bluuebunny/crossref_metadata_2025_split huggingface dataset:
An essential role of ITK in T cell migration to the gut during mouse γ-herpesvirus 68 infection
Authors: Cho Hyoung-Soo , Shin Hyun Mu , Berg Leslie J. 
Published: January 2023

Abstract
IL-2-inducible T cell kinase (ITK) is the predominant Tec family kinase in T cells. ITK functions to regulate the magnitude of T cell receptor signaling. Deficiencies in ITK in humans leads to a fatal lymphoproilferative syndrome due to uncontrolled Epstein-Barr virus (EBV) infection. Yet, Itk−/− mice mount protective immune responses against several acute viral infections. To address this dichotomy, we have studied the role of ITK in anti-viral T cell responses to latent γ-herpesvirus infection using mouse γ-herpesvirus 68 (MHV68). Despite a delayed response at D7 post-infection, Itk−/− mice controlled MHV68 replication in the spleen at D14, and WT and Itk−/− mice showed no differences in their viral titers in the spleen during latency. However, MHV68-infected Itk−/− mice (n=11/24) spontaneously developed a severe gastrointestinal pathology after D100 post-infection, whereas WT mice (n=24/24) showed no disease symptoms. Following LPS-induced virus reactivation, Itk−/− mice also showed this pathology in the gut accompanied by a high viral titer in the intestine at various time points during viral latency, while WT mice remained healthy. Consistently, the number of gut-resident T cells were substantially less in Itk−/− mice than WT with or without infection. We also found that P-selectin binding on activated Itk−/− T cells or small molecule inhibitor of ITK (PRN694)-treated WT T cells is greatly impaired. Overall, these data suggest that ITK plays an important role in T cell migration to γ-herpesvirus-infected gut tissue following reactivation from latency.
Heterologous protective immunity can lead to impaired activation, memory differentiation and inflation of new naive responses
Authors: Agarwal Priti , Kraft Anke , Watkin Levi B , Kenney Laurie Lea , Daniels Keith , Chen Alex , Brehm Michael A , Selin Liisa K 
Published: January 2023

Abstract
Activation of crossreactive memory CD8 T cells leads to heterologous protective immunity. Here we question whether early activation of crossreactive memory CD8 T cells alters the activation of new naïve CD8 responses to the second virus. Pichinde virus (PICV) and lymphocytic choriomeningitis virus (LCMV) contain a crossreactive usually subdominant epitope, NP205, which differs in 6 of 8 amino acids. We find that during PICV infection of LCMV-immune mice the NP205 memory response becomes immunodominant and mediates protective immunity. PICV is cleared early and does not persist. The usually dominant non-crossreactive new naïve PICV NP38 CD8 T cell response appears to be subdominant day 7-post infection. Surprisingly over the next 100 days post infection the NP38 response continues to gradually expand and then stabilizes. These NP38-specific CD8 T cells appear to arrest in an intermediate effector T cell phenotype (KLRG1low, CD62Llow IL-7Rhi), with altered functionality (decreased TNFalpha production). They present with altered expression of apoptotic genes on gene array. This NP38 inflationary response did not occur when mice were immunized with an LCMV mutant that did not present the crossreactive NP205 epitope. Our data shows that the strong crossreactive response kills DC’s expressing NP205 early (day 2–3) in PICV infection. This may result in an early termination of antigen-presentation leading to inadequate activation of the naïve NP38 response and inflation. (NIH U19 AI109858)
IL-2 and IL-15 on CD8 T cells during chronic viral infection: The exhaustion program awakens
Authors: Beltra Jean-Christophe , Bourbonnais Sara , Bédard Nathalie , Charpentier Tania , Boulangé Moana , Michaud Eva , Boufaied Ines , Bruneau Julie , Shoukry Naglaa H , Lamarre Alain , Decaluwe Helene 
Published: January 2023

