144 results on '"Shayakhmetov, Dmitry M."'
Search Results
2. Targeted, safe, and efficient gene delivery to human hematopoietic stem and progenitor cells in vivo using the engineered AVID adenovirus vector platform
- Author
-
Yao, Jia, Atasheva, Svetlana, Wagner, Nicole, Di Paolo, Nelson C., Stewart, Phoebe L., and Shayakhmetov, Dmitry M.
- Published
- 2024
- Full Text
- View/download PDF
3. A paracrine circuit of IL-1[beta]/IL-1R1 between myeloid and tumor cells drives genotype-dependent glioblastoma progression
- Author
-
Chen, Zhihong, Giotti, Bruno, Kaluzova, Milota, Vallcorba, Montse Puigdelloses, Rawat, Kavita, Price, Gabrielle, Herting, Cameron J., Pinero, Gonzalo, Cristea, Simona, Ross, James L., Ackley, James, Maximov, Victor, Szulzewsky, Frank, Thomason, Wes, Marquez-Ropero, Mar, Angione, Angelo, Nichols, Noah, Tsankova, Nadejda M., Michor, Franziska, Shayakhmetov, Dmitry M., Gutmann, David H., Tsankov, Alexander M., and Hambardzumyan, Dolores
- Subjects
Cytokines -- Genetic aspects ,T cells -- Genetic aspects ,Platelet-derived growth factor -- Genetic aspects ,Macrophages ,Glioblastoma multiforme -- Genetic aspects -- Development and progression ,Health care industry ,Emory University. School of Medicine - Abstract
Monocytes and monocyte-derived macrophages (MDMs) from blood circulation infiltrate glioblastoma (GBM) and promote growth. Here, we show that PDGFS-driven GBM cells induce the expression of the potent proinflammatory cytokine IL- 1[beta] in MDM, which engages IL-1R1 in tumor cells, activates the NF-[kappa]B pathway, and subsequently leads to induction of monocyte chemoattractant proteins (MCPs). Thus, a feedforward paracrine circuit of IL-1[beta]/IL-1R1 between tumors and MDM creates an interdependence driving PDGFS-driven GBM progression. Genetic loss or locally antagonizing IL-1[beta]/IL-1R1 leads to reduced MDM infiltration, diminished tumor growth, and reduced exhausted [CD8.sup.+] T cells and thereby extends the survival of tumor-bearing mice. In contrast to IL-1[beta], IL-1[alpha] exhibits antitumor effects. Genetic deletion of ll1a/b is associated with decreased recruitment of lymphoid cells and loss-of-interferon signaling in various immune populations and subsets of malignant cells and is associated with decreased survival time of PDGFS-driven tumor-bearing mice. In contrast to PDGFB-driven GBM, Nf1-silenced tumors have a constitutively active NF-[kappa]B pathway, which drives the expression of MCPs to recruit monocytes into tumors. These results indicate local antagonism of IL-1[beta] could be considered as an effective therapy specifically for proneural GBM., Introduction Glioblastoma (GBM) is the most prevalent and aggressive primary brain tumor in children and adults and has poor overall survival rates. Numerous therapies have entered clinical trials, with only [...]
- Published
- 2023
- Full Text
- View/download PDF
4. p38MAPK guards the integrity of endosomal compartments through regulating necrotic death
- Author
-
Yao, Jia, Atasheva, Svetlana, Toy, Randall, Blanchard, Emmeline L., Santangelo, Philip J., Roy, Krishnendu, Mocarski, Edward S., and Shayakhmetov, Dmitry M.
- Published
- 2022
- Full Text
- View/download PDF
5. Modification of primary amines to higher order amines reduces in vivo hematological and immunotoxicity of cationic nanocarriers through TLR4 and complement pathways
- Author
-
Toy, Randall, Pradhan, Pallab, Ramesh, Vijayeetha, Di Paolo, Nelson C., Lash, Blake, Liu, Jiaying, Blanchard, Emmeline L., Pinelli, Christopher J., Santangelo, Philip J., Shayakhmetov, Dmitry M., and Roy, Krishnendu
- Published
- 2019
- Full Text
- View/download PDF
6. Inflammasome components caspase-1 and adaptor protein apoptosis-associated speck-like proteins are important in resistance to Cryptosporidium parvum
- Author
-
McNair, Nina N., Bedi, Chetna, Shayakhmetov, Dmitry M., Arrowood, Michael J., and Mead, Jan R.
- Published
- 2018
- Full Text
- View/download PDF
7. Caspase-8 and RIP kinases regulate bacteria-induced innate immune responses and cell death
- Author
-
Weng, Dan, Marty-Roix, Robyn, Ganesan, Sandhya, Proulx, Megan K., Vladimer, Gregory I., Kaiser, William J., Mocarski, Edward S., Pouliot, Kimberly, Chan, Francis Ka-Ming, Kelliher, Michelle A., Harris, Phillip A., Bertin, John, Gough, Peter J., Shayakhmetov, Dmitry M., Goguen, Jon D., Fitzgerald, Katherine A., Silverman, Neal, and Lien, Egil
- Published
- 2014
8. Inflammasome-independent IL-1β mediates autoinflammatory disease in Pstpip2-deficient mice
- Author
-
Cassel, Suzanne L., Janczy, John R., Bing, Xinyu, Wilson, Shruti P., Olivier, Alicia K., Otero, Jesse E., lwakura, Yoichiro, Shayakhmetov, Dmitry M., Bassuk, Alexander G., Abu-Amer, Yousef, Brogden, Kim A., Burns, Trudy L., Sutterwala, Fayyaz S., and Ferguson, Polly J.
- Published
- 2014
9. Structural Model for Factor X Inhibition of IgM and Complement-Mediated Neutralization of Adenovirus.
- Author
-
Wagner, Nicole, Shayakhmetov, Dmitry M., and Stewart, Phoebe L.
- Subjects
- *
ADENOVIRUSES , *COMPLEMENT (Immunology) , *STRUCTURAL models , *IMMUNOGLOBULIN M , *ONCOLYTIC virotherapy , *COMPLEMENT activation , *BLOOD proteins - Abstract
Adenovirus has strong therapeutic potential as an oncolytic virus and gene therapy vector. However, injecting human species C serotype 5 adenovirus, HAdv-C5, into the bloodstream leads to numerous interactions with plasma proteins that affect viral tropism and biodistribution, and can lead to potent immune responses and viral neutralization. The HAdv/factor X (FX) interaction facilitates highly efficient liver transduction and protects virus particles from complement-mediated neutralization after intravenous delivery. Ablating the FX interaction site on the HAdv-C5 capsid leaves the virus susceptible to neutralization by natural IgM followed by activation of the complement cascade and covalent binding of complement components C4b and C3b to the viral capsid. Here we present structural models for IgM and complement components C1, C4b, and C3b in complex with HAdv-C5. Molecular dynamics simulations indicate that when C3b binds near the vertex, multiple stabilizing interactions can be formed between C3b, penton base, and fiber. These interactions may stabilize the vertex region of the capsid and prevent release of the virally encoded membrane lytic factor, protein VI, which is packaged inside of the viral capsid, thus effectively neutralizing the virus. In a situation where FX and IgM are competing for binding to the capsid, IgM may not be able to form a bent conformation in which most of its Fab arms interact with the capsid. Our structural modeling of the competitive interaction of FX and IgM with HAdv-C5 allows us to propose a mechanistic model for FX inhibition of IgM-mediated virus neutralization. According to this model, although IgM may bind to the capsid, in the presence of FX it will likely retain a planar conformation and thus be unable to promote activation of the complement cascade at the virus surface. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
10. Coagulation Factor X Activates Innate Immunity to Human Species C Adenovirus
- Author
-
Doronin, Konstantin, Flatt, Justin W., Di Paolo, Nelson C., Khare, Reeti, Kalyuzhniy, Oleksandr, Acchione, Mauro, Sumida, John P., Ohto, Umeharu, Shimizu, Toshiyuki, Akashi-Takamura, Sachiko, Miyake, Kensuke, MacDonald, James W., Bammler, Theo M., Beyer, Richard P., Farin, Frederico M., Stewart, Phoebe L., and Shayakhmetov, Dmitry M.
- Published
- 2012
- Full Text
- View/download PDF
11. Interdependence between Interleukin-1 and Tumor Necrosis Factor Regulates TNF-Dependent Control of Mycobacterium tuberculosis Infection
- Author
-
Di Paolo, Nelson C., Shafiani, Shahin, Day, Tracey, Papayannoupoulou, Thalia, Russell, David W., Iwakura, Yoichiro, Sherman, David, Urdahl, Kevin, and Shayakhmetov, Dmitry M.
