bluuebunny/crossref_metadata_2025_split huggingface dataset:
NKG2D optimizes CD8+ T cell function while certifying them to become memory cells
Authors: Perez Cynthia , Prajapati Kushal , Burke Brianna , Plaza-Rojas Lourdes , Guevara-Patiño José A. 
Published: January 2023

Abstract
The ability to form memory is a major characteristic of the adaptive immune system. NKG2D, as a costimulatory molecule for CD8+ T cells, was shown to rescue memory formation in absence of CD4+ help. However, what exactly is the function of NKG2D on CD8+ T cells and how important is NKG2D for memory formation is not known. To recapitulate a full course of CD8+ T cell activation, from priming to memory recall, we immunized C57BL/6 mice with peptide-pulsed dendritic cells and followed the CD8+ T cell responses at priming, effector and memory phases. Using NKG2D deficient (NKG2D-KO) CD8+ T cells, we found that NKG2D signaling was not necessary to achieve CD8+ T cell priming. However, NKG2D-KO CD8+ T cells showed lower effector functions (reduced in vivo kill and reduced cytokine secretion upon in vitro restimulation) compared to wildtype CD8+ T cells. Blocking NKG2D signaling at the effector phase with a single injection of anti-NKG2D antibody (non-depleting) recapitulated the results observed with NKG2D-KO CD8+ T cells. Importantly, the CD8+ T cells that lacked NKG2D signaling at effector phase failed to differentiate into protective memory cells. These results indicate that NKG2D signaling certifies CD8+ T cells to become memory cells and that this certification occurs during the effector phase.
Beta-glucan-induced trained innate immunity mediates antitumor efficacy in the mouse lung
Authors: Alexander Matthew P , Fiering Steven N , Ostroff Gary R , Cramer Robert A , Mullins David W 
Published: January 2023

Abstract
The innate arm of the vertebrate immune system has classically been regarded as nonspecific and lacking the capacity for adaptive responses, in contrast to the adaptive arm defined by its robust memory responses. Recent high impact work has demonstrated enhanced non-specific responses of innate immune cells to pathogens and malignancy after priming with the fungal cell wall component beta-glucan, a phenomenon termed trained innate immunity (TII). In a pathogen model, the training effect was mediated by epigenetic modifications underlying a metabolic shift to mTOR- and HIF1α-mediated aerobic glycolysis. To investigate whether these same pathways play a role in non-specific enhancement of antitumor responses, we assessed beta-glucan-mediated anti-tumor efficacy in a mouse model of metastatic melanoma. We observed that systemic pretreatment with a particulate beta-glucan significantly diminished the growth of metastatic-like B16 melanoma in the lungs, but not initial tumor cell engraftment. Further, lungs in beta-glucan treated animals had a robust myeloid immune infiltrate, particularly neutrophils and monocytes. Interestingly, initial studies demonstrate that tumor suppressed HIF1α expression, and beta-glucan pretreatment prevented this suppression. These results are consistent with beta-glucan induction of TII, leading to suppression of cancer growth. By extension, TII may modulate the immunogenic nature of the tumor microenvironment, thereby acting as an adjuvant to enhance the efficacy of existing therapies.
Development of a Curative Immunotherapeutic Strategy for the Treatment of Big Established CT26 Tumors
Authors: Nie Yingjie , Oppenheim Joost J , Yang De , Trivett Anna , Xin Haiyun 
Published: January 2023

Abstract
Immunotherapy has demonstrated the ability to cause some complete clinical remissions even in patients with late stage cancers. However, currently used approaches fail in many cases. We hypothesize that a combination of promoting the generation of antitumor immune response and reverting the immunosuppressive environment of cancers would be more effective. In this study, we investigated the therapeutic effect of a combination of cytoxan and HMGN1 on established CT26 tumors. HMGN1 is a Th1-polarizing alarmin capable of promoting antitumor immune protection, whereas cytoxan (CY) reduces regulatory T cells. Treatment with one dose of CY and four doses of HMGN1 completely inhibited tumor growth in Balb/c mice bearing small CT26 tumors, but only partially in mice bearing big (≈1.0 cm diameter) tumors. To improve the efficiency, R848 was added to the treatment combination. The triple combination (‘TheraVac’) eradicated big CT26 tumors. Importantly, the resulting tumor-free mice were resistant to rechallenge with CT26, but not resistant to unrelated 4T1 tumors, indicating the development of protective anti-CT26 immunity. Profiling of tumor infiltrating leukocytes revealed that there was a reduction of dendritic cells and a concomitant increase in T cells in the tumor after TheraVac treatment. Thus, we have developed a curative immunotherapeutic strategy for big established CT26 tumors.
HPV-E1 protein immunization promotes anti-tumoral CTL immune response in C57BL-6 mice.
Authors: Amador Alfredo , Trejo-Moreno Cesar , Sada-Ovalle Isabel , Romero Damaris , Lizano Marcela 
Published: January 2023

