from the human genome that are homologous to cellular proteins that disrupt innate immune pathways. We have found a novel ORF in the KSHV genome that is homologous to the human IPS-1 protein. This viral IPS-1 proteins (vIPS-1) contains the PRO domain of IPS-1 but lacks the CARD and TM domains. We hypothesize that vIPS-1 is able to interact with the cellular TRAF3 protein. Western blot data shows that co-expression of vIPS-1 does not affect RIG-I, decreases the stability of human IPS-1 and TRIF, and increases the stability of TRAF3. Also, vIPS-1 interacts with TRAF3 via coimmunoprecipitation, and by co-expressing vIPS-1 with different TRAF3 mutants we found that vIPS-1 interacts with TRAF3 through the TRAF domain which is specifically required for TRAF3-mediated type I IFN production. Furthermore, our data shows that while expression of IPS-1 is able to inhibit VSV-GFP replication, expression of vIPS-1 significantly enhances the VSV-GFP replication, consistent with our hypothesis that vIPS-1 blocks the cellular immune responses. In conclusion, the KSHV vIPS-1 protein blocks antiviral immunity by interacting with TRAF3 leading to a decrease in antiviral activity.
Discrepancy between neurokinin 1 receptor antagonist treatment and neurokinin 1 receptor knockout mice in the CD8 T cell response to corneal HSV-1 infection
Authors: Jerome Andrew , Suvas Susmit 
Published: January 2023

Abstract
The purpose of this study is to evaluate the effect blocking neurokinin 1 receptor (NK1R) signaling has on the CD8 T cell response to corneal herpes simplex 1 (HSV-1) infection, using NK1R antagonist (L 760735) or NK1R−/− mice. Corneal HSV-1 infection was carried out on C57BL/6 and NK1R−/− mice. Our results showed an enhanced CD8 T cell response to corneal HSV-1 infection by NK1R−/− mice compared to C57BL/6 mice. The data illustrates that at day 5 post infection, draining lymph nodes (DLNs) in NK1R−/− mice exhibit an increased CD8 T cell IFN-γ response as measured by intracellular cytokine staining. In addition, an increased CD8 T cell proliferation is noted in NK1R−/− mouse DLNs. Interestingly, L 760735 treatment given to HSV-1 infected C57BL/6 mice did not show an increase in CD8 T cell proliferation or IFN-γ production; in comparison to infected, vehicle treated control mice. To determine the underlying cause of this discrepancy, we detected pre-existing LYVE+ vessels in unmanipulated corneas of NK1R−/− mice. This was associated with higher levels of VEGF-C protein in ocular tissue (cornea + limbal conjunctiva) of unmanipulated NK1R−/− compared to C57BL/6 mice, as measured via western blot. We recently reported a massive accumulation of CD11b and CD11c expressing cells near the limbal area in unmanipulated eyes of NK1R−/− mice [J Immunol., 2016, 197 (10) 4021–4033]. Confocal microscopy results showed VEGF-C colocalization with these cells near the limbal area in NK1R−/− mice. Since corneal lymphatics play an important role in generating the CD8 T cell response to corneal HSV-1 infection, our results suggest an enhanced CD8 T cell response in NK1R−/− mice, likely due to a notable increase in corneal lymphatics in NK1R−/− compared to C57BL/6 mice.
Female sex hormones regulate the IL-36 family members in the female reproductive tract and impact HSV-2 pathogenesis
Authors: Gardner Jameson K. , Alexander Thessaly , Herbst-Kralovetz Melissa M. 
Published: January 2023

Abstract
Recently, the contraceptive Depo-Provera (DMPA) has been linked to an increased risk of sexually transmitted infection acquisition (e.g. HSV-2 and HIV). However, a fundamental gap exists in the understanding of specific mechanisms whereby DMPA use increases risk for HSV-2 infection. IL-36γ is a novel regulator of mucosal inflammation in the female reproductive tract (FRT) and we have shown that IL-36γ is induced in response to HSV-2. One study reported that IL-36γ was the only cytokine upregulated in cervicovaginal secretions from HSV-2 seropositive DMPA users, suggesting that IL-36γ may be a link between DMPA use and HSV-2 infection. We hypothesize that DMPA suppresses IL-36γ expression in the vaginal environment, thereby increasing susceptibility to genital HSV-2 infection. We found that DMPA treatment in mice decreased expression of the IL-36 cytokines in the FRT, while β-estradiol treatment increased expression. To evaluate the impact of IL-36γ on HSV-2 pathogenesis, we used an intravaginal challenge model and treated mice with DMPA to enhance susceptibility to infection. Mice treated with IL-36γ 4h prior to lethal challenge exhibited a delay in disease onset, less severe disease, decreased vaginal viral replication, increased disease resolution and overall survival. IL-36γ treatment increased levels of pro-inflammatory cytokines in vaginal lavages 4h after treatment, and neutrophil recruitment was enhanced and corresponded with increased levels of chemokines. Together, we demonstrate that IL-36γ is hormonally regulated in the FRT and that treatment with IL-36γ promotes the transient induction of immune mediators and neutrophil recruitment in the vaginal microenvironment that result in increased HSV-2 clearance.
Ectromelia virus suppresses cathepsins and cystatins expression at both mRNA and protein levels in dendritic cells
Authors: Bossowska-Nowicka Magdalena , Mielcarska Matylda Barbara , Kaczmarek Monika M. , Romaniewicz Marta , Struzik Justyna , Toka Felix Ngosa , Szulc-Dabrowska Lidia 
Published: January 2023

Abstract
Cathepsins belong to cysteine proteases and play essential roles in processing of internalized peptides during antigen presentation in the context of the major histocompatibility complex (MHC) class II molecules. Function of cathepsins is regulated by cystatins – endogenous protein inhibitors. Viruses, such as poxviruses, have evolved many mechanisms to escape immune responses after infection. Our studies are focused on ectromelia virus (ECTV, a poxvirus closely related to variola virus, VARV – a causative agent of smallpox), and its influence on cathepsins or cystatins. Our results show that ECTV down-regulates gene and protein expression of selected cathepsins and cystatins in infected murine JAWS II dendritic cells (DCs) and GM-CSF-derived bone marrow cells (GM-BM), composed of conventional DCs and macrophages. Moreover, the ability to endocytose and process a soluble antigen is reduced in JAWS II and GM-BM cells during ECTV infection. After knockdown of cathepsins and cystatins using siRNA in JAWS II DCs, the virus titer increases when compared with control cells. The inhibition of cathepsins and cystatins together with elevated virus titers during the absence of these proteases may be a viral strategy to escape immune responses and simultaneously enable the virus to replicate effectively in infected cells. Importantly, defining the poxvirus-host interactions, including lysosomal proteases, may lead to development of potential therapeutic targets or vaccination strategies.