Immunology and Infectious Disease Commons^™

Discovery Of Exosomes From Tick Saliva And Salivary Glands Reveals Therapeutic Roles For Cxcl12 And Il-8 In Wound Healing At The Tick-Human Skin Interface, Wenshuo Zhou, Faizan Tahir, Joseph Che-Yen Wang, Michael Woodson, Michael B. Sherman, Shahid Karim, Girish Neelakanta, Hameeda Sultana

Biological Sciences Faculty Publications

Ticks secrete various anti-coagulatory, anti-vasoconstrictory, anti-inflammatory, and anti-platelet aggregation factors in their saliva at the bite site during feeding to evade host immunological surveillance and responses. For the first time, we report successful isolation of exosomes (small membrane-bound extracellular signaling vesicles) from saliva and salivary glands of partially fed or unfed ixodid ticks. Our data showed a novel role of these in vivo exosomes in the inhibition of wound healing via downregulation of C-X-C motif chemokine ligand 12 (CXCL12) and upregulation of interleukin-8 (IL-8). Cryo-electron microscopy (cryo-EM) analysis revealed that tick saliva and salivary glands are composed of heterogeneous populations …

A Simple, Inexpensive Method For Mark-Recapture Of Ixodid Ticks, Alexis White, Robin Minch, Lindsey Bidder, Holly Gaff

Biological Sciences Faculty Publications

Mark-recapture techniques have been widely used and specialized to study organisms throughout the field of biology. To mark-recapture ticks (Ixodida), we have created a simple method to mark ticks using nail polish applied with an insect pin secured in a pencil that allows for a variety of questions to be answered. For measuring tick control efficacy, estimating population estimates, or measuring movement of ticks, this inexpensive mark-recapture method has been easily applied in the field and in the lab to provide useful data to answer a variety of questions about ticks.

Molecular Investigation And Phylogeny Of Species Of The Anaplasmataceae Infecting Animals And Ticks In Senegal, Mustapha Dahmani, Bernard Davoust, Masse Sambou, Hubert Bassene, Pierre Scandola, Tinhinene Ameur, Didier Raoult, Florence Feenollar, Oleg Mediannikov

Biological Sciences Faculty Publications

Background: Our study aimed to assess the diversity of the species of Anaplasmataceae in Senegal that infect animals and ticks in three areas: near Keur Momar Sarr (northern region), Dielmo and Diop (Sine Saloum, central region of Senegal), and in Casamance (southern region of Senegal).

Methods: A total of 204 ticks and 433 blood samples were collected from ruminants, horses, donkeys and dogs. Ticks were identified morphologically and by molecular characterization targeting the 12S rRNA gene. Molecular characterization of species of Anaplasmataceae infecting Senegalese ticks and animals was conducted using the 23S rRNA, 16S rRNA, rpoB and groEL genes.

Results: …

Diagnosis Of Schistosoma Mansoni Infections: What Are The Choices In Brazilian Low-Endemic Areas?, Vanessa Silva-Moraes, Lisa M. Shollenberger, Liliane Maria Vidal Siqueira, William Castro-Borges, Donald A. Harn, Rafaella Fortini Queiroz E. Grenfell, Ana Lucia Teles Rabello, Paulo Marcos Zech Coelho

Biological Sciences Faculty Publications

The population of Brazil is currently characterised by many individuals harbouring low-intensity Schistosoma mansoni infections. The Kato-Katz technique is the diagnostic method recommended by the World Health Organization (WHO) to assess these infections, but this method is not sensitive enough in the context of low egg excretion. In this regard, potential alternatives are being employed to overcome the limits of the Kato-Katz technique. In the present review, we evaluated the performance of parasitological and immunological approaches adopted in Brazilian areas. Currently, the diagnostic choices involve a combination of strategies, including the utilisation of antibody methods to screen individuals and then …

Dynamics Of Two Pathogens In A Single Tick Population, Alexis White, Elsa Schaefer, Chelsea Wright-Thompson, Christopher M. Kribs, Holly Gaff

Biological Sciences Faculty Publications

A mathematical model for a two-pathogen, one-tick, one-host system is presented and explored. The goal of this model is to determine how long an invading pathogen persists within a tick population in which a resident pathogen is already established. The numerical simulations of the model demonstrate the parameter ranges that allow for coexistence of the two pathogens. Sensitivity analysis highlights the importance of vector-borne, tick-to-host, transmission rates on the invasion reproductive number and persistence of the pathogens over time. The model is then applied to a case study based on a reclaimed swampland field site in southeastern Virginia using field …