Abstract
Cellular immune response is the main immune response to eliminate human papillomavirus (HPV) infected cells. HPV vaccines are effective in preventing HPV infections through humoral response, but fail to protect against many HPV types and established infections. Approximately 15% of women infected with high risk HPV do not have an effective immune response.
In the natural history of HPV infection, the most conserved protein among human papillomavirus, helicase E1, is the first immunological target of HPV infected cells. Therefore, we characterized the cellular immune response against E1 protein in a mouse model.
Immunizations with E1 protein + αGalacsosyl Ceramide (αGalCer) promoted an E1-antigen specific CTL response in C57BL-6 mice. We measured a CD107a/b+Granzime-A kinetic to identify antigen specific populations through flow cytometry.
The anti-tumoral response is only present in those mice immunized along with the αGalCer adjuvant. Within the set of six mice, three of them did not have any tumors and the other three developed small tumors in comparison with the mice in the control group, which were not immunized. In conclusion, the E1-antigen specific CTL plays an important role against tumor growth expressing E1-Carboxyl terminal domain.
Adoptive cell transfer enhances antitumor response generated by Cytomegalovirus-based vaccine
Authors: Grenier Jeremy M , Qiu Zhijuan , Khanna Kamal Mohan 
Published: January 2023

Abstract
Methods for generating long-lived T cell responses to tumor antigen have been highly sought for potential therapy against many cancers. Our group has recently developed a novel vaccine against melanoma using recombinant Mouse Cytomegalovirus (MCMV) expressing a modified tumor antigen which generated life-long CD8+ T cell responses to the melanoma antigen, gp100. While this vaccine delays tumor growth in a mouse model of B16 metastatic melanoma, mice eventually succumb to the disease. Adoptive cell therapy has shown clinical efficacy in some patients with melanoma; however, it requires in vitro activation of large numbers of T cells to be effective. We hypothesized that combining these therapies would result in more effective tumor rejection by 1) increasing precursor frequency of tumor specific T cells, thus inducing a more robust T cell response following vaccination and 2) promoting life-long persistence of transferred cells following transfer. To test this, C57BL/6 mice received 105 CD8+ T cells expressing a transgenic T cell receptor recognizing gp100 (PMEL) followed by vaccination. Five days after transfer, PMEL cells constituted up to 20% of all CD8+ T cells in peripheral blood following MCMV vaccination in contrast to 7.5% following vaccination with another viral vaccine, Vesicular Stomatitis Virus. Our results also show that adoptive transfer and MCMV vaccination delays B16 tumor growth and improves survival in a solid tumor model. In summary, this study tests a novel method for stimulating adoptively transferred antitumor T cells, while avoiding the need for long-term in vitroculture of cells. Future work will also examine if immune checkpoint inhibitors or costimulatory agonists further improve this therapy.
IL-21 dampens NFkB activation to augment efficacy of CD8+ T cell mediated adoptive cell therapy (ACT)
Authors: Shrikant Protul A , Myles Arpita 
Published: January 2023

Abstract
Exploiting mechanisms integrating antigen and cytokine signals to generate tumor-antigen specific CD8+ T cells that are durable and produce low proinflammatory profile can boost efficacy of adoptive cell therapy (ACT) of cancer. Herein, we show that IL-21 via STAT3 induces the atypical IKK family member, Bcl3, to dampen antigen driven NF-kB (p65/p50) activation. The IL-21 induced Bcl3 decreases antigen driven CD8+ proliferation as well as apoptosis to increase clonal expansion. Strikingly, Bcl3 skews expression of master transcriptional factors to facilitate transition of polyfunctional CD8+ effector cells to memory precursor cells (MPCs) with cytotoxicity but reduced IFNg production. The IL-21 conditioned CD8+ T cells are persistent and produce greater tumor efficacy upon adoptive transfer. These studies have revealed a role for NF-kB to serve as a node for integrating antigen and cytokine signals that determine fate of antigen stimulated CD8+ T cells and encourage development of new approaches to target NFkB for T cell mediated immunity.
Targeting T cell senescence in the tumor microenvironment for tumor immunotherapy
Authors: Peng Guangyong , Mo Wei , Li Lingyun , Liu Xia , Zhong Xiaoli , Ye Jian , Hsueh Eddy C. 
Published: January 2023

