bluuebunny/crossref_metadata_2025_split huggingface dataset:
Optimizing adoptive cancer immunotherapy with Vγ2Vδ2 T cells through zoledronate pulse stimulation
Authors: Al-Janabi Mohanad Hameed Nada , Wang Hong , Workalemahu Grefachew , Tanaka Yoshimasa , Morita Craig T 
Published: January 2023

Abstract
Human γδ T cells expressing Vγ2Vδ2 TCRs monitor foreign- and self-prenyl pyrophosphate metabolites in isoprenoid biosynthesis to mediate immunity to microbes and tumors. Vγ2Vδ2 cells have been used for adoptive cancer immunotherapy with some partial and complete remissions. Most trials have used continuous zoledronate exposure to expand Vγ2Vδ2 cells. Zoledronate inhibits farnesyl pyrophosphate synthase causing isopentenyl pyrophosphate to accumulate that then stimulates Vγ2Vδ2 cells. Because zoledronate exposure is toxic, we hypothesized that a short period of exposure would reduce T cell toxicity but still be sufficient for monocytes uptake. Supporting this, pulse zoledronate exposure with IL-2 resulted in more uniform expansion of Vγ2Vδ2 cells with higher purity and cell numbers as compared with continuous exposure. These Vγ2Vδ2 cells also had higher levels of CD107a and perforin and slightly increased tumor cytotoxicity. Importantly, adoptive immunotherapy with Vγ2Vδ2 cells derived by pulse stimulation controlled human PC-3 prostate carcinoma cells in immunodeficient NSG mice significantly better than those derived by continuous stimulation. Zoledronate pulse stimulation of Vγ2Vδ2 cells in IL-15 also resulted in higher purity and cell numbers. Like with CD8 αβ T cells, IL-15 preserved early memory Vγ2Vδ2 T cell subsets better than IL-2. However, despite this fact, adoptive immunotherapy with Vγ2Vδ2 cells derived in IL-15 showed similar inhibition of PC-3 tumors as those derived in IL-2. Thus, zoledronate pulse stimulation maximizes the purity, quantity, and quality of expanded Vγ2Vδ2 cells. This simple modification to existing protocols could enhance the effectiveness of adoptively transferred Vγ2Vδ2 T cells.
Studying the Effect of β-estradiol on HCC Antigen-Specific T Cells
Authors: Navarro Flor C 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) has a 4:1 male-to-female overall incidence ratio. These statistics lead to the belief that there are gender specific mechanisms that affect the development of this cancer. Previous research studies demonstrated that the gender differences regarding HCC may be related to a protective effect from estrogen (E2). However, the antitumor immune response following cancer development is not defined. To test specific immune responses to HCC we are utilizing a known hepatitis C virus (HCV) antigen to generate Ag-specific T cells that target HCC expressing the HCV antigen since the chronic infection of accounts for approximately 50% of HCC. With this model, we tested the effect of E2 on the immune response during immunotherapy. We have identified a novel effect of E2 treatment of TCR-transduced Ag-specific T cells. E2 treatment induced an increase in their cytotoxicity and effector function. Treatment at super physiological concentrations of β-estradiol resulted in an up-regulation of CD107a expression, demonstrating degranulation and an increase in TNFα secretion. On the other hand, E2 treatment also caused a reduction in the number of cytokines produced simultaneously demonstrated in a polyfunctional analysis of the T cells. Polycytokine profiles of the T cells showed that as E2 increases, the number of T cells producing 3 or 4 cytokines simultaneously decreases while the number of T cells displaying a reduced number of functions increases. Previous studies on memory T cell development show that memory T cells result from cells with greater polyfunctional abilities. Our results suggest that while exposure to E2 increases rapid effector function, it is at the expense of the generation of memory T cells.
Optical control of Calcium influx improves intratumoral cytotoxic T lymphocyte effector functions.
Authors: Kim Kyun Do , Bae Seyeon , Nedelkovska Hristina , Capece Tara , Kim Minsoo 
Published: January 2023

