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Background: Severe coronavirus disease 2019 (COVID-19) and multisystem inflammatory syndrome (MIS-C) are characterized by excessive inflammatory cytokines/chemokines. In adults, disease severity is associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific immunoglobulin G (IgG) Fc afucosylation, which induces proinflammatory cytokine secretion from innate immune cells. This study aimed to define spike IgG Fc glycosylation following SARS-CoV-2 infection in adults and children and following SARS-CoV-2 vaccination in adults and the relationships between glycan modifications and cytokines/chemokines., Methods: We analyzed longitudinal (n = 146) and cross-sectional (n = 49) serum/plasma samples from adult and pediatric COVID-19 patients, MIS-C patients, adult vaccinees, and adult and pediatric controls. We developed methods for characterizing bulk and spike IgG Fc glycosylation by capillary electrophoresis and measured levels of 10 inflammatory cytokines/chemokines by multiplexed enzyme-linked immunosorbent assay., Results: Spike IgG was more afucosylated than bulk IgG during acute adult COVID-19 and MIS-C. We observed an opposite trend following vaccination, but it was not significant. Spike IgG was more galactosylated and sialylated and less bisected than bulk IgG during adult COVID-19, with similar trends observed during pediatric COVID-19/MIS-C and following SARS-CoV-2 vaccination. Spike IgG glycosylation changed with time following adult COVID-19 or vaccination. Afucosylated spike IgG exhibited inverse and positive correlations with inflammatory markers in MIS-C and following vaccination, respectively; galactosylated and sialylated spike IgG inversely correlated with proinflammatory cytokines in adult COVID-19 and MIS-C; and bisected spike IgG positively correlated with inflammatory cytokines/chemokines in multiple groups., Conclusions: We identified previously undescribed relationships between spike IgG glycan modifications and inflammatory cytokines/chemokines that expand our understanding of IgG glycosylation changes that may impact COVID-19 and MIS-C immunopathology., Competing Interests: Potential conflicts of interest. E. J. A. is currently employed by Moderna and owns stock or stock options in Moderna. C. A. R. has received grants or contracts to Emory from BioFire Inc, GSK, MedImmune, Micron, Merck, Novavax, PaxVax, Regeneron, Pfizer, Sanofi-Pasteur, Janssen, Moderna, NIH, and CDC; is a co-inventor of patented respiratory syncytial virus (RSV) vaccine technology that has been licensed to Meissa Vaccines Inc and has received royalties from Meissa Vaccines Inc; and has patents pending or issued: “Chimeric RSV, Immunogenic Compositions, and Methods of Use,” International PCT Application No. PCT/US2016/058976, filed 28 December 2016 by Emory University and “RSV Live-Attenuated Vaccine Candidates with Deleted G-Protein Mucin Domains,” US Patent Application No. 63/411,251, filed 29 September 2022 by Emory University. N. R. has received grants or contracts to Emory from NIH, Merck, Sanofi, Pfizer, Vaccine Company, Immorna, Quidel, and Lilly; has received consulting fees from Krog; has received payment or honoraria for lectures, presentations, speaker’s bureaus, manuscript writing, or educational events from Virology Education; has received support for attending meetings and/or travel from Sanofi and Moderna; has participated in data safety monitoring boards for Emmes, ICON, Biomedical Advanced Research and Development Authority, CyanVac, and Micron and advisory boards for Moderna, Sanofi, Seqirus, and Pfizer; has held advisory roles for the Antibacterial Resistance Leadership Group, TMRC, and CDC-Pertussis challenge; is Associate Editor of Clinical Infectious Diseases; and has received equipment, materials, drugs, medical writing, gifts, or other services to Emory from Georgia Research Alliance. E. M. S. has received grants from NIH, JC Kennedy Foundation, and Georgia Clinical & Translational Science Alliance; has received materials from Merck; has received honoraria for lectures/presentations from Merck; and has patents pending or issued: “Anti-ALPP/ALPP2 Antibodies,” US Patent Provisional Application Nos. 63/162,635, 63/301,574, and 18/282,232 filed 18 March 2021, 21 January 2022, and 15 September 2023 by Seagen, “Anti-ALPP/ALPP2 Antibodies,” International PCT Application No. PCT/US2022/020697 filed 17 March 2022 by Seagen, “Antibodies that Bind CD228,” US Patent Provisional Application No. 63/408,605, filed 21 September 2022 by Seagen, “Novel Fusion Protein Specific for CD137 and CD228,” US Patent Provisional Application Nos. 63/408,634, 63/413,174, and 63/496,463 filed 21 September 2022, 4 October 2022, and 17 April 2023 by Seagen. All other authors report no potential conflicts., (© The Author(s) 2024. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Background: Rabies is a zoonotic viral encephalitis that is endemic in many countries and confers a high mortality. Licensed vaccines require several doses to ensure efficacy. To investigate a logistically favorable approach, we assessed the safety and immunogenicity of ChAd155-RG, a novel investigational rabies vaccine using a replication-defective chimpanzee adenovirus vector., Methods: We conducted a first-in-human, phase 1, randomized, double-blind, dose-escalation trial comparing ChAd155-RG with a licensed inactivated vaccine (RabAvert) in healthy adults. Participants received either RabAvert at standard dosing or ChAd155-RG at a low dose for one immunization or a high dose for one or two immunizations. To assess safety, we evaluated reactogenicity, unsolicited adverse events, and thrombotic events. To measure immunogenicity, we measured rabies viral neutralizing antibody (VNA) titers and anti-ChAd155 neutralizing antibodies., Results: Mild to moderate systemic reactogenicity and transient lymphopenia and neutropenia were more common among recipients of ChAd155-RG compared with those who received RabAvert. No thrombotic events or serious adverse events were reported. Only the groups receiving RabAvert or two doses of high-dose ChAd155-RG achieved 100 % seroconversion, and seroprotection was most durable in the RabAvert group. Most participants had preexisting anti-vector antibodies, which were boosted by ChAd155-RG. Baseline and post-vaccination anti-vector antibody titers were negatively associated with post-vaccination rabies VNA titers., Conclusions: In this phase 1 clinical trial, a novel rabies vaccine using a simian adenovirus vector was safe and tolerable, but generated lower, less durable rabies VNA titers than a standard inactivated rabies virus vaccine, which may be due to preexisting, anti-vector immunity., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Varun K. Phadke reports financial support was provided by National Institute of Allergy and Infectious Diseases. Mark J. Mulligan reports financial support was provided by National Institute of Allergy and Infectious Diseases. Nadine Rouphael reports financial support was provided by National Institute of Allergy and Infectious Diseases. Jamie Retallick reports financial support was provided by National Institute of Allergy and Infectious Diseases. Janine Seetahal reports financial support was provided by National Institute of Allergy and Infectious Diseases. Mark Koller reports financial support was provided by National Institute of Allergy and Infectious Diseases. Nadine Rouphael reports a relationship with The Krog Group that includes: consulting or advisory. Nadine Rouphael reports a relationship with Virology Education that includes: speaking and lecture fees. Nadine Rouphael reports a relationship with Sanofi SA that includes: consulting or advisory and travel reimbursement. Nadine Rouphael reports a relationship with Moderna Inc. that includes: consulting or advisory and travel reimbursement. Nadine Rouphael reports a relationship with Seqirus Srl that includes: consulting or advisory. Nadine Rouphael reports a relationship with Pfizer Inc. that includes: consulting or advisory. Nadine Rouphael reports a relationship with The Emmes Company LLC that includes: consulting or advisory. Nadine Rouphael reports a relationship with ICON Clinical Research LLC that includes: consulting or advisory. Nadine Rouphael reports a relationship with Biomedical Advanced Research and Development Authority that includes: consulting or advisory. Nadine Rouphael reports a relationship with Cyanvac LLC that includes: consulting or advisory. Mark J. Mulligan reports a relationship with HilleVax, Inc. that includes: consulting or advisory. Mark J. Mulligan reports a relationship with Sanofi SA that includes: consulting or advisory. Mark J. Mulligan reports a relationship with Pfizer Inc. that includes: consulting or advisory. Mark J. Mulligan reports a relationship with Meissa Inc. that includes: consulting or advisory. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. GlaxoSmithKline was provided the opportunity to review a preliminary version of this manuscript for factual accuracy, but the authors are solely responsible for final content and interpretation. The authors have no conflicts of interest to disclose., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Severe COVID and multisystem inflammatory syndrome (MIS-C) are characterized by excessive inflammatory cytokines/chemokines. In adults, disease severity is associated with SARS-CoV-2-specific IgG Fc afucosylation, which induces pro-inflammatory cytokine secretion from innate immune cells. This study aimed to define spike IgG Fc glycosylation following SARS-CoV-2 infection in adults and children and following SARS-CoV-2 vaccination in adults and the relationships between glycan modifications and cytokine/chemokine levels., Methods: We analyzed longitudinal (n=146) and cross-sectional (n=49) serum/plasma samples from adult and pediatric COVID patients, MIS-C patients, adult vaccinees, and adult and pediatric healthy controls. We developed methods for characterizing bulk and spike IgG Fc glycosylation by capillary electrophoresis (CE) and measured levels of ten inflammatory cytokines/chemokines by multiplexed ELISA., Results: Spike IgG were more afucosylated than bulk IgG during acute adult COVID and MIS-C. We observed an opposite trend following vaccination, but it was not significant. Spike IgG were more galactosylated and sialylated and less bisected than bulk IgG during adult COVID, with similar trends observed during pediatric COVID/MIS-C and following SARS-CoV-2 vaccination. Spike IgG glycosylation changed with time following adult COVID or vaccination. Afucosylated spike IgG exhibited inverse and positive correlations with inflammatory markers in MIS-C and following vaccination, respectively; galactosylated and sialylated spike IgG inversely correlated with pro-inflammatory cytokines in adult COVID and MIS-C; and bisected spike IgG positively correlated with inflammatory cytokines/chemokines in multiple groups., Conclusions: We identified previously undescribed relationships between spike IgG glycan modifications and inflammatory cytokines/chemokines that expand our understanding of IgG glycosylation changes that may impact COVID and MIS-C immunopathology.

