bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased CD4+ T cell proliferation upon treatment correlates with improved overall survival of patients with metastatic melanoma upon anti-CTLA-4 therapy
Authors: Miller William L , Contrad Valerie , Gonzalez Iliana , Meeuwsen Tanisha , Howells-Ferreira Ana Ana , Poplonski Tomasz , Curti Brendan , Urba Walter , Bahjat Keith , Koguchi Yoshinobu 
Published: January 2023

Abstract
Blockade of cytotoxic T-lymphocyte antigen 4 (CTLA-4) augments anti-tumor immunity and has shown success in treating patients with metastatic melanoma. Mechanisms of anti-CTLA-4 efficacy have been proposed in animal models, yet the mechanisms in humans remain to be elucidated. Increased frequency of ICOS+ CD4+ T cells has been shown to be a marker of drug bioactivity, but discovery of prognostic indicators and baseline indicators of which patients are most likely to benefit from CTLA-4 blockade remains a priority.
PBMCs from 29 metastatic melanoma patients in a multicenter compassionate use trial of anti-CTLA-4 antibody were evaluated using flow cytometry. At baseline and 12 weeks following initiation of treatment, CTLA-4, HLA-DR, ICOS, PD-L1, PD-1, and Ki-67 frequencies were assessed on CD4+ and CD8+ T cells, as well as CD3+CD4+ regulatory T cells.
Independent of survival, anti-CTLA-4 treatment significantly increased the frequency of ICOS+ CD4+ T cells, corroborating previous findings. Interestingly, this increase was also observed in ICOS+ Tregs, but not in CD8+ T cells. Increased proliferative Ki67+ CD4+ T cells and Ki67+ Tregs upon CTLA-4 blockade were both associated with prolonged overall survival. Additionally, when the frequency of Ki67+ CD4+ T cells post-treatment was compared to the baseline frequency, patients with a ratio greater than 1.5 had greater overall survival. Increased post-treatment absolute lymphocyte count—a known prognostic biomarker—also correlated with clinical outcome. Our results suggest that anti-CTLA-4 therapy may be mediated by proliferation of CD4+ T cells, and we identify this proliferation as a novel prognostic biomarker in metastatic melanoma patients treated with anti-CTLA-4 antibody.
Hybrid Th1/17 cells possess unique immuno-metabolic phenotype and surpiror anti-tumor activity
Authors: Chatterjee Shilpak , Daenthanasanmak Anusara , Meek Megan , Toth Kyle , Chakraborty Paramita , Paulos Chrystal M , Yu Xuezhong , Mehrotra Shikhar 
Published: January 2023

Abstract
Immunotherapy of cancer is a promising approach for successful eradication of large and established tumor. Accumulating evidence indicates that CD4+ T helper (Th) cells including Th1 and Th17 subsets are also effective in tumor immunotherapy. While the anti-tumor potential of Th17 cells primarily depends upon IFN-γ secretion and persistence, a long-term tumor control has still remained elusive. In current study, we established an optimal protocol to program CD4 T cells ex vivo into a combinational phenotype of Th1 and Th17 subset (named hybrid Th1/17 cells), which express high levels of IFN-γ, CD107a, T-bet, Granzyme B, IL23R, IL22, Bcl6, and Tcf7. A detailed microaray analysis followed by Ingenutity Pathway Analysis for hybrid Th1/17 cells further established its unique molecular signature. Importantly, a comprehensive analysis for about 400 metabolites also established that hybrid Th1/17 cells have unique metaboliomic signature. Furthermore, using two types of TCR transgenic models and established B16 tumor model either administrated subcutaneously or intravenously, we demonstrated that the hybrid Th1/17 cells exhibit a superior activity in controlling tumor as compared to Th1 or Th17 alone. We observed that the unique immuno-metabolic phenotype rendered to hybrid Th1/17 cells corrected with their increased ability to survive, overcome tumor mediated immunosuppression, resulting the superior effectiveness to control tumor in long-term.
Serial monoclonal antibody administration demonstrates therapeutic effects via induction of apoptosis in HPV related cervical cancer
Authors: Bhadauria Preeti , Hockenberry Kristi , Jiang Zewei , Christensen Neil , Dadachova Ekaterina , Phaëton Rebecca 
Published: January 2023

Abstract

Introduction
All cervical cancer is caused by the persistent expression of human papillomavirus (HPV) E6 and E7 oncoproteins that interfere with p53 and Rb pathways, respectively. Virally specific treatments form an excellent basis for highly specific, low toxicity therapies.