Abstract
Regulatory T cells (Tregs) have critical roles in the maintenance of immunological self-tolerance and in the control of antitumor immune responses. In tumor microenvironment, Tregs use several mechanisms to suppress cytotoxic T cell responses, including suppression of TCR-induced Ca2+-NFAT signaling, secretion of inhibitory cytokines and inhibition of the stimulatory capacity of antigen presenting cells. Particularly, suppression of Ca2+ signals in cytotoxic T cell is a significant challenge because Ca2+ plays pivotal roles in tumor-specific CD8+ cytotoxic T lymphocytes (CTLs) function by regulating the degranulation of CTLs, their expression of Fas ligand and production of TNF-α and IFN-γ.
Here we developed a strategy for optically controlling calcium signal using Ca2+ translocating channelrhodopsin (CatCh). The Ca2+/NFAT pathway is activated upon blue light (480nm) illumination via increase in intracellular calcium in CatCh expressing CTLs. In vitro, light stimulation enhances CatCh expressing CTLs activation, IFN-γ production and tumor cytotoxicity. Importantly, CatCh expressing CTLs restore against Treg-induce immunosuppression by light stimulation. In vivo, blue LED illumination onto the melanoma improved the efficacy of adoptive T-cell transfer immunotherapy, with a significant reduction in tumor growth. Together, our findings indicate that strong intracellular calcium signals can boost T cell functions by overcoming immunosuppressive Treg responses at tumor sites.
Fatty acid metabolism is essential to maintain functions of CD8+T cells within a hypoxic and hypoglycemic tumor microenvironment
Authors: Zhang Ying , Liu Ling , Kurupati Raj , Zhou Xiangyang , Rabinowitz Joshua , Ertl Hildegund 
Published: January 2023

Abstract
The efficacy of immunotherapy for solid tumors is dampened by functional declines of tumor-infiltrating T lymphocytes (TILs), which is generally viewed as the result of their exhaustion due to persistent antigenic stimulation. Disputing this notion, we observed in a mouse melanoma model that bystander CD8+TILs increase expression of co-inhibitors and lose effector functions, suggesting alternative mechanisms. Our data show that metabolic stresses within tumor microenvironment (TME) profoundly affect differentiation and functions of TILs. Hypoxia through HIF-1α drives increases of co-inhibitor LAG-3 while lack of glucose (Glu) enhances PD-1 levels on activated CD8+T cells. Both conditions impair functions of CD8+T cells. When deprived of Glu, activated CD8+T cells enhance fatty acid (FA) catabolism and this is further increased under hypoxia. Using 13C-stable isotope tracing in vivo and liquid chromatography-mass spectrometry (LC-MS) analysis, our study show that CD8+TILs in late stage tumors increasingly depend on catabolism of FAs including ketone bodies fueled by exogenous FA uptake and triacylglycerol (TG) turnover to meet their energy demand. Promoting FA catabolism of CD8+TILs increases their PD-1 expression but preserves some effector functions and improves their anti-tumor efficacy. Our findings show that lack of Glu and oxygen plays a critical role in driving the metabolic reprograming and functional exhaustion of CD8+TILs. They further indicate that metabolic interventions that promote FA catabolism of adoptively transferred or vaccine-induced CD8+T cells may improve the efficacy of cancer immunotherapy.
Toll-like receptor agonists targeted to cancer cell metabolism
Authors: Mancini Rock , Nielsen Amy , Hantho Joseph 
Published: January 2023

Abstract
Imidazoquinoline Toll-Like Receptor (TLR) agonists such as Imiquimod are effective immunotherapeutics in the treatment of basal skin cell carcinoma, and more recently, melanoma. However, direct injection of imidazoquinoline TLR agonists causes systemic inflammation and autoimmune side-effects thereby limiting clinical use to topical applications. Here we use a metabolic caging strategy to confine the immunostimulatory effects of imidazoquinoline TLR agonists to tumor microenvironment. We have covalently attached a galactopyranoside metabolic cage to the primary amino group, critical for activity, of the TLR7 agonist Imiquimod. Addition of the metabolic cage abrogates TLR agonism as measured by NFκB transcription in RAW-Blue cells or inflammatory cytokines produced by primary murine Bone Marrow Derived Dendritic Cells (BMDCS). The galactopyranoside cage is efficiently removed via enzymatic cleavage from exogenously added β-galactosidase resulting in activation of immune cells that is comparable to native agonist. Enzymatic cleavage also occurs using β-galactosidase that is overexpressed in B16 melanoma cells. This results in increased immunostimulatory effects in co-cultures of BMDCs and B16 melanoma cells relative to BMDCs co-cultured with healthy melanocytes. We anticipate this will provide a general framework upon which TLR agonists can be targeted to the tumor microenvironment.
Increased leptin during obesity abrogates antitumor immunity
Authors: Murphy Katherine A , James Britnie R , Brincks Erik L , Kucaba Tamara A , Griffith Thomas S 
Published: January 2023