Introduction: Establishing the safety and immunogenicity of a hepatitis E virus vaccine in multiple populations could facilitate broader access and prevent maternal and infant mortality., Methods: We conducted a phase 1, randomized, double-blinded, placebo-controlled (4:1 vaccine: placebo) trial of 30 µg HEV-239 (Hecolin®, Xiamen Innovax Biotech Company Limited, China) administered intramuscularly in healthy US adults aged 18-45 years. Participants were vaccinated on days 1, 29, and 180. Participants reported solicited local and systemic reactions for 7 days following vaccination and were followed through 12 months after enrollment for safety and immunogenicity (IgG, IgM)., Results: Solicited local and systemic reactions between treatment and placebo group were similar and overall mild. No participants experienced serious adverse events related to HEV-239. All participants receiving HEV-239 seroconverted at one month following the first dose and remained seropositive throughout the study. HEV-239 elicited a robust hepatitis E IgG response that peaked one month following the second dose (Geometric Mean Concentration (GMC) 6.16; 95% CI 4.40-8.63), was boosted with the third dose (GMC 11.50; 95% CI 7.90-16.75) and persisted through 6 months., Conclusions: HEV-239 is safe and elicits a durable immune response through at least 6 months after the third dose in healthy US adults., Clinical Trials Registration: NCT03827395. Safety Study of Hepatitis E Vaccine (HEV239) - Full Text View - ClinicalTrials.gov., (© The Author(s) 2024. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

