Virology tidbits

Virology tidbits

Sunday, 31 January 2016

Zika Virus (ZIKV): similarities to other arboviruses

Zika Virus (ZIKV) is an arbovirus belonging to the Flaviviridae transmitted primarily by mosquitoes (including Aedes Agypti and Aedes albopictus). Although first identified in a rhesus monkey from the forests in Uganda in the year 1947 -with an estimated first emergence probably in 1920- the first human case was only reported in 1952 in Nigeria. ZIKV has been shown to be distributed Northern Africa as well as in Southeast Asia and the Pacific; however only a few human cases in Africa and Asia were identified until 2007, when a ZIKV outbreak was reported in Yap/Micronesia followed by an outbreak in French Polynesia and New Caledonia 2013 and currently in Central and South America as well as the Caribbean.
Most epidemiological studies that are based on the seroprevalence of neutralizing antibodies, suggest that up to 73% of the population (6.1-73%) have been exposed to ZIKV.

Table 1: Outbreaks of ZIKV in Africa, Asia and the Pacific prior 2016 and seroprevalence of
ZIKV 



Being a Flavivirus, ZIKV -like the related West Nile Virus (WNV), Japanese Encephalitis Virus (JEV), Chikungunya (CHIKV) and Dengue Virus (DENGV)- has a single strand, positive sense RNA genome, encoding for a polyprotein that ultimately is processed into three structural proteins, the Capsid (C), precursor of Membrane protein (prM), and Envelope (E) protein as well as seven non-structural proteins (NS1-5).

Figure: ZIKV structure

Figure: ZIKV polyprotein
Table 2: ZIKV proteins



Similar to CHIKV, the infection with ZIKV is a relative mild disease, characterised symptoms ranging from mild fever, headaches, conjunctivitis, maculopapular rashes, vertigo, or myalgia with low mortality and often is asymptomatic.
Since the clinical presentation is similar to infections with other arboviruses, in particular CHIKV and DENGV, diagnostics is difficult and mostly done by using RT-PCR of samples that are CHIKV and DENGV negative. Although serological tests (ELISA, Immunofluorescence) have been used in the past, they are less reliable due to cross-reactivity with other flaviviruses such as DENGV or Yellow Fever Virus. Currently no commercial kit is available to test for ZIKV in patients.

Being transmitted by infected mosquitoes, the initial target cells are in the skin compartment, both skin fibroblasts, epidermal keratinocytes and dendritic cells are highly permissible for ZIKV which enters the host cell a number cellular receptors, in particular DC-SIGN, AXL, Tyro-3, and (albeit to a lesser extent) TIM-1. Primary skin fibroblasts infected with ZIKV support viral replication whereas in infected primary epidermal keratinocytes, similar to keratinocytes infected with DENV, exhibit large cytoplasmic vacuolation and pyknotic nuclei can be observed, suggesting that ZIKV induces apoptosis in these cells despite supporting viral replication.
Similar to JEV and CHIKV infected cell, ZIKV induces the formation of autophagosome-like structure that form the scaffold for the assembly of viral replication centres. If the expression of the viral preE2/E1 and Capsid proteins also induces ER stress (similar to CHIKV) and if the ER stress induced by ZIKV contributes to the induction of autophagy and/or caspase dependent apoptosis has not been demonstrated yet, but seems to be very likely. 

Figure: CHIKV and the ER stress response

Figure: JEV and the ER stress response

Figure: JEV and the induction of autophagosome formation via non-structural and structural
proteins located at the ER

Likewise it has not been demonstrated if the formation of autophagosomes by ZIKV in primary skin fibroblasts is -as in the case of CHIKV- dependent on LC3-C and NDP52. Given that only the Asian lineage, in contrast to the African lineage, of ZIKV has been able to cause prolonged epidemics in the human populations (in 2007 and currently in the Americas) it has been speculated that the viral NS1 gene adaptions increases viral fitness in humans. Therefore, it might be necessary to investigate the ability of different isolates from different lineages to induce the formation of replication centres in primary human cells. If the decrease in autophagic flux or viral induced ER stress in infected primary dermal and skin cells or in keratinocytes induces apoptosis has not been demonstrated.


Figure: ZIKV induction of the antiviral response

Antiviral response induced by ZIKV : role of the antiviral interferon response

In primary skin cells, following viral entry and release of the viral genome, the viral RNA induces the transcription of Interferon stimulated genes (ISG) such as OAS2, MX1, and ISG15, as well as the transcription of RIG-1, MDA-5, and Toll-like receptor (TLR-3) in a Interferon-β (IFN-β).As a result, viral replication is inhibited.

