المؤلفون: Xiao-Tang Yin, Alexis Hartman, Nadia Sirajuddin, Deepak Shukla, Anthony St. Leger, Tammie L. Keadle, Patrick M. Stuart

المصدر: Scientific Reports, Vol 14, Iss 1, Pp 1-13 (2024)

مصطلحات موضوعية: Medicine, Science

الوصف: Abstract Reactivation of latent herpes simplex type 1 results in virus returning to the cornea leading to recurrent herpetic stromal keratitis (rHSK). We compare two competing models to reactivate viruses from latency, UV-B irradiation and cyclophosphamide (CP). Results revealed that while both result in corneal recrudescence, only UV-B irradiation results in rHSK. To better understand the dynamics of reactivation, we analyzed corneas for both the presence of infectious viruses and the dynamics of exposure to multiple reactivations using UV-B. We noted that multiple reactivations result in progressively worse corneal disease. We also noted that expression of IFNα and STING, surragate markers for the presence of virus, are induced by the presence of reactivated virus. Studies to determine the importance of STING to the development of HSK revealed that in the absence of STING, mice do not develop significant HSK and the magnitude of the infiltrate of CD45+ cells in these corneas is significantly reduced. The resulting paucity of CD45+CD11b+GR-1+F4/80-neutrophils, and to a lesser extent CD45+CD11b+GR-1-F4/80+ macrophages in B6-STING KO mice following reactivation is likely the underlying cause for lack of rHSK as has been noted by ourselves and others. These results underscore the critical importance of STING’s role in developing rHSK.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2045-2322Test

الوصول الحر: https://doaj.org/article/f1ec81674d424adba6b32ee87a214526Test

المؤلفون: Ilina Bhattacharya, Ipsita Volety, Deepak Shukla

المصدر: mBio, Vol 14, Iss 6 (2023)

مصطلحات موضوعية: HSV-1, eye infection, OPTN, TBK1, PLK1, antiviral defense, Microbiology, QR1-502

الوصف: ABSTRACTTank-binding kinase 1 (TBK1) plays a pivotal role as a cellular factor in regulating the immune response against herpes simplex virus type 1 (HSV-1). TBK1’s role in defending against HSV-1 is attributed to its capacity to induce the type I interferon response. Recent discoveries emphasize TBK1’s involvement in the regulation of optineurin (OPTN), an autophagy adaptor protein that plays a direct role in enhancing the antiviral response against HSV-1 through selective autophagy. Intriguingly, OPTN can also reciprocally regulate TBK1, leading to a limited understanding of the precise role of the OPTN-TBK1 axis in HSV-1 infection. In our study, we shed light on the relationship between OPTN and TBK1 in influencing the outcome of HSV-1 infection. Surprisingly, when TBK1 or OPTN was absent in cells, the spread and infectivity of HSV-1 did not align as expected. Furthermore, it was unexpected to note that cells lacking OPTN showed higher interferon expression compared to normal cells, yet they exhibited enhanced viral growth. Our investigations revealed that OPTN can directly degrade HSV-1 through autophagy even in the absence of active TBK1. Interestingly, we identified that another protein, Polo-like kinase 1 (PLK1), can compensate for the absence of TBK1 and trigger autophagy to restrict HSV-1. In summary, our findings highlight that OPTN significantly impacts the outcome of HSV-1 infection, regardless of the conventional antiviral response mediated by TBK1. Additionally, by introducing a new role for PLK1, this research provides valuable insights into the intricate interplay among OPTN, TBK1, and autophagy in shaping the course of HSV-1 infection.IMPORTANCEHerpes simplex virus type 1 (HSV-1) is globally prevalent, with latent infections observed in up to 80% of the population. The virus is known for subverting host defense mechanisms and infiltrating the nervous system to establish latency in peripheral ganglia. Multiple stressors can reactivate the virus, and recurrent herpes has been linked to vision loss and neurodegeneration. Identifying critical host factors that limit the spread of HSV-1 and the subsequent establishment of latent infection holds the potential to drive new intervention strategies for eradicating the virus. Numerous pieces of evidence underscore the significance of Tank-binding kinase 1 (TBK1) in restricting HSV-1. Reports have also suggested that phosphorylation of optineurin (OPTN) by TBK1 is required for triggering OPTN-mediated autophagy for HSV degradation. This report adds new insights into the roles of OPTN and TBK1 in HSV-1 infection and provides proof of a TBK1-independent HSV-1 restriction through OPTN. It confirms that TBK1 activation can be substituted by PLK1 to provide protection against HSV-1. In contrast, the activation of OPTN is likely an indispensable host defense mechanism for optimal defense against HSV-1.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2150-7511Test