Methods
Serial doses of anti-HPV 16/18 E6 C1P5 (Abcam) were compared against Cisplatin in the treatment of human cervical cancer derived tumors (CaSki). C1P5 and Cisplatin were administered on days 1, 4, and 7 after CaSki tumor size reached 3–5mm on the flank of athymic nude mice. Measurements were performed every 3 days during the 40 day observation period. After which the tumors were procured, paraffin embedded, and stained with H&E. SiHa cells which compare to HPV copies in patient explants (previously published) were used in mechanistic studies. Dual staining flow cytometry with PI and Annexin was performed to evaluate cell death and MTT assay was used to confirm findings.

Results
Tumor growth was inhibited in all treatment groups compared to untreated controls (p &lt;0.05). No statistically significant differences between C1P5 treatment groups were noted when compared to cisplatin. However, on H&E we noted increased necrosis proportional to the number of doses: 1 dose 30%, 2 doses 60%, and 3 doses 85%. Dual-staining of Annexin and PI on flow cytometry demonstrated increased apoptosis as a result of C1P5 administration suggesting this as the underlying mechanism of increased necrosis. These results were corroborated by MTT assay (p&lt;0.05).

Conclusion
Virally specific therapy is promising for the treatment of women with cervical cancer. To extend the paucity of options, further mechanistic understanding must be developed to derive maximal therapeutic benefit.
Tumor specific epitopes on Podocalyxin as a target for ADC and CAR T cell therapy
Authors: von Rossum Anna , Samudio Ismael , Bergqvist Peter , Cruz Eric , Wee Kathlee , Hughes Michael , Canals Diana , Babcook John , Roskelly Calvin , McNagny Kelly , Bond Chris 
Published: January 2023

Abstract
Podocalyxin (Podxl) is a heavily glycosylated sialomucin type 1 membrane protein that has been shown to be involved in cell adhesion, migration, and polarity. Although Podxl is found on a variety of normal cell types, it is aberrantly expressed in acute myeloid leukemia (AML) blasts, and its presence in breast, colorectal and glial malignancies is correlated with poor clinical outcome. Using the human triple negative breast cancer line MDA-MB-231, we have demonstrated a functional role for Podxl in tumor growth and metastasis in xenografted mice, and have recently developed high affinity antibodies to a tumor-specific post-translational modification of Podxl. Preliminary IHC data suggests that this post-translational modification on Podxl may found in glioblastoma, melanoma, and female reproductive tract tumors, but not on matched normal tissue. Staining intensity in melanoma samples correlates with stage of disease and is strongest in tissues collected from metastatic lesions. Preliminary experiments indicate that expression of this glyco-epitope is increased in myeloid leukemia cells lines by co-culture with bone marrow stromal cells or hypoxia, suggesting that the tumor microenvironment may contribute to the observed reactivity. Our lead antibody, Podo447, binds this tumor-associated glycoform of Podxl with high affinity and induces cytotoxicity in various tumor cell lines as an antibody drug conjugate (ADC), and when expressed as a human IgG1 Fc chimera stimulates antibody dependent cellular cytotoxicity. With its exquisite specificity for malignant tissues, and the number of potential indications, Podo447 is an ideal targeting arm for an antibody drug conjugate or CAR-T enabled therapeutic.
Modeling anti-CD20 chimeric antigen receptor therapy in canine B cell lymphoma patients
Authors: Panjwani M. Kazim , Gnanandarajah Josephine S. , MaloneyHuss Martha A. , Mason Nicola J. 
Published: January 2023

Abstract
Chimeric antigen receptor (CAR) therapy has demonstrated great promise in treating human leukemias, but preclinical murine models are limited in their ability to predict safety and efficacy in humans. Given the rapid and on-going advances in CAR T cell technology in the laboratory, it now becomes necessary to identify and develop an outbred, large animal spontaneous cancer model in which the safety of novel targets and therapeutic effectiveness of re-directed T cells can be evaluated and optimized.
Canines naturally develop spontaneous B cell lymphoma, and we have previously shown the feasibility of evaluating CAR T cell therapy in dogs using CD20-targeting RNA CAR T cells. To parallel human CAR T cell approaches, we have now optimized primary canine T cell transduction with lentivirus. Once peak T cell activation and artificial antigen presenting cell elimination timeframes were established, transduction yielded up to 26% CAR+ canine T cells. Canine T cells transduced with a second generation CD20-targeting CAR (CD20-28-ζ) demonstrated CAR-mediated, antigen-specific proliferation and efficient cytolysis of a CD20+ canine B cell lymphoma line in vitro.
Three relapsed canine B cell lymphoma patients were treated with autologous CD20-28-ζ CAR T cells. Despite the administration of low numbers of CAR T cells, transient anti-tumor effects were observed in the first 2 patients. Following administration of ~700,000 CAR T cells/kg intravenously to the third patient, CAR T cells were found to persist and expand in the periphery and malignant nodes and disease progression was halted. This work establishes the feasibility of using the dogs with spontaneous cancer as pre-clinical models for advancing human CAR T cell therapy.
Small-molecule probe targeting viral PPxY L-domain inhibits filovirus and arenavirus egress.
Authors: Prugar Laura I , Dye John M , Loughran Marie , Wrobel Jay E , Reitz Allen B , Freedman Bruce D , Harty Ronald N , Herbert Andrew S 
Published: January 2023