Since the infection of primary human dermal fibroblasts and primary human foreskin fibroblast cells (HFF) with ZIKV also induces the formation of LC3-II positive membrane vesicles , it might be possible that TLR-3 mediated NF-κB signaling induces autophagy in a DRAM-1 dependent manner.
If these vesicles however represent viral replication centres or are involved in antiviral signaling by degrading viral components (or are even involved in the presentation of viral antigens  via the MHC- Class I and II complexes) is not clear.

Figure: TLR-3 and autophagy; formation of autophagosomes via DRAM-1



The intracelebral inoculation of mice with ZIKV results in productive viral replication in both neuronal and astroglial cells, with autophagosome-like structures present that resemble viral replication centres. A subset of infected cells however has been shown to undergo necrosis, suggesting that the infection of neuronal cells with ZIKV -similar to human medullablastoma TE-671 cells, astrocytes and neuronal cells infected with JEV- induces both the formation of autophagosome-like structures as well as cell death. If however ZIKV induced cell death is dependent on the induction of caspase-3 and -9 in addition to ROS dependent activation of apoptosis signaling kinase (ASK)-1 and p38 MAPK signaling pathways has not been demonstrated. Despite the close proximity of ZIKV replication centres, an involvement of the ER stress response in the formation of ZIKV RC has not been demonstrated as well.

                           ZIKV and the nucleolus

As discussed in a previous post, a number of viral proteins localises to the nucleolus, thus (potentially) inducing the formation of autophagosomes and/or inducing via activation of p53. In the case of the small isoform of West Nile Virus (WNV) Capsid protein for instance, p53 dependent apoptosis is induced by sequestration of Hdm2 to the nucleolus whilst the large isoform inhibits apoptosis and activates mTOR-dependent p70S6K, thus inducing translation of viral genes. If either isoform, affects also the formation of LC3-II positive vesicles has not been demonstrated; it should be noted however that the NS4A and NS4B proteins from all WNV isolates (except NY99) induce autophagy as a result of ER stress. Since the NS4B protein derived from the WNV Kunjin subtype also localises to the nucleolus, an involvement of the nucleolus in the formation of WNV RC cannot be ruled out.

Table3: Flavivirus proteins that localise to the nucleolus
It might therefore possible that the nucleolar localisation of ZIKV protein(s) might induce a cell cycle delay, promote apoptosis or induce autophagy.        

Figure: ZIKV might promote autophagy or induce apoptosis by sequestering p53 


      ZIKV, Microcephaly, and Guillan-Barre Syndrome

In the case of viral induced microcephaly, in utero infection with Human Cytomegalovirus (HCMV) has been associated with cases of microcephaly in newborn infants, but if ZIKV is a causative agent of microcephaly as well or if other factors -such as co-infection of pregnant women with other neurotrophic viruses such as DENGV or CHIKV, and/or parasites such as malaria as well as environmental factors despite the isolation of viral RNA from the placenta and amniotic fluid also play a role is not clear.
During the current outbreak of ZIKV in the Americas as well as during the outbreak of ZIKV in French Polynesia, an increase in patients exhibiting Guillan-Barre Syndrome (GBS), a rare neurological disorder caused by an autoimmune response, has been reported. Although so far no direct link between ZIKV and GBS has been reported, it might be possible that the decrease in autophagic flux in ZIKV infected neuronal cells induces apoptosis or if the observed increase is related to an autoimmune response as a result of the immune response (as in cases of rheumatoid arthritis in CHIKV positive patients) remains to be seen.


In summary, based on results published on other members of the Flaviviridae, ZIKV might exhibit similar features regarding the interaction with host cell, including the formation of replication centres by inducing the ER stress response, the localisation of viral proteins to the nucleolus and -probably most importantly-the induction of antiviral Interferon signaling via the induction of TLR-3 mediated NF-κB signaling (that might also induce the formation of autophagosomes, supporting viral replication and/or degrading viral proteins and RNA) and the induction of Interferon stimulated genes (ISG), thus inhibiting viral replication.