People living with HIV (PLH) experience higher rates of HPV infection as well as an increased risk of HPV-related disease, including malignancies. Although they are considered a high-priority group for HPV vaccination, there are limited data regarding the long-term immunogenicity and efficacy of HPV vaccines in this population. Seroconversion rates and geometric mean titers elicited by vaccination are lower in PLH compared to immunocompetent participants, especially in individuals with CD4 counts below 200 cells/mm 3 and a detectable viral load. The significance of these differences is still unclear, as a correlate of protection has not been identified. Few studies have focused on demonstrating vaccine efficacy in PLH, with variable results depending on the age at vaccination and baseline seropositivity. Although waning humoral immunity for HPV seems to be more rapid in this population, there is evidence that suggests that seropositivity lasts at least 2-4 years following vaccination. Further research is needed to determine the differences between vaccine formulations and the impact of administrating additional doses on durability of immune protection., Competing Interests: E.M.S. received honoraria from Merck for lectures delivered. N.R. is a paid consultant for ICON and EMMES as well as a safety consultant for clinical trials and serves on the advisory boards for GSK and Moderna. Emory receives funds for N.R. to conduct research from Sanofi, Lilly, Merck, Quidel and Pfizer. C.L., H.S., B.K., L.K. and I.O. declare no conflicts of interest.

The coronavirus disease 2019 (COVID-19) pandemic and associated increase in family care responsibilities resulted in unsustainable personal and professional workloads for infectious diseases (ID) faculty on the front lines. This was especially true for early-stage faculty (ESF), many of whom had caregiving responsibilities. In addition, female faculty, underrepresented in medicine and science faculty and particularly ESF, experienced marked declines in research productivity, which significantly impacts career trajectories. When combined with staffing shortages due to an aging workforce and suboptimal recruitment and retention in ID, these work-life imbalances have brought the field to an inflection point. We propose actionable recommendations and call on ID leaders to act to close the gender, racial, and ethnic gaps to improve the recruitment, retention, and advancement of ESF in ID. By investing in systemic change to make the ID workforce more equitable, we can embody the shared ideals of diversity and inclusion and prepare for the next pandemic., Competing Interests: Potential conflicts of interest. The authors disclose the following relationships unrelated to the manuscript: grants or contracts: NIH, Sanofi, and GSK (A. W.); Centers for Disease Control and Prevention for vaccine safety studies (K. M. E.); NIH (1UM1AI148372-01HHSN272201600016C, HHSN272201600014C, 5UL1TR000077, and 5U101AI163099 to L. D. I.); Ansun BioPharma, Astellas, Pfizer, Takeda, and AiCuris (L. D. I.); Merck (L. D. I. and I. O.); National Cancer Institute at the NIH (R01 CA213130-S), Department of Medicine Diversity Academic Development Scholar Award at the University of Washington, and Bristol-Myers Squibb Foundation–National Medical Fellowships (H. S. K.); Pfizer and AstraZeneca (F. S. H.). Royalties or licenses: UpToDate (A. W.). Consulting fees: Bionet and IBM (K. M. E.); Takeda (L. D. I.); and Aicuris, Crozet, Auritec, Dxnow, and Gilead (A. W.). Payment or honoraria: RMEI (L. D. I.). Support for attending meetings/travel: ID SAFE: Swiss Infectious Disease Meeting (L. D. I.); Innovative Molecules (A. W.). Participation on a data and safety monitoring board or advisory board: Pfizer, Sanofi, Seqirus, Merck, Moderna, Roche, X-4 Pharma (K. M. E.); Merck (L. D. I.); Merck, X-Vax, Vir, and Curevo (A. W.). Leadership or fiduciary role: Executive Board and Past President, International Society of Heart and Lung Transplantation (L. D. I.); Chair for Disease Transmission Ad Hoc committee, Organ Procurement and Transplantation Network/United Network for Organ Sharing (L. D. I.); Research Affairs Committee, Pediatric Infectious Diseases Society (L. M. H.). Receipt of equipment, materials, drugs, medical writing, gifts, or other services: Merck vaccine for a clinical trial (A. W.). All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2022. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Grandparenting activities are of increasing interest to researchers seeking to understand reduced social engagement and depression among aging adults. Heterogeneity in the population and caretaking roles complicate its measurement. We piloted a measure of grandparenting activities among 79 grandparents (aged 55+) in Sri Lanka and correlated those activity levels with psychological distress. Second, we explored whether the aforementioned correlation varied by grandparent functional limitations. We found that greater engagement in generative grandparenting activities was correlated with lower distress, and that association was stronger among grandparents with more functional limitations. We discuss possible explanations and implications of these findings.