- Published
- 2015
- Full Text
- View/download PDF
12. Cytokine Responses to Adenovirus and Adenovirus Vectors.
- Author
-
Atasheva, Svetlana and Shayakhmetov, Dmitry M.
- Subjects
- *
ADENOVIRUSES , *CYTOKINE release syndrome , *CYTOKINES , *INTRAVENOUS injections , *GENE therapy , *IMMUNE system - Abstract
The expression of cytokines and chemokines in response to adenovirus infection is tightly regulated by the innate immune system. Cytokine-mediated toxicity and cytokine storm are known clinical phenomena observed following naturally disseminated adenovirus infection in immunocompromised hosts as well as when extremely high doses of adenovirus vectors are injected intravenously. This dose-dependent, cytokine-mediated toxicity compromises the safety of adenovirus-based vectors and represents a critical problem, limiting their utility for gene therapy applications and the therapy of disseminated cancer, where intravenous injection of adenovirus vectors may provide therapeutic benefits. The mechanisms triggering severe cytokine response are not sufficiently understood, prompting efforts to further investigate this phenomenon, especially in clinically relevant settings. In this review, we summarize the current knowledge on cytokine and chemokine activation in response to adenovirus- and adenovirus-based vectors and discuss the underlying mechanisms that may trigger acute cytokine storm syndrome. First, we review profiles of cytokines and chemokines that are activated in response to adenovirus infection initiated via different routes. Second, we discuss the molecular mechanisms that lead to cytokine and chemokine transcriptional activation. We further highlight how immune cell types in different organs contribute to synthesis and systemic release of cytokines and chemokines in response to adenovirus sensing. Finally, we review host factors that can limit cytokine and chemokine expression and discuss currently available and potential future interventional approaches that allow for the mitigation of the severity of the cytokine storm syndrome. Effective cytokine-targeted interventional approaches may improve the safety of systemic adenovirus delivery and thus broaden the potential clinical utility of adenovirus-based therapeutic vectors. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
13. Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and present them to antiviral B cells
- Author
-
Junt, Tobias, Moseman, E. Ashley, Iannacone, Matteo, Massberg, Steffen, Lang, Philipp A., Boes, Marianne, Fink, Katja, Henrickson, Sarah E., Shayakhmetov, Dmitry M., Di Paolo, Nelson C., van Rooijen, Nico, Mempel, Thorsten R., Whelan, Sean P., and von Andrian, Ulrich H.
- Published
- 2007
- Full Text
- View/download PDF
14. Nano-Emulsion Adjuvant Enhances Cross-presentation by Lipid Body Formation Independent of Glycolysis
- Author
-
Suresh, M, Lee, Woojong, Kingstad-Bakke, Brock, Paulson, Brett, Larsen, Autumn, Overmyer, Katherine, Marinaik, Chandranaik, Dull, Kelly, Toy, Randall, Vogel, Gabriela, Mueller, Katherine, Tweed, Kelsey, Walsh, Alex, Russell, Jason, Saha, Krishanu, Reyes, Leticia, Skala, Melissa, Sauer, John-Demian, Shayakhmetov, Dmitry M, Coon, Joshua, and Roy, Krishnendu
- Abstract
Here, we report that a carbomer-based adjuvant, Adjuplex® (ADJ), stimulated robust CD8 T-cell responses to subunit antigens by modulating multiple steps in the cytosolic pathway of cross-presentation, and afforded effective immunity against virus and intracellular bacteria. Cross-presentation induced by TLR agonists requires a critical switch to anabolic metabolism, but ADJ enhanced cross presentation without this metabolic switch in DCs and NLRP3-driven caspase 1 activity. Instead, ADJ induced in DCs, an unique metabolic state, typified by dampened oxidative phosphorylation and basal levels of glycolysis. In the absence of increased glycolytic flux, induction of ROS and lipid bodies (LBs) and alterations in LB composition mediated by ADJ were critical for DC cross-presentation. These findings challenge the prevailing metabolic paradigm by suggesting that DCs can perform effective DC cross-presentation, independent of glycolysis to induce robust T cell-dependent protective immunity to intracellular pathogens. These findings have implications in the rational development of novel adjuvants that promote cross-presentation and elicit potent cell-mediated immunity.
- Published
- 2020
- Full Text
- View/download PDF
15. Interleukin-1 Receptor 2 Keeps the Lid on Interleukin-1α
- Author
-
Di Paolo, Nelson C. and Shayakhmetov, Dmitry M.
- Published
- 2013
- Full Text
- View/download PDF
16. CD46 is a cellular receptor for group B adenoviruses
- Author
-
Gaggar, Anuj, Shayakhmetov, Dmitry M, and Lieber, Andre
- Abstract
Author(s): Anuj Gaggar [1, 2]; Dmitry M Shayakhmetov [2]; Andre Lieber (corresponding author) [1, 2] Group B adenoviruses, a subgenus of human Adenoviridae, are associated with a variety of often-fatal [...]
- Published
- 2003
- Full Text
- View/download PDF
17. Virus Binding to a Plasma Membrane Receptor Triggers Interleukin-1α-Mediated Proinflammatory Macrophage Response In Vivo
- Author
-
Di Paolo, Nelson C., Miao, Edward A., Iwakura, Yoichiro, Murali-Krishna, Kaja, Aderem, Alan, Flavell, Richard A., Papayannopoulou, Thalia, and Shayakhmetov, Dmitry M.
- Published
- 2009
- Full Text
- View/download PDF
18. Carbomer-based adjuvant elicits CD8 T-cell immunity by inducing a distinct metabolic state in cross-presenting dendritic cells.
- Author
-
Lee, Woojong, Kingstad-Bakke, Brock, Paulson, Brett, Larsen, Autumn, Overmyer, Katherine, Marinaik, Chandranaik B., Dulli, Kelly, Toy, Randall, Vogel, Gabriela, Mueller, Katherine P., Tweed, Kelsey, Walsh, Alex J., Russell, Jason, Saha, Krishanu, Reyes, Leticia, Skala, Melissa C., Sauer, John-Demian, Shayakhmetov, Dmitry M., Coon, Joshua, and Roy, Krishnendu
- Subjects
DENDRITIC cells ,IMMUNITY ,CELLULAR immunity ,AIDS ,CYTOSOL ,T cells ,GLYCOLYSIS - Abstract
There is a critical need for adjuvants that can safely elicit potent and durable T cell-based immunity to intracellular pathogens. Here, we report that parenteral vaccination with a carbomer-based adjuvant, Adjuplex (ADJ), stimulated robust CD8 T-cell responses to subunit antigens and afforded effective immunity against respiratory challenge with a virus and a systemic intracellular bacterial infection. Studies to understand the metabolic and molecular basis for ADJ's effect on antigen cross-presentation by dendritic cells (DCs) revealed several unique and distinctive mechanisms. ADJ-stimulated DCs produced IL-1β and IL-18, suggestive of inflammasome activation, but in vivo activation of CD8 T cells was unaffected in caspase 1-deficient mice. Cross-presentation induced by TLR agonists requires a critical switch to anabolic metabolism, but ADJ enhanced cross presentation without this metabolic switch in DCs. Instead, ADJ induced in DCs, an unique metabolic state, typified by dampened oxidative phosphorylation and basal levels of glycolysis. In the absence of increased glycolytic flux, ADJ modulated multiple steps in the cytosolic pathway of cross-presentation by enabling accumulation of degraded antigen, reducing endosomal acidity and promoting antigen localization to early endosomes. Further, by increasing ROS production and lipid peroxidation, ADJ promoted antigen escape from endosomes to the cytosol for degradation by proteasomes into peptides for MHC I loading by TAP-dependent pathways. Furthermore, we found that induction of lipid bodies (LBs) and alterations in LB composition mediated by ADJ were also critical for DC cross-presentation. Collectively, our model challenges the prevailing metabolic paradigm by suggesting that DCs can perform effective DC cross-presentation, independent of glycolysis to induce robust T cell-dependent protective immunity to intracellular pathogens. These findings have strong implications in the rational development of safe and effective immune adjuvants to potentiate robust T-cell based immunity. Author summary: An adjuvant is the pharmacological agent that is added to vaccines to boost immune responses. Currently, there are only seven FDA-approved adjuvants for human use, and vaccines based on these adjuvants have mainly been evaluated for elicitation of antibody-based immunity. However, vaccines need to also stimulate T cell-mediated immunity to protect against diseases such as AIDS, TB and Malaria. Hence, there is a critical need to develop adjuvants that stimulate protective T cell immunity. Here, we identified an adjuvant (Adjuplex; ADJ) that safely induces strong T cell immunity and protects against virus and intracellular bacteria. We also found that ADJ stimulated T cell immunity by unique mechanisms that did not include metabolic activation of antigen-presenting dendritic cells. Instead, ADJ induced a low metabolic state and engaged mechanisms including lipid pathways and induction of reactive oxygen species to promote activation of T cells by dendritic cells, following vaccination. These data not only provide new mechanistic insights into the mechanisms driving activation of T cells by ADJ, it provides a blue print for what adjuvants need to do to induce protection against infections that require T cell immunity. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
19. Systemic cancer therapy with engineered adenovirus that evades innate immunity.
- Author
-
Atasheva, Svetlana, Emerson, Corey C., Yao, Jia, Young, Cedrick, Stewart, Phoebe L., and Shayakhmetov, Dmitry M.