ResearchBlogging.org




Further reading


Ioos S, Mallet HP, Leparc Goffart I, Gauthier V, Cardoso T, & Herida M (2014). Current Zika virus epidemiology and recent epidemics. Medecine et maladies infectieuses, 44 (7), 302-7 PMID: 25001879


Jan C, Languillat G, Renaudet J, & Robin Y (1978). [A serological survey of arboviruses in Gabon]. Bulletin de la Societe de pathologie exotique et de ses filiales, 71 (2), 140-6 PMID: 743766


Grard G, Caron M, Mombo IM, Nkoghe D, Mboui Ondo S, Jiolle D, Fontenille D, Paupy C, & Leroy EM (2014). Zika virus in Gabon (Central Africa)--2007: a new threat from Aedes albopictus? PLoS neglected tropical diseases, 8 (2) PMID: 24516683


Kuno G, & Chang GJ (2007). Full-length sequencing and genomic characterization of Bagaza, Kedougou, and Zika viruses. Archives of virology, 152 (4), 687-96 PMID: 17195954


Hamel R, Dejarnac O, Wichit S, Ekchariyawat P, Neyret A, Luplertlop N, Perera-Lecoin M, Surasombatpattana P, Talignani L, Thomas F, Cao-Lormeau VM, Choumet V, Briant L, Desprès P, Amara A, Yssel H, & Missé D (2015). Biology of Zika Virus Infection in Human Skin Cells. Journal of virology, 89 (17), 8880-96 PMID: 26085147



Choi SH, Gonen A, Diehl CJ, Kim J, Almazan F, Witztum JL, & Miller YI (2015). SYK regulates macrophage MHC-II expression via activation of autophagy in response to oxidized LDL. Autophagy, 11 (5), 785-95 PMID: 25946330


Gannage M, da Silva RB, & Münz C (2013). Antigen processing for MHC presentation via macroautophagy. Methods in molecular biology (Clifton, N.J.), 960, 473-88 PMID: 23329508


Chemali M, Radtke K, Desjardins M, & English L (2011). Alternative pathways for MHC class I presentation: a new function for autophagy. Cellular and molecular life sciences : CMLS, 68 (9), 1533-41 PMID: 21390546


Gannage M, & Münz C (2010). MHC presentation via autophagy and how viruses escape from it. Seminars in immunopathology, 32 (4), 373-81 PMID: 20857294


Yang TC, Shiu SL, Chuang PH, Lin YJ, Wan L, Lan YC, & Lin CW (2009). Japanese encephalitis virus NS2B-NS3 protease induces caspase 3 activation and mitochondria-mediated apoptosis in human medulloblastoma cells. Virus research, 143 (1), 77-85 PMID: 19463724

   
Bell TM, Field EJ, & Narang HK (1971). Zika virus infection of the central nervous system of mice. Archiv fur die gesamte Virusforschung, 35 (2), 183-93 PMID: 5002906 Tetro JA (2016). Zika and microcephaly: Causation, correlation, or coincidence? Microbes and infection / Institut Pasteur PMID: 26774330

Foy BD, Kobylinski KC, Chilson Foy JL, Blitvich BJ, Travassos da Rosa A, Haddow AD, Lanciotti RS, & Tesh RB (2011). Probable non-vector-borne transmission of Zika virus, Colorado, USA. Emerging infectious diseases, 17 (5), 880-2 PMID: 21529401


Salvetti A, & Greco A (2014). Viruses and the nucleolus: the fatal attraction. Biochimica et biophysica acta, 1842 (6), 840-7 PMID: 24378568 

Xu Z, Anderson R, & Hobman TC (2011). The capsid-binding nucleolar helicase DDX56 is important for infectivity of West Nile virus. Journal of virology, 85 (11), 5571-80 PMID: 21411523

Katoh H, Mori Y, Kambara H, Abe T, Fukuhara T, Morita E, Moriishi K, Kamitani W, & Matsuura Y (2011). Heterogeneous nuclear ribonucleoprotein A2 participates in the replication of Japanese encephalitis virus through an interaction with viral proteins and RNA. Journal of virology, 85 (21), 10976-88 PMID: 21865391 

Buckley A, & Gould EA (1988). Detection of virus-specific antigen in the nuclei or nucleoli of cells infected with Zika or Langat virus. The Journal of general virology, 69 ( Pt 8), 1913-20 PMID: 2841406 

Westaway EG, Khromykh AA, Kenney MT, Mackenzie JM, & Jones MK (1997). Proteins C and NS4B of the flavivirus Kunjin translocate independently into the nucleus. Virology, 234 (1), 31-41 PMID: 9234944 

Buckley A, Gaidamovich S, Turchinskaya A, & Gould EA (1992). Monoclonal antibodies identify the NS5 yellow fever virus non-structural protein in the nuclei of infected cells. The Journal of general virology, 73 ( Pt 5), 1125-30 PMID: 1534119 


Yang MR, Lee SR, Oh W, Lee EW, Yeh JY, Nah JJ, Joo YS, Shin J, Lee HW, Pyo S, & Song J (2008). West Nile virus capsid protein induces p53-mediated apoptosis via the sequestration of HDM2 to the nucleolus. Cellular microbiology, 10 (1), 165-76 PMID: 17697133 