Background: Antibodies against SARS-CoV-2 can be detected by various testing platforms, but a detailed understanding of assay performance is critical. Methods: We developed and validated a simple enzyme-linked immunosorbent assay (ELISA) to detect IgG binding to the receptor-binding domain (RBD) of SARS-CoV-2, which was then applied for surveillance. ELISA results were compared to a set of complimentary serologic assays using a large panel of clinical research samples. Results: The RBD ELISA exhibited robust performance in ROC curve analysis (AUC> 0.99; Se = 89%, Sp = 99.3%). Antibodies were detected in 23/353 (6.5%) healthcare workers, 6/9 RT-PCR-confirmed mild COVID-19 cases, and 0/30 non-COVID-19 cases from an ambulatory site. RBD ELISA showed a positive correlation with neutralizing activity ( p = <0.0001, R 2 = 0.26). Conclusions: We applied a validated SARS-CoV-2-specific IgG ELISA in multiple contexts and performed orthogonal testing on samples. This study demonstrates the utility of a simple serologic assay for detecting prior SARS-CoV-2 infection, particularly as a tool for efficiently testing large numbers of samples as in population surveillance. Our work also highlights that precise understanding of SARS-CoV-2 infection and immunity at the individual level, particularly with wide availability of vaccination, may be improved by orthogonal testing and/or more complex assays such as multiplex bead assays., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Sherman, Smith, Zhu, Taibl, Howard-Anderson, Landay, Pisanic, Kleinhenz, Simon, Espinoza, Edupuganti, Hammond, Rouphael, Shen, Fairley, Edupuganti, Cardona-Ospina, Rodriguez-Morales, Premkumar, Wrammert, Tarleton, Fridkin, Heaney, Scherer and Collins.)

We discovered 3 invasive, multidrug-resistant Streptococcus pneumoniae isolates of vaccine-refractory capsular serotype 3 that recently arose within the successful sequence type 271 complex through a serotype switch recombination event. Mapping genomic recombination sites within the serotype 3/sequence type 271 progeny revealed a 55.9-kb donated fragment that encompassed cps3, pbp1a, and additional virulence factors.

The factors that control the development of an effective immune response to the recently emerged SARS-CoV-2 virus are poorly understood. In this study, we provide a cross-sectional analysis of the dynamics of B cell responses to SARS-CoV-2 infection in hospitalized COVID-19 patients. We observe changes in B cell subsets consistent with a robust humoral immune response, including significant expansion of plasmablasts and activated receptor-binding domain (RBD)-specific memory B cell populations. We observe elevated titers of Abs to SARS-CoV-2 RBD, full-length Spike, and nucleoprotein over the course of infection, with higher levels of RBD-specific IgG correlating with increased serum neutralization. Depletion of RBD-specific Abs from serum removed a major portion of neutralizing activity in most individuals. Some donors did retain significant residual neutralization activity, suggesting a potential Ab subset targeting non-RBD epitopes. Taken together, these findings are instructive for future vaccine design and mAb strategies., (Copyright © 2021 by The American Association of Immunologists, Inc.)

The isolation of human monoclonal antibodies (mAbs) arising from natural infection with human pathogens has proven to be a powerful technology, facilitating the understanding of the host response to infection at a molecular level. mAbs can reveal sites of vulnerability on pathogens and illuminate the biological function of the antigenic targets. Moreover, mAbs have the potential to be used directly for therapeutic applications such as passive delivery to prevent infection in susceptible target populations, and as treatment of established infection. The isolation of antigen-specific B cells from vaccine trials can also assist in deciphering whether the desired B cells are being targeted by a given vaccine. Several different processes have been developed to isolate mAbs, but all are generally labor-intensive and result in varying degrees of efficiency. Here, we describe the development of a cost-effective feeder cell line that stably expresses CD40-ligand, interleukin-2 and interleukin-21. Sorting of single B cells onto a layer of irradiated feeder cells sustained antibody production that permits functional screening of secreted antibodies in a manner that enables subsequent recovery of B cells for recombinant antibody cloning. As a proof of concept, we show that this approach can be used to isolate B cells that secrete antibodies that neutralize human papilloma virus (HPV) from participants of an HPV vaccine study., (Copyright © 2020. Published by Elsevier B.V.)