Abstract
CD8 T cell exhaustion is an immunosuppressive mechanism by which chronic viruses escape adaptive immune response. Yet, the direct implication of cytokines as well as the precise candidates involved in this process have remained elusive. Here we demonstrate the direct contribution of IL-2 and IL-15 on several aspects of CD8 T cell exhaustion. Using a mouse model of chronic viral infection (LCMV clone-13), we show that IL2Rβ (CD122; the receptor chain common to IL-2 and IL-15) is maintained on CD8 T cells during chronicity. Its expression correlates with deeper exhaustion and selectively marks terminally exhausted CD8 T cells in mice and humans. Adoptive transfer of IL2Rβ−/− P14 CD8 T cells (unresponsive to combined IL-2 and IL-15-dependent signals) revealed that IL-2 and IL-15 directly control the expression of a specific pattern of inhibitory receptors and are mandatory for the development of PD-1hi effectors. Further, IL-2 and IL-15 instigate the arrested development of central memory cells seen during antigen persistence and preclude responsiveness to IL-7-dependent signals. Together, we ascribe a novel role to IL-2 and IL-15 as instigators of CD8 T cell exhaustion during chronic viral infection.
TCR modifications that increase proper chain pairing also enhance reactivity against naturally occurring HCV mutant peptides
Authors: Foley Kendra C , Spear Timothy , Nagato Kaoru , Murray David , Garrett-Mayer Elizabeth , Nishimura Michael I. 
Published: January 2023

Abstract
TCR gene modified T cells in adoptive cell transfer have been shown to have clinical success in treating melanoma and other malignancies. One of the challenges in using TCR gene modified T cells include proper expression of the introduced TCR and function. Mispairing between endogenous and introduced TCR chains allows for the potential of unanticipated off-target reactivity, autoimmunity, and impaired therapeutic efficacy against targeted antigens. One approach to improve TCR expression, pairing, and T cell function involves modifying the introduced TCR genes to promote proper pairing by introducing a disulfide bridge in the alpha/beta constant regions, murine constant regions, codon optimization, TCR chain leucine zipper fusions, or a single chain TCR. To compare these modifications, we used a TCR we previously cloned from a HLA-A2 restricted, HCV NS3:1406-1415-reactive T cell clone. Results revealed that the murine Cβ2 and the leucine zipper TCR have the highest levels of expression on transduced T cells when compared to the wild type TCR and on a transduced cell by cell basis, these cells have the highest percentages of bi-functionality. Our data showed proper chain pairing can have an impact on the T cell’s ability to be reactive against naturally occurring HCV NS3 mutant peptides and lead to loss of CD8 dependence. Although this could be therapeutically beneficial for viral infections with unstable genomes, cross reactivity against nonspecific antigens due to high TCR density on the cell surface could be detrimental. These studies have given us a better understanding of how TCR modifications can impact expression and function that may allow for optimization of adoptive T cell transfer to treat viral infections and malignancies.
Aminobisphosphonates reveal a selective anti-viral potential of Vγ9Vδ2 T cells
Authors: Champagne Eric , Barragué Hugo , Daguzan Charline , Abravanel Florence , Bellvert Floriant , Peyrottes Suzanne , Izopet Jacques 
Published: January 2023

Abstract
Vγ9Vδ2 cells are activated by stressed cells producing endogenous phosphoantigens (PAgs) following tumoral transformation or treatment with aminobisphosphonates (ABP) such as zoledronate (Zol). Alternatively they respond to exogenous PAgs produced by bacteria. Their potential interest in viral infections is not well established. We analyzed in vitro their activation and activity against HCMV-infected cells. Although they were not activated by HCMV-infected fibroblasts, following treatment of targets with ABPs they were preferentially and more potently activated by infected cells, resulting in a stronger IFNγ production and induction of TNF. In conditions where uninfected cells were not affected, they limited viral replication through an IFNγ and TNF-dependent mechanism, whereas cytotoxicity did not play a significant role. This could be explained by an increased uptake of a fluorescent Zol derivative by infected cells. In addition, HCMV infection stimulated the transcription of mevalonate pathway enzymes HMG-CoA reductase and synthase evaluated by RT PCR. As a result, the production of endogenous PAgs IPP and ApppI following low-dose treatment with Zol was also boosted. Cytokines such as IL18, putatively produced by infected cells did not play a significant role in the synergistic effect of HCMV and Zol. In order to examine whether this was true for other viruses, we infected hepatocytoma cells with the hepatitis E virus and treated them with Zol. In this case, viral infection only moderately increased Zol-induced IFNγ production, but not TNF, by Vγ9Vδ2 cells. Our results suggest that ABPs could be used to target the anti-viral potential of Vγ9Vδ2 T cells against selected viruses responsible for chronic infections.
Deep-sequencing of longitudinal samples from RSV-infected adults and infants reveals heterogeneous dynamics of within-host viral diversification
Authors: Kim-Hoehamer Young-In , Lau Jessica , Murphy Ryan , Newman Ruchi , Yang Xiao , Zody Michael , Grad Yonatan H , DeVincenzo John 
Published: January 2023