Shives KD, Beatman EL, Chamanian M, O'Brien C, Hobson-Peters J, & Beckham JD (2014). West nile virus-induced activation of mammalian target of rapamycin complex 1 supports viral growth and viral protein expression. Journal of virology, 88 (16), 9458-71 PMID: 24920798 

Urbanowski MD, & Hobman TC (2013). The West Nile virus capsid protein blocks apoptosis through a phosphatidylinositol 3-kinase-dependent mechanism. Journal of virology, 87 (2), 872-81 PMID: 23115297 

Blázquez AB, Martín-Acebes MA, & Saiz JC (2014). Amino acid substitutions in the non-structural proteins 4A or 4B modulate the induction of autophagy in West Nile virus infected cells independently of the activation of the unfolded protein response. Frontiers in microbiology, 5 PMID: 25642225 

Martín-Acebes MA, & Saiz JC (2011). A West Nile virus mutant with increased resistance to acid-induced inactivation. The Journal of general virology, 92 (Pt 4), 831-40 PMID: 21228127 Martín-Acebes MA, Blázquez AB, & Saiz JC (2015). Reconciling West Nile virus with the autophagic pathway. Autophagy, 11 (5), 861-4 PMID: 25946067 

Naing ZW, Scott GM, Shand A, Hamilton ST, van Zuylen WJ, Basha J, Hall B, Craig ME, & Rawlinson WD (2016). Congenital cytomegalovirus infection in pregnancy: a review of prevalence, clinical features, diagnosis and prevention. The Australian & New Zealand journal of obstetrics & gynaecology, 56 (1), 9-18 PMID: 26391432



19 comments:

  1. I am currently writing a graded unit on the topic of ZIKV. This information provided me with a great amount of detail and understanding. Do you possess any other information which would be useful to me and is maybe more simplified?

    ReplyDelete
    Replies
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  2. Dear Alana,
    I am more than happy to assist you. From what I gather you are interested in the differences and similarities between ZIKV and other arboviruses as part of a school or university project?
    During the past year, we have a seen a tremendous of research being published identifying cells that are infected by ZIKV but not other arboviruses such as West Nile Virus (WNV), Dengue Virus (DENV) or Yellow Fever Virus (YFV). Cells that are infected by ZIKV and support ZIKV replication but not DENV include human placental cells, foetal neural precursor cells (neural progenitor cells and foetal neural stem cells) and mice testicular cells, thus explaining not only the development of microcephaly but also male to female transmission. In general, microcephaly caused by ZIKV is believed due to the induction of cell death either by the virus directly or indirectly by inducing inflammation. One of the factors distinguishing ZIKV from DENV or WNV is the cellular receptor that is required for entry of the viral particle into the host cell, but there may be additional factors as well.

    A good start is http://www.scielo.br/pdf/rsbmt/v49n5/0037-8682-rsbmt-49-05-00537.pdf I also recommend https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4859708/ to understand the transmission cycle as well as http://cmr.asm.org/content/29/3/487.long All of these articles are free of charge.
    Please feel free to contact me if you have any further questions and good luck !

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  8. I have been suffering from (HERPES) disease for the last four years
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    ReplyDelete
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    ReplyDelete
  11. I am so happy, i never believe i will be this happy again in life, I was working as an air-hoster ( cabby crew ) for 3years but early this year, i loose my job because of this deadly disease called Herpes virus (HSV), I never felt sick or have any symptom, till all workers were ask to bring their doctors report, that was how i got tested and i found out that am HSV positive that make me loose my job, because it was consider as an STD and is incurable disease, i was so depress was thinking of committing suicide, till i explain to a friend of mine, who always said to me a problem share is a problem solved, that was how she directed me to Dr Isibor, that was how i contacted him and get the medication from this doctor and i got cured for real, I just went back to my work and they also carry out the test to be real sure and i was negative. Please contact this doctor if you are herpes positive diseases his email is: drisiborspellhome@gmail.com. or you can call or whatsApp his mobile number +2348107855231.

    ReplyDelete
  12. i have been suffering from (HSV) disease for the last four years and had constant pain, especially in my knees. During the first year,I had faith in God that i would be healed someday.This disease started circulate all over my body and i have been taking treatment from my doctor, few weeks ago i came on search on the internet if i could get any information concerning the prevention of this disease, on my search i saw a testimony of someone who has been healed from (Hepatitis B and Cancer) by this Man Dr Isibor and she also gave the email address of this man and advise we should contact him for any sickness that he would be of help, so i wrote to Dr Isibor telling him about my (HPV) he told me not to worry that i was going to be cured!! hmm i never believed it,, well after all the procedures and remedy given to me by this man few weeks later i started experiencing changes all over me as the Dr assured me that i have cured,after some time i went to my doctor to confirmed if i have be finally healed behold it was TRUE, So friends my advise is if you have such sickness or any other at all you can email Dr Isibor on his email :drisiborspellhome@gmail.com or his whatsApp contact +2348107855231.