الوصول الحر: https://doaj.org/article/a9a62baf4084410c9ef74c7a11451771Test

المؤلفون: James Elste, Nicole Cast, Shalini Udawatte, Kabita Adhikari, Shannon Harger Payen, Subhash C. Verma, Deepak Shukla, Michelle Swanson-Mungerson, Vaibhav Tiwari

المصدر: Biomedicines, Vol 12, Iss 4, p 821 (2024)

مصطلحات موضوعية: viral entry, virus cell fusion, ACE2, NPC1L1, Biology (General), QH301-705.5

الوصف: The entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into human embryonic kidney (HEK293T) cells has been shown to be a cholesterol-rich, lipid raft-dependent process. In this study, we investigated if the presence of a cholesterol uptake receptor Niemann-pick type c1-like1 (NPC1L1) impacts SARS-CoV-2 cell entry. Initially, we utilized reporter-based pseudovirus cell entry assays and a spike (S) glycoprotein-mediated cell-to-cell fusion assay. Using Chinese hamster ovary (CHO-K1) cells, which lack endogenous receptors for SARS-CoV-2 entry, our data showed that the co-expression of NPC1L1 together with the ACE2 receptor synergistically increased SARS-CoV-2 pseudovirus entry even more than the cells expressing ACE-2 receptor alone. Similar results were also found with the HEK293T cells endogenously expressing the ACE2 receptor. Co-cultures of effector cells expressing S glycoprotein together with target cells co-expressing ACE-2 receptor with NPC1L1 significantly promoted quantitative cell-to-cell fusion, including syncytia formation. Finally, we substantiated that an elevated expression of NPC1L1 enhanced entry, whereas the depletion of NPC1L1 resulted in a diminished SARS-CoV-2 entry in HEK293T-ACE2 cells using authentic SARS-CoV-2 virus in contrast to their respective control cells. Collectively, these findings underscore the pivotal role of NPC1L1 in facilitating the cellular entry of SARS-CoV-2. Importance: Niemann-Pick type C1-like1 (NPC1L1) is an endosomal membrane protein that regulates intracellular cholesterol trafficking. This protein has been demonstrated to play a crucial role in the life cycle of several clinically important viruses. Although SARS-CoV-2 exploits cholesterol-rich lipid rafts as part of its viral entry process, the role of NPC1L1 in SARS-CoV-2 entry remains unclear. Our research represents the first-ever demonstration of NPC1L1’s involvement in facilitating SARS-CoV-2 entry. The observed role of NPC1L1 in human kidney cells is not only highly intriguing but also quite relevant. This relevance stems from the fact that NPC1L1 exhibits high expression levels in several organs, including the kidneys, and the fact that kidney damages are reported during severe cases of SARS-CoV-2. These findings may help us understand the new functions and mechanisms of NPC1L1 and could contribute to the identification of new antiviral targets.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2227-9059/12/4/821Test; https://doaj.org/toc/2227-9059Test

الوصول الحر: https://doaj.org/article/61b310de6e7f40a98b7a0fdeb26dca2fTest

المؤلفون: Tejabhiram Yadavalli, Sudhanshu Kumar Singh, Abhijit A. Date, Deepak Shukla

المصدر: Biomedicine & Pharmacotherapy, Vol 165, Iss , Pp 115056- (2023)

مصطلحات موضوعية: Herpesviruses, Oral medication, Pharmacokinetic, Acute toxicity, Antiviral, Therapeutics. Pharmacology, RM1-950

