Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Awasthi, Vibhudutta; Meinken, G.E.; Springer, Karen; Srivastava, Suresh C.; Freimuth, Paul

doi:10.1128/jvi.78.12.6431-6438.2004

Cited by 38 publications

(25 citation statements)

References 46 publications

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“…Alternatively, the inhibitory effect could be due to a reduction of a specific population of recycling CAR in LGAC that is actually responsible for Ad5 endocytosis. Such a population might exist in cells expressing a significant amount of CAR, such as liver and lacrimal gland, as mentioned above (2).…”

Section: Discussionmentioning

confidence: 99%

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

Xie

Chiang²,

Contreras³

et al. 2006

J Virol

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The established mechanism for infection of most cells with adenovirus serotype 5 (Ad5) involves fiber capsid protein binding to coxsackievirus-adenovirus receptor (CAR) at the cell surface, followed by penton base capsid protein binding to ␣ v integrins, which triggers clathrin-mediated endocytosis of the virus. Here we determined the identity of the capsid proteins responsible for mediating Ad5 entry into the acinar epithelial cells of the lacrimal gland. Ad5 transduction of primary rabbit lacrimal acinar cells was inhibited by excess Ad5 fiber or knob (terminal region of the fiber) but not excess penton base. Investigation of the interactions of recombinant Ad5 penton base, fiber, and knob with lacrimal acini revealed that the penton base capsid protein remained surface associated, while the knob domain of the fiber capsid protein was rapidly internalized. Introduction of rabbit CAR-specific small interfering RNA (siRNA) into lacrimal acini under conditions that reduced intracellular CAR mRNA significantly inhibited Ad5 transduction, in contrast to a control (nonspecific) siRNA. Preincubation of Ad5 with excess heparin or pretreatment of acini with a heparinase cocktail each inhibited Ad5 transduction by a separate and apparently additive mechanism. Functional and imaging studies revealed that Ad5, fiber, and knob, but not penton base, stimulated macropinocytosis in acini and that inhibition of macropinocytosis significantly reduced Ad5 transduction of acini. However, inhibition of macropinocytosis did not reduce Ad5 uptake. We propose that internalization of Ad5 into lacrimal acini is through a novel fiber-dependent mechanism that includes CAR and heparan sulfate glycosaminoglycans and that the subsequent intracellular trafficking of Ad5 is enhanced by fiber-induced macropinocytosis.

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Section: Discussionmentioning

confidence: 99%

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

Xie

Chiang²,

Contreras³

et al. 2006

J Virol

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“…The tissues and the organs were excised and the activity associated with each organ/tissue was measured in a flat-type NaI(Tl) scintillation counter. Activity accumulated in per gram of femur was considered for obtaining the total skeletal uptake assuming skeletal weight to be 10% of the total body weight [34,35]. Distribution of the activity in different organs was calculated as percentage of injected activity (dose) (%ID) per organ.…”

Section: Biodistribution Studies In Ratsmentioning

confidence: 99%

¹⁷⁷Lu-DOTMP: A viable agent for palliative radiotherapy of painful bone metastasis