Abstract
Cancer immunotherapy offers the promise of long–lived protection against a variety of tumors; yet, the efficacy seen in laboratory models has not always translated into clinical success. One reason may be the preclinical use of young, healthy mice lacking common co-morbidities found in adult humans. Obesity is associated with an increased risk of developing cancer and correlates with a poor prognosis. Further, obesity related factors, such as leptin, can alter the behavior of tumor and immune cells. Renal cell carcinoma (RCC) more closely correlates with obesity than cancer at any other anatomical site and obesity-related cytokines play important roles in the progression of metastatic RCC. We investigated the therapeutic efficacy of a recombinant adenovirus encoding TRAIL in combination with CpG in an orthotopic mouse model of RCC. Ad5-TRAIL/CpG induced systemic T cell-mediated antitumor immunity capable of reducing primary and metastatic tumor burden, as well as reduced MDSC within the tumor-bearing kidney. In contrast, Ad5-TRAIL/CpG failed to induce antitumor immunity or reduce tumor burden in diet-induced obese (DIO) mice and MDSC levels remained high in the tumor-bearing kidney. Leptin reduced Ad infection of the tumor cells, decreased TRAIL receptor (DR5) expression, and Ad5-TRAIL-induced death of RCC cells. DC activation by CpG was also decreased in the presence of leptin. Leptin neutralization in DIO mice restored Ad5-TRAIL/CpG efficacy, and leptin-deficient (ob/ob) obese mice had reduced tumor burden. These data suggest the elevated leptin present during diet-induced obesity affects both tumor and immune cells that leads to reduced antitumor immunity.
Targeting of a novel signaling program in Philadelphia chromosome-like (Ph-like) acute lymphoblastic leukemia (ALL)
Authors: Thomas Kerri R , Allenspach Eric J , James Richard , Tasian Sarah , Teachey David , Rawlings David J 
Published: January 2023

Abstract
Molecular crosstalk between the interleukin-7 receptor (IL-7R) and the pre-B cell receptor (pre-BCR) represents a key developmental checkpoint regulating early B cell proliferation and differentiation. Dysregulation of these pathways has been linked to early lymphoid malignancies. While Philadelphia chromosome-like acute lymphoblastic leukemias (Ph-like ALL) lack the canonical chromosome translocation present in BCR-ABL1+ (Ph+) ALL, these tumors share a similar gene expression profile. Notably, however, Ph-like ALL exhibits a poor clinical prognosis and is refractory to most available therapies. A subset of Ph-like ALL results from alterations in the IL-7R signaling pathway, with gain-of-function mutations in IL-7Rα and loss-of-function mutations in the adaptor, SH2B3, in numerous samples. In this study we directly assess the signaling program downstream of the IL-7R and SH2B3 and its role in disease pathogenesis. Primary human Ph-like ALL tumors display constitutive STAT5 activation, a phenotype recapitulated by in vitro analysis of transduced cell lines expressing an activating IL-7Rα mutation or loss of SH2B3. Our results suggest that both IL-7Rα and SH2B3 mutations can individually contribute to this altered signaling program and may be capable of inducing oncogenic transformation. We have also developed a mouse model that allows for inducible expression of an activating IL-7Rα mutation for future in vivo analysis. Current work is focused on using mass spectrometry and phosphoproteomics to determine downstream signaling aberrations in both primary tumors and our stably transduced cell lines. The results from these studies may lead to new therapeutic targets for high-risk ALL patients.
Oxidative phosphorylation controls regulatory T cell suppressor activity of in the tumor microenvironment
Authors: Maj Tomasz , Wang Wei , Kryczek Ilona , Zou Weiping 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are important elements of the tumor-associated immunosuppresive network. Here we show that Tregs in the tumor microenvironment are highly apoptotic. Although apoptotic Tregs express classical immunosuppressive proteins including PD-L1, CTLA-4, TGF-β and IL-35, blockade of these mediators does not abolish their suppressive effect. Apoptotic Tregs release a large amount of ATP and suppress T cell responses via adenosine A2A receptor. Furthermore, Tregs have higher mitochondrial load and their survival is associated with OxPhos and fatty acid β-oxidation. Apoptosis of Tregs leads to pannexin-1-dependent ATP release and provide a substrate for immunosuppression. The data support a novel notion that Tregs amplify their suppressor capacity via apoptosis and OxPhos may be a central metabolic pathway controlling Treg functionality in tumor.
Suboptimal influenza vaccination results in higher numbers of pulmonary macrophages upon influenza challenge in mice
Authors: Schotsaert Michael , Garcia-Sastre Adolfo 
Published: January 2023