Multistate Survey Of American Dog Ticks (Dermacentor Variabilis) For Rickettsia Species, Joy A. Hecht, Michelle E.J. Allerdice, Elizabeth A. Dykstra, Laura Mastel, Rebecca J. Eisen, Tammi L. Johnson, Holly D. Gaff, Andrea S. Varela-Stokes, Jerome Goddard, Benedict B. Pagac, Christopher D. Paddock, Sandor E. Karpathy

Biological Sciences Faculty Publications

Dermacentor variabilis, a common human-biting tick found throughout the eastern half and along the west coast of the United States, is a vector of multiple bacterial pathogens. Historically, D. variabilis has been considered a primary vector of Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever. A total of 883 adult D. variabilis, collected between 2012 and 2017 from various locations in 12 states across the United States, were screened for rickettsial DNA. Tick extracts were evaluated using three real-time PCR assays; an R. rickettsii-specific assay, a Rickettsia bellii-specific assay, and a Rickettsia genus-specific …

Serological Proteomic Screening And Evaluation Of A Recombinant Egg Antigen For The Diagnosis Of Low-Intensity Schistosoma Mansoni Infections In Endemic Area In Brazil, Vanessa Silva-Moraes, Lisa Marie Shollenberger, William Castro-Borges, Ana Lucia Teles Rabello, Donald A. Harn, Lia Carolina Soares Medeiros, Wander De Jesus Jeremias, Liliane Maria Vidal Siqueira, Caroline Stephane Salviano Pereira, Maria Luysa Camargos Pedrosa, Nathalie Bonatti Franco Almeida, Aureo Almeida, Jose Roberto Lambertucci, Nidia Francisca De Figueiredo Carneiro, Paulo Marcos Zech Coelho, Refaella Fortini Queiroz Grenfell

Biological Sciences Faculty Publications

Background

Despite decades of use of control programs, schistosomiasis remains a global public health problem. To further reduce prevalence and intensity of infection, or to achieve the goal of elimination in low-endemic areas, there needs to be better diagnostic tools to detect low-intensity infections in low-endemic areas in Brazil. The rationale for development of new diagnostic tools is that the current standard test Kato-Katz (KK) is not sensitive enough to detect low-intensity infections in low-endemic areas. In order to develop new diagnostic tools, we employed a proteomics approach to identify biomarkers associated with schistosome-specific immune responses in hopes of developing …

Prime-Boost Vaccine Regimen For Sjtpl And Sjc23 Schistosome Vaccines, Increases Efficacy In Water Buffalo In A Field Trial In China, Akram A. Da'dara, Changlin Li, Xinling Yu, Mao Zheng, Jie Zhou, Lisa M. Shollenberger, Yue-Sheng Li, Donald A. Ham

Biological Sciences Faculty Publications

Schistosomiasis remains a serious zoonotic disease in China and the Philippines. Water buffalo and cattle account for the majority of transmission. Vaccination of water buffalo is considered a key strategy to reduce disease prevalence. Previously, we showed that vaccination of water buffalo with SjC23 or SjCTPI plasmid DNA vaccines, induced 50% efficacy to challenge infection. Here, we evaluated several parameters to determine if we can develop a two dose vaccine that maintains the efficacy of the three dose vaccine. We performed four trials evaluating: (1) lab produced vs. GLP grade vaccines, (2) varying the time between prime and boost, (3) …

Parasitic Dinoflagellate Hematodinium Perezi Prevalence In Larval And Juvenile Blue Crabs Callinectes Sapidus From Coastal Bays Of Virginia, H. J. Small, J. P. Huchin-Mian, K. S. Reece, K. M. Pagenkopp Lohan, Mark J. Butler Iv, J. D. Shields

Biological Sciences Faculty Publications

The parasitic dinoflagellate Hematodinium perezi infects the American blue crab Callinectes sapidus and other decapods along the Eastern seaboard and Gulf of Mexico coast of the USA. Large juvenile and adult blue crabs experience high mortality during seasonal outbreaks of H. perezi, but less is known about its presence in the early life history stages of this host. We determined the prevalence of H. perezi in megalopae and early benthic juvenile crabs from multiple locations along the Virginia portion of the Delmarva Peninsula. The DNA of H. perezi was not detected in any megalopae collected from several locations within …

