bluuebunny/crossref_metadata_2025_split huggingface dataset:
Harnessing violet and ultraviolet fluorophores for flexibility in T cell panel design
Authors: Meng Lihao , Sternberg Luise , Ruiz Natalie , Funatake Castle J 
Published: August 2023

Abstract
The cytotoxic capabilities of T cells are being leveraged for immunotherapeutic approaches in the fight against cancer. These treatments have demonstrated significant promise thus far and warrant refinement to increase efficacy, reduce unwanted effects, and expand their capabilities to more diverse tumor types. As such, continuing to broaden our understanding of T cell biology at a basic research level is critical for developing the next generation of T cell immunotherapeutics.
Spectral flow cytometry enables recognition of the unique spectral signature of individual fluorophores and facilitates the addition of more antibodies by accommodating fluorophore combinations which are incompatible on conventional cytometers. An increased availability of fluorophore-conjugated antibodies for surface and intracellular antigens in the ultraviolet and violet laser range enable researchers to deepen their understanding of T cell activation and function with more flexibility in adding markers of interest. Here, we developed an optimized panel for the investigation of human T cell differentiation, activation, proliferation, and function. This panel demonstrates consistent and robust characterization of in vitro-activated human T cells, while leaving most channels on the blue, yellow-green, and red lasers available for additional markers of interest. This approach lends itself additional customization, facilitating deeper insights from a single, limited sample enabled by the broad availability of fluorophore-conjugated antibodies supporting these laser lines.
Thermo fisher Scientific
Adenosine deaminase augments SARS-CoV-2 specific cellular and humoral responses in aged mouse models of immunization and challenge
Authors: Cusimano Gina , Gary Ebony N , Tursi Nicholas J , Warner Bryce M , Connors Jennifer , Parzych Elizabeth M , Griffin Bryan D , Bell Matthew , Ali Ali R , Frase Drew , Hojecki Casey , Canziani Gabriela , Chaiken Irwin , Kannan Toshitha , Moffat Estella , Embury-Hyatt Carissa , Wootton Sarah , Kossenkov Andrew , Patel Ami , Kobasa Darwyn , Kutzler Michele A , Haddad Elias , Weiner David B 
Published: August 2023

Abstract
Despite numerous clinically available vaccines and therapeutics, aged patients remain at increased risk for COVID-19 morbidity and mortality. Furthermore, the aged patient population has suboptimal responses to SARS-CoV-2 vaccine antigens. Here, we characterized vaccine-induced responses to SARS-CoV-2 synthetic DNA vaccine antigens in aged mouse models. Aged mice had altered cellular responses, including decreased IFNγ secretion and increased TNFα secretion as compared to young mice. Aged mice had significantly decreased binding and neutralizing antibodies in their serum compared to their young counterparts. Co-immunization with the plasmid-encoded molecular adjuvant adenosine deaminase-1 (pADA) enhanced cellular responses and expanded the breadth and affinity of humoral responses in aged mice. pADA co-delivery also altered the gene expression profiles of lymph node lymphocytes in aged animals. scRNAseq analysis of aged lymph nodes revealed that pADA co-immunization supported a strong TH1 gene profile and decreased FoxP3 gene expression. Upon challenge pADA co-immunization decreased viral loads and age-associated morbidity and mortality in mouse-adapted and ACE2 transgenic SARS-CoV-2 challenge models. These data support the use of aged mice as a model for age-associated decreases in vaccine immunogenicity and increased in infection-mediated morbidity and mortality in the context of SARS-CoV-2. These studies provide further support for the use of adenosine deaminase as a molecular adjuvant in the elderly.
This work was supported by NIH NCI award T32 CA09171 (ENG) and NIH NIAID award T32-AI-055400 (EMP) and The Wistar Institute coronavirus discovery fund with additional support from CEPI/Inovio Pharmaceuticals.
Culturally-specific education can change perception of risk of Human Papillomavirus infection and need for vaccination
Authors: Poole Brian D , Redd David S , Altman Jessica D , Jensen Jamie L , Sloan-Aagard Chantel D , Crook Triston B , Asay Aaron E , Nielson Bryce U , Bodily Ruth J , Miner Dashiell S 
Published: August 2023