- Subjects
IMMUNOGLOBULIN M ,NATURAL immunity ,CANCER treatment ,LIVER cells ,BLOOD coagulation factors ,ADENOVIRUSES - Abstract
De-livering an oncolytic virus: Oncolytic viruses, optimized for killing tumor cells, are a promising modality in cancer immunotherapy. However, there are some challenges that these viruses must overcome, including antiviral immune responses that can affect both the efficacy and the safety of these viruses. For example, naturally occurring immunoglobulin M (IgM) antibodies bind to human species C adenovirus, resulting in trapping of the virus in the liver. To overcome this, Atasheva et al. engineered a modified version of this adenovirus, with mutations in the binding site for IgM. This modified virus was not trapped in the liver and effectively suppressed tumor growth in mouse models without safety concerns. Oncolytic virus therapy is a cancer treatment modality that has the potential to improve outcomes for patients with currently incurable malignancies. Although intravascular delivery of therapeutic viruses provides access to disseminated tumors, this delivery route exposes the virus to opsonizing and inactivating factors in the blood, which limit the effective therapeutic virus dose and contribute to activation of systemic toxicities. When human species C adenovirus HAdv-C5 is delivered intravenously, natural immunoglobulin M (IgM) antibodies and coagulation factor X rapidly opsonize HAdv-C5, leading to virus sequestration in tissue macrophages and promoting infection of liver cells, triggering hepatotoxicity. Here, we showed that natural IgM antibody binds to the hypervariable region 1 (HVR1) of the main HAdv-C5 capsid protein hexon. Using compound targeted mutagenesis of hexon HVR1 loop and other functional sites that mediate virus-host interactions, we engineered and obtained a high-resolution cryo–electron microscopy structure of an adenovirus vector, Ad5-3M, which resisted inactivation by blood factors, avoided sequestration in liver macrophages, and failed to trigger hepatotoxicity after intravenous delivery. Systemic delivery of Ad5-3M to mice with localized or disseminated lung cancer led to viral replication in tumor cells, suppression of tumor growth, and prolonged survival. Thus, compound targeted mutagenesis of functional sites in the virus capsid represents a generalizable approach to tailor virus interactions with the humoral and cellular arms of the immune system, enabling generation of "designer" viruses with improved therapeutic properties. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
20. Innate immunity to adenovirus: lessons from mice.
- Author
-
Atasheva, Svetlana, Yao, Jia, and Shayakhmetov, Dmitry M.
- Subjects
ADENOVIRUSES ,NATURAL immunity ,GENETIC vectors ,DISSEMINATED intravascular coagulation ,ANIMALS ,MICE ,INTRAVENOUS injections - Abstract
Adenovirus is a highly evolutionary successful pathogen, as it is widely prevalent across the animal kingdom, infecting hosts ranging from lizards and frogs to dolphins, birds, and humans. Although natural adenovirus infections in humans rarely cause severe pathology, intravenous injection of high doses of adenovirus‐based vectors triggers rapid activation of the innate immune system, leading to cytokine storm syndrome, disseminated intravascular coagulation, thrombocytopenia, and hepatotoxicity, which individually or in combination may cause morbidity and mortality. Much of the information on exactly how adenovirus activates the innate immune system has been gathered from mouse experimental systems. Intravenous administration of adenovirus to mice revealed mechanistic insights into cellular and molecular components of the innate immunity that detect adenovirus particles, activate pro‐inflammatory signaling pathways and cytokine production, sequester adenovirus particles from the bloodstream, and eliminate adenovirus‐infected cells. Collectively, this information greatly improved our understanding of mechanisms of activation of innate immunity to adenovirus and may pave the way for designing safer adenovirus‐based vectors for therapy of genetic and acquired human diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
21. Tumour-associated macrophage-derived interleukin-1 mediates glioblastoma-associated cerebral oedema.
- Author
-
Herting, Cameron J, Chen, Zhihong, Maximov, Victor, Duffy, Alyssa, Szulzewsky, Frank, Shayakhmetov, Dmitry M, and Hambardzumyan, Dolores
- Subjects
VASCULAR endothelial growth factors ,INTERLEUKIN-1 ,PLATELET-derived growth factor ,GLIOBLASTOMA multiforme ,INTERLEUKIN-1 receptors ,CEREBELLAR tumors - Abstract
Glioblastoma is the most common and uncompromising primary brain tumour and is characterized by a dismal prognosis despite aggressive treatment regimens. At the cellular level, these tumours are composed of a mixture of neoplastic cells and non-neoplastic cells, including tumour-associated macrophages and endothelial cells. Cerebral oedema is a near-universal occurrence in patients afflicted with glioblastoma and it is almost exclusively managed with the corticosteroid dexamethasone despite significant drawbacks associated with its use. Here, we demonstrate that dexamethasone blocks interleukin-1 production in both bone marrow-derived and brain resident macrophage populations following stimulation with lipopolysaccharide and interferon gamma. Additionally, dexamethasone is shown to inhibit downstream effectors of interleukin-1 signalling in both macrophage populations. Co-culture of bone marrow-derived macrophages with organotypic tumour slices results in an upregulation of interleukin-1 cytokines, an effect that is absent in co-cultured microglia. Genetic ablation of interleukin-1 ligands or receptor in mice bearing RCAS/tv-a-induced platelet-derived growth factor B-overexpressing glioblastoma results in reduced oedema and partial restoration of the integrity of the blood-brain barrier, respectively; similar to results obtained with vascular endothelial growth factor neutralization. We establish that tumours from dexamethasone-treated mice exhibit reduced infiltration of cells of the myeloid and lymphoid compartments, an effect that should be considered during clinical trials for immunotherapy in glioblastoma patients. Additionally, we emphasize that caution should be used when immune profiling and single-cell RNA sequencing data are interpreted from fresh glioblastoma patient samples, as nearly all patients receive dexamethasone after diagnosis. Collectively, this evidence suggests that interleukin-1 signalling inhibition and dexamethasone treatment share therapeutic efficacies and establishes interleukin-1 signalling as an attractive and specific therapeutic target for the management of glioblastoma-associated cerebral oedema. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
22. Interdependence between interleukin-1 and tumor necrosis factor controls TNF-dependent effector functions during Mycobacterium tuberculosis infection
- Author
-
Di Paolo, Nelson C., Shafiani, Shahin, Day, Tracey, Papayannoupoulou, Thalia, Russell, David W., Iwakura, Yoichiro, Sherman, David, Urdahl, Kevin, and Shayakhmetov, Dmitry M.
- Subjects
Mice, Knockout ,Receptors, Interleukin-1 Type I ,Tumor Necrosis Factor-alpha ,Fluorescent Antibody Technique ,Cell Separation ,Mycobacterium tuberculosis ,Flow Cytometry ,Article ,Mice, Inbred C57BL ,Disease Models, Animal ,Mice ,Interleukin-1alpha ,Animals ,Tuberculosis - Abstract
The interleukin-1 receptor I (IL-1RI) is critical for host resistance to Mycobacterium tuberculosis (Mtb), yet the mechanisms of IL-1RI-mediated pathogen control remain unclear. Here, we show that without IL-1RI, Mtb-infected newly recruited Ly6Ghigh myeloid cells failed to up-regulate tumor necrosis factor receptor I (TNF-RI) and to produce reactive oxygen species, resulting in compromised pathogen control. Furthermore, simultaneous ablation of IL-1RI and TNF-RI signaling on either stroma or hematopoietic cells led to early lethality, indicating non-redundant and synergistic roles of IL-1 and TNF in mediating macrophage-stroma cross-talk that was critical for optimal control of Mtb infection. Finally, we show that even in the presence of functional Mtb-specific adaptive immunity, the lack of IL-1α and not IL-1β led to an exuberant intracellular pathogen replication and progressive non-resolving inflammation. Our study reveals functional interdependence between IL-1 and TNF in enabling Mtb control mechanisms that are critical for host survival.
- Published
- 2015
23. Influence of Fat on Differential Receptor Interacting Serine/Threonine Protein Kinase 1 Activity Leading to Apoptotic Cell Death in Murine Liver Ischemia Reperfusion Injury Through Caspase 8.
