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John F Anderson

Publications and source records attributed to John F Anderson.

At least 19 recordsLinked to original sources
PubMed2026

Evolutionary history of Jamestown Canyon virus reveals complex multi-vector ecology.

Jamestown Canyon virus (JCV) is a historically understudied mosquito-borne virus of increasing concern in North America. We generated 658 whole-genome JCV sequences from northeast United States, including 84% (500/597) of all JCV-positive mosquitoes detected in Connecticut from 1997 to 2022. Then, we applied phylodynamic methods to demonstrate how mosquito phenology structures the maintenance and evolution of JCV. Our phylogenetic analyses estimate that JCV was introduced in the Northeast by at least the early 1700s, and the primary introductions of lineages A and B into Connecticut occurred during the mid-1800s to mid-1900s. Further, we estimate that JCV evolves at a rate of ∼3 × 10-5 substitutions per site per year (s/s/y), making it one of the slowest-evolving known RNA viruses, because the virus spends ∼10 months per year in evolutionary stasis while overwintering in mosquito eggs. To investigate ecological drivers of JCV spread in Connecticut, we paired discrete trait and continuous phylogeographic reconstructions with mosquito surveillance data. We estimate that JCV has a low diffusion rate of ∼30-60 km2/year, which is more similar to slow-moving tick-borne viruses than to other mosquito-borne viruses. We found that univoltine Aedes mosquitoes were likely to maintain the virus across years through overwintering in eggs, accounting for its slow evolution and dispersal, while multivoltine mosquitoes contributed to periodic bursts of spatial diffusion and amplification within seasons. We demonstrate the utility of dense sequencing and phylodynamics to disentangle complex transmission cycles, offering a framework for rapidly advancing our evolutionary and ecological knowledge of understudied viruses.

Animals↗
PubMed2006

The Lyme disease agent Borrelia burgdorferi requires BB0690, a Dps homologue, to persist within ticks.

Borrelia burgdorferi survives in an enzootic cycle, and Dps proteins protect DNA against damage during starvation or oxidative stress. The role of a Dps homologue encoded by Borrelia in spirochaete survival was assessed. Dps-deficient spirochaetes were infectious in mice via needle-inoculation at the dose of 10(5) spirochaetes. Larval ticks successfully acquired Dps-deficient spirochaetes via a blood meal on mice. However, after extended periods within unfed nymphs, the Dps-deficient spirochaetes failed to be transmitted to a new host when nymphs fed. Our data suggest that Dps functions to protect the spirochaetes during dormancy in unfed ticks, and in its absence, the spirochaetes become susceptible during tick feeding. dps is differentially expressed in vivo- low in mice and high in ticks - but constitutively expressed in vitro, showing little change during growth or in response to oxidative stress. Borrelia Dps forms a dodecameric complex capable of sequestering iron. The Dps-deficient spirochaetes showed no defect in starvation and oxidative stress assays, perhaps due to the lack of iron in spirochaetes grown in vitro. Dps is critical for spirochaete persistence within ticks, and strategies to interfere with Dps could potentially reduce Borrelia populations in nature and thereby influence the incidence of Lyme disease.

Amino Acid Sequence↗
PubMed2006

Antiviral peptides targeting the west nile virus envelope protein.

West Nile virus (WNV) can cause fatal murine and human encephalitis. The viral envelope protein interacts with host cells. A murine brain cDNA phage display library was therefore probed with WNV envelope protein, resulting in the identification of several adherent peptides. Of these, peptide 1 prevented WNV infection in vitro with a 50% inhibition concentration of 67 muM and also inhibited infection of a related flavivirus, dengue virus. Peptide 9, a derivative of peptide 1, was a particularly potent inhibitor of WNV in vitro, with a 50% inhibition concentration of 2.6 muM. Moreover, mice challenged with WNV that had been incubated with peptide 9 had reduced viremia and fatality compared with control animals. Peptide 9 penetrated the murine blood-brain barrier and was found in the brain parenchyma, implying that it may have antiviral activity in the central nervous system. These short peptides serve as the basis for developing new therapeutics for West Nile encephalitis and, potentially, other flaviviruses.