Abstract
Understanding the molecular mechanisms involved in creating and sustaining the tumor suppressive microenvironment is critical for the development of novel anti-tumor therapeutic strategies. Regulatory T cells (Treg) are a key player for the tumor immunosuppressive microenvironment. However, the molecular processes and functions of suppressed responder T cells mediated by Treg cells remain largely unknown. We discovered a novel suppressive mechanism whereby human Treg cells induce senescence in naïve and effector T cells that then exhibit potent suppressive activity and amplify immune suppression. Treg-induced senescent T cells were distinctly different from the exhausted or anergic T cells based on the transcriptome analyses and phenotypic profiles. We further identified that the nuclear kinase ataxia-telangiectasia mutated protein (ATM)-induced DNA damage was critical and the main cause for the induction of responder T cell senescence and dysfunction mediated by human Treg cells. In addition, MAPK ERK1/2 and p38 were involved in the regulation of senescence process in responder T cells. Importantly, we further revealed that human Treg-induced senescence and suppressor function could be blocked by specific ATM signaling and/or ERK1/2 and p38 signaling inhibition in vitro and in vivo in animal models. Our studies identify a novel molecular mechanism responsible for human Treg cell suppression, and provide new insights relevant for the development of strategies capable of preventing and/or reversing Treg-induced immune suppression for tumor immunotherapy.
Funding Support: Supported by the grants from the Melanoma Research Alliance, American Cancer Society and the National Institutes of Health.
Mice with highly metastatic breast cancer transfer express exosomes in serum which increase metastatic capacity of a poorly metastatic tumor
Authors: Gorczynski R. M , Erin Nuray , Zhu Fang 
Published: January 2023

Abstract
Altered interaction between CD200 and CD200R, represents an example of “checkpoint blockade” disrupting an effective, tumor-directed, host response in murine breast cancer cells. In CD200R1KO mice, long-term cure of EMT6 breast cancer, including metastatic spread to lung and liver, was achieved in BALB/c mice. The reverse was observed with 4THM tumors, an aggressive, inflammatory breast cancer, with increased tumor metastasis in CD200R1KO.
To explore possible explanations for this difference we measured the frequency of circulating tumor cells (CTCs) in peripheral blood of tumor bearers, as well as in lung/liver and draining lymph nodes, and in addition infused exosomes from non-tumor controls, or tumor bearers, with/without additional infusions of anti-cytokine antibodies into tumor bearing mice.
The measured frequency of circulating tumor cells (CTCs) in peripheral blood was equivalent in the two models in both WT and CD200R1KO mice. Increased metastasis in EMT6 tumor bearers was seen following adoptive transfer of serum, or serum-derived exosomes, from 4THM tumor bearers, an effect which was attenuated by anti-IL-6, and anti-IL-17, but not anti-TNFa, antibody. Anti-IL-6 also attenuated enhanced migration of EMT6 cells in vitro induced by 4THM serum or exosomes, or recombinant IL-6. Exosome cytokine proteomic profiles responses in 4THM and EMT6 tumor bearer mice were regulated by CD200:CD200R interactions, with attenuation of both IL-6 and IL-17 in 4THM CD200tg mice, and enhanced levels in 4THM CD200R1KO mice.
We hypothesize that these cytokines act on the microenvironment at sites within the host, and/or directly on tumor cells themselves, to increase metastatic potential.
Mucosal vaccine adjuvant cyclic di-GMP induces STING-dependent pulmonary dendritic cell activation
Authors: Mansouri Samira , Blaauboer Steven , Jin Lei 
Published: January 2023