Characterization Of Tick Organic Anion Transporting Polypeptides (Oatps) Upon Bacterial And Viral Infections, Vikas Taank, Wenshou Zhou, Xuran Zhuang, John F. Anderson, Utpal Pal, Hameeda Sultana, Girish Neelakanta

Biological Sciences Faculty Publications

Background: Ixodes scapularis organic anion transporting polypeptides (OATPs) play important roles in tickrickettsial pathogen interactions. In this report, we characterized the role of these conserved molecules in ticks infected with either Lyme disease agent Borrelia burgdorferi or tick-borne Langat virus (LGTV), a pathogen closely related to tick-borne encephalitis virus (TBEV).

Results: Quantitative real-time polymerase chain reaction analysis revealed no significant changes in oatps gene expression upon infection with B. burgdorferi in unfed ticks. Synchronous infection of unfed nymphal ticks with LGTV in vitro revealed no significant changes in oatps gene expression. However, expression of specific oatps was significantly downregulated upon …

Macrophages But Not Astrocytes Harbor Hiv Dna In The Brains Of Hiv-1-Infected Aviremic Individuals On Suppressive Antiretroviral Therapy, Allen Ko, Guobin Kang, Julian B. Hattler, Hadiza I. Galadima, Junfeng Zhang, Qingsheng Li, Woong-Ki Kim

Community & Environmental Health Faculty Publications

The question of whether the human brain is an anatomical site of persistent HIV-1 infection during suppressive antiretroviral therapy (ART) is critical, but remains unanswered. The presence of virus in the brains of HIV patients whose viral load is effectively suppressed would demonstrate not only the potential for CNS to act as an anatomical HIV reservoir, but also the urgent need to understand the factors contributing to persistent HIV behind the blood-brain barrier. Here, we investigated for the first time the presence of cells harboring HIV DNA and RNA in the brains from subjects with undetectable plasma viral load and …

The Mechanisms Shaping The Repertoire Of Cd4(+)Foxp3(+) Regulatory T Cells, Piotr Kraj, Leszek Ignatowicz

Biological Sciences Faculty Publications

Regulatory T (Treg) cells expressing Foxp3 transcription factor control homeostasis of the immune system, antigenic responses to commensal and pathogenic microbiota, and immune responses to self and tumour antigens. The Treg cells differentiate in the thymus, along with conventional CD4⁺ T cells, in processes of positive and negative selection. Another class of Treg cells is generated in peripheral tissues by inducing Foxp3 expression in conventional CD4⁺ T cells in response to antigenic stimulation. Both thymic and peripheral generation of Treg cells depends on recognition of peptide/MHC ligands by the T-cell receptors (TCR) expressed on thymic Treg precursors or …

Range Expansion Of Tick Disease Vectors In North America: Implications For Spread Of Tick-Borne Disease, Daniel E. Sonenshine

Biological Sciences Faculty Publications

Ticks are the major vectors of most disease-causing agents to humans, companion animals and wildlife. Moreover, ticks transmit a greater variety of pathogenic agents than any other blood-feeding arthropod. Ticks have been expanding their geographic ranges in recent decades largely due to climate change. Furthermore, tick populations in many areas of their past and even newly established localities have increased in abundance. These dynamic changes present new and increasing severe public health threats to humans, livestock and companion animals in areas where they were previously unknown or were considered to be of minor importance. Here in this review, the geographic …

Using An In Vitro System For Maintaining Varroa Destructor Mites On Apis Mellifera Pupae As Hosts: Studies Of Mite Longevity And Feeding Behavior, Noble I. Egekwu, Francisco Posada, Daniel E. Sonenshine, Steven Cook

Biological Sciences Faculty Publications

Varroa destructor mites (varroa) are ectoparasites of Apis mellifera honey bees, and the damage they inflict on hosts is likely a causative factor of recent poor honey bee colony performance. Research has produced an arsenal of control agents against varroa mites, which have become resistant to many chemical means of their control, and other means have uncertain efficacy. Novel means of control will result from a thorough understanding of varroa physiology and behavior. However, robust knowledge of varroa biology is lacking; mites have very low survivability and reproduction away from their natural environment and host, and few tested protocols of …

Do Avian Blood Parasites Influence Hypoxia Physiology In A High Elevation Environment?, Farah Ishtiaq, Sahas Barve