Abstract
With high mutation rates, large population sizes, and rapid turnover, RNA viruses can evolve over the course of individual infections. While the dynamics of viral evolution under selective pressures from host immunity has been described for chronic infections, such as those from HIV and HCV, the extent and nature of viral diversification in acute viral infections remains uncharacterized. Here, we investigate the within-host diversification of human respiratory syncytial virus (RSV), a highly prevalent cause of respiratory infections, in healthy adults who were experimentally infected with identical virus and in infants hospitalized with severe manifestations of naturally acquired infections. In aggregate, viral diversification in the adults peaked at day 3, with statistically significant overrepresentation of diversity in the matrix protein 2 (M2) and non-structural protein 2 (NS2) genes. A later peak in diversity at day 10 was attributable to a single study subject, with diversity predominantly in known and predicted B and T cell epitopes. In the infant infections, much less diversity was observed, possibly related to sampling starting at the time of diagnosis and hospitalization, and trended towards overrepresentation in the G surface protein. Our findings raise the possibility of multiple mechanisms for control of RSV infection that vary over the course of an infection and by host, and establish a framework for further investigation into the basis of heterogeneous host response to infection. Moreover, the results have broad implications for investigations into immune response and the use of deep sequencing in viral pathogenicity and the molecular epidemiology of pathogen spread.
Novel antibody-like single-chain TCR antibody Fc fusion protein
Authors: Wong Hing C , Newman Robert G , Marcus Warren D , Liu Bai , Jeng Emily K , Alter Sarah , Rhode Peter R 
Published: January 2023

Abstract
We have previously reported the construction of a fusion protein composed of a soluble single-chain T cell receptor genetically linked to the constant domain of the human IgG1 heavy chain (TCR-Ig). The antigen recognition portion of the protein binds to an unmutated peptide derived from human p53 (amino acids 264–272) presented in the context of HLA-A2.1, whereas the IgG1 Fc provides effector functions. The protein is capable of forming dimers, specifically staining tumor cells, and promoting target and effector cell conjugation. The protein also has potent antitumor effects against p53+/HLA-A2.1+ human tumor xenografts in athymic nude mice and can mediate cell killing by antibody-dependent cellular cytotoxicity. Therefore, TCR-Ig behaves like an antibody, but possesses the ability to recognize antigens derived from intracellular targets. To test TCR-Ig in fully immunocompetent models, we have generated murine tumor cells stably expressing the human p53 epitope by constructing a single chain trimer composed of the human p53 peptide genetically linked to murine beta 2 microglobulin genetically linked to HLA-A2.1, with the inclusion of a disulfide trap to enhance stability. Utilizing haNK (NK-92 cells stably expressing high affinity Fc receptor and IL-2) as effector cells in the presence of TCR-Ig we demonstrated specific killing of murine single chain trimer expressing tumor cells. We are currently evaluating the antitumor activities and vaccinal effects of TCR-Ig in an HLA-A2.1 transgenic mouse model. TCR-Ig may represent a novel group of immunotherapeutics that has the potential to expand the range of tumors available for targeted therapies beyond those currently addressed by conventional antibody-based approaches.
Variegated gene expression and Sox-13 mediated regulation of WC1, a hybrid PRR/Co-receptor exclusive to memory gd T cells
Authors: Damani-Yokota Payal , Telfer Janice C , Baldwin Cynthia L 
Published: January 2023