- Author
-
Kolachala, Vasantha L., Palle, Sirish K., Shen, Ming, Shenoi, Asha, Shayakhmetov, Dmitry M., and Gupta, Nitika A.
- Subjects
LIVER injuries ,PROTEIN kinases ,REPERFUSION injury - Abstract
Current understanding is that receptor interacting serine/threonine protein kinase 1 (RIPK1) can lead to two distinct forms of cell death: RIPK3‐mediated necroptosis or caspase 8 (Casp8)‐mediated apoptosis. Here, we report that RIPK1 signaling is indispensable for protection from hepatocellular injury in a steatotic liver undergoing ischemia reperfusion injury (IRI) but not in the lean liver. In lean liver IRI, RIPK1‐mediated cell death is operational, leading to protection in RIP1 kinase‐dead knock‐in (RIPK1K45A) mice and necrostatin‐1s (Nec1s)‐treated lean wild‐type (WT) mice. However, when fed a high‐fat diet (HFD), RIPK1K45A‐treated and Nec1s‐treated WT mice undergoing IRI demonstrate exacerbated hepatocellular injury along with decreased RIPK1 ubiquitylation. Furthermore, we demonstrate that HFD‐fed RIPK3–/–/Casp8–/– mice show protection from IRI, but HFD‐fed RIPK3–/–/Casp8–/+ mice do not. We also show that blockade of RIPK1 leads to increased Casp8 activity and decreases mitochondrial viability. Conclusion: Although more studies are required, we provide important proof of concept for RIPK1 inhibition leading to distinctive outcomes in lean and steatotic liver undergoing IRI. Considering the rising incidence of nonalcoholic fatty liver disease (NAFLD) in the general population, it will be imperative to address this critical difference when treating patients with RIPK1 inhibitors. This study also presents a new target for drug therapy to prevent hepatocellular injury in NAFLD. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
24. Adenovirus sensing by the immune system.
- Author
-
Atasheva, Svetlana and Shayakhmetov, Dmitry M.
- Abstract
The host immune system developed multiple ways for recognition of viral pathogens. Upon disseminated adenovirus infection, the immune system senses adenovirus invasion from the moment it enters the bloodstream. The soluble blood factors, FX, antibodies, and complement, can bind and activate plethora of host-protective immune responses. Adenovirus binding to the cellular β3 integrin and endosomal membrane rupture trigger activation of IL-1α/IL-1R1 proinflammatory cascade leading to attraction of cytotoxic immune cells to the site of infection. Upon cell entry, adenovirus exposes its DNA genome in the cytoplasm and triggers DNA sensors signaling. Even when inside the nucleus, the specialized cellular machinery that recognizes the double-strand DNA breaks become activated and triggers viral DNA replication arrest. Thus, the host employs very diverse mechanisms to prevent viral dissemination. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
25. Immune Complexes Indirectly Suppress the Generation of Th17 Responses In Vivo.
- Author
-
Ciraci, Ceren, Janczy, John R., Jain, Nidhi, Haasken, Stefanie, Pecli e Silva, Cyntia, Benjamim, Claudia F., Sadler, Jeffrey J., Olivier, Alicia K., Iwakura, Yoichiro, Shayakhmetov, Dmitry M., Sutterwala, Fayyaz S., and Cassel, Suzanne L.
- Subjects
T helper cells ,NATURAL immunity ,CD4 antigen ,IMMUNOLOGICAL adjuvants ,INFLAMMASOMES ,DENDRITIC cells - Abstract
The precise context in which the innate immune system is activated plays a pivotal role in the subsequent instruction of CD4
+ T helper (Th) cell responses. Th1 responses are downregulated when antigen is encountered in the presence of antigen-IgG immune complexes. To assess if Th17 responses to antigen are subject to similar influences in the presence of immune complexes we utilized an inflammatory airway disease model in which immunization of mice with Complete Freund’s Adjuvant (CFA) and ovalbumin (Ova) induces a powerful Ova-specific Th1 and Th17 response. Here we show that modification of that immunization with CFA to include IgG-Ova immune complexes results in the suppression of CFA-induced Th17 responses and a concurrent enhancement of Ova-specific Th2 responses. Furthermore, we show the mechanism by which these immune complexes suppress Th17 responses is through the enhancement of IL-10 production. In addition, the generation of Th17 responses following immunization with CFA and Ova were dependent on IL-1α but independent of NLRP3 inflammasome activation. Together these data represent a novel mechanism by which the generation of Th17 responses is regulated. [ABSTRACT FROM AUTHOR]- Published
- 2016
- Full Text
- View/download PDF
26. The Analysis of Innate Immune Response to Adenovirus Using Antibody Arrays.
- Author
-
Di Paolo, Nelson C. and Shayakhmetov, Dmitry M.
- Published
- 2014
- Full Text
- View/download PDF
27. Construction of Targeted and Armed Oncolytic Adenoviruses.
- Author
-
Doronin, Konstantin and Shayakhmetov, Dmitry M.
- Published
- 2012
- Full Text
- View/download PDF
28. Efficacy, Toxicity, and Immunogenicity of Adenoviral Vectors.
- Author
-
Teicher, Beverly A., Hunt, Kelly K., Vorburger, Stephan A., Swisher, Stephen G., and Shayakhmetov, Dmitry M.
- Abstract
To date, over 60% of all gene therapy clinical trials in the United States have focused on the development and validation of new therapies for cancer. Many of these trials utilize Ad vectors as novel anticancer therapeutics. In recent years, however, initial enthusiasm and high expectations for successful clinical application of Ad-based vectors as efficient anticancer therapeutics has been dampened based on the data obtained during a series of clinical trials. Along with the major concerns over the safety of systemic Ad application, which was found to be associated with immediate innate and inflammatory host responses and can also lead to fatalities, such issues as rapid clearance of the bulk of administered vector by cells of the reticulo-endothelial system, neutralization of virus particles by highly prevalent pre-existing antibodies, and poor transduction of primary tumors resulting from low-level Ad receptor expression and/or anatomical barriers, including extracellular matrix surrounding tumors, have established a great need for research to further improve existing Ad vectors and unravel their true therapeutic potential as anticancer agents. This chapter reviews and discusses the current status, limitations and future challenges for the Ad vector development field with respect to their efficacy, toxicity and immunogenicity. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
29. Generation of Adenovirus Vectors Devoid of All Viral Genes by Recombination Between Inverted Repeats.
- Author
-
Walker, John M., Machida, Curtis A., Stecher, Hartmut, Carlson, Cheryl A., Shayakhmetov, Dmitry M., and Lieber, André
- Abstract
Adenovirus (Ad) vector-mediated gene transfer is useful in applications where transient, high-level transgene expression is required. In contrast to onco-retroviral vectors, Ad vectors efficiently infect nondividing cells. Most recombinant Ad vectors currently used for in vitro and in vivo gene transfer are usually deleted for the E1 and/or E3 region (first-generation Ad vectors) and are based on serotype 5 (Ad5). First-generation Ad vectors can be produced and purified at high titers. Disadvantages of these gene transfer vehicles include their episomal nature that only allows for short-term gene expression, cytotoxicity from expressed viral proteins, and elicitation of immune responses. In order to circumvent these problems, new recombinant Ad vectors devoid of all viral genes have been developed (1). These "gutless" Ad (ΔAd) vectors show significantly less cytotoxic and immunogenic side effects. [ABSTRACT FROM AUTHOR]