Biological Sciences Faculty Publications

BACKGROUND: Montane birds which engage in elevational movements have evolved to cope with fluctuations in environmental hypoxia, through changes in physiological parameters associated with blood oxygen-carrying capacity such as haemoglobin concentration (Hb) and haematocrit (Hct). In particular, elevational migrants which winter at low elevations, encounter varying intensities of avian haemosporidian parasites as they traverse heterogeneous environments. Whilst high intensity parasite infections lead to anaemia, one can expect that the ability to cope with haemosporidian infections should be a key trait for elevational migrants that must be balanced against reducing the oxygen-carrying capacity of blood in response to high elevation. In …

Modelling The Effects Of Habitat And Hosts On Tick Invasions, Robyn M. Nadolny, Holly D. Gaff

Biological Sciences Faculty Publications

Many tick species are invading new areas because of anthropogenic changes in the landscape, shifting climatic variables and increasing populations of suitable host species and tick habitat. However, the relative influences of habitat and hosts in tick dispersal and tick population establishment remain in question. A spatially explicit agent-based model was developed to explore the spatio-temporal dynamics of a generic tick population in the years immediately following the introduction of ticks into a novel environment. The general model was then adapted to investigate a case study of two recent tick species invasions into the Mid-Atlantic United States. The recent simultaneous …

Towards Understanding The Role Of Small Mammals In The Transmission Of Lyme Disease In Virginia, Usa, Robert K. Rose, Laura Bitzer, Wayne Hynes, Holly D. Gaff, Robyn Nadolny, Jory Brinkerhoff

Biological Sciences Faculty Publications

Larvae and nymphs of many species of ticks select small mammals, reptiles, and birds as hosts. Since 2011, we have examined more than 1,000 small mammals of 10 species and collected a small tissue sample as well as all visible ticks; rates of infestation range from 18% in harvest mice to 53% in meadow voles. Identification of immature ticks is challenging and requires molecular or genetic methods beyond classification as being from the genus Ixodes or another genus. Ixodes ticks include Ixodes scapularis, the species that transmits the Lyme disease-causing bacterium Borrelia burgdorferi to humans. Of the approximately 2,300 ticks …

Ticks, Ixodes Scapularis, Feed Repeatedly On White-Footed Mice Despite Strong Inflammatory Response: An Expanding Paradigm For Understanding Tick-Host Interactions, Jennifer M. Anderson, Ian N. Moore, Bianca M. Nagata, José M.C. Ribeiro, Jesus G. Valenzuela, Daniel E. Sonenshine

Biological Sciences Faculty Publications

Ticks transmit infectious agents including bacteria, viruses and protozoa. However, their transmission may be compromised by host resistance to repeated tick feeding. Increasing host resistance to repeated tick bites is well known in laboratory animals, including intense inflammation at the bite sites. However, it is not known whether this also occurs in wild rodents such as white-footed mice, Peromyscus leucopus, and other wildlife, or if it occurs at all. According to the "host immune incompetence" hypothesis, if these mice do not have a strong inflammatory response, they would not reject repeated tick bites by Ixodes scapularis. To test …

Effects Of Carboxylated Nanodiamonds On Macrophages During And After Differentiation, Maisoun E. Bani Hani

Biological Sciences Theses & Dissertations

Nanodiamonds (ND) are a carbon-based nanomaterial that are increasingly being proposed for developing novel imaging techniques, as carriers of biomolecules and therapeutic drugs, as coatings for implants, and for other biomedical applications. The exceptional chemical, mechanical, and optical properties of ND make this material suitable in a wide range of fields. The application of ND in the biomedical field is attractive but requires more in-depth investigation into the safety of ND and its interactions with different cells and systems. The effects of ND on the immune system are not fully understood or investigated and there are several controverting reports regarding …

Microbial Invasion Vs. Tick Immune Regulation, Daniel E. Sonenshine, Kevin R. Macaluso

Biological Sciences Faculty Publications

Ticks transmit a greater variety of pathogenic agents that cause disease in humans and animals than any other haematophagous arthropod, including Lyme disease, Rocky Mountain spotted fever, human granulocytic anaplasmosis, babesiosis, tick-borne encephalitis, Crimean Congo haemorhagic fever, and many others (Gulia-Nuss et al., 2016). Although diverse explanations have been proposed to explain their remarkable vectorial capacity, among the most important are their blood feeding habit, their long term off-host survival, the diverse array of bioactive molecules that disrupt the host's natural hemostatic mechanisms, facilitate blood flow, pain inhibitors, and minimize inflammation to prevent immune rejection (Hajdušek et al., 2013). Moreover, …