SARS-CoV-2, the virus responsible for COVID-19, is causing a devastating worldwide pandemic, and there is a pressing need to understand the development, specificity, and neutralizing potency of humoral immune responses during acute infection. We report a cross-sectional study of antibody responses to the receptor-binding domain (RBD) of the spike protein and virus neutralization activity in a cohort of 44 hospitalized COVID-19 patients. RBD-specific IgG responses are detectable in all patients 6 days after PCR confirmation. Isotype switching to IgG occurs rapidly, primarily to IgG1 and IgG3. Using a clinical SARS-CoV-2 isolate, neutralizing antibody titers are detectable in all patients by 6 days after PCR confirmation and correlate with RBD-specific binding IgG titers. The RBD-specific binding data were further validated in a clinical setting with 231 PCR-confirmed COVID-19 patient samples. These findings have implications for understanding protective immunity against SARS-CoV-2, therapeutic use of immune plasma, and development of much-needed vaccines., Competing Interests: The authors declare no competing interests., (© 2020 The Author(s).)

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that first emerged in late 2019 is responsible for a pandemic of severe respiratory illness. People infected with this highly contagious virus can present with clinically inapparent, mild, or severe disease. Currently, the virus infection in individuals and at the population level is being monitored by PCR testing of symptomatic patients for the presence of viral RNA. There is an urgent need for SARS-CoV-2 serologic tests to identify all infected individuals, irrespective of clinical symptoms, to conduct surveillance and implement strategies to contain spread. As the receptor binding domain (RBD) of the spike protein is poorly conserved between SARS-CoVs and other pathogenic human coronaviruses, the RBD represents a promising antigen for detecting CoV-specific antibodies in people. Here we use a large panel of human sera (63 SARS-CoV-2 patients and 71 control subjects) and hyperimmune sera from animals exposed to zoonotic CoVs to evaluate RBD's performance as an antigen for reliable detection of SARS-CoV-2-specific antibodies. By day 9 after the onset of symptoms, the recombinant SARS-CoV-2 RBD antigen was highly sensitive (98%) and specific (100%) for antibodies induced by SARS-CoVs. We observed a strong correlation between levels of RBD binding antibodies and SARS-CoV-2 neutralizing antibodies in patients. Our results, which reveal the early kinetics of SARS-CoV-2 antibody responses, support using the RBD antigen in serological diagnostic assays and RBD-specific antibody levels as a correlate of SARS-CoV-2 neutralizing antibodies in people., (Copyright © 2020, The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Current guidance recommends that adolescents receive a 2-dose human papillomavirus (HPV) vaccine, whereas young adults and immunocompromised persons receive 3 doses. We examined secondary responses of vaccine-elicited memory B cells (Bmem) in naive women receiving 3 doses of the quadrivalent HPV vaccine to understand the quality of B-cell memory generated by this highly effective vaccine. Unexpectedly, we observed a lower Bmem response rate and magnitude of Bmem responses to the third dose than to a booster dose administered at month 24. Moreover, high titers of antigen-specific serum antibody at vaccination inversely correlated with Bmem responses. As the purpose of additional doses/boosters is to stimulate Bmem to rapidly boost antibody levels, these results indicate the timing of the third dose is suboptimal and lend support to a 2-dose HPV vaccine for young adults. Our findings also indicate more broadly that multidose vaccine schedules should be rationally determined on the basis of Bmem responses., (© The Author(s) 2017. Published by Oxford University Press for the Infectious Diseases Society of America.)

Although licensed human papillomavirus (HPV) vaccines are most efficacious in persons never infected with HPV, they also reduce infection and disease in previously infected subjects, indicating natural immunity is not entirely protective against HPV re-infection. The aim of this exploratory study was to examine the B cell memory elicited by HPV infection and evaluate whether vaccination merely boosts antibody (Ab) levels in previously infected subjects or also improves the quality of B cell memory. Toward this end, the memory B cells (Bmem) of five unvaccinated, HPV-seropositive subjects were isolated and characterized, and subject recall responses to a single HPV vaccine dose were analyzed. Vaccination boosted Ab levels 24- to 930-fold (median 77-fold) and Bmem numbers 3- to 27-fold (median 6-fold). In addition, Abs cloned from naturally elicited Bmem were generally non-neutralizing, whereas all those isolated following vaccination were neutralizing. Moreover, Ab and plasmablast responses indicative of memory recall responses were only observed in two subjects. These results suggest HPV vaccination augments both the magnitude and quality of natural immunity and demonstrate that sexually active persons could also benefit from HPV vaccination. This study may have important public policy implications, especially for the older 'catch-up' group within the vaccine's target population., (Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.)

