Virology tidbits

Virology tidbits

Friday 4 April 2014

MERS-CoV orf4a -an antagonist of antiviral signaling

As mentioned in a previous entry MERS-CoV is capable to inhibit antiviral signaling by antagonizing interferon signaling pathways which are induced by the recognition of dsRNA by intracellular pathogen-associated molecular pattern (PAMP) receptors. Although having a ssRNA genome, dsRNA is produced during Coronavirus replication as an intermediate and thus can be recognized by Toll-like receptors -3 and -9 (TLR-3/TLR-9). Imaging of newly synthesized RNA in cells infected with MHV revealed the formation of spherules tethered to the cellular membrane shielding these intermediates from the recognition by TLR-3 and/or TLR-9. If however these spherules also exist in MERS-CoV remains to be investigated.
In addition to TLR mediated signaling, following the infection of cells with a Coronavirus, cytoplasmic viral RNAs are recognised by two RIG-1 like proteins, RIG-1 and MDA-5. Both RIG-1 and MDA-5 require PACT, which binds to RIG-1 and MDA-5, followed by intracellular signaling involving the mitochondrial adaptor protein MAVS and two protein kinases (TBK1 and IKKε).  This in turn allows the nuclear translocation of two transcription factors, IRF3 and IRF7 and subsequent induction of recruited to the IFN promoters to activate transcription of Interferon-β.
In the case of MERS-CoV, the 4a protein was shown previously to be capable of not only binding dsRNA but also to inhibit MDA5 mediated signaling - and thus to block antiviral signaling. A result from another group however favors a different mechanism. They observed that MERS-CoV 4a fails to suppress MDA5 mediated activation of the IFN-β promoter and does not form a complex with RIG-1or MDA5 but forms a complex with PACT in a RNA-dependent manner. Both groups however agree that MERS-CoV 4a inhibits the expression of IFN-β. If and how 4b contributes to the antiviral signaling is still debated. Importantly, BtCoV-HKU-4 4a and a point mutation of MERS-CoV 4a incapable of binding dsRNA prevented PACT mediated signaling thus emphasizing the role of PACT - in contrast to BtCoV HKU-5 4a. 


As always , these results need to be confirmed in other cell lines, besides the Human Airway Epithelia (Calu3) cell line used, such as monocyte derived cell lines (THP-1); the only camel derived cell line I am aware of are Dubca cells (skin fibroblast).
If confirmed, MERS-CoV 4a will be not the only viral protein inhibiting PACT signaling. Other examples of viral proteins inhibiting the activation of RIG-1 by binding to PACT include Influenza NS1, Herpes Virus Us11 and Ebola VP35 and the list might be growing once new viruses are discovered and old ones are revisited.
Finally it remains to be seen if the inhibition of PACT by MERS-CoV also has an impact on the activation of Dicer, which can cleave viral RNA. In cells infected with Influenza A, the inactivation of Dicer has been associated with a modest increase in virus production. Ebola virus VP30 and VP35 proteins have also been postulated to prevent the formation of siRNA by inhibition of the activation of Dicer or inhibiting the contact of Dicer with PACT.
From the present data it is not clear if the viral ssRNA or dsRNA intermediates are responsible for the activation of the antiviral response in absence of orf 4a. Indeed the present data only indicate that 4a is capable of inhibiting experimentally, Poly (I:C), induced dsRNA antiviral response, but the use of the reverse genetic system available should allow to characterize the role of MERS-CoV orf4a in the context of viral replication.


Last but not least a few words about MERS-CoV 4b protein. In a recent paper it was shown that the protein encoded by MERS-CoV orf4b as well as BtCoV-HKU-4 orf4b and BtCoV HKU-5 orf 4b inhibit the IFN type 1 response by localizing to the nucleus. In this case, the protein inhibits RIG-1 mediated IFN-β signaling while only having a moderate effect on NF-κB signalling. 


