Medicine and Health Sciences Commons^™

Eliminating Contamination In Umbilical Cord Blood Culture Sampling For Early-Onset Neonatal Sepsis., Vilmaris Quinones Cardona, Vanessa Lowery, David Cooperberg, Endla K Anday, Alison J Carey

Department of Pediatrics Faculty Papers

Introduction: Despite the advantages of umbilical cord blood culture (UCBC) use for diagnosis of early onset sepsis (EOS), contamination rates have deterred neonatologists from its widespread use. We aimed to implement UCBC collection in a level III neonatal intensive care unit (NICU) and apply quality improvement (QI) methods to reduce contamination in the diagnosis of early onset sepsis. Methods: Single-center implementation study utilizing quality improvement methodology to achieve 0% contamination rate in UCBC samples using the Plan-Do-Study-Act (PDSA) model for improvement. UCBC was obtained in conjunction with peripheral blood cultures (PBC) in neonates admitted to the NICU due to maternal …

The Mrna-Lnp Platform's Lipid Nanoparticle Component Used In Preclinical Vaccine Studies Is Highly Inflammatory, Sonia Ndeupen, Zhen Qin, Sonya Jacobsen, Aurélie Bouteau, Henri Estanbouli, Botond Z. Igyártó

Department of Microbiology and Immunology Faculty Papers

Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against COVID-19. Clinical trials and ongoing vaccinations present with varying degrees of protection levels and side effects. However, the drivers of the reported side effects remain poorly defined. Here we present evidence that Acuitas' LNPs used in preclinical nucleoside-modified mRNA vaccine studies are highly inflammatory in mice. Intradermal and intramuscular injection of these LNPs led to rapid and robust inflammatory responses, characterized by massive neutrophil infiltration, activation of diverse inflammatory pathways, and production of various inflammatory cytokines and chemokines. The same dose of …

A Strategy To Detect Emerging Non-Delta Sars-Cov-2 Variants With A Monoclonal Antibody Specific For The N501 Spike Residue, Rama Devudu Puligedda, Fetweh H Al-Saleem, Christoph Wirblich, Chandana Devi Kattala, Marko Jović, Laura Geiszler, Himani Devabhaktuni, Giora Z Feuerstein, Matthias J. Schnell, Markus Sack, Lawrence L Livornese, Scott K Dessain

Department of Microbiology and Immunology Faculty Papers

Efforts to control SARS-CoV-2 have been challenged by the emergence of variant strains that have important implications for clinical and epidemiological decision making. Four variants of concern (VOCs) have been designated by the Centers for Disease Control and Prevention (CDC), namely, B.1.617.2 (delta), B.1.1.7 (alpha), B.1.351 (beta), and P.1 (gamma), although the last three have been downgraded to variants being monitored (VBMs). VOCs and VBMs have shown increased transmissibility and/or disease severity, resistance to convalescent SARS-CoV-2 immunity and antibody therapeutics, and the potential to evade diagnostic detection. Methods are needed for point-of-care (POC) testing to rapidly identify these variants, protect …

Cancer Vaccines And Immunotherapy For Tumor Prevention And Treatment, Jagmohan Singh, Wilbur Bowne, Adam E. Snook

Kimmel Cancer Center Faculty Papers

In this editorial, we highlight articles published in this Special Issue of Vaccines on "Cancer Vaccines and Immunotherapy for Tumor Prevention and Treatment", recent developments in the field of cancer vaccines, and the potential for immunotherapeutic combinations in cancer care. This issue covers important developments and progress being made in the cancer vaccine field and possible future directions for exploring new technologies to produce optimal immune responses against cancer and expand the arena of prophylactic and therapeutic cancer vaccines for the treatment of this deadly disease.

Development Of A Recombinant Vaccine Against Human Onchocerciasis., David Abraham, John Graham-Brown, Darrick Carter, Sean A. Gray, Jessica A. Hess, Benjamin L. Makepeace, Sara Lustigman

Department of Microbiology and Immunology Faculty Papers

Introduction: Human onchocerciasis caused by the filarial nematode parasite Onchocerca volvulus remains a major cause of debilitating disease infecting millions primarily in Sub-Saharan Africa. The development of a prophylactic vaccine, along with mass drug administration, would facilitate meeting the goal of onchocerciasis elimination by 2030.