Abstract
The mucosal surface is the major entry site for many human pathogens, however there are only a dozen of mucosal vaccines approved for human use. The challenge for mucosal vaccines is to develop safe and effective mucosal adjuvants. 3′, 5′-Cyclic di-GMP (CDG) is an attractive mucosal vaccine adjuvant candidate that activates STING (stimulator of interferon genes). However, the in vivo mechanism of action of CDG is not completely understood. Our previous studies found that intranasal immunization with CDG directly activates pulmonary dendritic cells (DCs). We found that CDG adjuvant activity depends on TNFα and not type I interferon. Deletion of STING in pulmonary DCs resulted in the reduction of antigen-specific antibodies and a decrease in Th1/Th2/Th17 responses. Here, we investigated the role of different pulmonary DCs subsets in mediating the mucosal adjuvant activity of CDG. Our results demonstrate that CDG targets unique and specialized functions of pulmonary DC subsets. Using Batf3−/− mice to deplete CD103+ DCs, we found that CD103+ DCs do not mediate the CDG-induced antibody production, but are needed for Th responses as well as the recruitment of the novel CD103+CD11b+ pulmonary DC population. The population of pulmonary CD11b+ DCs was intact in the Batf3−/− mice. CDG-induced humoral immunity completely depends on pulmonary DCs. Our data, thus, suggested that CD11b+ DCs, which were the only pulmonary DCs not depleted in Batf3−/− mice, were sufficient for CDG-induced humoral immunity. How STING mediates CDG responses in these DC subsets will be discussed. Understanding the role of different DC subsets will reveal insight in the mode of action of CDG in vivo and advance the development of CDG as an efficacious mucosal vaccine adjuvant.
An oral prime/boost pediatric vaccine strategy for the prevention of mother-to-child transmission of HIV by breast-feeding
Authors: Curtis Alan Dale , Phillips Bonnie , Choudhary Neelima , Tuck Ryan , Lifson Jeff , Van Rompay Koen , Kozlowski Pamela , De Paris Kristina 
Published: January 2023

Abstract
In 2014, an estimated 220,000 children became newly HIV-infected, with 50% being transmitted via breast-feeding. The present study assessed whether inclusion of an oral route of immunization would increase pediatric vaccine efficacy in the rhesus macaque model of oral SIV infection.
Two groups of 6 neonatal macaques received 2 DNA-SIV immunizations at weeks 0 and 3, followed by 2 boosts with modified vaccinia Ankara virus (MVA)-SIVgag/pol/env at weeks 6 and 9. Group A was immunized by the intramuscular (IM) route. Infants in Group B received the DNA-prime both per-orally (PO) and IM and MVA-SIV was administered both by the sublingual (SL) and IM route. Mock control animals received saline immunizations PO+IM at weeks 0 and 3 and mock MVA vector SL+IM at weeks 6 and 9. At week 12, all infants were challenged once weekly using a low-dose (102 TCID50) oral SIVmac251 challenge regimen until systemic infection was confirmed by PCR. Statistical significance was determined using nonparametric Mann-Whitney or Kruskal-Wallis tests with p&lt;0.05.
Infants vaccinated by the SL/IM route had a significantly lower per-exposure risk of oral SIV infection (p&lt;0.0001; log-ranked Mantel-Cox test) compared to mock- and IM/IM-vaccinated animals and lower peak viremia (p=0.03) compared to mock controls. Control of viremia was associated with higher fecal IgG responses to SIV V1V2 (p=0.0087) and p55 gag (p=0.0087) at the time of oral challenge initiation. In contrast, plasma SIV-specific antibodies did not influence challenge outcome.
These data indicate that the inclusion of an oral route in a pediatric vaccine to prevent oral SIV infection is superior to an IM only regimen and emphasizes the importance of enhancing mucosal immune responses via local immunization.
Deciphering the Role of Suppressor of Cytokine Signaling (SOCS) 1 and 3 in the IL-10 Regulation of Inflammation Induced by Live Chlamydia trachomatis in mouse macrophages
Authors: Duncan Skyla A. , Dixit Saurabh , Sahu Rajnish , Singh Shree R. , Dennis Vida A 
Published: January 2023

Abstract
Chlamydia trachomatis (CT), a bacterial pathogen responsible for sexually transmitted infections worldwide, causes severe inflammation including but not limited to, pelvic inflammatory disease, ectopic pregnancy and infertility in women. Secretion of inflammatory mediators plays a pivotal role in its pathogenesis of disease. Our laboratory published that IL-10 can down-regulate the secretion of inflammatory cytokines triggered by CT in mouse macrophages. Our goal in this study is to decipher the mechanisms of IL-10 down-regulation of CT-induced cytokines by focusing on the intracellular negative regulatory mediators, suppressor of cytokine signaling (SOCS) 1 and 3. We show that live CT and its recombinant major outer membrane protein (rMOMP), in a dose-dependent fashion, markedly induced the transcriptional upregulation of SOCS 1 and 3 in macrophages. In the presence of exogenously added recombinant IL-10, SOCS1 expression was reduced while that of SOCS3 was enhanced, suggesting differential regulation of these regulators by IL-10 to mediate its inhibitory effect. Experiments using various concentrations of exogenously added IL-10 over several time-points further revealed the differential regulation of SOCS 1 and 3 by IL-10 in macrophages. Signal transduction pathway inhibition studies demonstrated that the MAPK pathways including p38, MEK and JNK, particularly the latter are possibly involved in inducing SOCS 1 and 3 by CT and added IL-10, correlating with down-regulation of the cytokines, IL-6 and IL-12p40. Overall, our data helps in understanding the mechanisms of IL-10 regulation of inflammation in CT-infected macrophages by shedding light on the roles of SOCS 1 and 3 in mediating this IL-10 anti-inflammatory effect.
Characterizing the germinal center response to electroporated DNA immunogens
Authors: Gary Ebony Nicole , Bernui Mariana , Kutzler Michele A. 
Published: January 2023