ResearchBlogging.org




Further reading

Siu KL, Yeung ML, Kok KH, Yuen KS, Kew C, Lui PY, Chan CP, Tse H, Woo PC, Yuen KY, & Jin DY (2014). Middle East respiratory syndrome coronavirus 4a protein is a double-stranded RNA-binding protein that suppresses PACT-induced activation of RIG-I and MDA5 in innate antiviral response. Journal of virology PMID: 24522921


Hagemeijer MC, Vonk AM, Monastyrska I, Rottier PJ, & de Haan CA (2012). Visualizing coronavirus RNA synthesis in time by using click chemistry. Journal of virology, 86 (10), 5808-16 PMID: 22438542


Kok KH, Lui PY, Ng MH, Siu KL, Au SW, & Jin DY (2011). The double-stranded RNA-binding protein PACT functions as a cellular activator of RIG-I to facilitate innate antiviral response. Cell host & microbe, 9 (4), 299-309 PMID: 21501829


Niemeyer D, Zillinger T, Muth D, Zielecki F, Horvath G, Suliman T, Barchet W, Weber F, Drosten C, & Müller MA (2013). Middle East respiratory syndrome coronavirus accessory protein 4a is a type I interferon antagonist. Journal of virology, 87 (22), 12489-95 PMID: 24027320



Yang Y, Zhang L, Geng H, Deng Y, Huang B, Guo Y, Zhao Z, & Tan W (2013). The structural and accessory proteins M, ORF 4a, ORF 4b, and ORF 5 of Middle East respiratory syndrome coronavirus (MERS-CoV) are potent interferon antagonists. Protein & cell, 4 (12), 951-61 PMID: 24318862



Yoneyama M, Kikuchi M, Natsukawa T, Shinobu N, Imaizumi T, Miyagishi M, Taira K, Akira S, & Fujita T (2004). The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nature immunology, 5 (7), 730-7 PMID: 15208624



Kok KH, Ng MH, Ching YP, & Jin DY (2007). Human TRBP and PACT directly interact with each other and associate with dicer to facilitate the production of small interfering RNA. The Journal of biological chemistry, 282 (24), 17649-57 PMID: 17452327



Matskevich AA, & Moelling K (2007). Dicer is involved in protection against influenza A virus infection. The Journal of general virology, 88 (Pt 10), 2627-35 PMID: 17872512


Fabozzi G, Nabel CS, Dolan MA, & Sullivan NJ (2011). Ebolavirus proteins suppress the effects of small interfering RNA by direct interaction with the mammalian RNA interference pathway. Journal of virology, 85 (6), 2512-23 PMID: 21228243

Matthews KL, Coleman CM, van der Meer Y, Snijder EJ, & Frieman MB (2014). The ORF4b-encoded accessory proteins of Middle East respiratory syndrome coronavirus and two related bat coronaviruses localize to the nucleus and inhibit innate immune signalling. The Journal of general virology, 95 (Pt 4), 874-82 PMID: 24443473

20 comments:

  1. 2 WEEKS AGO I ENDED NATURAL HERBAL TREATMENT FROM TRADITIONAL HEALER WHO IS IN AFRICAN, I GOT RID OF HPV VIRUS OUT OF MY BODY WITH HIS HERBAL TREATMENT. It SO AMAZING I'M FREE FROM THIS VIRUS FOREVER. YOU CAN REACH HIM ON; DRONOKUNHERBALCURE@GMAIL.COM

    ReplyDelete
    Replies
    1. I have been using this herbal cure remedy for 3 months now. So far, it has cured my HERPES, eliminated my COLD SORE, improved energy and metabolism, in the morning I feel recharged and I no longer feel sleepy after a meal. As for the scent, it does smell like motor oil but I chase it down with 100% Cranberry Juice 1tsp. twice a day AFTER a meal. I also experienced vivid dreams for about 5 days now it's diminishing to during the first 2 or 3 hours of sleep . contact dr Emmanuel: EMAIL: nativehealthclinic@gmail.com or call/whatsapp +2348140073965. you can get the cure from him and be free from all kinds of diseases. HIV/AIDS,Diabtetes,Cancer,hepatitis 

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    2. Good Day everyone, just want to share my experience here so you all should be more careful when contacting any herbalist for solution cos majority of the testimonies are fake and were the ones that fooled me.they all uses the same local code number (+234) cos i have been screwed by some of them.My Husband was suffering from Hsv for years and i went everywhere to get useful information that will help cure or suppress the disease.then i came across testimonies of those that took my hard earn money without any useful information or cure but they keep telling stories.about two months ago, a new staff was employed in my place of work and when He heard about my Husband Disease He was so remorseful and told me about Dr Emmanuel.but i felt it was still one of those Internet guys so i didn't call or text Dr Emmanuel. later that week, He asked me if i`ve heard from Dr Emmanuel and he gave me proof and reasons to try.well, lots of thanks to Dr Emmanuel cos as im writing this message, my Husband is CURED through the herbs that Dr Emmanuel sent to us.so after my experience, i decided to save some people from falling to the hands of SCAMS.thx for your time and if you require the service of Dr Emmanuel, you have to reach him via email: nativehealthclinic@gmail.com  or WhatsApp/Call +2348140073965
      you can also reach me on  +1(862) 260-4433  
      he have cure to all kinds of virus and disease,thank you 