Defining The Risk Of Zika And Chikungunya Virus Transmission In Human Population Centers Of The Eastern United States, Carrie A. Manore, Richard S. Ostfeld, Folashade B. Agusto, Holly Gaff, Shannon L. Ladeau

Biological Sciences Faculty Publications

The recent spread of mosquito-transmitted viruses and associated disease to the Americas motivates a new, data-driven evaluation of risk in temperate population centers. Temperate regions are generally expected to pose low risk for significant mosquito-borne disease; however, the spread of the Asian tiger mosquito (Aedes albopictus) across densely populated urban areas has established a new landscape of risk. We use a model informed by field data to assess the conditions likely to facilitate local transmission of chikungunya and Zika viruses from an infected traveler to Ae. albopictus and then to other humans in USA cities with variable human …

Abiotic Factors Contributing To The Survival Of Three Tick Species In Southeastern Virginia, Amblyomma Americanum (Lone Star Tick), Dermacentor Variabilis (American Dog Tick), And Amblyomma Maculatum (Gulf Coast Tick), Lindsey A. Bidder

Biological Sciences Theses & Dissertations

Amblyomma americanum, Amblyomma maculatum, and Dermacentor variabilis are hard-bodied ticks in the Hampton Roads area of southeastern Virginia. This study consisted of two field projects focused on these tick species. To estimate the off-host survival of local tick species, a capture-mark-recapture (CMR) study was performed. An environmental survival study was performed to quantify the ability of these three tick species to survive in situ. Four field sites were used in the Hampton Roads region covering a variety of habitat types and vegetation; specifically two drier, upland field sites and two flood-prone sites. CMR was conducted from May through …

Genomic Insights Into The Ixodes Scapularis Tick Vector Of Lyme Disease, Monika Gulia-Nuss, Andrew B. Nuss, Jason M. Meyer, Daniel E. Sonenshine, R. Michael Roe, Robert M. Waterhouse, David B. Sattelle, Jose De La Fuente, Jose M. Ribeiro, Karine Megy

Biological Sciences Faculty Publications

Ticks transmit more pathogens to humans and animals than any other arthropod. We describe the 2.1 Gbp nuclear genome of the tick, Ixodes scapularis (Say), which vectors pathogens that cause Lyme disease, human granulocytic anaplasmosis, babesiosis and other diseases. The large genome reflects accumulation of repetitive DNA, new lineages of retro-transposons, and gene architecture patterns resembling ancient metazoans rather than pancrustaceans. Annotation of scaffolds representing ~57% of the genome, reveals 20,486 protein-coding genes and expansions of gene families associated with tick-host interactions. We report insights from genome analyses into parasitic processes unique to ticks, including host 'questing', prolonged feeding, cuticle …

Investigating Cholera Using An Sir Model With Age-Class Structure And Optimal Control, K. Renee Fister, Holly Gaff, Elsa Schaefer, Glenna Buford, Bryce C. Norris

Biological Sciences Faculty Publications

The use of systems of differential equations in mathematical modeling in conjunction with epidemiology continues to be an area of focused research. This paper briefly acquaints readers with epidemiology, cholera, and the need for effective control strategies; discusses cholera dynamics through a variation on the SIR epidemiological model in which two separate age classes exist in a population; finds the numeric value for R₀ to be approximately 1.54 using estimated parameters for Bangladesh; and employs an optimal control resulting in a suggestion that a protection control be implemented at the end of the monsoon season.