- Published
- 2003
- Full Text
- View/download PDF
30. Caspase-8 and RIP kinases regulate bacteria-induced innate immune responses and cell death.
- Author
-
Dan Weng, Marty-Roix, Robyn, Ganesan, Sandhya, Proulx, Megan K., Vladimer, Gregory I., Kaiser, William J., Mocarski, Edward S., Pouliot, Kimberly, Francis Ka-Ming Chan, Kelliher, Michelle A., Harris, Phillip A., Bertin, John, Gough, Peter J., Shayakhmetov, Dmitry M., Goguen, Jon D., Fitzgerald, Katherine A., Silverman, Neal, and Lien, Egil
- Subjects
CASPASES ,YERSINIA pestis ,CELL-mediated cytotoxicity ,RECEPTOR-interacting proteins ,MACROPHAGES ,NECROSIS - Abstract
A number of pathogens cause host cell death upon infection, and Yersinia pestis, infamous for its role in large pandemics such as the "Black Death" in medieval Europe, induces considerable cytotoxicity. The rapid killing of macrophages induced by Y. pestis, dependent upon type III secretion system effector Yersinia outer protein J (YopJ), is minimally affected by the absence of caspase-1, caspase- 11, Fas ligand, and TNF. Caspase-8 is known to mediate apoptotic death in response to infection with several viruses and to regulate programmed necrosis (necroptosis), but its role in bacterially induced cell death is poorly understood. Here we provide genetic evidence for a receptor-interacting protein (RIP) kinasecaspase- 8-dependent macrophage apoptotic death pathway after infection with Y. pestis, influenced by Toll-like receptor 4-TIR-domain- containing adapter-inducing interferon-β (TLR4-TRIF). Interestingly, macrophages lacking either RIP1, or caspase-8 and RIP3, also had reduced infection-induced production of IL-lβ, IL-18, TNF, and IL-6; impaired activation of the transcription factor NF-KB; and greatly compromised caspase-1 processing. Cleavage of the proform of caspase-1 is associated with triggering inflammasome activity, which leads to the maturation of IL-lβ and IL-18, cytokines important to host responses against Y. pestis and many other infectious agents. Our results identify a RIP1-caspase-8/RIP3-dependent caspase- 1 activation pathway after Y. pestis challenge. Mice defective in caspase-8 and RIP3 were also highly susceptible to infection and displayed reduced proinflammatory cytokines and myeloid cell death. We propose that caspase-8 and the RIP kinases are key regulators of macrophage cell death, NF-κB and inflammasome activation, and host resistance after Y. pestis infection. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
31. IL-1α and Complement Cooperate in Triggering Local Neutrophilic Inflammation in Response to Adenovirus and Eliminating Virus-Containing Cells.
- Author
-
Di Paolo, Nelson C., Baldwin, Lisa K., Irons, Eric E., Papayannopoulou, Thalia, Tomlinson, Stephen, and Shayakhmetov, Dmitry M.
- Subjects
INFLAMMATION ,HOMEOSTASIS ,ADENOVIRUSES ,MACROPHAGES ,CHEMOKINES - Abstract
Inflammation is a highly coordinated host response to infection, injury, or cell stress. In most instances, the inflammatory response is pro-survival and is aimed at restoring physiological tissue homeostasis and eliminating invading pathogens, although exuberant inflammation can lead to tissue damage and death. Intravascular injection of adenovirus (Ad) results in virus accumulation in resident tissue macrophages that trigger activation of CXCL1 and CXCL2 chemokines via the IL-1α-IL-1RI signaling pathway. However, the mechanistic role and functional significance of this pathway in orchestrating cellular inflammatory responses to the virus in vivo remain unclear. Resident metallophilic macrophages expressing macrophage receptor with collagenous structure (MARCO
+ ) in the splenic marginal zone (MZ) play the principal role in trapping Ad from the blood. Here we show that intravascular Ad administration leads to the rapid recruitment of Ly-6G+ 7/4+ polymorphonuclear leukocytes (PMNs) in the splenic MZ, the anatomical compartment that remains free of PMNs when these cells are purged from the bone marrow via a non-inflammatory stimulus. Furthermore, PMN recruitment in the splenic MZ resulted in elimination of virus-containing cells. IL-1α-IL-1RI signaling is only partially responsible for PMN recruitment in the MZ and requires CXCR2, but not CXCR1 signaling. We further found reduced recruitment of PMNs in the splenic MZ in complement C3-deficient mice, and that pre-treatment of IL-1α-deficient, but not wild-type mice, with complement inhibitor CR2-Crry (inhibits all complement pathways at C3 activation) or CR2-fH (inhibits only the alternative complement activation pathway) prior to Ad infection, abrogates PMN recruitment to the MZ and prevents elimination of MARCO+ macrophages from the spleen. Collectively, our study reveals a non-redundant role of the molecular factors of innate immunity – the chemokine-activating IL-1α-IL-1RI-CXCR2 axis and complement – in orchestrating local inflammation and functional cooperation of PMNs and resident macrophages in the splenic MZ, which collectively contribute to limiting disseminated pathogen spread via elimination of virus-containing cells. [ABSTRACT FROM AUTHOR]- Published
- 2014
- Full Text
- View/download PDF
32. The Transcription Factor IRF3 Triggers “Defensive Suicide” Necrosis in Response to Viral and Bacterial Pathogens.
- Author
-
Di Paolo, Nelson C., Doronin, Konstantin, Baldwin, Lisa K., Papayannopoulou, Thalia, and Shayakhmetov, Dmitry M.
- Abstract
Summary: Although molecular components that execute noninflammatory apoptotic cell death are well defined, molecular pathways that trigger necrotic cell death remain poorly characterized. Here, we show that in response to infection with adenovirus or Listeria monocytogenes, macrophages in vivo undergo rapid proinflammatory necrotic death that is controlled by interferon-regulatory factor 3 (IRF3). The transcriptional activity of IRF3 is, surprisingly, not required for the induction of necrosis, and it proceeds normally in mice deficient in all known regulators of necrotic death or IRF3 activation, including RIPK3, caspases 1, 8, or 11, STING, and IPS1/MAVS. Although L. monocytogenes triggers necrosis to promote the infection, IRF3-dependent necrosis is required for reducing pathogen burden in the models of disseminated infection with adenovirus. Therefore, our studies implicate IRF3 as a principal and nonredundant component of a physiologically regulated necrotic cell-death pathway that operates as an effective innate immune mechanism of host protection against disseminated virus infection. [Copyright &y& Elsevier]
- Published
- 2013
- Full Text
- View/download PDF
33. Recognition of Virus Infection and Innate Host Responses to Viral Gene Therapy Vectors.
- Author
-
Shayakhmetov, Dmitry M., Di Paolo, Nelson C., and Mossman, Karen L.
- Subjects
- *
VIRUS diseases , *GENE therapy , *INFLAMMATION , *CELL membranes , *ENDOSOMES , *CYTOSOL , *HOMEOSTASIS - Abstract
The innate immune and inflammatory response represents one of the key stumbling blocks limiting the efficacy of viral-based therapies. Numerous human diseases could be corrected or ameliorated if viruses were harnessed to safely and effectively deliver therapeutic genes to diseased cells and tissues in vivo. Recent studies have shown that host cells recognize viruses using an elaborate network of sensor proteins localized at the plasma membrane, in endosomes, or in the cytosol. Three classes of sensors have been implicated in sensing viruses in mammalian cells—Toll-like receptors (TLRs), retinoid acid-inducible gene (RIG)-I-like receptors (RLRs), and nucleotide oligomerization domain (NOD)-like receptors (NLRs). The interaction of virus-associated nucleic acids with these sensor molecules triggers a signaling cascade that activates the principal host defense program aimed to limit or eliminate virus infection and restore tissue homeostasis. In addition, recent data strongly suggest that host cells can mount innate immune responses to viruses without prior recognition of their nucleic acids. To deliver therapeutic genes into the nuclei of diseased cells, viral gene therapy vectors must be efficient at penetrating either the plasma or endosomal membrane. The therapeutic use of high numbers of virus particles disturbs cellular homeostasis, triggering cell damage and stress pathways, or “sensing of modified self”. Accumulating data indicate that the sensing of modified self might represent a powerful framework explaining the innate immune response activation by viral gene therapy vectors. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
34. Virus Infection Recognition and Early Innate Responses to Non-Enveloped Viral Vectors.
- Author
-
Shayakhmetov, Dmitry M.
- Subjects
- *
NATURAL immunity , *VIRUS diseases , *GENE therapy , *GENETIC engineering , *DNA viruses , *ADENOVIRUSES , *HUMAN genetics , *IMMUNE response , *IMMUNOLOGY - Abstract
Numerous human genetic and acquired diseases could be corrected or ameliorated if viruses are harnessed to safely and effectively deliver therapeutic genes to diseased cells and tissues in vivo. Innate immune and inflammatory response represents one of the key stumbling blocks during the development of viral-based therapies. In this review, current data on the early innate immune responses to viruses and to the most commonly used gene therapy vectors (using adenovirus and adeno-associated virus) will be discussed. Recent findings in the field may help develop new approaches to moderate these innate immune anti-viral responses and thus improve the safety of viral vectors for human gene therapy applications. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
35. A Capsid-Modified Helper-Dependent Adenovirus Vector Containing the β-Globin Locus Control Region Displays a Nonrandom Integration Pattern and Allows Stable, Erythroid-Specific Gene Expression.