الوصف: Herpes simplex viruses type-1 (HSV-1) and type-2 (HSV-2) are ubiquitous human pathogens causing serious pathologies in the ocular, orofacial and anogenital regions. While current treatments such as nucleoside analogs are effective in most cases, the emergence of drug resistance necessitates the development of newer antivirals with different mechanisms of action. In this regard, BX795, a small molecule inhibitor has shown significant benefit in the treatment of herpesvirus infections previously when dosed topically. However, the efficacy of BX795′s systemic dosage remains to be tested. In this study, we evaluated acute and short-term toxicity of orally administered BX795 at a concentration of 400 and 100 mg/kg respectively in mice. This was followed by an evaluation of pharmacokinetics and tissue distribution of BX795 on intravenous and oral administration. Based on these studies, we performed an in vivo antiviral study using murine models of ocular HSV-1 and genital HSV-2 infection. Our results indicate that orally administered BX795 is very well tolerated, had oral bioavailability of 56%, and reached ocular and genital tissues within the first 15 min of dosing. Our studies indicate that BX795 administered orally can significantly reduce herpesvirus replication in the ocular and genital tissue.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S0753332223008466Test; https://doaj.org/toc/0753-3322Test

الوصول الحر: https://doaj.org/article/419b5188165d4b979cfa61f124878acaTest

المؤلفون: James Elste, Angelica Chan, Chandrashekhar Patil, Vinisha Tripathi, Daniel M. Shadrack, Dinesh Jaishankar, Andrew Hawkey, Michelle Swanson Mungerson, Deepak Shukla, Vaibhav Tiwari

المصدر: Computational and Structural Biotechnology Journal, Vol 21, Iss , Pp 1030-1040 (2023)

مصطلحات موضوعية: Heparan sulfare, Virus entry, Virus cell-to-cell spread, Virus cell fusion, Sulphated heparan sulfate, Biotechnology, TP248.13-248.65

الوصف: The structural diversity of metazoic heparan sulfate (HS) composed of unique sulfated domains is remarkably preserved among various vertebrates and invertebrate species. Interestingly the sulfated moieties of HS have been known as the key determinants generating extraordinary ligand binding sites in the HS chain to regulate multiple biological functions and homeostasis. One such ligand for 3-O sulfation in the HS chain is a glycoprotein D (gD) from an ancient herpesvirus, herpes simplex virus (HSV). This interaction between gD and 3-O sulfated HS leads to virus-cell fusion to promote HSV entry. It is quite astonishing that HSV-1, which infects two-thirds of the world population, is also capable of causing severe diseases in primates and non-primates including primitive zebrafish. Supporting evidence that HSV may cross the species barrier comes from the fact that an enzymatic modification in HS encoded by 3-O sulfotransferase-3 (3-OST-3) from a vertebrate zoonotic species enhances HSV-1 infectivity. The latter phenomenon suggests the possible role of sulfated-HS as an entry receptor during reverse zoonosis, especially during an event when humans encounter domesticated animals in proximity. In this mini-review, we explore the possibility that structural diversity in HS may have played a substantial role in species-specific adaptability for herpesviruses in general including their potential role in promoting cross-species transmission.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S2001037023000053Test; https://doaj.org/toc/2001-0370Test

الوصول الحر: https://doaj.org/article/56528fbe8e3f401f97c53e9003dfd746Test

المؤلفون: Raghuram Koganti, Tejabhiram Yadavalli, Deepak Shukla

المصدر: STAR Protocols, Vol 4, Iss 2, Pp 102342- (2023)

مصطلحات موضوعية: Cell Biology, Cell culture, Cell-based Assays, Microscopy, Science (General), Q1-390

الوصف: Summary: The study of inflammation is of key interest to biomedical research; however, techniques to induce inflammation in vitro are difficult to implement. Here, we present a protocol that optimizes the induction and measurement of NF-kB-mediated inflammation in vitro using a human macrophage cell line. We describe steps for growing, differentiating, and inducing inflammation in THP-1 cells. We detail the process of staining and grid-based confocal imaging. We also discuss approaches to test anti-inflammatory drug efficacy in inhibiting inflammatory milieu.For complete details on the use and execution of this protocol, please refer to Koganti et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S266616672300309XTest; https://doaj.org/toc/2666-1667Test