Abstract
Viral matrix proteins containing a highly conserved PPxY Late (L) domain motif are critical for efficient viral budding of several emerging RNA viruses. The L domain motif functions by recruiting host cell proteins necessary for separation of virus particles from the cell surface during the budding process. The conserved nature of the PPxY motif and its importance for viral egress make it an attractive drug target with potential broad spectrum antiviral activity. Here we describe a PPxY inhibitor capable of reducing viral budding of four highly lethal, category A RNA viruses: Ebola, Marburg, Lassa, and Junín viruses.
Genetic background influences the humoral responses to vaccines
Authors: Nouri-Shirazi Mahyar A , Abu-Nuwar Emily , Nourishirazi Erika 
Published: January 2023

Abstract

Background
The assessment of TLR agonists as promising candidate adjuvants for the induction of effective Th1 immunity continues to rely on the use of inbred mouse. However, the genetic variation among commonly used inbred mice may influence the efficacy of candidate adjuvants and bias the study conclusion. Indeed, we have found significant differences in the recruitment, phenotypes and cytokines production of DCs and NK cells and the generation of effector T cell in lymphoid tissues of two genetically non-identical mouse strains immunized with a protein based vaccine formulated with TLR agonists.

Objective
In this study, we further evaluated the nature and amplitude of humoral immune responses of B6 and BALB/c strains immunized with protein antigen ovalbumin plus Alum, TLR3, TLR4, TLR7/8, or TLR9 agonists.

Results
Our data suggest that compared to the Alum, all other adjuvants induced more effectively the recruitment of B cells and production of OVA-specific antibodies in both strains. We also observed significant differences between two strains. BALB/c strain recruited less B cells but produced significantly higher amounts of IgG2a in response to any of the TLR agonists. In addition, both strains produced similar amounts of IgG1 and IgG2b in response to TLR agonist except for MPLA which induced significantly more of IgG2b production in B6 strain.

Conclusion
Thus, our data suggests that genetic background should be taken into consideration when evaluating and comparing the activities of adjuvants for use in new prophylactic and therapeutic vaccines. NIH-NIAID 1R03AI103750-01A1
Type-1 T cell responses and gene signatures driven by a novel virus-like particle (VLP) influenza A (H1N1) vaccine
Authors: Jones Leigh A , Skibinski David A , Xie Lin Wu , Au Bijin , Lee Bernett , Zhu Yuan , Fairhurst Anna-Marie , Lee Audrey , Kaliaperumal Nivashini , Poidinger Michael , Hibberd Martin , Ooi Eng Eong , Hanson Brendon J , Diermayr Veronica , Matter Alex , Bachmann Martin , Saudan Phillipe , Connolly John E 
Published: January 2023