Carbopol is a polyanionic carbomer used in man for topical application and drug delivery purposes. However parenteral administration of Carbopol in animal models results in systemic adjuvant activity including strong pro-inflammatory type-1 T-cell (Th1) polarization. Here we investigated potential pathways of immune activation by Carbopol by comparison with other well-characterized adjuvants. Carbopol administration triggered rapid and robust leukocyte recruitment, pro-inflammatory cytokine secretion and antigen capture largely by inflammatory monocytes. The induction of antigen specific Th1 cells by Carbopol was found to occur via a non-canonical pathway, independent of MyD88/TRIF signaling and in the absence of pattern-recognition-receptor (PRR) activation typically associated with Th1/Ig2a induction. Using multispectral fluorescence imaging (Imagestream) and electron microscopy we demonstrated that phagocytic uptake of Carbopol particles followed by entry into the phagosomal/lysosomal pathway elicited conformational changes to the polymer and reactive oxygen species (ROS) production. We therefore conclude that Carbopol may mediate its adjuvant activity via novel mechanisms of antigen presenting cell activation and Th1 induction, leading to enhanced IgG2a responses independent of microbial pattern recognition., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Licensed human papillomavirus (HPV) vaccines provide near complete protection against the types of HPV that most commonly cause anogenital and oropharyngeal cancers (HPV 16 and 18) when administered to individuals naive to these types. These vaccines, like most other prophylactic vaccines, appear to protect by generating antibodies. However, almost nothing is known about the immunological memory that forms following HPV vaccination, which is required for long-term immunity. Here, we have identified and isolated HPV 16-specific memory B cells from female adolescents and young women who received the quadrivalent HPV vaccine in the absence of pre-existing immunity, using fluorescently conjugated HPV 16 pseudoviruses to label antigen receptors on the surface of memory B cells. Antibodies cloned and expressed from these singly sorted HPV 16-pseudovirus labeled memory B cells were predominantly IgG (>IgA>IgM), utilized diverse variable genes, and potently neutralized HPV 16 pseudoviruses in vitro despite possessing only average levels of somatic mutation. These findings suggest that the quadrivalent HPV vaccine provides an excellent model for studying the development of B cell memory; and, in the context of what is known about memory B cells elicited by influenza vaccination/infection, HIV-1 infection, or tetanus toxoid vaccination, indicates that extensive somatic hypermutation is not required to achieve potent vaccine-specific neutralizing antibody responses.

Protection against mucosally transmitted infections probably requires immunity at the site of pathogen entry, yet there are no mucosal adjuvant formulations licensed for human use. Polyethyleneimine (PEI) represents a family of organic polycations used as nucleic acid transfection reagents in vitro and DNA vaccine delivery vehicles in vivo. Here we show that diverse PEI forms have potent mucosal adjuvant activity for viral subunit glycoprotein antigens. A single intranasal administration of influenza hemagglutinin or herpes simplex virus type-2 (HSV-2) glycoprotein D with PEI elicited robust antibody-mediated protection from an otherwise lethal infection, and was superior to existing experimental mucosal adjuvants. PEI formed nanoscale complexes with antigen, which were taken up by antigen-presenting cells in vitro and in vivo, promoted dendritic cell trafficking to draining lymph nodes and induced non-proinflammatory cytokine responses. PEI adjuvanticity required release of host double-stranded DNA that triggered Irf3-dependent signaling. PEI therefore merits further investigation as a mucosal adjuvant for human use.

The broadly neutralizing anti-HIV antibody 4E10 recognizes an epitope very close to the virus membrane on the glycoprotein gp41. It was previously shown that epitope recognition improves in a membrane context and that 4E10 binds directly, albeit weakly, to lipids. Furthermore, a crystal structure of Fab 4E10 complexed to an epitope peptide revealed that the centrally placed, protruding H3 loop of the antibody heavy chain does not form peptide contacts. To investigate the hypothesis that the H3 loop apex might interact with the viral membrane, two Trp residues in this region were substituted separately or in combination with either Ala or Asp by site-directed mutagenesis. The resultant IgG variants exhibited similar affinities for an epitope peptide as WT 4E10 but lower apparent affinities for both viral membrane mimetic liposomes and Env(-) virus. Variants also exhibited lower apparent affinities for Env(+) virions and failed to significantly neutralize a number of 4E10-sensitive viruses. For the extremely sensitive HXB2 virus, variants did neutralize, but at 37- to >250-fold lower titers than WT 4E10, with Asp substitutions exerting a greater effect on neutralization potency than Ala substitutions. Because reductions in lipid binding reflect trends in neutralization potency, we conclude that Trp residues in the antibody H3 loop enable membrane proximal epitope recognition through favorable lipid interactions. The requirement for lipophilic residues such as Trp adjacent to the antigen binding site may explain difficulties in eliciting 4E10-like neutralizing antibody responses by immunization and helps define a unique motif for antibody recognition of membrane proximal antigens.