Areas covered: Models used to study immunity to Onchocerca include natural infection of cattle with Onchocerca ochengi and O. volvulus infective third-stage larvae implanted within diffusion chambers in mice. A vaccine, comprised of two adjuvanted recombinant antigens, induced protective immunity in genetically diverse mice suggesting that it will function similarly in diverse human populations. These …

Multiple Autonomous Cell Death Suppression Strategies Ensure Cytomegalovirus Fitness, Pratyusha Mandal, Lynsey Nagrani, Liliana Hernandez, Anita Louise Mccormick, Christopher Dillon, Heather Koehler, Linda Roback, Emad S Alnemri, Douglas Green, Edward Mocarski

Department of Biochemistry and Molecular Biology Faculty Papers

Programmed cell death pathways eliminate infected cells and regulate infection-associated inflammation during pathogen invasion. Cytomegaloviruses encode several distinct suppressors that block intrinsic apoptosis, extrinsic apoptosis, and necroptosis, pathways that impact pathogenesis of this ubiquitous herpesvirus. Here, we expanded the understanding of three cell autonomous suppression mechanisms on which murine cytomegalovirus relies: (i) M38.5-encoded viral mitochon-drial inhibitor of apoptosis (vMIA), a BAX suppressor that functions in concert with M41.1-encoded viral inhibitor of BAK oligomerization (vIBO), (ii) M36-encoded viral inhibitor of caspase-8 activation (vICA), and (iii) M45-encoded viral inhibitor of RIP/RHIM activation (vIRA). Following infection of bone marrow-derived macrophages, the virus initially …

Loss Of N1-Methylation Of G37 In Trna Induces Ribosome Stalling And Reprograms Gene Expression, Isao Masuda, Jae-Yeon Hwang, Thomas Christian, Sunita Maharjan, Fuad Mohammad, Howard Gamper, Allen R. Buskirk, Ya-Ming Hou

Department of Biochemistry and Molecular Biology Faculty Papers

N¹-methylation of G37 is required for a subset of tRNAs to maintain the translational reading-frame. While loss of m¹G37 increases ribosomal +1 frameshifting, whether it incurs additional translational defects is unknown. Here, we address this question by applying ribosome profiling to gain a genome-wide view of the effects of m¹G37 deficiency on protein synthesis. Using E coli as a model, we show that m¹G37 deficiency induces ribosome stalling at codons that are normally translated by m¹G37-containing tRNAs. Stalling occurs during decoding of affected codons at the ribosomal A site, indicating …

Targeting Human Langerin Promotes Hiv-1 Specific Humoral Immune Responses., Jérôme Kervevan, Aurélie Bouteau, Juliane S Lanza, Adele Hammoudi, Sandra Zurawski, Mathieu Surenaud, Lydie Dieudonné, Marion Bonnet, Cécile Lefebvre, Hakim Hocini, Romain Marlin, Aurélie Guguin, Barbara Hersant, Oana Hermeziu, Elisabeth Menu, Christine Lacabaratz, Jean-Daniel Lelièvre, Gerard Zurawski, Véronique Godot, Sandrine Henri, Botond Z. Igyártó, Yves Levy, Sylvain Cardinaud

Department of Microbiology and Immunology Faculty Papers

The main avenue for the development of an HIV-1 vaccine remains the induction of protective antibodies. A rationale approach is to target antigen to specific receptors on dendritic cells (DC) via fused monoclonal antibodies (mAb). In mouse and non-human primate models, targeting of skin Langerhans cells (LC) with anti-Langerin mAbs fused with HIV-1 Gag antigen drives antigen-specific humoral responses. The development of these immunization strategies in humans requires a better understanding of early immune events driven by human LC. We therefore produced anti-Langerin mAbs fused with the HIV-1 gp140z Envelope (αLC.Env). First, we show that primary skin human LC and …

Inhibitory Molecules Pd-1, Cd73 And Cd39 Are Expressed By Cd8+ T Cells In A Tissue-Dependent Manner And Can Inhibit T Cell Responses To Stimulation, Corinne J. Smith, Christopher M. Snyder