Das¹,

Chakraborty²,

Sarma³

et al. 2008

Radiochimica Acta

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The suitable nuclear decay characteristics [T 1/2 = 6.73 d, E β(max) = 497 keV, E γ = 113 keV (6.4%), 208 keV (11%)] as well as the feasibility of large-scale production with adequate specific activity and radionuclidic purity using a moderate flux reactor are important attributes towards 177 Lu to be considered as a promising radionuclide for palliative care in painful bone metastasis. The present study describes the preparation of 177 Lu complex of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetramethylene phosphonic acid (DOTMP) and its preliminary biological evaluation in animal models with an aim to proposing it as a viable radiopharmaceutical for bone pain palliation. The choice DOTMP as the polyaminophosphonic acid carrier ligand is based on the enhanced thermodynamic stability and kinetic inertness of the metal-ligand complexes with macrocyclic chelators. 177 Lu was produced with a specific activity of ∼ 12 GBq/mg (∼ 324 mCi/mg) and radionuclidic purity of 99.98% by irradiation of natural Lu 2 O 3 target at a thermal neutron flux of ∼ 6 × 10 13 n/cm 2 s for 21 d. 177 Lu-DOTMP complex was prepared in high yield and excellent radiochemical purity (> 99%) using DOTMP synthesized and characterized in-house. The complex exhibited excellent invitro stability at room temperature. Biodistribution studies in Wistar rats showed rapid skeletal accumulation of the injected activity [(1.60 ± 0.19)% per gram in femur at 3 h post-injection] with fast clearance from blood and minimal uptake in any of the major organs. Scintigraphic studies carried out in normal Wistar rats and New Zealand white rabbits also demonstrated significant accumulation of the agent in skeleton and almost no retention in any other vital organs.

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“…S4, available at www.jneurosci.org as supplemental material). Expression and isolation of fiber knob Ad2, Ad2C428N, or Ad5 were performed, with slight modifications, as detailed previously Kirby et al, 2000;Awasthi et al, 2004). The laminin-neurexinsex hormone (LNS) domain of rat neurexin ␤1 was expressed as a GSTfusion protein followed by cleavage of the GST portion using PreScission Protease (GE Healthcare) and further purified by ion exchange chromatography (Comoletti et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

The Coxsackievirus–Adenovirus Receptor Reveals Complex Homophilic and Heterophilic Interactions on Neural Cells

Patzke¹,

Max²,

Behlke³

et al. 2010

J. Neurosci.

119

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The coxsackievirus-adenovirus receptor (CAR) is a member of the Ig superfamily strongly expressed in the developing nervous system. Our histological investigations during development reveal an initial uniform distribution of CAR on all neural cells with a concentration on membranes that face the margins of the nervous system (e.g., the basal laminae and the ventricular side). At more advanced stages, CAR becomes downregulated and restricted to specific regions including areas rich in axonal and dendritic surfaces.To study the function of CAR on neural cells, we used the fiber knob of the adenovirus, extracellular CAR domains, blocking antibodies to CAR, as well as CAR-deficient neural cells. Blocking antibodies were found to inhibit neurite extension in retina organ and retinal explant cultures, whereas the application of the recombinant fiber knob of the adenovirus subtype Ad2 or extracellular CAR domains promoted neurite extension and adhesion to extracellular matrices.We observed a promiscuous interaction of CAR with extracellular matrix glycoproteins, which was deduced from analytical ultracentrifugation experiments, affinity chromatography, and adhesion assays. The membrane proximal Ig domain of CAR, termed D2, was found to bind to a fibronectin fragment, including the heparin-binding domain 2, which promotes neurite extension of wild type, but not of CAR-deficient neural cells. In contrast to heterophilic interactions, homophilic association of CAR involves both Ig domains, as was revealed by ultracentrifugation, chemical cross-linking, and adhesion studies. The results of these functional and binding studies are correlated to a U-shaped homodimer of the complete extracellular domains of CAR detected by x-ray crystallography.

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Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Cited by 38 publications

References 46 publications

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

¹⁷⁷Lu-DOTMP: A viable agent for palliative radiotherapy of painful bone metastasis

The Coxsackievirus–Adenovirus Receptor Reveals Complex Homophilic and Heterophilic Interactions on Neural Cells

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Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Cited by 38 publications

References 46 publications

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in LacrimalAcini

177Lu-DOTMP: A viable agent for palliative radiotherapy of painful bone metastasis

The Coxsackievirus–Adenovirus Receptor Reveals Complex Homophilic and Heterophilic Interactions on Neural Cells

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Product

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¹⁷⁷Lu-DOTMP: A viable agent for palliative radiotherapy of painful bone metastasis