- Author
-
Hongjie Wang, Shayakhmetov, Dmitry M., Leege, Tobias, Harkey, Michael, Qiliang Li, Papayannopoulou, Thalia, Stamatoyannopolous, George, and Lieber, André
- Subjects
- *
HEMOGLOBINOPATHY , *GENETIC transformation , *HEMATOPOIETIC stem cells , *GENE expression , *ADENOVIRUSES , *CELL lines , *VIROLOGY - Abstract
Gene therapy for hemoglobinopathies requires efficient gene transfer into hematopoietic stem cells and high-level erythroid-specific gene expression. Toward this goal, we constructed a helper-dependent adenovirus vector carrying the β-globin locus control region (LCR) to drive green fluorescent protein (GFP) expression, whereby the LCR-GFP cassette is flanked by adeno-associated virus (AAV) inverted terminal repeats (Ad.LCR-β-GFP). This vector possesses the adenovirus type 35 fiber knob that allows efficient infection of hematopoietic cells. Transduction and vector integration studies were performed in MO7e cells, a growth factor-dependent CD34+ erythroleukemic cell line, and in cord blood-derived human CD34+ cells. Stable transduction of MO7e cells with Ad.LCR-β-GFP was more efficient and less subject to position effects and silencing than transduction with a vector that did not contain the β-globin LCR. Analysis of integration sites indicated that Ad.LCR-β-GFP integration in MO7e cells was not random but tethered to chromosome 11, specifically to the globin LCR. More than 10% of analyzed integration sites were within the chromosomal β-globin LCR. None of the Ad.LCR-β-GFP integrations occurred in exons. The integration pattern of a helper-dependent vector that contained X-chromosomal stuffer DNA was different from that of the β-globin LCR-containing vector. Infection of primary CD34+ cells with Ad.LCR-β-GFP did not affect the clonogenic capacity of CD34+ cells. Transduction of CD34+ cells with Ad.LCR-β-GFP resulted in vector integration and erythroid lineage-specific GFP expression. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
36. Adenovirus Binding to Blood Factors Results in Liver Cell Infection and Hepatotoxicity.
- Author
-
Shayakhmetov, Dmitry M., Gaggar, Anuj, Shaoheng Ni, Zong-yi Li, and Lieber, André
- Subjects
- *
ADENOVIRUSES , *ADENOVIRUS diseases , *LIVER cells , *HEPATOTOXICOLOGY , *VIROLOGY , *VIRUS diseases , *VIRUSES , *DISEASES - Abstract
Adenoviruses (Ad) are efficient vehicles for gene delivery in vitro and in vivo. Therefore, they are a promising tool in gene therapy, particularly in the treatment of cancer and cardiovascular diseases. However, preclinical and clinical studies undertaken during the last decade have revealed a series of problems that limit both the safety and efficacy of Ad vectors, specifically after intravenous application. Major obstacles to clinical use include innate toxicity and Ad sequestration by nontarget tissues. The factors and mechanisms underlying these processes are poorly understood. The majority of intravenously injected Ad particles are sequestered by the liver, which in turn causes an inflammatory response characterized by acute transaminitis and vascular damage. Here, we describe a novel pathway that is used by Ad for infection of hepatocytes and Kupffer cells upon intravenous virus application in mice. We found that blood factors play a major role in targeting Ad vectors to hepatic cells. We demonstrated that coagulation factor IX and complement component C4-binding protein can bind the Ad fiber knob domain and provide a bridge for virus uptake through cell surface heparan sulfate proteoglycans and low-density lipoprotein receptor-related protein. An Ad vector, Ad5mut, which contained mutations in the fiber knob domain ablating blood factor binding, demonstrated significantly reduced infection of liver cells and liver toxicity in vivo. This study contributes to a better understanding of adenovirus-host interactions for intravenously applied vectors. It also provides a rationale for novel strategies to target adenovirus vector to specific tissues and to reduce virus-associated toxicity after systemic application. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
37. Deletion of Penton RGD Motifs Affects the Efficiency of both the Internalization and the Endosome Escape of Viral Particles Containing Adenovirus Serotype 5 or 35 Fiber Knobs.
- Author
-
Shayakhmetov, Dmitry M., Eberly, Andrea M., Zong-Yi Li, and Lieber, Andre´
- Subjects
- *
VIRUSES , *VIRUS diseases , *ADENOVIRUSES , *GLYCOPROTEINS , *GENES , *VIROLOGY - Abstract
Adenovirus (Ad) vectors are widely used for gene delivery in vitro and in vivo. A solid understanding of the biology of this virus is imperative for the development of novel, effective, and safe vectors. For the group C adenovirus serotypes 2 and 5 that use CAR as a primary attachment receptor, it is known that the penton base RGD motifs interact with cellular integrins and that this interaction promotes virus internalization. However, the RGD motif's impact on the efficiency of postinternalization steps, such as the escape of the virus particle from the endosome, is less defined. Furthermore, the role of penton-integrin interactions remains unknown for new vectors possessing group B Ad fiber knobs that use CD46 as a primary virus attachment receptor. In this study, we used vectors with the RGD motif deleted that contained Ad5 and B-group Ad35 fiber knobs and long fiber shafts and studied the role of RGD-integrin interactions in virus internalization and endosome escape. The deletion of the RGD motif in the penton base did not affect virus attachment, regardless of the type of cellular receptor used for attachment. RGD motif deletion, however, significantly reduced the rate of virus internalization for both the Ad5 and Ad35 fiber knob-containing vectors. This study also demonstrates the role of penton RGD motifs in facilitating the endosome escape step of virus infection and indicates that penton-integrin interactions are involved in internalization of capsid-chimeric CD46-interacting Ads with long fiber shafts. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
38. Genome Size and Structure Determine Efficiency of Postinternalization Steps and Gene Transfer of Capsid-Modified Adenovirus Vectors in a Cell-Type-Specific Manner.
- Author
-
Shayakhmetov, Dmitry M., Zong-Yi Li, Gaggar, Anuj, Gharwan, Helen, Ternovoi, Vladimir, Sandig, Volker, and Lieber, André
- Subjects
- *
ADENOVIRUSES , *GENETIC transformation , *TRANSGENES , *VIRAL genetics , *GENE expression , *HEMATOPOIETIC stem cells , *VIROLOGY - Abstract
Adenovirus serotype 5 (Ad5) vectors containing Ad B-group fibers have become increasingly popular as gene transfer vectors because they efficiently transduce human cell types that are relatively refractory to Ad5 infection. So far, most B-group fiber-containing vectors have been first-generation vectors, deleted of E1 and/or E3 genes. Transduction with these vectors, however, results in viral gene expression and is associated with cytotoxicity and immune responses against transduced cells. To circumvent these problems, we developed fiber-chimeric Ad vectors devoid of all viral genes that were produced either by the homologous recombination of first-generation vectors or by using the Cre/lox-based helper virus system. In this study we compared early steps of infection between first-generation (35-kb genome) and Ad vectors devoid of all viral genes with genome sizes of 28 kb and 12.6 kb. All vectors possessed an Ad35-derived fiber knob domain, which uses CD46 as a primary attachment receptor. Using immortalized human hematopoietic cell lines and primary human CD34-positive hematopoietic cells, we found that the Ad genome size did not affect the efficiency of virus attachment to and internalization into cells. Furthermore, independently of the genome length and structure, all vectors migrated to the nucleus through late endosomal and lysosomal cellular compartments. However, the vector containing the short 12.6-kb genome was unable to efficiently escape from endosomes and deliver its DNA into the nucleus. Moreover, compared to other vectors, these Ad particles were less stable and had an abnormal capsid protein composition, including a lack of capsid-stabilizing protein IX. Our data indicate that the size and structure of the packaged viral genomes can affect the integrity of Ad particles, which in turn results in lower infectivity of Ad vectors. [ABSTRACT FROM AUTHOR]
- Published
- 2004
- Full Text
- View/download PDF
39. Analysis of Adenovirus Sequestration in the Liver, Transduction of Hepatic Cells, and Innate Toxicity after Injection of Fiber-Modified Vectors.
- Author
-
Shayakhmetov, Dmitry M., Zong-Yi Li, Shaoheng Ni, and Lieber, André
- Subjects
- *
ADENOVIRUSES , *SEQUESTRATION (Chemistry) , *KUPFFER cells , *GENETIC transduction , *MACROPHAGES , *MICROBIAL genetics , *NATURAL immunity - Abstract
After intravenous administration, adenovirus (Ad) vectors are predominantly sequestered by the liver. Delineating the mechanisms for Ad accumulation in the liver is crucial for a better understanding of Ad clearance and Ad-associated innate toxicity. To help address these issues, in this study, we used Ad vectors with different fiber shaft lengths and either coxsackievirus-Ad receptor (CAR)-interacting Ad serotype 9 (Ad9) or non-CAR-interacting Ad35 fiber knob domains. We analyzed the kinetics of Ad vector accumulation in the liver, uptake into hepatocytes and Kupffer cells, and induction of cytokine expression and release in response to systemic vector application. Immediately after intravenous injection, all Ad vectors accumulated equally efficiently in the liver; however, only genomes of long-shafted Ads were maintained in the liver tissue over time. We found that Kupffer cell uptake of long-shafted Ads was mediated by the fiber knob domain and was CAR independent. The short-shafted Ads were unable to efficiently interact with hepatocellular receptors and were not taken up by Kupffer cells. Moreover, our studies indicated that Kupffer cells were not the major reservoir for the observed accumulation of Ads (used in this study) in the liver within the first 30 min after virus infusion. The lower level of liver cell transduction by short-shafted Ads correlated with a significantly reduced inflammatory anti-Ad response as well as liver damage induced by the systemic administration of these vectors. This study contributes to a better understanding of the biology of systemically applied Ad and will help in designing safer vectors that can efficiently transduce target tissues. [ABSTRACT FROM AUTHOR]
- Published
- 2004
- Full Text
- View/download PDF
40. The Interaction between the Fiber Knob Domain and the Cellular Attachment Receptor Determines the Intracellular Trafficking Route of Adenoviruses.