Objective: In a recent report [Haynes et al. Science 2005; 308:1906-1908], difficulties in eliciting broadly neutralizing antibodies to HIV were linked to the binding of prototypic broadly neutralizing monoclonal antibodies to autoantigens and in particular, to the binding of two antigp41 antibodies, 2F5 and 4E10, to the autoantigen cardiolipin. We used a number of assays to understand whether 2F5 and 4E10 are autoreactive, polyreactive, or have a generalized affinity for lipids that may facilitate recognition of their membrane proximal epitopes., Methods: 2F5 and 4E10 were evaluated for autoreactivity using diagnostic assays developed to detect serum antibodies associated with antiphospholipid syndrome (APS). As an indication of polyreactivity, we measured the binding of 2F5 and 4E10 to liposomal bilayers of differing composition using surface plasmon resonance (SPR) spectroscopy and to protein microarrays using biochip technology., Results: 2F5 showed completely negative results in the APS and SPR studies, indicating that it is neither autoreactive nor absolutely requires phospholipid binding for epitope recognition. In contrast, 4E10 bound to more than one lipid and showed weak activity in the APS studies. The activity displayed by 4E10 more closely resembles that of antiphospholipid antibodies elicited during many infections than that of autoimmune APS antibodies, at variance with the notion that difficulites in eliciting 4E10-like antibodies can be attributed to tolerance mechanisms. The microarray studies further indicated that broadly neutralizing anti-HIV mAb are not exceptionally polyreactive., Conclusion: These results suggest that autoantigen mimicry cannot be reliably invoked as a general mechanism for HIV immune evasion.

Launched in the early 1980s, Magnet Recognition has been the focus of nurse administrators for over 2 decades in their quest for excellence in nursing practice and outcomes of patient care. The authors describe a comprehensive evaluation of the appraisal process within the Magnet Recognition Program commissioned by the American Nurses Credentialing Center and discusses recommendations for change in the program.

We have used ab initio calculations to compute all of the tensor elements of the electric field gradient for each carbon-deuterium bond in the ring of deuterated 3-methyl-indole. Previous analyses have ignored the smaller tensor elements perpendicular to principal component Vzz which is aligned with the C-2H bond (local bond z-axis). At each ring position, the smallest element Vxx is in the molecular plane and Vyy is normal to the plane of the ring. The asymmetry parameter = (Vyy - Vxx)/Vzz ranges from 0.07 at C4 to 0.11 at C2. We used the perpendicular (off-bond) tensor elements, in concert with an improved understanding of the indole ring geometry, to analyze prototype 2H NMR spectra from well-oriented, hydrated peptide/lipid samples. For each of the four tryptophans of membrane-spanning gramicidin A (gA) channels, the inclusion of the perpendicular elements changes the deduced ring tilt by nearly 10 and increases the ring principal order parameter Szz for overall "wobble" with respect to the membrane normal (molecular z-axis). With the improved analysis, the magnitude of Szz for the outermost indole rings of Trp13 and Trp15 is indistinguishable from that observed previously for backbone atoms (0.93 +/- 0.03). For the Trp9 and Trp11 rings, which are slightly more buried within the membrane, Szz is slightly lower (0.86 +/- 0.03). The results show that the perpendicular elements are important for the detailed analysis of 2H NMR spectra from aromatic ring systems.

An advanced oxidation process, anodic Fenton treatment (AFT), and a mixed microbial culture were used to degrade metribuzin [4-amino-6-tert-butyl-3-methylthio-1,2,4- triazin-5(4H)-one], a broad-use triazinone herbicide. Complete and rapid removal of metribuzin was demonstrated. The appearance and subsequent degradation of metribuzin oxidation products--deaminated metribuzin (DA), diketo metribuzin (DK), as well as the production of deaminated diketo metribuzin (DADK)--were observed. To support the use of AFT as a chemical pretreatment, the ratio of 5-day biochemical oxygen demand (BOD5) as measured in the standard test to chemical oxygen demand (COD) was investigated, and an increase from 0.03, a nonbiodegradable solution, to 0.35, a biodegradable solution, was observed. This increase in biodegradability was associated with decreased metribuzin, DA, and DK concentrations and increased DADK concentration. AFT effluent was inoculated with either an enriched microbial culture or Polyseed, a commercially available inoculum. Although there was minimal biodegradation of the remaining metribuzin, there was a significant decrease in DA concentration in inoculated incubations compared with sterile controls after 5- and 10-minute AFT treatment. The enrichment inoculate appeared more adapted toward the less-oxidized, 5-minute-treated effluent, whereas the Polyseed culture, developed to degrade complex waste solutions, appeared to be more effective in a moreoxidized, 10-minute-treated, and potentially more complex effluent. This research supports the continued investigation of AFT and biodegradation as a binary treatment of aqueous pesticide wastes.