Abstract
Macrophages are innate immune cells present in the airways and are important for the initial immune answer against invading pathogens. Alveolar macrophages (AM) are transiently lost in the mouse-influenza infection model during the first days of infection. During later stages of infection, AM recover to similar numbers as before challenge. We investigated if AM were protected during experimental sublethal influenza challenge after vaccination with an alum-adjuvanted trivalent inactivated influenza vaccine (TIV) that antigenically matches the challenge virus. TIV vaccination could not prevent morbidity, however, it correlated with lower viral lung titers and faster recovery after challlenge. AM were completely abolished at seven days post infection in negative control mice, but not in TIV-vaccinated mice. Taken together, using the mouse influenza infection model we observed that suboptimal vaccination with TIV results in higher levels of AM in lungs upon sublethal challenge with homologous virus. The possible impact of higher levels of pulmonary macrophages on antigen presentation and disease modulation are further investigated.
A combinatory mucosal vaccine against influenza and botulism
Authors: Yan Yongyong , Li Junwei , Chen Yanping , Wu Lihong , Wang Haiyan , Diaz-Arevalo Diana , Zeng Mingtao 
Published: January 2023

Abstract
Botulinum neurotoxins (BoNTs) are the most poisonous substances known in nature and cause the life-threatening botulism. Currently, there is no FDA licensed botulism vaccine available for public use. Influenza is still an important problem for public health and current influenza vaccines are not optimal for protection against seasonal and potential pandemic influenza. Influenza virus is a negative, segmented RNA virus without DNA intermediate. This makes it safer as a vaccine delivery vector than most DNA viruses that have potential to integrate their genetic elements into host genomes. We hypothesize that a live-attenuated influenza virus expressing botulinum antigens can be developed to a dual use vaccine against both botulism and influenza. Based on influenza virus H1N1 PR8, an engineered influenza viral vector with a 2A cleavage site on NA, was constructed. Using this vector, we generated a recombinant influenza virus, PR8/HC50-rbsd, expressing the receptor binding subdomain of the heavy chain C-terminal receptor binding domain (HC50-rbsd) from BoNT/A. We tested the growth characters of the recombinant PR8/HC50-rbsd virus in chicken eggs and Madin-Darby Canine Kidney epithelial cells (MDCK), and showed that it can be produced to a titer of 5×10E6 plaque forming units/ml in chicken eggs and MDCK cells. Subsequently, mice intranasally vaccinated with the PR8/HC50-rbsd virus conferred protection against challenge with lethal doses of active BoNT/A toxin and wild type H1N1 PR8 influenza virus. Our results demonstrated that it is possible to develop a dual purpose nasal vaccine against both botulism and influenza.
Comparison of influenza vaccine regimens in pediatric patients:immunity and efficacy in 2014–15
Authors: Archibald Crystal A , Elbourne Vicki , Moehling Krissy , Susick Michael , Nowalk Patricia , Zimmerman Richard , Zimmerman Richard , Cole Kelly Stefano 
Published: January 2023