Abstract
Safe and effective vaccines have been developed that protect against high-risk strains of HPV, but uptake is relatively low. We previously identified factors such as sexual attitudes and HPV knowledge that impact the intent of Christian parents to vaccinate their children against HPV. One of the major factors that decreased vaccine utilization was the belief that they did not need to vaccinate their children because of their moral beliefs. We hypothesized that culturally specific interventions in the form of short videos would be effective at improving HPV vaccine attitudes. We made three short educational videos, one with a Christian focus, one informational about HPV, and one control. Videos were distributed electronically with accompanying surveys, and attitudes were measured before and after watching a randomly selected video. The religious-focused and educational interventions significantly (P=0.001) improved attitudes about HPV vaccination. The religiously-focused video also significantly diminished the belief that the HPV vaccine is unnecessary because of a family’s values (p=0.023). Parents significantly credited both interventions with improving their intent to vaccinate their children against HPV (p&lt;0.001 for both). These results suggest that culturally focused educational interventions are effective at influencing vaccine attitudes, even when those attitudes are based on religious or cultural feelings. Highly specific interventions are likely to be necessary for optimal improvement in vaccine hesitancy.
Supported in part by a research grant from Investigator-Initiated Studies Program of Merck Sharp & Dohme LLC.
Altered antibody profiles after receipt of third dose SARS-CoV-2 mRNA vaccines in solid organ transplant recipients
Authors: Lee Jenny MinChen , Shapiro Janna R , Lee John S , Sitaris Ioannis , Peralta Stephanie R , Li Maggie , Wouters Camille , Karaba Andrew H , Blankson Joel N , Tobian Aaron A R , Werbel William A , Pekosz Andrew , Klein Sabra L 
Published: August 2023

Abstract
Immunosuppressed patients, including solid organ transplant recipients (SOTRs), developed significantly lower antibody responses and had more breakthrough infections after two doses of COVID-19 mRNA vaccines, as compared with immunocompetent individuals. The aim of this study was to characterize vaccine-induced antibody responses, including IgG binding antibodies and live virus neutralizing antibodies (NAb) against the vaccine strain and Omicron variants, BA.1 and BA.5, as well as the levels of the IgG subclasses, IgG1-4, in SOTRs. Serum samples from SOTRs were analyzed at pre-vaccination, 30-days, and 90-days following the third dose COVID-19 mRNA vaccines (D3). For several measures, antibody responses were compared with healthy controls (HC). A third dose vaccination increased anti-spike (S) IgG titers in response to the SARS-CoV-2 vaccine and Omicron variants from pre- to post-D3 in SOTRs. Anti-S IgG responses to Omicron variants, however, were significantly lower than to the vaccine strain. SOTRs also had low live virus NAb titers to the vaccine and Omicron BA.1 variant, with many individuals having no detectable NAb. The subclass-specific anti-S IgG antibodies against the SARS-CoV-2 vaccine strain predominantly consisted of IgG1, IgG3, and IgG4 in SOTRs, which resembled the subclass profiles in HC at post-D3. Together, our findings illustrate that while a booster dose of mRNA vaccine induces antibody responses, including IgG1 and IgG3, which are the dominant antiviral IgG subclasses, in SOTRs, these responses are still lower than immunocompetent individuals. Therefore, consideration should be given to different formulations and doses of COVID-19 vaccines in people with severely compromised immune systems.
Boosting location of COVID-19 mRNA vaccine regulates the germinal center reaction
Authors: Jiang Wenxia , Yuan Xue , Renshaw Scott , Luo Wei 
Published: August 2023

Abstract
Germinal centers (GCs) are the specialized sites for B cells to differentiate into long-lived plasma cells or memory B cells during vaccinations. COVID-19 mRNA vaccines require boosting to mount high levels of neutralizing antibody responses. mRNA vaccines induce persistent GCs in the draining lymph nodes (dLNs), which might change the microenvironment for subsequent immunizations. It is unclear whether the site of booster immunization (ipsilateral versus contralateral) will affect the GC responses in the dLNs. To test this, we compared GC responses induced by ipsilateral and contralateral booster vaccination three weeks post the primary immunization in mice. Although these two boosting strategies induce similar numbers of GC B cells in the dLNs, we discovered that ipsilateral boosting elicits more SARS-CoV-2 RBD-specific GC B cells than contralateral boosting. However, very few RBD-specific GC B cells were differentiating into plasma cells as probed by BCL6low IRF4high PC precursors within the GC B cell population. Serum antibody levels were comparable between these boosting strategies, suggesting the memory recall response was not different. These studies suggest boosting location of COVID19 mRNA vaccines might affect RBD-specific GC B cell responses. Future studies are needed to investigate whether boosting location may impact the persistence of neutralizing antibody response induced by COVID19 mRNA vaccines.
Visualizing the early immune response to Mycobacterium tuberculosis infection in macaques immunized with intravenous BCG
Authors: Bucsan Allison N , Darrah Patricia A , Dahlvang James D , Lehman Chelsea C , Robertson Molly A , Kamenyeva Olena , Kabat Juraj , Fabozzi Giulia , March Kylie , Via Laura , Walker April , Roederer Mario , Seder Robert 
Published: August 2023