Abstract
γδ T cells have broader reactivity and respond more rapidly compared to αβ T cells which are restricted to peptide bound to MHC. Some γδ T cells bear lineage-specific glycoproteins known as WC1 that are coded by a multigenic family composed of 13 members. Structurally, WC1 molecules have 6–11 SRCR domains, followed by a conserved transmembrane and a variable cytoplasmic tail. γδ T cells expressing these molecules can be further divided into subsets that express either WC1-3 or WC1-4, a dichotomy analogous to CD4 and CD8 molecules and similarly set up in the thymus. While WC1-3+ and WC1-4+ γδ T cells share a restricted repertoire of the γδ TCR genes, expression of WC1-3 vs. WC1-4 determines whether the γδ T cell is activated by Leptospira; reactivity correlates with expression of WC1-3 and the ability of its SRCR domains to bind to Leptospira. ShRNA-mediated knockdown of WC1-3 significantly reduces the ability of WC1-3 expressing γδ T cells to respond to the bacteria, guiding us to a hypothesis that WC1 is essential for recognition of bacterial pathogens by γδ T cells and that the WC1 family members expressed by a particular cell will determine its ability to respond to an infection. Evaluation of WC1 gene expression among individual γδ T cells indicates that WC1-3 and WC1-4 are always expressed reciprocally, establishing functional subsets. These γδ T cell subsets appear to be set in the thymus, alluding to the transcription factor occupancy of WC1-3 vs WC1-4. Occupancy of candidate Sox13 binding sites differ between WC1-3 and WC1-4 loci, suggesting that Sox13 may play an important role in determining the ability of mature γδ T cells to respond to bacteria.
Assessing the serological antibody repertoire to Flaviviruses in the endemic population of the Zika forest in Uganda
Authors: Demina Anna Vladimirovna , Lutwama Julius J , Hertz Tomer , Lobel Leslie 
Published: January 2023

Abstract
While the Zika virus has been suspected to cause complications in newborns of women infected during pregnancy, it has circulated in Africa for more --than half a century without any reported cases of microcephaly in children. To assess the prevalence of Zika infections in the Zika forest region we collected human sera samples from 81 people (17–56 years old), who live in Zika forest region, Entebbe, Uganda in 2016, and included both individuals hospitalized with acute febrile illness, as well as health individuals. Elisa assays were used to profile the IgM, IgG serological repertoire to Zika, Dengue 1-4 (DENV1-4) and Yellow Fever (YFV) viruses. We found that 17 individuals (21%) were positive for Zika, 7 of which were hospitalized with acute infection. The hospitalized patients had overall higher antibody titers to Zika. There also were serologically positive samples for: DENV1 n=11 (13.5%); DENV2 n=9 (11%), DENV3 n=4 (5%); DENV 4 n=4 (5%). 11 subjects (13.5%) were positive for YFV. Overall, 49% of the subjects had antibodies to at least one of these Flaviviruses, and 17% had antibodies to two or more Flaviviruses. While serological responses cannot accurately diagnose the specific Flavirvirus infections in these individuals due to significant cross reactivity within this viruses our data suggests that Flavivirus infections are very common in the Zika forest region. While a recent study reported that previous dengue infections may cause antibody dependant enhancement upon subsequent Zika infection, our data suggests that the Zika forest region in which many different Flaviviruses have been circulating for over 50 years may an important region to study such effects in the human population.
Development of a rapid diagnostic kit for antigen and antibody of Middle East respiratory syndrome coronavirus
Authors: Lee Eun-Young , Ko Hae Li , Jung Seo-Yeon , Nam Jae-Hwan 
Published: January 2023

Abstract
In 2015, there was an outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) in Korea. Therefore, rapid diagnostic kits for the viral antigen and antibodies against MERS-CoV are urgently required. First, we produced and purified a spike nanoparticle of MERS-CoV by the insect expression system. It was highly soluble and intact for immunization. The spike nanoparticle was used as a coating antigen for enzyme-linked immunosorbent assay (ELISA) to detect MERS-CoV-specific antibodies in the serum. The spike nanoparticle, at least with 1.5 ng/well as a coating antigen, can detect serially diluted standard antibodies purified from rabbits immunized with inactivated MERS-CoV, and from rats immunized with spike nanoparticle. Moreover, it can successfully detect MERS-CoV in patient serum. Next, to develop sandwich ELISA to detect MERS-CoV antigens, we generated 616 hybridoma clones and selected 37 monoclonal antibodies (MAbs), which exclusively reacted against the spike nanoparticle. Among them, three MAbs (S-15, S-16, and S-34) showed about 40% plaque reduction against MERS-CoV. Of the 37 MAbs, we selected the two that showed the optimal combinations to develop sandwich ELISA. This sandwich ELISA also can detect serially diluted MERS-CoV antigen. In this study, we developed two MERS-CoV-specific ELISA kits. One is for detecting MERS antibodies in patient serum and the other is for detecting MERS-CoV itself. Both ELISA kits can be reliably used for diagnosis of MERS-CoV.
Adaptive immune responses to Zika virus are important for controlling virus infection and preventing infection in brain and testes
Authors: Winkler Clayton W , Myers Lara M , Woods Tyson A , Messer Ron J , Carmody Aaron B , McNally Kristin L , Scott Dana P , Hasenkrug Kim J , Best Sonja M , Peterson Karin E 
Published: January 2023