Amino Acid Sequence↗
PubMed2006

Importance of vertical and horizontal transmission of West Nile virus by Culex pipiens in the Northeastern United States.

West Nile virus (WNV) has become established in the northeastern United States, where mosquitoes are inactive during winter. There have been no documented studies to explain how this virus survives winter and reinitiates infection in spring. We report that WNV was vertically transmitted to 2 F(1) female Culex pipiens from a naturally infected female collected in Stratford, Connecticut. One vertically infected F(1) female, which was 168 days old, fed on a hamster that died 8 days later of West Nile disease. This suggests that WNV survives winter in unfed, vertically infected C. pipiens with amplification initiated in spring by horizontal transmission.

Animals↗
PubMed2006

Gamma delta T cells facilitate adaptive immunity against West Nile virus infection in mice.

West Nile (WN) virus causes fatal meningoencephalitis in laboratory mice, and gammadelta T cells are involved in the protective immune response against viral challenge. We have now examined whether gammadelta T cells contribute to the development of adaptive immune responses that help control WN virus infection. Approximately 15% of TCRdelta(-/-) mice survived primary infection with WN virus compared with 80-85% of the wild-type mice. These mice were more susceptible to secondary challenge with WN virus than the wild-type mice that survived primary challenge with the virus. Depletion of gammadelta T cells in wild-type mice that survived the primary infection, however, does not affect host susceptibility during secondary challenge with WN virus. Furthermore, gammadelta T cells do not influence the development of Ab responses during primary and at the early stages of secondary infection with WN virus. Adoptive transfer of CD8(+) T cells from wild-type mice that survived primary infection with WN virus to naive mice afforded partial protection from lethal infection. In contrast, transfer of CD8(+) T cells from TCRdelta(-/-) mice that survived primary challenge with WN virus failed to alter infection in naive mice. This difference in survival correlated with the numeric and functional reduction of CD8 memory T cells in these mice. These data demonstrate that gammadelta T cells directly link innate and adaptive immunity during WN virus infection.

Adoptive Transfer↗
PubMed2006

An Ixodes scapularis protein required for survival of Anaplasma phagocytophilum in tick salivary glands.

Anaplasma phagocytophilum is the agent of human anaplasmosis, the second most common tick-borne illness in the United States. This pathogen, which is closely related to obligate intracellular organisms in the genera Rickettsia, Ehrlichia, and Anaplasma, persists in ticks and mammalian hosts; however, the mechanisms for survival in the arthropod are not known. We now show that A. phagocytophilum induces expression of the Ixodes scapularis salp16 gene in the arthropod salivary glands during vector engorgement. RNA interference-mediated silencing of salp16 gene expression interfered with the survival of A. phagocytophilum that entered ticks fed on A. phagocytophilum-infected mice. A. phagocytophilum migrated normally from A. phagocytophilum-infected mice to the gut of engorging salp16-deficient ticks, but up to 90% of the bacteria that entered the ticks were not able to successfully infect I. scapularis salivary glands. These data demonstrate the specific requirement of a pathogen for a tick salivary protein to persist within the arthropod and provide a paradigm for understanding how Rickettsia-like pathogens are maintained within vectors.

Anaplasma phagocytophilum↗
PubMed2006

West Nile virus from female and male mosquitoes (Diptera: Culicidae) in subterranean, ground, and canopy habitats in Connecticut.