Abstract
Tuberculosis (TB) causes 1.6 million deaths annually and the standard human vaccine, intradermal (ID) Bacille-Calmette Guérin (BCG), does not prevent pulmonary TB. Previously, we showed that intravenous (IV) BCG immunization elicited robust TB-specific lung-resident T cell responses compared to ID BCG in rhesus macaques (RMs). Following Mycobacterium tuberculosis (Mtb) challenge, 6 out of 10 RMs had no detectable disease as demonstrated by bacterial burden, granuloma formation, and primary Mtb-specific immune responses. Together, these data suggest that Mtb is cleared rapidly from the lungs of IV BCG-vaccinated RMs. To investigate the vaccine-induced lung environment and anamnestic response that might mediate protection against Mtb early after challenge, we developed a high-dose Mtb infection model in RMs that allowed us to locate and live-image areas of Mtb infection in the lungs using confocal microscopy. RMs immunized for 5 months with either IV or ID BCG were challenged with Mtb Erdman mCherry and euthanized 4 or 11 days later to characterize the lung microenvironment. We observed reductions in bacterial burdens in the lungs of RMs that received IV BCG compared to ID BCG-vaccinated and unvaccinated controls by CFU plating, flow cytometry, and confocal microscopy despite using a high challenge dose. Increased T cells were quantified in the lungs of both IV and ID BCG-vaccinated RMs compared to unvaccinated controls by flow cytometry and confocal microscopy. This research provides an opportunity to mine immune correlates of protection in a preclinical model, at the site of infection, and at a post-challenge time point that has been minimally characterized.
NIH Intramural Program
Intramuscular mRNA prime plus intranasal adenoviral vector booster elicit robust respiratory mucosal IgA responses against SARS-CoV-2
Authors: Tang Jinyi , Wu Yue , Zeng Cong , Taylor Justin J , Zhu Guizhi , Weissman Drew , Liu Shan-Lu , Sun Jie 
Published: August 2023

Abstract
Current SARS-CoV-2 mRNA vaccines induce robust humoral and cellular immunity in the circulation, but its ability in eliciting respiratory mucosal immunity is less characterized. Here, we demonstrated that systemic mRNA expressing ancestral spike (mRNA-S) vaccination alone induced weak respiratory mucosal neutralizing antibody and cellular immunity, particularly against SARS-CoV-2 Omicron strain. In contrast, an immunization strategy combining systemic mRNA-S administration plus mucosal adenoviral vector expressing ancestral spike (Ad5-S) booster induced strong T cell, B cell, and antibody responses in the respiratory tract, which can last for a long time. Furthermore, we found that Ad5-S mucosal booster promoted robust IgA-producing B cells in the respiratory tract, which were correlated with the levels of mucosal S-specific IgA levels. Strikingly, these local IgA-producing B cells were more cross-reactive to the Delta and Omicron Spike proteins compared to those IgG-producing B cells. We further showed that CD4 T cell help was required for the development of IgA-producing B cell in the respiratory mucosa. Taken together, our study identified a vaccination strategy and its associated mechanisms for the induction of strong cross-reactive IgA responses, which were shown to correlate with optimal protection against breakthrough infection, especially by Omicron sub-lineages. Hence, our data provide insights into the rational design of next-generation SARS-CoV-2 mucosal vaccines required for the protection against infection by SARS-CoV-2 Omicron sub-lineages or future variants.
Mechanistic insight into immune amnesia and immunity induced by wild-type measles virus compared to live-attenuated measles vaccine
Authors: Chan Kwan Pui , Bartlett Maggie , Rubens Jessica , Brockhurst Jacqueline , Ghimire Shristi , Nguyen Benjamin H , Hauer Debbie , Griffin Diane E 
Published: August 2023