- Author
-
Shayakhmetov, Dmitry M., Zong-Yi Li, Ternovoi, Vladimir, Gaggar, Anuj, Gharwan, Helen, and Lieber, André
- Subjects
- *
ADENOVIRUSES , *TROPISMS , *GENETIC vectors , *CELL receptors - Abstract
Most of the presently used adenovirus (Ad) vectors are based on serotype 5. However, the application of these vectors is limited by the native tropism of Ad5. To address this problem, a series of fiber chimeric vectors were produced to take advantage of the different cellular receptors used by Ad of different subgroups. In this study we utilize an Ad5-based chimeric vector containing sequences encoding the Ad35 fiber knob domain instead of the Ad5 knob (Ad5/35L) to analyze factors responsible for selection of intracellular trafficking routes by Ads. By competition analysis with recombinant Ad5 and Ad35 knobs we showed that the Ad5/35L vector infected cells through a receptor different from the Ad5 receptor. Intracellular trafficking of Ad5 and Ad5/35L viruses was analyzed in HeLa cells by tracking fluorophore-conjugated Ad particles, by immunostaining for capsid hexon protein, by electron microscopy, and by Southern blotting for viral DNA. These studies showed that the interaction with the Ad35 receptor(s) predestines Ad5/35L vector to intracellular trafficking pathways different from those of Ad5. Ad5 efficiently escaped from the endosomes early after infection. In contrast, Ad5/35L remained longer in late endosomal/lysosomal compartments and used them to achieve localization to the nucleus. However, a significant portion of Ad5/35L particles appeared to be recycled back to the cell surface. This phenomenon resulted in significantly less efficient Ad5/35L-mediated gene transfer compared to that of Ad5. We also demonstrated that the selection of intracellular trafficking routes was determined by the fiber knob domain and did not depend on the length of the fiber shaft. This study contributes to a better understanding of the mechanisms that govern the infection of retargeted, capsid-modified vectors which have potential application for hematopoietic stem cell and tumor gene therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2003
- Full Text
- View/download PDF
41. An Adenoviral Expression System for AAV Rep78 Using Homologous Recombination
- Author
-
Carlson, Cheryl A., Shayakhmetov, Dmitry M., and Lieber, André
- Subjects
- *
ADENOVIRUSES , *GENETIC vectors - Abstract
The construction and amplification of adenoviral (Ad) vectors expressing biologically active transgenes that are cytotoxic or inhibit Ad replication can be extremely difficult, if not impossible. In this study, we harnessed the ability of Ad genomes to undergo efficient homologous recombination to reconstitute the adeno-associated virus (AAV) rep78 gene, a cytotoxic gene that strongly inhibits Ad replication, which was divided between two parental, first-generation Ad vectors. A functional open reading frame was generated by recombination only upon co-infection of both parental vectors and after the onset of viral DNA replication. We were able to amplify both parental rep78 vectors to normal titers without any signs of inhibition or toxicity and could use them to generate progeny vectors containing a functional rep78 gene without any Ad genes. Using this vector recombination system in AAV rescue assays demonstrated that no Ad protein was essential for Rep78 mediated rescue of AAV ITR flanked DNA from plasmid or Ad backbones; the amount of rescue product generated was substantially greater in the presence of Ad infection; neither cellular nor viral DNA replication was necessary for rescue to occur; and progeny vector genomes were efficiently co-replicated along with conventional, first-generation Ad vectors. [Copyright &y& Elsevier]
- Published
- 2002
- Full Text
- View/download PDF
42. Restoration of a Functional Open Reading Frame by Homologous Recombination between Two Adenoviral Vectors
- Author
-
Carlson, Cheryl A., Shayakhmetov, Dmitry M., and Lieber, André
- Subjects
- *
GENETIC vectors , *ADENOVIRUSES - Abstract
In this study, we examined the ability of adenoviral (Ad) vectors to undergo homologous recombination. The lacZ gene was divided between two parental, first-generation vectors such that neither encoded a functional product but both shared 494 bp in common. The open reading frame could only be restored by homologous recombination. We observed β-galactosidase activity only upon co-infection of both parental vectors and after the onset of viral DNA replication, creating a delay in expression of 24–36 hours in HeLa cells. At peak efficiency, this recombination vector system resulted in β-galactosidase activity levels 100× above background and just 18× less than a conventional, first-generation vector in HeLa cells. After recombination, the resultant progeny vector genomes containing reconstituted expression cassettes were devoid of all viral genes and contained two packaging signals. These progeny genomes were efficiently packaged, could be separated from their parental vectors based on their lighter buoyant densities in CsCl gradients, and were subsequently used as functional gene transfer vectors. This novel recombination vector system should be useful for transferring large transgenes (because the carrying capacity of two Ad vectors can be exploited) or expressing any cytotoxic or Ad replication inhibitory protein (because the parental vectors exhibit no background expression). [Copyright &y& Elsevier]
- Published
- 2002
- Full Text
- View/download PDF
43. A paracrine circuit of IL-1ß/IL-1R1 between myeloid and tumor cells drives genotype-dependent glioblastoma progression.
- Author
-
Zhihong Chen, Giotti, Bruno, Kaluzova, Milota, Vallcorba, Montse Puigdelloses, Rawat, Kavita, Price, Gabrielle, Herting, Cameron J., Pinero, Gonzalo, Cristea, Simona, Ross, James L., Ackley, James, Maximov, Victor, Szulzewsky, Frank, Thomason, Wes, Marquez-Ropero, Mar, Angione, Angelo, Nichols, Noah, Tsankova, Nadejda M., Michor, Franziska, and Shayakhmetov, Dmitry M.
- Subjects
- *
MYELOID cells , *T-cell exhaustion , *MONOCYTE chemotactic factor , *BLOOD circulation , *GLIOBLASTOMA multiforme , *TUMOR growth , *MYELOID differentiation factor 88 - Abstract
Monocytes and monocyte-derived macrophages (MDMs) from blood circulation infiltrate glioblastoma (GBM) and promote growth. Here, we show that PDGFB-driven GBM cells induce the expression of the potent proinflammatory cytokine IL-1ß in MDM, which engages IL-1R1 in tumor cells, activates the NF-κB pathway, and subsequently leads to induction of monocyte chemoattractant proteins (MCPs). Thus, a feedforward paracrine circuit of IL-1ß/IL-1R1 between tumors and MDM creates an interdependence driving PDGFB-driven GBM progression. Genetic loss or locally antagonizing IL-1ß/IL-1R1 leads to reduced MDM infiltration, diminished tumor growth, and reduced exhausted CD8+ T cells and thereby extends the survival of tumor-bearing mice. In contrast to IL-1ß, IL-1α exhibits antitumor effects. Genetic deletion of Il1a/b is associated with decreased recruitment of lymphoid cells and loss-of-interferon signaling in various immune populations and subsets of malignant cells and is associated with decreased survival time of PDGFB-driven tumor-bearing mice. In contrast to PDGFB-driven GBM, Nf1-silenced tumors have a constitutively active NF-κB pathway, which drives the expression of MCPs to recruit monocytes into tumors. These results indicate local antagonism of IL-1ß could be considered as an effective therapy specifically for proneural GBM. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
44. 138. Blood Factors Affect Adenovirus Infectivity and Bio-Distribution In Vivo
- Author
-
Shayakhmetov, Dmitry M.
- Subjects
- *
ADENOVIRUSES , *BLOOD - Abstract
An abstract of the article "Blood Factors Affect Adenovirus Infectivity and Bio-Distribution In Vivo," by Dmitry M. Shayakhmetov is presented.