In total, 93,532 female mosquitoes (Diptera: Culicidae) were captured in traps placed in subterranean (catch basin), ground (approximately 1.5 m above ground), and canopy (approximately 7.0 m above ground) habitats in Stamford and Stratford, CT, during 2003-2005. Culex pipiens L. was the most abundant (64.8%) of the 31 species identified. Significantly greater numbers of Cx. pipiens were captured in canopy-placed mosquito magnet experimental traps, and significantly greater numbers were collected in catch basin-placed (Centers for Disease Control) CDC traps than in CDC traps placed elsewhere. Culex restuans Theobald was captured in significantly greater numbers in traps placed in catch basins. Aedes vexans (Meigen), Aedes cinereus Meigen, and Aedes cantator (Coquillett) were significantly more abundant in ground traps. In total, 429 isolations of West Nile virus (WNV) were made from seven species of mosquitoes from late June through the end of October during 2003 through 2005. Three hundred ninety-eight (92.8%) isolates were from Cx. pipiens. Others were from Cx. restuans (n = 16), Culex salinarius Coquillett (n = 5), Ae. vexans (n = 4), Ae. cantator (n = 3), Aedes triseriatus (Say) (n = 2), and Ae. cinereus (n = 1). Multiple isolates from Cx. pipiens were made each week, primarily during the later part of July through the end of September. Weekly minimum infection rates (MIRs) were lower in 2004 (highest weekly MIR = 7.1) when no human cases were reported in Connecticut in comparison with 2003 and 2005 (highest weekly MIR = 83.9) when human cases were documented. Frequencies of infected pools were significantly higher in Cx. pipiens captured in traps in the canopy and significantly higher in catch basin placed traps than in traps at ground level. The physiological age structure of Cx. pipiens captured in the canopy was significantly different from that of Cx. pipiens collected in catch basins. Invariably, Cx. pipiens captured in the canopy were nulliparous or parous with ovaries in Christophers' stage 2, whereas 58.7% of the females captured in catch basins possessed ovaries filled with mature oocytes in Christophers' stage 5. Our results suggest that females in the canopy are seeking hosts, and after digestion of the bloodmeal and development of mature oocytes, they descend to catch basins for shelter and deposition of eggs. WNV was isolated from three, one, and two pools of male Cx. pipiens captured in catch basin-, ground-, and canopy-placed traps, respectively, and from six nulliparous Cx. pipiens females collected in the canopy. Weekly MIR ranged from 1.2 to 31.1 per 1,000 male specimens. These data show that mosquitoes become infected by means other than by blood feeding, possibly by transovarial transmission. The placement of traps in tree canopies and in catch basins can be used to augment current practices of placement of traps near the ground for surveillance of mosquitoes infected with WNV and for studies of the ecology of WNV.

Aedes↗
PubMed2006

Host feeding patterns of Culex mosquitoes and West Nile virus transmission, northeastern United States.

To evaluate the role of Culex mosquitoes as enzootic and epidemic vectors for WNV, we identified the source of vertebrate blood by polymerase chain reaction amplification and sequencing portions of the cytochrome b gene of mitochondrial DNA. All Cx. restuans and 93% of Cx. pipiens acquired blood from avian hosts; Cx. salinarius fed frequently on both mammals (53%) and birds (36%). Mixed-blood meals were detected in 11% and 4% of Cx. salinarius and Cx. pipiens, respectively. American robin was the most common source of vertebrate blood for Cx. pipiens (38%) and Cx. restuans (37%). American crow represented <1% of the blood meals in Cx. pipiens and none in Cx. restuans. Human-derived blood meals were identified from 2 Cx. salinarius and 1 Cx. pipiens. Results suggest that Cx. salinarius is an important bridge vector to humans, while Cx. pipiens and Cx. restuans are more efficient enzootic vectors in the northeastern United States.

Animals↗
PubMed2006

Typhus fever. 1915.

Explore the source record for details and available documents.

Communicable Disease Control↗
PubMed2005

The Lyme disease agent exploits a tick protein to infect the mammalian host.