Abstract
Despite the availability of a safe and effective vaccine, measles still causes significant mortality and morbidity among children in developing countries. As a disease that induces immune amnesia with increased susceptibility to other infections but also life-long immunity to measles, little is known about the immunogenetics of B cells that elicit measles-specific antibodies (Abs) in natural infection vs vaccination. We therefore conducted single-cell RNA sequencing (scRNA-seq) of longitudinal peripheral blood mononuclear cells (PBMC) and lymph node (LN) samples from wild-type (WT) measles virus (MeV) and live-attenuated measles vaccine (LAMV)-infected rhesus macaques (RMs) and characterized their gene expression, B cell repertoires and clonotype changes via bioinformatic pipelines. Preliminary analysis of two RMs across four timepoints (one WT MeV- and one LAMV-infected RM) revealed similarities and differences in immune cell populations and B cell repertoires, providing important mechanistic insights into B cell clonal development in natural infection as compared to vaccination.
Supported by grants from NIH R01 AI153140 and the Jeffrey Richardson Fellowship
Serial intravascular staining reveals altered T cell trafficking in aged and SIV+ rhesus macaques
Authors: Potter E Lake , Brammer Daniel , Foulds Kathryn , Roederer Mario 
Published: August 2023

Abstract
Lymphocyte trafficking is required for lymphocyte development and maturation, immune surveillance and response, and for immunological memory. While trafficking is integral for a functional immune system, it may also play a role in immune dysfunction. Patients with HIV and the elderly have immune dysfunction characterized by increased inflammation, immune activation, and susceptibility to infections and cancer. As trafficking is implicated in these processes, we hypothesized that these cohorts may have altered trafficking kinetics, resulting in immunosenescence.
To determine the relationship between trafficking kinetics and immune dysfunction, we performed serial intravascular staining (SIVS) in healthy, aged, and SIV-infected rhesus macaques. SIVS uses sequential infusions of fluorescent antibody to label cells in vivo, enabling us to follow them as they migrate over time. In all animals, we found that naïve and central memory T cells underwent more and faster trafficking into tissues while effector memory and terminal effector T cells were more likely to remain in circulation. In SIV+ and aged animals, we observed increased trafficking of all subsets into tissues. A possible result of increased trafficking is that naïve and central memory cells may spend less time in lymphoid tissue and may therefore be less likely to initiate an immune response. Increased trafficking of effector cells may result in cytotoxic cells entering tissues and potentially increasing inflammation, a hallmark of the immunosenesence found in both aged and HIV+ patients. Using SIVS to evaluate trafficking in aged and SIV+ rhesus provides new insights into the crucial role of lymphocyte trafficking in the immune system.
Supported by Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Disease, National Institutes of Health
Bystander CD8+ T cells are Important Antiviral Effectors in the Brain
Authors: Berger Jennifer , Monogue Brendan , Heiden Dustin , Klarquist Jared , Beckham John David , Berg Leslie J 
Published: August 2023

Abstract
The central nervous system (CNS) presents a unique immunological environment with restricted infiltration of immune cells from the periphery. Since much is unknown about CNS-infiltrating immune cells during infection, we utilized the attenuated Venezuelan equine encephalitis virus vaccine strain TC83 (VEEV-TC83) to study T cell responses to this viral infection. VEEV-TC83 intracranial (IC) infection of wild-type mice produces high viral titers in the brain without lethality, enabling the study of the complete antiviral immune response in the CNS. Many T cells rapidly (5dpi) infiltrate the CNS, in a seemingly antigen-independent manner, upon IC VEEV-TC83 infection. During infection, CD8+ T cells (~40% of CNS-infiltrating T cells) in the brain express high levels of innate-memory markers. These bystander CD8+ T cells can produce IFNγ when treated with IL-12 and IL-18, cytokines known to induce innate signaling. IL-12, IL-18, and IL-15 (another innate-memory T cell supporter) are upregulated upon IC VEEV-TC83 infection. IC infection of IL-15 knockout (KO) mice results in decreased CNS infiltration by bystander CD8+ T cells. CD8+ T cells that do infiltrate the brains of IL-15 KO mice express fewer activation markers and cytokines, and are therefore likely not fully functional. Additionally, microglia express innate-memory cytokines during infection and likely support the recruitment and function of bystander CD8+ T cells in the CNS during infection. Data deposited in GEO indicate that infection with many neurotropic viruses induces Il15, Il12, and Il18 expression in CNS cells and tissues. Bystander CD8+ T cells are likely an important first line of defense during viral infection of the CNS.
Supported by grants from the NIH (Training grant T32 AI07405 and RO1 NS123431)
Oral and intranasal Ad5 influenza vaccination induces hemagglutinin-specific and influenza neutralizing antibodies in serum and milk of pregnant and lactating ferrets
Authors: Langel Stephanie N , Beverly Christopher , Byrne Jake , Sun Francis , Gurley Sarah Anne , Tomaras Georgia D , Tucker Sean N , Johnson Susan 
Published: August 2023