Metribuzin, a widely used herbicide and a frequently detected pollutant in the environment, was studied as a target compound for membrane anodic Fenton treatment (AFT), a Fenton technology with application potential for on-site treatment of pesticide wastewater. It was found that the degradation kinetics of metribuzin do not obey the AFT model, a previously developed model that fit AFT degradation kinetics of all previously investigated pesticides. The lack of fit for metribuzin data was determined to result from a weak interaction between metribuzin and the ferric ion, resulting in a significant reduction in availability of metribuzin for reaction with hydroxyl radicals during AFT, thus slowing degradation. A revised kinetic model was developed based on the original AFT model with the addition of this interaction. Results demonstrate that the new kinetic model fits metribuzin degradation data quite well at different delivery rates of Fenton reagent and at different temperatures. This weak interaction is also found to exist between ferric ion and several other triazinone/triazine herbicides during membrane AFT. The interaction intensity correlates with the electron-withdrawing/-donating property of substituents on the triazine/triazinone ring. The stronger the electron-donating ability of substituents, the stronger the interaction.

Chloral hydrate is widely used as a sedative in pediatric medicine and is a by-product of water chlorination and a metabolic intermediate in the biotransformation of trichloroethylene. Chloral hydrate and its major metabolite, trichloroacetic acid, induce liver tumors in B6C3F1 mice, a strain that can exhibit high rates of background liver tumor incidence, which is associated with increased body weight. This report describes the influence of diet and body weight on the acute toxicity, hepatic enzyme response, and toxickinetics of chloral hydrate as part of a larger study investigating the carcinogenicity of chloral hydrate in ad libitum-fed and dietary controlled mice. Dietary control involves moderate food restriction to maintain the test animals at an idealized body weight. Mice were dosed with chloral hydrate at 0, 50, 100, 250, 500, and 1000 mg/kg daily, 5 days/week, by aqueous gavage for 2 weekly dosing cycles. Three diet groups were used: ad libitum, dietary control, and 40% caloric restriction. Both dietary control and caloric restriction slightly reduced acute toxicity of high doses of chloral hydrate and potentiated the induction of hepatic enzymes associated with peroxisome proliferation. Chloral hydrate toxicokinetics were investigated using blood samples obtained by sequential tail clipping and a microscale gas chromatography technique. It was rapidly cleared from serum within 3 h of dosing. Trichloroacetate was the major metabolite in serum in all three diet groups. Although the area under the curve values for serum trichloroacetate were slightly greater in the dietary controlled and calorically restricted groups than in the ad libitum-fed groups, this increase did not appear to completely account for the potentiation of hepatic enzyme induction by dietary restriction.

We have used experimental deuterium NMR spectra from labeled tryptophans in membrane-spanning gramicidin A (gA)(1) channels to refine the geometry of the indole ring and, specifically, the C2-(2)H bond direction. By using partial exchange in a cold organic acid, we were able to selectively deuterate ring positions C2 and C5 and, thereby, define unambiguous spectral assignments. In a backbone-independent analysis, the assigned spectra from four distinct labeled tryptophans were used to assess the geometry of the planar indole ring. We found that the C2-(2)H bond makes an angle of about 6 degrees with respect to the normal to the indole ring bridge, and the experimental geometry was confirmed by density functional calculations using a 6-311G** basis set. The precisely determined ring geometry and the experimental spectra in turn are the foundation for calculations of the orientation of each tryptophan indole ring, with respect to the bilayer membrane normal, and of a principal order parameter S(zz) for each ring. The results have general significance for revising the tryptophan ring geometry that is used in protein molecular modeling, as well as for the analysis of tryptophan ring orientations in membrane-spanning proteins. The experimental precision in the definition of the indole ring geometry demonstrates yet another practical application emanating from fundamental research on the robust gramicidin channel.

As an approach to gene therapy for the respiratory manifestations of cystic fibrosis (CF), in vivo plasmid-mediated direct transfer of the normal CF transmembrane conductance regulator (CFTR) gene to the airway epithelium was investigated in mice. To evaluate the feasibility of this strategy, pRSVL, a plasmid composed of a firefly luciferase gene driven by the Rous sarcoma virus long terminal repeat (RSV-LTR), along with cationic liposomes was instilled into the trachea of C57BI/6NCR mice. With administration of 200-400 micrograms plasmid DNA, luciferase expression could be detected in the mouse lung homogenates for at least 4 wk. With this background, a CFTR expression plasmid vector (pRSVCFTR) constructed by replacing the luciferase cDNA from pRSVL with the normal human CFTR cDNA was evaluated in vivo in mice. Intratracheal instillation of pRSVCFTR with cationic liposomes followed by analysis of mouse lung RNA by polymerase chain reaction amplification (after conversion of mRNA to cDNA) using a RSV-LTR specific sense primer and a human CFTR-specific antisense primer demonstrated human CFTR mRNA transcripts from one day to 4 wk after instillation. Further, in vivo evaluation of beta-galactosidase activity after intratracheal administration of an E. coli lacZ gene expression plasmid vector directed by the cytomegalovirus promoter (pCMV beta) demonstrated that the airway epithelium was the major target of transfer and expression of the exogenous gene. These observations demonstrate successful plasmid-mediated gene transfer to the airway epithelium in vivo. This strategy may be feasible as a form of gene therapy to prevent the pulmonary manifestations of CF.