الوصول الحر: https://doaj.org/article/2e114cbcf34a46789a34cd7a1e224ed8Test

المؤلفون: Tarun Bhatnagar, Sirshendu Chaudhuri, Manickam Ponnaiah, Pragya D Yadav, R Sabarinathan, Rima R Sahay, Faheem Ahmed, S Aswathy, Pankaj Bhardwaj, Anil Bilimale, M Santhosh Kumar, M. Logaraj, Uday Narlawar, C Palanivel, Prakash Patel, Sanjay K Rai, Vartika Saxena, Arvind Singh, Jeromie WV Thangaraj, Ashwini Agarwal, Yasir Alvi, Amoghashree, P Ashok, Dinesh Babu, Yogesh Bahurupi, Sangita Bhalavi, Priyamadhaba Behera, Priyanka Pandit Biswas, Jaykaran Charan, Nishant Kumar Chauhan, KB Chetak, Lalit Dar, Ayan Das, R Deepashree, Minakshi Dhar, Rahul Dhodapkar, TS Dipu, Mridu Dudeja, Manisha Dudhmal, Ravisekhar Gadepalli, Mahendra Kumar Garg, AV Gayathri, Akhil Dhanesh Goel, H Basavana Gowdappa, Randeep Guleria, Manoj Kumar Gupta, Farzana Islam, Mannu Jain, Vineet Jain, M Lanord Stanley Jawahar, Rajendra Joshi, Shashi Kant, Sitanshu Sekhar Kar, Deepjyoti Kalita, Meenakshi Khapre, Satyendra Khichar, Sarika Prabhakar Kombade, Sunil Kohli, Abhinendra Kumar, Anil Kumar, Deepak Kumar, Kiran G Kulirankal, KV Leela, Triparna Majumdar, Baijayantimala Mishra, Puneet Misra, Sanjeev Misra, Prasanta Raghab Mohapatra, M Narayana Murthy, Dimpal A Nyayanit, Manish Patel, Monika Pathania, Savita Patil, Binod Kumar Patro, Ramniwas Jalandra, Pragati Rathod, Naimesh Shah, Anita Shete, Deepak Shukla, M Shwethashree, Smita Sinha, MN Sumana, Ashish Surana, Anjan Trikha, A Tejashree, Mahalingam Venkateshan, G Vijaykrishnan, Sarita Wadhava, Naveet Wig, Nivedita Gupta, Priya Abraham, Manoj V Murhekar

المصدر: International Journal of Infectious Diseases, Vol 122, Iss , Pp 693-702 (2022)

مصطلحات موضوعية: COVID-19 vaccines, BBV152/Covaxin, AZD1222/Covishield, Delta variant, Effectiveness, India, Infectious and parasitic diseases, RC109-216

الوصف: Objectives: India introduced BBV152/Covaxin and AZD1222/Covishield vaccines in January 2021. We estimated the effectiveness of these vaccines against severe COVID-19 among individuals aged ≥45 years. Methods: We did a multi-centric, hospital-based, case-control study between May and July 2021. Cases were severe COVID-19 patients, and controls were COVID-19 negative individuals from 11 hospitals. Vaccine effectiveness (VE) was estimated for complete (2 doses ≥ 14 days) and partial (1 dose ≥ 21 days) vaccination; interval between two vaccine doses and vaccination against the Delta variant. We used the random effects logistic regression model to calculate the adjusted odds ratios (aOR) with a 95% confidence interval (CI) after adjusting for relevant known confounders. Results: We enrolled 1143 cases and 2541 control patients. The VE of complete vaccination was 85% (95% CI: 79-89%) with AZD1222/Covishield and 71% (95% CI: 57-81%) with BBV152/Covaxin. The VE was highest for 6-8 weeks between two doses of AZD1222/Covishield (94%, 95% CI: 86-97%) and BBV152/Covaxin (93%, 95% CI: 34-99%). The VE estimates were similar against the Delta strain and sub-lineages. Conclusion: BBV152/Covaxin and AZD1222/Covishield were effective against severe COVID-19 among the Indian population during the period of dominance of the highly transmissible Delta variant in the second wave of the pandemic. An escalation of two-dose coverage with COVID-19 vaccines is critical to reduce severe COVID-19 and further mitigate the pandemic in the country.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S1201971222004271Test; https://doaj.org/toc/1201-9712Test

الوصول الحر: https://doaj.org/article/ae9a4d01bbc948f5903a25216c64b295Test

المؤلفون: Hemant Borase, Deepak Shukla

المصدر: Viruses, Vol 15, Iss 11, p 2195 (2023)

مصطلحات موضوعية: genital herpes, herpes simplex virus-2, vaccines, antivirals, immune response, microbiome, Microbiology, QR1-502