Abstract
The recent association between Zika virus (ZIKV) and neurological complications including Guillain-Barré Syndrome (GBS) in adults and CNS abnormalities in fetuses highlights the urgency to understand the immunological mechanisms controlling this emerging infection. Studies have indicated that ZIKV evades the human type I IFN response suggesting a role for the adaptive immune response in resolving infection. However, the inability of ZIKV to antagonize the mouse IFN response renders the virus highly susceptible to circulating IFN in murine models. Thus, as we show here, although wild type C57BL/6 mice mount both cell-mediated and humoral adaptive immune responses to ZIKV, these responses were not required to prevent disease. However, when the type I IFN response of mice was suppressed, then the adaptive immune responses became critical. For example, when type I IFN signaling was blocked by antibodies in Rag1−/− immunodeficient mice, the mice showed dramatic weight loss and ZIKV infection in the brain and testes. This phenotype was not observed in Rag1−/− mice or mice treated with anti-IFNAR alone. Furthermore, we found that the CD8+ T cell responses of pregnant mice to ZIKV infection were diminished compared to non-pregnant mice. It is possible that diminished cell-mediated immunity during pregnancy could increase virus spread to the fetus. These results demonstrate an important role for the adaptive immune response in control of ZIKV infection, and imply that vaccination may prevent ZIKV-related disease, particularly when the type I IFN response is suppressed as it is in humans.
Dissecting the polyclonal antibody response of nurse sharks (Ginglymostoma cirratum) to immunization with influenza A virus suggests different roles for IgNAR and IgM, but a common immunodominance hierarchy with hemagglutinin on top
Authors: Altman Meghan O , Angeletti Davide , Yewdell Jonathon W , Flajnik Martin F 
Published: January 2023

Abstract
Cartilaginous fishes are the earliest extant phylogenic group to harness germline recombination and somatic hypermutation of immunoglobulins antibodies (Igs) for immunity. In this study we immunized and 1-year-later boosted, two adult nurse sharks (Ginglymostoma cirratum) with inactivated H1 influenza A virus (IAV). After immunization, polyclonal sera from these sharks had strong antiviral properties. Primary immune and recall sera neutralized viral growth in cell culture, caused hemagglutination inhibition, and inhibited neuraminidase (NA) activity. By ELISA and protein microarray, we examined the contribution of two shark Ig isotypes, IgNAR and IgM. Unique to cartilaginous fishes, IgNAR are heavy-chain-only Igs. IgNAR from immunized sharks bound only the immunizing strain of virus. But, flow cytometry of spleen and blood revealed ~1% of IgNAR-positive cells bound both H1 and H5 HA subtypes. While these crossreactive cells exist, any crossreactive antibodies made by cells in these lineages were below our limit of detection. By contrast, both preimmune and immune serum IgM was highly crossreactive, binding H1, H3, H5, and even headless HA. In cartilaginous fish, secreted IgM comes in two independently-produced forms high affinity monomeric (7s) and pentameric (19s). Serum fractionation determined these crossreactive IgM antibodies were 19s. While capable of binding virus, we found 19s alone could not neutralize virus. Despite these differences, regardless of isotype, and in-line with our previous work with mice and lamprey, the immunodominance hierarchy of shark vaccine-induced IgNAR and IgM antibodies to IAV is consistent—HA is dominant, followed by NA, and then core proteins, such as, nucleoprotein and matrix.