The Lyme disease agent, Borrelia burgdorferi, is maintained in a tick-mouse cycle. Here we show that B. burgdorferi usurps a tick salivary protein, Salp15 (ref. 3), to facilitate the infection of mice. The level of salp15 expression was selectively enhanced by the presence of B. burgdorferi in Ixodes scapularis, first indicating that spirochaetes might use Salp15 during transmission. Salp15 was then shown to adhere to the spirochaete, both in vitro and in vivo, and specifically interacted with B. burgdorferi outer surface protein C. The binding of Salp15 protected B. burgdorferi from antibody-mediated killing in vitro and provided spirochaetes with a marked advantage when they were inoculated into naive mice or animals previously infected with B. burgdorferi. Moreover, RNA interference-mediated repression of salp15 in I. scapularis drastically reduced the capacity of tick-borne spirochaetes to infect mice. These results show the capacity of a pathogen to use a secreted arthropod protein to help it colonize the mammalian host.

Animals↗
PubMed2005

A recombinant envelope protein vaccine against West Nile virus.

West Nile (WN) virus is a flavivirus that first appeared in North America in 1999. Since then, more than 600 human deaths and 22,000 equine infections have been attributed to the virus in the United States. We expressed a truncated form of WN virus envelope (E) protein in Drosophila S2 cells. This soluble recombinant E protein was recognized by antibodies from naturally infected horses, indicating that it contains native epitopes. Mice and horses produced high-titer antibodies when immunized with recombinant E protein combined with aluminum hydroxide. Immunized mice were resistant to challenge with a lethal viral dose. Sera from immunized horses, administered to naive mice, conferred resistance against a lethal WN viral challenge. In addition, sera of immunized horses neutralized West Nile virus in vitro, as demonstrated by plaque reduction assays. This recombinant form of E protein, combined with aluminum hydroxide, is a candidate vaccine that may protect humans and horses against WN virus infections.

Adjuvants, Immunologic↗
PubMed2005

Isolations of Potosi virus from mosquitoes (Diptera: Culicidae) collected in Connecticut.

Potosi virus (POTV) (Bunyaviridae: Orthobunyavirus) was first isolated from Aedes albopictus (Skuse) collected in Potosi, MO, in 1989, and subsequent isolations were reported from Illinois, Michigan, Ohio, and the Carolinas. To determine whether the distribution of this virus extends into the northeastern United States, we analyzed arboviruses acquired from mosquitoes collected in Connecticut from 1998 to 2004. In 2001, a bunyavirus was isolated from Aedes vexans (Meigen) that was different from other arboviruses known to occur in Connecticut by cross-neutralization and reverse transcription-polymerase chain reaction (RT-PCR) assays. Nucleotide and encoded amino acid sequences of a portion of the G2 envelope gene were 99 and 100% similar to POTV, respectively, yet distinct from indigenous strains of Jamestown Canyon (JCV), Cache Valley (CVV), and Trivittatus virus (TVTV). Viral isolates obtained from the statewide surveillance program were retested by RT-PCR coupled with restriction enzyme analysis to distinguish POTV from other bunyaviruses. POTV isolates, previously typed by neutralization, were correctly identified by RT-PCR; however, many isolates classified as JCV or CVV by enzyme-linked immunosorbent assay proved to be POTV by molecular assays. In total, 92 strains of POTV were isolated from 12 mosquito species in 2000, 2001, and 2003, whereas POTV was not detected in mosquitoes sampled during 1998, 1999, 2002, and 2004. Viral isolation rates were highest for Anopheles punctipennis (Say) (3.2-11.3 infection rate per 1,000 mosquitoes), whereas the greatest number of isolates came from Ochlerotatus trivittatus (Coquillett) (8-16 isolates). This finding represents the first detection of POTV in the northeastern United States where it infects a diverse array of mosquito species.

Animals↗
PubMed2005

Protective and therapeutic capacity of human single-chain Fv-Fc fusion proteins against West Nile virus.

West Nile virus has spread rapidly across the United States, and there is currently no approved human vaccine or therapy to prevent or treat disease. Passive immunization with antibodies against the envelope protein represents a promising means to provide short-term prophylaxis and treatment for West Nile virus infection. In this study, we identified a panel of 11 unique human single-chain variable region antibody fragments (scFvs) that bind the envelope protein of West Nile virus. Selected scFvs were converted to Fc fusion proteins (scFv-Fcs) and were tested in mice for their ability to prevent lethal West Nile virus infection. Five of these scFv-Fcs, 11, 15, 71, 85, and 95, protected 100% of mice from death when given prior to infection with virus. Two of them, 11 and 15, protected 80% of mice when given at days 1 and 4 after infection. In addition, four of the scFv-Fcs cross-neutralized dengue virus, serotype 2. Binding assays using yeast surface display demonstrated that all of our scFvs bind to sites within domains I and II of West Nile virus envelope protein. These recombinant human scFvs are potential candidates for immunoprophylaxis and therapy of flavivirus infections.