Department of Microbiology and Immunology Faculty Papers

The salivary gland is an important tissue for persistence and transmission of multiple viruses. Previous work showed that salivary gland tissue-resident CD8+ T cells elicited by viruses were poorly functional ex vivo. Using a model of persistent murine cytomegalovirus (MCMV) infection, we now show that CD8+ T cells in the salivary gland and other non-lymphoid tissues of mice express multiple molecules associated with T cell exhaustion including PD-1, CD73 and CD39. Strikingly however, these molecules were expressed independently of virus or antigen. Rather, PD-1-expressing T cells remained PD-1+ after migration into tissues regardless of infection, while CD73 was activated on …

Sars-Cov-2 Viral Proteins Nsp1 And Nsp13 Inhibit Interferon Activation Through Distinct Mechanisms, Christine Vazquez, Sydnie E Swanson, Seble G Negatu, Mark Dittmar, Jesse Miller, Holly Ramage, Sara Cherry, Kellie A Jurado

Department of Microbiology and Immunology Faculty Papers

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a devastating global pandemic, infecting over 43 million people and claiming over 1 million lives, with these numbers increasing daily. Therefore, there is urgent need to understand the molecular mechanisms governing SARS-CoV-2 pathogenesis, immune evasion, and disease progression. Here, we show that SARS-CoV-2 can block IRF3 and NF-κB activation early during virus infection. We also identify that the SARS-CoV-2 viral proteins NSP1 and NSP13 can block interferon activation via distinct mechanisms. NSP1 antagonizes interferon signaling by suppressing host mRNA translation, while NSP13 downregulates interferon and NF-κB promoter signaling by limiting TBK1 …

Intracranial Mycoplasma Hominis Infection Following Emergent Craniectomy, Zahra Qamar, Stavropoula Tjoumakaris, Matthew Pattengill, Maliha Ahmed, Bryan Hess

Division of Infectious Diseases and Environmental Medicine Faculty Papers

We present a case of a young healthy female who developed recurrent cranial wound infections after a traumatic injury, the etiologic organism finally identified as Mycoplasma hominis, an uncommon and difficult to isolate bacterium.

Resistance To Lethal Ectromelia Virus Infection Requires Type I Interferon Receptor In Natural Killer Cells And Monocytes But Not In Adaptive Immune Or Parenchymal Cells., Carolina R Melo-Silva, Pedro Alves-Peixoto, Natasha Heath, Lingjuan Tang, Brian Montoya, Cory J Knudson, Colby Stotesbury, Maria Ferez, Eric B. Wong, Luis J. Sigal

Department of Microbiology and Immunology Faculty Papers

Type I interferons (IFN-I) are antiviral cytokines that signal through the ubiquitous IFN-I receptor (IFNAR). Following footpad infection with ectromelia virus (ECTV), a mouse-specific pathogen, C57BL/6 (B6) mice survive without disease, while B6 mice broadly deficient in IFNAR succumb rapidly. We now show that for survival to ECTV, only hematopoietic cells require IFNAR expression. Survival to ECTV specifically requires IFNAR in both natural killer (NK) cells and monocytes. However, intrinsic IFNAR signaling is not essential for adaptive immune cell responses or to directly protect non-hematopoietic cells such as hepatocytes, which are principal ECTV targets. Mechanistically, IFNAR-deficient NK cells have reduced …

Inactivated Rabies Virus Vectored Sars-Cov-2 Vaccine Prevents Disease In A Syrian Hamster Model., Drishya Kurup, Delphine C Malherbe, Christoph Wirblich, Rachael Lambert, Adam J Ronk, Leila Zabihi Diba, Alexander Bukreyev, Matthias J. Schnell

Department of Microbiology and Immunology Faculty Papers

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emergent coronavirus that has caused a worldwide pandemic. Although human disease is often asymptomatic, some develop severe illnesses such as pneumonia, respiratory failure, and death. There is an urgent need for a vaccine to prevent its rapid spread as asymptomatic infections accounting for up to 40% of transmission events. Here we further evaluated an inactivated rabies vectored SARS-CoV-2 S1 vaccine CORAVAX in a Syrian hamster model. CORAVAX adjuvanted with MPLA-AddaVax, a TRL4 agonist, induced high levels of neutralizing antibodies and generated a strong Th1-biased immune response. Vaccinated hamsters were protected from …

Viral Infection Modulates Qa-1b In Infected And Bystander Cells To Properly Direct Nk Cell Killing., Maria Ferez, Cory J. Knudson, Avital Lev, Eric B. Wong, Pedro Alves-Peixoto, Lingjuan Tang, Colby Stotesbury, Luis J. Sigal