Abstract
Pregnant women and young children are at increased risk for severe influenza infection. While there are intramuscular (IM) inactivated influenza vaccines available to pregnant individuals, IM immunization may not be an ideal route to boost neutralizing antibodies in breastmilk (BM). Therefore, innovative vaccine strategies are needed to increase BM antibodies that promote neonatal protection against influenza. We hypothesize that maternal mucosal immunization stimulates a protective influenza-specific antibody response, by generating antibodies in both blood and BM. To test this, we immunized pregnant and lactating ferrets at two weeks prepartum and two weeks postpartum with an H1N1 hemagglutinin (HA) adenovirus type 5 (Ad5) vaccine via oral or intranasal routes. We detected HA-specific IgG antibodies in dam serum and HA-specific IgG and IgA antibodies in secreted milk after oral and intranasal immunization by ELISA and chemiluminescence assays. We demonstrated that serum and BM from oral and intranasally immunized dams neutralized influenza virus in vitro. Additionally, ferret kits born to oral and intanasally immunized dams had HA-specific IgG antibodies in circulation at 5–6 weeks of life. These preclinical data led to the announcement by Vaxart, Inc. that they will test their oral tablet vaccine in human lactating mothers to determine if vaccination induces BM antibodies. Future work includes using our ferret model to determine the relationship between BM antibody quantity and function and neonatal protection against influenza. An easy-to-deliver mucosal vaccine that boosts systemic and mucosal protective antibodies in the maternal-neonatal dyad is an ideal strategy for protection of these vulnerable populations.
Bill and Melinda Gates Foundation grant INV-22595D
Antiviral and autoimmune antibody levels for two years post SAR-CoV-2 infection show uniquely durable vaccination responses and partial waning of self-reactive IgG
Authors: McConney Christina , Ayuso Maria-Jose , Smith Erika , Volfson-Sedletsky Victoria , Sagar Manish , Olson Alex , Bhadelia Nahid , Snyder-Cappione Jennifer 
Published: August 2023

Abstract
As temporal distance from the start of the COVID-19 pandemic increases, there is mounting concern of the long-term immunological consequences of SARS-CoV-2 infection(s). Abnormally high levels of a panoply of autoantibodies are found during acute COVID-19, and to date it is still unclear if these antibodies are sustained for a year or longer post-infection. Additionally, if there is improved longevity of vaccine-induced antibodies when administered against the immune backdrop of previous SARS-CoV-2 infection remains to be elucidated. To address these questions, we measured a series of anti-viral and autoimmune antibodies from samples taken at intervals from seven subjects infected with SARS-CoV-2 in the spring of 2020 (five outpatient and two hospitalized). Blood draws were performed beginning at acute disease and for an additional 10–25 months. Plasma IgG and IgA antibodies reactive with SARS-CoV-2 spike, RBD, and nucleocapsid, as well as CMV Mosaic, EBV GP 350, and Flu H1 were measured using the BU ELISA protocol, autoantibodies to self-antigens associated with connective tissue disease were measured using a commercial kit. Anti-viral antibody responses induced by vaccination were sustained for more than one year post SARS-CoV-2 infection in multiple subjects. Autoantibody trajectories varied, with many positive ‘hits’ falling below baseline after 1 year, and others remaining high for all time points measured. Future work will include increasing subject numbers to a larger cohort with differing clinical presentations and outcomes to define connections between serological signatures, protection from re-infection, and PASC.
Supported by grants from CTSI Pilot grant (NIH/NCATS 5UL1TR001430-05), BU Internal Funding, MassCPR PASC Subaward #FP-0034311, and NIH R01 AG065050-01A1