الوصف: Genital herpes, primarily caused by herpes simplex virus-2 (HSV-2), remains a pressing global health concern. Its remarkable ability to intertwine with cellular processes, from harnessing host machinery for replication to subverting antiviral defenses like autophagy and programmed cell death, exemplifies the intricate interplay at the heart of its pathogenesis. While the biomedical community has extensively researched antiviral interventions, the efficiency of these strategies in managing HSV-2 remains suboptimal. Recognizing this, attention has shifted toward leveraging host cellular components to regulate HSV-2 replication and influence the cell cycle. Furthermore, innovative interventional strategies—including drug repurposing, microbivacs, connecting the host microbiome, and exploiting natural secondary metabolites—are emerging as potential game changers. This review summarizes the key steps in HSV-2 pathogenesis and newly discovered cellular interactions, presenting the latest developments in the field, highlighting existing challenges, and offering a fresh perspective on HSV-2’s pathogenesis and the potential avenues for its treatment by targeting cellular proteins and pathways.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/1999-4915/15/11/2195Test; https://doaj.org/toc/1999-4915Test

الوصول الحر: https://doaj.org/article/19e312e87c0a47ebb4fd44afd5a8592fTest

المؤلفون: Raghuram Koganti, Tejabhiram Yadavalli, Yogesh Sutar, Sudipta Mallick, Abhijit Date, Deepak Shukla

المصدر: iScience, Vol 25, Iss 12, Pp 105682- (2022)

مصطلحات موضوعية: Ophthalmology, Molecular medicine, Immune response, Science

الوصف: Summary: Chronic inflammation of the immune privileged cornea originating from viral or nonviral conditions results in significant vision loss. Topical corticosteroids are the common treatments for corneal inflammation, but the drugs cause serious and potentially blinding side effects in the long term. Therefore, new standalone and/or synergistic anti-inflammatory therapies with lower side effects are desperately needed. Here, we show that the aromatic fatty acid phenylbutyrate (PBA) acts as a potent inhibitor of inflammation in preclinical ocular-inflammation models. PBA prevents the transcription as well as translation of pro-inflammatory cytokines by LPS and poly(I:C) via persistent inhibition of NF-κB signaling. PBA quickens the resolution of ocular inflammation in mice by decreasing corneal thickness and immune cell infiltration. More importantly, PBA can synergize with the dexamethasone to antagonize NF-κB signaling at lower drug concentrations. Our results demonstrate that PBA therapy exerts previously unreported anti-inflammatory effects in the eye and facilitates corneal healing during persistent inflammation.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S258900422201954XTest; https://doaj.org/toc/2589-0042Test

الوصول الحر: https://doaj.org/article/b952d891e3f747bd9eecb01fd851d328Test

المؤلفون: Raza Ali Naqvi, Deepak Shukla, Afsar R. Naqvi

المصدر: Immunity, Inflammation and Disease, Vol 10, Iss 1, Pp 22-25 (2022)

مصطلحات موضوعية: complement, gene regulation processes, infections, molecules, viral/retroviral, Immunologic diseases. Allergy, RC581-607

الوصف: Abstract Background The lack of knowledge about the specific preventive measures and limited scientific information on severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) led to an excruciating onset and progression of coronavirus disease 2019 (COVID‐19). Swift development of various successful vaccines around the globe is striving to contain the exponential surges of COVID‐19 cases. However, the ongoing struggle to vaccinate the global population and alarming spread of highly transmissible variants may thwart global initiatives to contain SARS‐CoV‐2 as observed by less robust protective immunity. Methods In this perspective, we propose a thought‐provoking, two‐pronged strategy involving RNA interference approach to degrade essential SARS‐CoV‐2 ORFs required for replication and entry in conjunction with a complement inhibitor (compstatin) to stymie the detrimental proinflammatory cytokine storm that exacerbate disease progression and severity. Results We provide supporting evidence suggesting that concurrent targeting of viral and host components will be a superior strategy to effectively suppress viral spread and clinical manifestations of COVID‐19. Conclusion SARS‐CoV‐2 specific RNAi in conjunction with systemic delivery of compstatin will be an effective two‐pronged strategy to combat local and systemic immune responses in both symptomatic and asymptomatic COVID‐19 patients.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2050-4527Test

الوصول الحر: https://doaj.org/article/b96a275ac2ef49f6b868d9d58b0cd06eTest