Abstract
Currently, there are two types of influenza vaccines available to children and adolescents, Influenza Inactivated Vaccine (IIV) which is administered intramuscularly, and Live Attenuated Influenza Vaccine (LAIV) which is administered intranasally. The present study was designed to compare the efficacy of the two vaccines in pediatric patients in eliciting immune responses and protecting against infection in the 2014–15 influenza season. To conduct the study, patients ranging from ages 3 to 17 years were recruited for vaccination and evaluation. Patients were segregated by age (3 to 8 vs. 9 to17) and vaccine (IIV vs. LAIV). Peripheral blood samples were collected between the months of August and December, 2014 and processed for collection and storage of serum, plasma and peripheral blood mononuclear cells (PBMC). Each child who agreed to participate in the study was prescreened, consented and provided blood samples at three time points: D0 (blood drawn prior to vaccination), D7 and D21 following influenza vaccination. At the end of the vaccination season and once all enrolled patient samples had been collected, antibody responses were evaluated for reactivity to the vaccine strains as well as against additional circulating or heterogonous strains of influenza. Early in the 2014–15 season, it became apparent that the LAIV strain in the vaccine was not well matched to the circulating strain of influenza, resulting in less-than-effective immunity. Therefore, our analyses focused on IIV only. Our results showed that all participants in the study demonstrated higher titers and increased breadth of reactivity to diverse influenza strains when vaccinated with IIV, regardless of prior vaccination history.
Mucosal Vaccination Primes NK Cell-Dependent Development of CD8+ T Cells for Protection Against Pulmonary Brucella Infection
Authors: wang hongbin , Yang Xinghong , Hoffman Carol , Clapp Beata , Pascual David W. 
Published: January 2023

Abstract
Brucellosis is the most common zoonotic disease worldwide, usually transmitted from Brucella-infected livestock after consumption of contaminated foods or by aerosol exposure. Since no vaccines for humans are available, we developed a live, double-mutant Brucella abortus (BADM) strain, that after mucosal vaccination, confers complete protection, but abated in IFN-γ−/− mice. Pulmonary infection of naive mice with wild-type BA fails to recruit innate and adaptive lymphocytes to the lungs. Thus, we hypothesize that BADM’s protection is tied to innate cell stimulation. To test this hypothesis, 5 days after nasal BADM vaccination, mice showed increased lung IFN-γ-producing ILCs and EOMES+ NK cells by 3- and 6-fold, respectively; CD4+ and CD8+ T cells increased 3- and 5-fold, respectively. By 2 wks, CD8+ T cells were the dominant IFN-γ source, being 15-fold &gt; naive lungs. After pulmonary challenge, BADM-vaccinated lungs showed fewer CD4+ T cells, but a net increase in CD8+ T cells unlike naive mice, showed no differences in their numbers of CD4+ and CD8+ T cells. To assess if NK cells impact CD8+ T cells, vaccinated mice treated with anti-asialo GM1 Ab to deplete their NK cells, resulted in increased splenic weights, but reduced lung neutrophil influx 5 days after vaccination relative to isotype-treated, vaccinated mice. By 14 days, NK cell depletion reduced by 4-fold (P&lt;0.001) the IFN-γ-producing CD8+ T cells. Thus, NK cell recruitment enhances protection to pulmonary BA in BADM-vaccinated, but not naive mice, which fail to activate NK cells. Supported by NIH AI123244.
Recombinant spore delivered M. tuberculosis antigens elicit immune response in mice
Authors: Das Kishore , Thomas Tima , Garnica Omar , Dhandayuthapani Subramanian 
Published: January 2023

Abstract
BCG, the vaccine against tuberculosis (TB), is one of the mandatory vaccines administered to infants in several parts of the world. However, BCG conferred immunity wanes when children reach adulthood. To develop booster vaccines, we engineered recombinant Bacillus subtilis spores to deliver Mycobacterium tuberculosis (Mtb) antigens. MTAG1 recombinant strain was designed to express a fusion protein of CotC-Ag85B-CFP10 on the spore coat, while MTAG2 and MTAG3 strains were designed to express Ag85B-CFP10 fusion protein in vegetative cells. The difference between MTAG2 and MTAG3 strains is that the latter expresses secreted listeriolysin (LLO) in addition to Ag85B-CFP10. Immunoblot analysis revealed that MTAG1 strain showed positive signals for Ag85B and CFP10 in the spore-coat and other strains in the vegetative cell extracts, indicating that antigens are expressed in these strains as expected. Splenocytes of mice immunized with recombinant spores, through intranasal route, displayed significantly higher antigen specific proliferation of IFN-γ producing cells compared to the splenocytes of control mice. Also, the culture fluids of splenocytes from recombinant spores immunized mice exhibited higher antigen specific release of Th1 cytokines than the culture fluids of splenocytes from control mice. These results indicate that antigens delivered via recombinant spores can be processed and presented to the immune cells. Additional in vivo studies may reveal whether these recombinant spores can be used to boost immunity in BCG vaccinated individuals.