    ReplyDelete
  13. Thanks to great Dr Isibor that i met on this site. i am dana guerrie from California. i was an HPV victim. i had it for over four years i do visit hospital always before. I was just at the point of death with shame after i have lost everything in life even my love one until my close relative direct me to this site to look for great Dr Isibor . It so amazing to see my self back to my feet with everything i have lost. i am very happy because Dr Isibor did not only cure my sickness but he restructure my life by bringing back my ex and every thing i lost to the course of this sickness(HPV). i do not really know where to start from in appreciating you Dr Isibor. please my people out there do not play with your problem go out and seeks for the help of great Dr Isibor herbal healing, he healed me without future problem or hidden charges. now i am free from HPV and My husband, my joy and happiness are back. please with out a second choice go contact great Dr Isibor for the betterment of your life. CONTACT HIM VIA EMAIL;drisiborspellhome@gmail.comor whatsapp number +2348107855231

    ReplyDelete
  14. Hello everyone out there, I'm here to give my testimony about a herbalist doctor who helped me. I was infected with HERPES SIMPLEX VIRUS in 2011, I went to many hospitals to heal myself but there was no solution, so I was thinking how I can get a solution so that my body can be well. One day I was in the river thinking about where I can go to get a solution. so a lady walked towards me telling me why I'm so sad and I open everything by telling her my problem, she told me she could help me, she introduced me to a doctor who uses herbal medicines to cure the SIMPLEX HERPES VIRUS and gave me your email, so I sent you an email. He told me everything I had to do and also gave me instructions to take, which I followed correctly. Before I knew what was happening after two weeks, the SIMPLEX HERPES VIRUS that was in my body disappeared. therefore, if you also have a broken heart and need help, you can also send an email to {oliha.miraclemedicine@gmail.com}or whatsapp him number: +2349038382931 or Facebook page https://www.facebook.com/drolihamiraclemedicine/
    Contact him today and he will have a testimony ... Good luck!

    Dr. OLIHA also cures:
    1. HIV / AIDS
    2. HERPES 1/2
    3. CANCER
    4. ALS (Lou Gehrig's disease)
    5. Hepatitis B
    6. chronic pancreatic
    7. emphysema
    8. COPD (chronic obstructive pulmonary disease).../

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  15. My name is Gwen Stacy Wailker , am here to appreciate Dr isiramen for using his herbal medicine to cure my Herpes virus. Is about 3 years and 6 months now I have been living with this virus and it has been a serious problem to me, I was so confused cause i have been taking several drugs to be cured but all of my effort was in vain,one morning I was browsing through the internet then I saw several testimonies about Dr. isiramen curing people from Herpes virus and immediately i contacted Dr. isiramen on his email: oyimeanelisiramen@gmail.com i told him about my troubles and he told me that i must be cured, he gave me some instructions and which i rightly followed. so he prepared a herbal medicine and sent it to me which i used for 2 weeks and i was cured everything was like a dream to me and my Herpes virus was totally gone, dr .isiramen, God bless you and give you more power and ability for more cure.I don't know if there is any one out there suffering for herpes virus or any of these diseases..DIABETES, CANCER, HIV/AIDS, HERPES HEPATITiS A AND B DISEASES etc. why don't you contact dr.isiramen today and be free from your diseases because he is very good and honest Doctor and he is also called the godfather of herbalism contact him via email; oyimeanelisiramen@gmail.com or whatsApp him on +2348117963221

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  16. I has suffered for Human papillomavirus HPV) for 2years, I was given some tablets at the hospital but I refused to take it, They said I have to be on it for life so I don't want take a drugs everyday for life. No point in taking medicine everyday when u won't get cure from it and I was advice to seek for natural herbal cure, after some time I found dr onokun is the most trustful herbalist that have herbs to cure wicked symptom's,I emailed dr onokun, for 2weeks been his patient he cured my (HPV) with his herbal. I only used his natural herbs for two weeks it was 100% cure. I'm not (HPV) patient anymore. I'm happy about it i finally got cured out of this mess been in my body for 2years. I also recommend you if you're living with (HPV) or herpes symptoms i also want you to be free contact dr onokun with the email attach to my post. dronokunherbalcure@gmail.com

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