Department of Microbiology and Immunology Faculty Papers

Natural killer (NK) cell activation depends on the signaling balance of activating and inhibitory receptors. CD94 forms inhibitory receptors with NKG2A and activating receptors with NKG2E or NKG2C. We previously demonstrated that CD94-NKG2 on NK cells and its ligand Qa-1b are important for the resistance of C57BL/6 mice to lethal ectromelia virus (ECTV) infection. We now show that NKG2C or NKG2E deficiency does not increase susceptibility to lethal ECTV infection, but overexpression of Qa-1b in infected cells does. We also demonstrate that Qa-1b is down-regulated in infected and up-regulated in bystander inflammatory monocytes and B cells. Moreover, NK cells activated …

Hematopoietic Cell-Mediated Dissemination Of Murine Cytomegalovirus Is Regulated By Nk Cells And Immune Evasion., Shunchuan Zhang, Lauren E Springer, Han-Zhi Rao, Renee G Espinosa Trethewy, Lindsey M Bishop, Meaghan H Hancock, Finn Grey, Christopher M. Snyder

Department of Microbiology and Immunology Faculty Papers

Cytomegalovirus (CMV) causes clinically important diseases in immune compromised and immune immature individuals. Based largely on work in the mouse model of murine (M)CMV, there is a consensus that myeloid cells are important for disseminating CMV from the site of infection. In theory, such dissemination should expose CMV to cell-mediated immunity and thus necessitate evasion of T cells and NK cells. However, this hypothesis remains untested. We constructed a recombinant MCMV encoding target sites for the hematopoietic specific miRNA miR-142-3p in the essential viral gene IE3. This virus disseminated poorly to the salivary gland following intranasal or footpad infections but …

Dysregulated Anti-Viral Innate Immune Cascade During Aging., Colby Stotesbury, Luis J. Sigal

Department of Microbiology and Immunology Faculty Papers

Aging predisposes to increased morbidity and lethality to infectious diseases, which becomes apparent with the high mortality rates suffered by older people when infected with influenza virus or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). At the root of this increased susceptibility to infections, is the wide-spread deterioration of the immune system.

Onchocerca Volvulus Bivalent Subunit Vaccine Induces Protective Immunity In Genetically Diverse Collaborative Cross Recombinant Inbred Intercross Mice, Nathan M Ryan, Jessica A. Hess Ligas, Fernando Pardo-Manuel De Villena, Benjamin E Leiby, Ayako Shimada, Lei Yu, Amir Yarmahmoodi, Nikolai Petrovsky, Bin Zhan, Maria Elena Bottazzi, Benjamin L Makepeace, Sara Lustigman, David Abraham

Department of Microbiology and Immunology Faculty Papers

This study tests the hypothesis that an Onchocerca volvulus vaccine, consisting of two recombinant antigens (Ov-103 and Ov-RAL-2) formulated with the combination-adjuvant Advax-2, can induce protective immunity in genetically diverse Collaborative Cross recombinant inbred intercross mice (CC-RIX). CC-RIX lines were immunized with the O. volvulus vaccine and challenged with third-stage larvae. Equal and significant reductions in parasite survival were observed in 7 of 8 CC-RIX lines. Innate protective immunity was seen in the single CC-RIX line that did not demonstrate protective adaptive immunity. Analysis of a wide array of immune factors showed that each line of mice have a unique …

Manganese Exposure In Juvenile C57bl/6 Mice Increases Glial Inflammatory Responses In The Substantia Nigra Following Infection With H1n1 Influenza Virus., Collin M Bantle, C Tenley French, Jason E Cummings, Shankar Sadasivan, Kevin Tran, Richard A Slayden, Richard Jay Smeyne, Ronald B Tjalkens

Farber Institute for Neuroscience Faculty Papers

Infection with Influenza A virus can lead to the development of encephalitis and subsequent neurological deficits ranging from headaches to neurodegeneration. Post-encephalitic parkinsonism has been reported in surviving patients of H1N1 infections, but not all cases of encephalitic H1N1 infection present with these neurological symptoms, suggesting that interactions with an environmental neurotoxin could promote more severe neurological damage. The heavy metal, manganese (Mn), is a potential interacting factor with H1N1 because excessive exposure early in life can induce long-lasting effects on neurological function through inflammatory activation of glial cells. In the current study, we used a two-hit model of neurotoxin-pathogen …