Animals↗
PubMed2004

Toll-like receptor 3 mediates West Nile virus entry into the brain causing lethal encephalitis.

West Nile virus (WNV), a mosquito-borne single-stranded (ss)RNA flavivirus, causes human disease of variable severity. We investigated the involvement of Toll-like receptor (Tlr) 3, which recognizes viral double-stranded (ds)RNA, on WNV infection. Tlr3-deficient (Tlr3(-/-)) mice were more resistant to lethal WNV infection and had impaired cytokine production and enhanced viral load in the periphery, whereas in the brain, viral load, inflammatory responses and neuropathology were reduced compared to wild-type mice. Peripheral WNV infection led to a breakdown of the blood-brain barrier and enhanced brain infection in wild-type but not in Tlr3(-/-) mice, although both groups were equally susceptible upon intracerebroventricular administration of the virus. Tumor necrosis factor-alpha receptor 1 signaling is vital for blood-brain barrier compromise upon Tlr3 stimulation by dsRNA or WNV. Collectively, WNV infection leads to a Tlr3-dependent inflammatory response, which is involved in brain penetration of the virus and neuronal injury.

Animals↗
PubMed2004

TROSPA, an Ixodes scapularis receptor for Borrelia burgdorferi.

The Lyme disease agent Borrelia burgdorferi naturally persists in a cycle that primarily involves ticks and mammals. We have now identified a tick receptor (TROSPA) that is required for spirochetal colonization of Ixodes scapularis. B. burgdorferi outer surface protein A, which is abundantly expressed on spirochetes within the arthropod and essential for pathogen adherence to the vector, specifically bound to TROSPA. TROSPA mRNA levels in ticks increased following spirochete infestation and decreased in response to engorgement, events that are temporally linked to B. burgdorferi entry into and egress from the vector. The blockade of TROSPA by TROSPA antisera or by the repression of TROSPA expression via RNA interference reduced B. burgdorferi adherence to the I. scapularis gut in vivo, thereby preventing efficient colonization of the vector and subsequently reducing pathogen transmission to the mammalian host. Identification of an I. scapularis receptor for B. burgdorferi is the first step toward elucidating arthropod ligands that are required for survival of spirochetes in nature.

Amino Acid Sequence↗
PubMed2004

Disruption of Ixodes scapularis anticoagulation by using RNA interference.

Ixodes scapularis ticks transmit many pathogens, including Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. Vaccines directed against arthropod proteins injected into the host during tick engorgement could prevent numerous infectious diseases. Salp14, a salivary anticoagulant, poses a key target for such intervention. Salp14 is the prototypic member of a family of potential I. scapularis anticoagulants, expressed and secreted in tick saliva during tick feeding. RNA interference was used to assess the role of Salp14 in tick feeding. Salp14 and its paralogs were silenced, as demonstrated by the reduction of mRNA and protein specific for these antigens. Tick salivary glands lacking Salp14 had reduced anticoagulant activity, as revealed by a 60-80% reduction of anti-factor Xa activity. Silencing the expression of salp14 and its paralogs also reduced the ability of I. scapularis to feed, as demonstrated by a 50-70% decline in the engorgement weights. Because ticks have several anticoagulants, it is likely that the expression of multiple anticoagulants in I. scapularis saliva would have to be ablated simultaneously to abolish tick feeding. These studies demonstrate that RNA interference can silence I. scapularis genes and disrupt their physiologic function in vivo, and they identify vaccine candidates that can alter vector engorgement.

Actins↗