Abstract
Effective influenza vaccination is currently assessed by the induction of antibodies directed against the influenza virus envelope glycoprotein hemagglutinin (HA; HAI test) otherwise known as seroconversion. However, protective antibody responses are sub-optimal in some vaccinated populations, particularly in the elderly. Furthermore, many clinical studies highlight the importance of T cells in driving protection. Therefore, a need exists for vaccine strategies which can engage both the humoral and cell mediated arms of the immune response. Herein, we describe the induction of anti-influenza T cell responses following vaccination with a novel VLP vaccine. Peripheral blood mononuclear cells (PBMCs) stimulated ex vivo with either influenza or vaccine specific antigens led to increased CD4+ and CD8+ T cell proliferation and increased production of cytokines such as IL-17A, IL-17F, IL-5, IL-13, IL-9, IL-10, IL-21 and, most significantly, IFN-γ. A subset of individuals demonstrated a shift from a pre-vaccination IL-5- and IL-13- driven type-2 response to a more protective IFN-γ driven type-1 response following vaccination with VLP. Microarray of whole blood samples collected pre- and post-vaccination with VLP revealed significantly different transcriptional profiles. The top most significantly downregulated gene following VLP vaccination, DEAD Box Helicase 17 (DDX17), was also found to have reduced protein expression in PBMCs by flow cytometry. Finally, weighted gene correlation network analysis (WGCNA) revealed that the post vaccination reduction in protein expression of DDX17, increase in HAI titres and increase in T cell proliferation correlates with particular gene modules.
TGF-β2 decreases surface expression of MHC I and MHC II on equine bone marrow-derived mesenchymal stromal cells.
Authors: Berglund Alix K , Cameron Kristin K , Poole Emma J , Harris Elizabeth A , Schnabel Lauren V 
Published: January 2023

Abstract
Allogeneic bone marrow-derived mesenchymal stromal cells (MSCs) are a promising cell source for treatment of inflammatory diseases, but elicit immune responses in vivo due to recipient recognition of mismatched Major Histocompatibility Complex (MHC) molecules. The purposes of this study were to evaluate the effectiveness of TGF-β2 in decreasing and stabilizing MHC expression on equine bone marrow-derived MSCs before and after IFN-γ stimulation. FACS analysis showed that MSCs treated with 1, 5, or 10 ng/ml of TGF-β2 had significantly reduced MHC I expression. MHC I expression levels on TGF-β2-treated MSCs were also quantified and determined to be similar to fetal fibroblasts. MHC II expression in two positive animals was greatly reduced compared to untreated MSCs. MSCs pre-treated and continuously treated with 1 ng/ml TGF-β2 had significantly lower MHC I expression following stimulation with IFN-γ for 72 hours compared with untreated. Continuously treated MSCs that were baseline positive for MHC II had significantly lower MHC II expression following IFN-γ stimulation, but there was no significant difference in MHC II expression between TGF-β2-treated and untreated MHC II-negative MSCs after IFN-γ stimulation. These results indicate TGF-β2 treatment has significant promise for reducing recipient immune recognition of MHC-mismatched molecules on allogeneic MSCs, but further work is needed to maintain MHC expression stability.
Induction of robust cytotoxic T lymphocytes using adenovirus-vectored multivalent antigen cocktail
Authors: Mwangi Waithaka , Lokhandwala Shehnaz , Waghela Surya 
Published: January 2023

Abstract
Development of subunit vaccines against complex pathogens entails inclusion of multiple targets to improve efficacy and or broaden protection. Importantly, testing of an immunogen in the actual host that require protection can hasten vaccine development and generate knowledge needed to generate other similar vaccines. The African Swine Fever Virus (ASFV) causes a fatal hemorrhagic disease in domestic swine and, at present, no treatment or vaccine is available. Previous studies suggest that induction of ASFV-specific cytotoxic T lymphocytes (CTLs) is critical for protection. We evaluated immunogenicity of adenovirus-vectored ASFV multivalent cocktail formulated in two adjuvants and at two doses in swine. The immunogen induced unprecedented CTL responses against all the antigens in the cocktail. Notably, CTL responses were detected in swine immunized with a low dose (1 × 1010 ifu/each construct) and a high dose (1 × 1011 ifu/each construct). In addition, both adjuvants elicited strong CTL responses. Significant antigen-specific IFN-γ+ responses were detected in PBMCs of vaccinees post-priming and post-boosting. Furthermore, the cocktail induced robust antigen-specific antibody responses against all the antigens and the antibody responses underwent rapid isotype-switching within one week post-priming. The primed antibody responses underwent rapid recall upon boost four months post-priming. Taken together, this study showed for the first time that the adenovirus-vectored ASFV multivalent cocktail safely induced robust cellular and antibody responses in swine following homologous prime-boost immunization. The relevance of the induced immune responses in regards to protection will be evaluated in a challenge study.
Functional differences exist in contribution of VH and VL from polyspecific IgM and monospecific IgG antibodies in antigen recognition and virus neutralization functions.
Authors: Kaushik Azad Kumar , Pasman Yfke 
Published: January 2023