Donor Platelet Plasma Components Inactivate Sensitive And Multidrug Resistant Acinetobacter Baumannii Isolates, Chelsea M. Edelblute, Olga N. Pakhomova, Fanying Li, Barbara Y. Hargrave, Loree C. Heller

Bioelectrics Publications

Acinetobacter baumannii is an environmentally resilient healthcare-associated opportunistic pathogen responsible for infections at many body sites. In the last 10 years, clinical strains resistant to many or all commonly used antibiotics have emerged globally. With few antimicrobial agents in the pharmaceutical pipeline, new and alternative agents are essential. Platelets secrete a large number of proteins, including proteins with antimicrobial activity. In a previous study, we demonstrated that donor platelet supernatants and plasma significantly inhibited the growth of a reference strain of A. baumannii in broth and on skin. This inhibition appeared to be unrelated to the platelet activation state. In …

A Study Of A Simulated Infectious Disease On Healthcare Workers' Reaction, Knowledge, Attitudes, And Performance Towards Hand Hygiene, Lydia Wigglesworth-Ballard

Health Services Research Dissertations

Problem Statement. The high rate of healthcare worker-patient contact provides many opportunities for exposure to pathogens, which creates challenges in assessing healthcare workers' success at preventing healthcare associated infections caused by these dangerous pathogens.

Methods. This study explored the effects of a simulated infectious disease on healthcare workers' hand hygiene knowledge, attitudes, performance, and reactions using Kirkpatrick's Four levels of Evaluation. The study utilized a mixed method pre-test – 2 post-test design. The dependent variables were hand hygiene knowledge, attitudes toward hand hygiene guidelines, hand hygiene performance, and reaction to the overall experience. Data was collected three times over a …

Behavioral Immunity Suppresses An Epizootic In Caribbean Spiny Lobsters, Mark J. Butler Iv, Donald C. Behringer Jr., Thomas W. Dolan Iii, Jessica Moss, Jeffrey D. Shields

Biological Sciences Faculty Publications

Sociality has evolved in a wide range of animal taxa but infectious diseases spread rapidly in populations of aggregated individuals, potentially negating the advantages of their social interactions. To disengage from the coevolutionary struggle with pathogens, some hosts have evolved various forms of "behavioral immunity"; yet, the effectiveness of such behaviors in controlling epizootics in the wild is untested. Here we show how one form of behavioral immunity (i.e., the aversion of diseased conspecifics) practiced by Caribbean spiny lobsters (Panulirus argus) when subject to the socially transmitted PaV1 virus, appears to have prevented an epizootic over a large seascape. We …

Comparative Population Genetics Of Two Invading Ticks: Evidence Of The Ecological Mechanisms Underlying Tick Range Expansions, Robyn Nadolny, Holly Gaff, Jens Carlsson, David Gauthier

Biological Sciences Faculty Publications

Two species of ixodid tick, Ixodes affinis Neumann and Amblyomma maculatum Koch, are simultaneously expanding their ranges throughout the mid-Atlantic region of the US. Although we have some understanding of the ecology and life history of these species, the ecological mechanisms governing where and how new populations establish and persist are unclear. To assess population connectivity and ancestry, we sequenced a fragment of the 16S mitochondrial rRNA gene from a representative sample of individuals of both species from populations throughout the eastern US. We found that despite overlapping host preferences throughout ontogeny, each species exhibited very different genetic and geographic …

Transmission-Blocking Vaccines: Focus On Anti-Vector Vaccines Against Tick-Borne Diseases, Girish Neelakanta, Hameeda Sultana

Biological Sciences Faculty Publications

Tick-borne diseases are a potential threat that account for significant morbidity and mortality in human population worldwide. Vaccines are not available to treat several of the tick-borne diseases. With the emergence and resurgence of several tick-borne diseases, emphasis on the development of transmission-blocking vaccines remains increasing. In this review, we provide a snap shot on some of the potential candidates for the development of anti-vector vaccines (a form of transmission-blocking vaccines) against wide range of hard and soft ticks that include Ixodes, Haemaphysalis, Dermacentor, Amblyomma, Rhipicephalus and Ornithodoros species.

Altered Connexin 43 Expression Underlies Age-Dependent Decrease Of Regulatory T Cell Suppressor Function In Nonobese Diabetic Mice, Michel Kuczma, Cong-Yi Wang, Leszek Ignatowicz, Robert Gourdi, Piotr Kraj

Biological Sciences Faculty Publications

Type 1 diabetes is one of the most extensively studied autoimmune diseases, but the cellular and molecular mechanisms leading to T cell–mediated destruction of insulin-producing β cells are still not well understood. In this study, we show that regulatory T cells (T_regs) in NOD mice undergo age-dependent loss of suppressor functions exacerbated by the decreased ability of activated effector T cells to upregulate Foxp3 and generate T_regs in the peripheral organs. This age-dependent loss is associated with reduced intercellular communication mediated by gap junctions, which is caused by impaired upregulation and decreased expression of connexin 43. Regulatory …