Abstract
Most licensed vaccines rely on the production of antibodies that mediate protection from disease. The germinal center (GC) is a dynamic cellular substructure found in lymphoid organs where follicular helper T cells (TFH) provide the necessary proliferation stimuli and cytokine signaling that promote antibody class-switching, somatic hypermutation, and affinity maturation of B cell receptors. Here, mature B cells differentiate into long-lived antibody-secreting cells and memory B cells, instituting the characteristic humoral memory response to vaccination. Although HIV-1 DNA vaccines have been shown to be highly immunogenic, leading to enhanced HIV-1 env epitope breadth and titers in pre-clinical and clinical studies, many questions remain regarding the quality of the humoral immune response and the ability of the DNA antigen/molecular adjuvant to induce long-lived anti-viral antibody that seeds mucosal secondary immune tissues. However, the magnitude and kinetics of the GC response to EP DNA has not been elucidated. Our results indicate that DNA antigens induce a GC response. Antigen-specific GC B cells (B220+CD138−IgM−IgD−PNA+CXCR5+gp120+) are successfully detected by flow cytometry using fluorophore-conjugated HIV-1 gp120 protein after two immunizations in the spleens and draining lymph nodes of vaccinated animals and by antigen-specific ELISpot assay. The presence of antigen-specific antibodies of multiple isotypes is successfully detected in the serum of vaccinated animals via ELISA. These results represent, to our knowledge, the first reports on the magnitude and kinetics of the germinal center response to EP DNA. This work is funded by a W.W. Smith Charitable Foundation grant to MK.
Mature B cell deficient-BAFFR−/− mice can be protected from a lethal WNV-Tx (WNV) infection by vaccination with an anti-CD180 (RP105) antibody - WNVEnv protein conjugate (αCD180-WNVE)
Authors: Giordano Daniela , Draves Kevin E , Bryan Marianne A , Dresch Christiane , Young Lucy B , Roe Kelsey , Richner Justin M , Diamond Michael S , Gale Michael J , Clark Edward A 
Published: January 2023

Abstract
BAFFR−/− mice are a model of humoral immunodeficiency since they lack mature B cells. We found that in contrast to WT mice BAFFR−/− mice were highly susceptible to WNV. BAFFR−/− mice infected with 100pfu WNV succumbed to infection between 8 to 12 days post-infection and higher virus titers were found in their brains compared to WT mice. Infected BAFFR−/− mice had substantially reduced WNVE-specific IgG responses and delayed but still detectable WNV neutralizing Ab compared to WT mice. Passive transfer of immune sera from previously infected WT mice rescued BAFFR−/− mice as well as μMT mice lacking all B cells, but in contrast to μMT mice that all died around 30 days post-infection, BAFFR−/− mice survived and were able to overcome a second WNV challenge. Furthermore, rescued BAFFR−/− mice receiving immune sera later produced their own WNVE-specific IgG, although at much lower levels than WT mice. These data suggested that protective immunity could be induced in these mature B cell-deficient mice. Indeed, one injection of αCD180-WNVE 30 days prior to WNV infection induced strong neutralizing Ab responses in BAFFR−/− mice and significant protective immunity against a normally lethal infection. In contrast, μMT mice could not be protected by αCD180-WNVE immunization confirming that the vaccination requires B cells to induce protective immunity to WNV infection. BAFFR−/− mice have a profound reduction in mature B cells but, unlike μMT mice, have normal numbers of T1 B cells, which our laboratory has shown previously can make IgG Abs. Interestingly, αCD180 immunization rapidly expands T1/T2 B cells. Thus a CD180-based vaccine may be useful to induce protective immunity in immune compromised individuals. (Supported by NIH grants AI83109 and AI52203).