Abstract
We analyzed role of individual variable heavy (FdVH) and variable light (FdVL) domains in comparison with VH+VL pair (scFv) originating from a polyspecific bovine IgM, with an exceptionally long CDR3H (61 amino acids), and a monospecific IgG1 antibody in antigen (Ag) recognition and virus neutralization functions. To this end, recombinant FdVH, FdVL and scFv were constructed and expressed in Pichia pastoris from bovine polyspecific IgM and IgG1 encoding cDNA. The scFv1H12 showed polyspecific antigen recognition similar to parent IgM antibody with minor differences. Unlike variable light domain FdVL1H12, variable heavy domain FdVH1H12 recognized multiple antigens that differed from scFv1H12 and the parent IgM antibody. Nevertheless, role of FdVL1H12 in providing structural support to FdVH in antigen recognition is noted. By contrast, the individual FdVH073 and FdVL074, originating from induced BoHV-1 neutralizing IgG1 antibody, recognized target epitope on BoHV-1 relatively weakly when compared to VH+VL pair as scFv3-18L. Both VH and VL domains of induced IgG antibody are required to achieve BoHV-1 neutralization function. To conclude, there exist subtle functional differences in relative contribution of VH and VL from polyspecific IgM and monospecific IgG antibodies in antigen recognition and virus neutralization functions. [Supportd by NSERC Canada]
Genome wide association study identifies loci associated with immune cell influx following experimental mammary challenge with Streptococcus uberis
Authors: Pighetti Gina M , Siebert Lydia , Staton Meg E , Oliver Stephen P 
Published: January 2023

Abstract
Mastitis, an inflammation of the mammary gland, significantly impairs animal health, productivity, and milk quality. A common mastitis-causing pathogen, Streptococcus uberis, is observed in 14–26% of subclinical and clinical cases. To identify potential candidate genes and regulatory regions that influence immune cell influx and clearance from the gland, Holstein dairy cows (n=40) were challenged intramammarily with S. uberis. Somatic cell counts (SCC), a measure of immune cells present in milk, were determined prior to and for 28 days following experimental challenge. Traditional GWAS have used SCC (cells/ml) to identify loci of interest. Our approach uses SCC to generate three novel phenotypes: area under the curve (AUC) of SCC for 0–7 d, AUC for 0–28 d, and resolution of infection based on SCC return to less than 200,000 cells/ml in milk (grouped as &lt;21 d, 21–28 d, or &gt;28 d). SNP were identified using the BovineSNP50 v2 DNA Analysis BeadChip (Illumina) and associations with SCC phenotypes tested using Plink. Analyses revealed 16 highly significant (p&lt;1.0×10−5) SNP. The BTA20 region associated with SCC AUC for the first 7 days corresponded to a previously identified QTL for SCC. This region contains 8 genes and 1 miRNA, with two genes linked to migration and apoptosis. A significant association with the recovery or return to SCC &lt;200,000 cells/ml was observed on BTA9, with the greatest association (p=8.5×10−7) observed within an immune related gene. The identified loci will be evaluated for potential contribution to the observed phenotypes. Such investigations could lead to novel treatment or preventive strategies for controlling mastitis, as well as identify markers for genetic selection of cows with greater resistance to infection.
Expression of T cell genes in adult Xenopus laevis and TCR gene expression in the Xenopus tadpole tail.
Authors: Foulkrod Ashley M , Geibel Gretchen M , Kongprachaya Yuthana , Appasamy Pierette M 
Published: January 2023

Abstract
Xenopus laevis is a unique and useful model for studies of the evolution and development of the adaptive immune response. The Xenopus immune system shares much similarity with the immune system of higher animals, with some unique exceptions, is relatively easy to maintain, and it’s larval period can be manipulated since it exists outside the female. This study explored the distribution of T cell specific genes, as markers for the presence of T cells, in adult X. laevis as well as X. laevis tadpoles. Transcripts for all four TCR genes (γ, δ, α and β) were detected in the adult Xenopus thymus and spleen and TCR δ and β were detected in adult Xenopus skin, intestine and lung, using RT-PCR of total cellular RNA extracted from these tissues. These data suggest that γδ and αβ T cells are plentiful in X. laevis and that the development and distribution of these cells is similar to that of humans and mice. In addition, because epidermal T cells are increasingly appreciated as having a significant role in wound healing, the mammalian equivalent of regeneration, we have begun to examine the expression of T cell-specific genes in the parts of the tadpole that are able to regenerate, specifically the tadpole tail and limbs. We have observed the presence of TCR β transcripts in the tadpole tail, and are extending our studies to an evaluation of the other TCR genes and other T cell-specific genes. These findings point to the utility of X. laevis in studies of the role of T cells in regeneration.