- Published
- 2005
- Full Text
- View/download PDF
45. 137. Localization of Regions in CD46 That Interact with Adenovirus
- Author
-
Gaggar, Anuj, Shayakhmetov, Dmitry M., Liszewski, M. Kathryn, Atkinson, John P., and Lieber, Andre
- Subjects
- *
ADENOVIRUSES - Abstract
An abstract of the article "Localization of Regions in CD46 That Interact with Adenovirus," by Anuj Gaggar, Dmitry M. Shayakhmetov, M. Kathryn Liszewski, John P. Atkinson, and Andre Lieber is presented.
- Published
- 2005
- Full Text
- View/download PDF
46. Redundant and Synergistic Mechanisms Control the Sequestration of Blood-born Adenovirus in the Liver.
- Author
-
Di Paolo, Nelson C., van Rooijen, Nico, and Shayakhmetov, Dmitry M.
- Subjects
- *
ADENOVIRUSES , *LIVER cells , *LIVER , *GENETIC mutation , *BLOOD - Abstract
Human adenovirus (Ad) is a ubiquitous pathogen causing a wide range of diseases. Although the interactions of human Ad serotype 5 (Ad5) with susceptible cells in vitro are known in great detail, host factors controlling the tissue specificity of Ad5 infection in vivo remain poorly understood. Here, we analyzed the mechanisms of sequestration by the liver for blood-born human Ads and Ad5-based vectors. Our data suggest that several known mechanisms that lead to Ad5 sequestration by the liver become engaged in a redundant, sequential, and synergistic manner to ensure the rapid clearance of circulating virus particles from the blood. These mechanisms include (i) trapping of the virus by liver residential macrophages, Kupffer cells; (ii) Ad5 hepatocyte infection via blood factor–hexon interactions; and (iii) Ad5 penton RGD motif–mediated interactions with liver endothelial cells and hepatocytes, mediating virus retention in the space of Disse. More important, we show that when all of these mechanisms are simultaneously inactivated via mutations of Ad5 capsid proteins and pharmacological interventions, virus sequestration by the liver is markedly reduced. Therefore, our study is the first demonstration of the principal possibility of ablating the sequestration of blood-born Ad in the liver via specific inactivation of a defined set of mechanisms that control this process.Molecular Therapy (2009) 17 4, 675–684 doi:10.1038/mt.2008.307 [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
47. FX and Host Defense Evasion Tactics by Adenovirus.
- Author
-
Baker, Andrew H, Nicklin, Stuart A, and Shayakhmetov, Dmitry M
- Subjects
- *
ADENOVIRUSES , *IMMUNOGLOBULINS , *IMMUNOGLOBULIN M , *LIVER cells , *BLOOD coagulation - Abstract
The article discusses research being done on adenovirus-mediated evasion against host-derived natural antibodies. It references the study "Coagulation Factor X Shields Adenovirus Type 5 From Attack by Natural Antibodies and Complement," by Z. Xu et al. in the 2013 issue of "Nature Medicine." The researcher observed that host-derived coagulation factor X (FX) has linked to the adenovirus capsid and entered hepatocytes in mice without natural immunoglobulin M (IgM) antibodies.
- Published
- 2013
- Full Text
- View/download PDF
48. Hepatocyte Heparan Sulfate Is Required for Adeno-Associated Virus 2 but Dispensable for Adenovirus 5 Liver Transduction In Vivo.
- Author
-
Zaiss, Anne K., Foley, Erin M., Lawrence, Roger, Schneider, Lina S., Hoveida, Hamidreza, Secrest, Patrick, Catapang, Arthur B., Yamaguchi, Yu, Alemany, Ramon, Shayakhmetov, Dmitry M., Esko, Jeffrey D., and Herschman, Harvey R.
- Subjects
- *
ADENO-associated virus , *ADENOVIRUSES , *GENE therapy , *PROTEOGLYCANS , *LIVER cells - Abstract
Adeno-associated virus 2 (AAV2) and adenovirus 5 (Ad5) are promising gene therapy vectors. Both display liver tropism and are currently thought to enter hepatocytes in vivo through cell surface heparan sulfate proteoglycans (HSPGs). To test directly this hypothesis, we created mice that lack Ext1, an enzyme required for heparan sulfate biosynthesis, in hepatocytes. Ext1HEP mutant mice exhibit an 8-fold reduction of heparan sulfate in primary hepatocytes and a 5-fold reduction of heparan sulfate in whole liver tissue. Conditional hepatocyte Ext1 gene deletion greatly reduced AAV2 liver transduction following intravenous injection. Ad5 transduction requires blood coagulation factor X (FX); FX binds to the Ad5 capsid hexon protein and bridges the virus to HSPGs on the cell surface. Ad5.FX transduction was abrogated in primary hepatocytes from Ext1HEP mice. However, in contrast to the case with AAV2, Ad5 transduction was not significantly reduced in the livers of Ext1HEP mice. FX remained essential for Ad5 transduction in vivo in Ext1HEP mice. We conclude that while AAV2 requires HSPGs for entry into mouse hepatocytes, HSPGs are dispensable for Ad5 hepatocyte transduction in vivo. This study reopens the question of how adenovirus enters cells in vivo. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
49. Coagulation Factor Binding Orientation and Dimerization May Influence Infectivity of Adenovirus-Coagulation Factor Complexes.
- Author
-
Irons, Eric E., Flatt, Justin W., Doronin, Konstantin, Fox, Tara L., Acchione, Mauro, Stewart, Phoebe L., and Shayakhmetov, Dmitry M.
- Subjects
- *
BLOOD coagulation factors , *DIMERIZATION , *ADENOVIRUS diseases , *VITAMINS , *LIVER cells , *MEDICAL simulation , *SERINE proteinases - Abstract
Adenoviruses (Ads) are promising vectors for therapeutic interventions in humans. When injected into the bloodstream, Ad vectors can bind several vitamin K-dependent blood coagulation factors, which contributes to virus sequestration in the liver by facilitating transduction of hepatocytes. Although both coagulation factors FVII and FX bind the hexon protein of human Ad serotype 5 (HAdv5) with a very high affinity, only FX appears to play a role in mediating Ad-hepatocyte transduction in vivo. To understand the discrepancy between efficacy of FVII binding to hexon and its apparently poor capacity for supporting virus cell entry, we analyzed the HAdv5-FVII complex by using high-resolution cryo-electron microscopy (cryo-EM) followed by molecular dynamic flexible fitting (MDFF) simulations. The results indicate that although hexon amino acids T423, E424, and T425, identified earlier as critical for FX binding, are also involved in mediating binding of FVII, the FVII GLA domain sits within the surface-exposed hexon trimer depression in a different orientation from that found for FX. Furthermore, we found that when bound to hexon, two proximal FVII molecules interact via their serine protease (SP) domains and bury potential heparan sulfate proteoglycan (HSPG) receptor binding residues within the dimer interface. In contrast, earlier cryo-EM studies of the Ad-FX interaction showed no evidence of dimer formation. Dimerization of FVII bound to Ad may be a contributing mechanistic factor for the differential infectivity of Ad-FX and Ad-FVII complexes, despite high-affinity binding of both these coagulation factors to the virus. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
50. Fiber Shaft-Chimeric Adenovirus Vectors Lacking the KKTK Motif Efficiently Infect Liver Cells In Vivo.
- Author
-
Di Paolo, Nelson C., Kalyuzhniy, Oleksandr, and Shayakhmetov, Dmitry M.
- Subjects
- *
ADENOVIRUSES , *LIVER cells , *PROTEOGLYCANS , *INFECTION , *AMINO acids - Abstract
The molecular mechanisms governing the infectivity of adenovirus (Ad) toward specific cell and tissue types in vivo remain poorly understood. The direct Ad binding to hepatic heparan sulfate proteoglycans via the KKTK motif within the fiber shaft domain was suggested to be the major mechanism of Ad liver cell infection in vivo. Here, we describe the generation and in vitro and in vivo infectivity studies of Ad5-based vectors possessing long Ad31- or Ad41-derived fiber shaft domains, which lack the KKTK motif. We found that all the critical early steps of Ad infection, including attachment to the cellular receptor, internalization, and virus genome transfer into the nucleus, occurred with similar levels of efficiency for fiber shaft-chimeric vectors and unmodified Ad5. Upon intravenous delivery into mice, fiber shaft-chimeric vectors accumulated in liver tissue, transduced liver cells, and induced the production of proinflammatory cytokines (tumor necrosis factor alpha and interleukin-6) and the chemokine monocyte chemoattractant protein 1 at levels indistinguishable from those observed for Ad5. Thus, our data provide evidence that the Ad5 fiber shaft amino acid sequence does not play any substantial role in determining adenovirus infectivity toward hepatic cells in vivo. The data obtained contribute to improving our understanding of the molecular mechanisms determining Ad infectivity and biodistribution in vivo and may aid in designing novel Ad-based vectors for gene therapy applications. [ABSTRACT FROM AUTHOR]
- Published
- 2007
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.