      Delete
    3. Hello I'm cambell by name, I'm giving a testimony about Dr.Emmanuel the great Herbalist, he has the cure to all manner of diseases, he cured my herpes disease, though I went through different website I saw different testimonies about different spell casters and herbalist, I was like: 'Many people have the herpes cure why are people still suffering from it?' I though of it, then I contact Dr. Emmanuel via email, I didn't believe him that much, I just wanted to give him a try, he replied my mail and Needed some Information about me, then I sent them to him, he prepared it (CURE) and sent it to Airfreight Online Courier Service for delivery, he gave my details to the Courier Office, they told me that 3-5 days I will receive the package and i took the medicine as prescribed by him and I went for check-up 2 weeks after finishing the medicine, I was tested herpes negative, if you are herpes positive do me a favor for you to contact him and I will try my possible best to make sure you get cured, when you contact him, make sure you tell him that I referenced you.. contact him via: nativehealthclinic@gmail.com or WhatsApp +2348140073965 

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    5. I am sharing this testimony to partners suffering in their relationships because there is an enduring solution. My husband left me and our 4 kids for another woman for 1 year. I tried to be strong just for my kids but I could not control the pains that torment my heart. I was hurt and confused. I needed a help, so i did a research on the internet and came across a site where I saw that Dr. Godday a spell caster,who can help get lovers back within few hours I contacted him and he did a special prayer and spells for me. To my surprises, after some days, my husband came back home. That was how we reunited again and there was a lot of love, joy and peace in the family. You can as well contact Dr. Godday a powerful spell-caster for solutions on his contact Email: goddayspiritualhome@gmail.com Whats app only Or Call/What’s-app +1{919}4956404

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  2. MY LIFE IS BACK, Am so happy to share this testimony on how Dr LUCKY , was able to heal me from HpV Disease, well i was detected positive on 23th may 2015, and ever since i have been looking for a way to treat and cure this disease from me, but all ways i try there was no solution, few weeks ago i saw a testimony of some people on how Dr LUCKY was able to cure them from HpV disease, still on i heard about it on media when a young lady also gave her testimony about this same Dr, well left with no hope i message Dr LUCKY , telling him all my problem, he told me what we were to do, that he was going to send me a portion which i am going to take, and after which i took this herb portion he sent to me, he told me to go back to the hospital for check up and after i have done that i should come and tell him the good news result, at first when i saw the message i was so shock and still did not believed i will be cured, well friend to make my story short i am HpV negative now after many time of sorrow, i am Negative, and my sickness are gone, i thank God for leading me to this man….you can email him on (drluckyherbalcure@gmail.com) or you can also call him +2348154637647, he can be of help to you on any problem like
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    ReplyDelete
  3. 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 uza is the most trustful herbalist that have herbs to cure wicked symptom's,I emailed dr uza, 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 uza with the email attach to my post. druzaherbalcure@gmail.com

    ReplyDelete
  4. Am here to appreciate Dr Oliha for using his herbal medicine to cure my Herpes virus. since 4years now I have been living with this virus and it has been giving me challenges, I was so perplexed 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. Oliha curing people from Herpes virus and immediately i contacted Dr. Oliha on his email: oliha.miraclemedicine@gmail.com , i told him about my troubles and he told me that i must be cured, gave me some instructions and which i rightly followed. so he prepared a herbal medicine for me and sent it to me which i used for 2 weeks and everything was like a dream to me and my Herpes virus was totally gone, why don't you contact him today and be free from your diseases because he is very good and honest Doctor. contact him via email; oliha.miraclemedicine@gmail.com or whatsApp him on +2349038382931

    ReplyDelete
  5. 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
  6. I have been using this herbal cure remedy for 3 months now. So far, it has cured my HERPES, eliminated my COLD SORE, improved energy and metabolism, in the morning I feel recharged and I no longer feel sleepy after a meal. As for the scent, it does smell like motor oil but I chase it down with 100% Cranberry Juice 1tsp. twice a day AFTER a meal. I also experienced vivid dreams for about 5 days now it's diminishing to during the first 2 or 3 hours of sleep . contact dr Emmanuel: EMAIL: nativehealthclinic@gmail.com or call/whatsapp +2348140073965. you can get the cure from him and be free from all kinds of diseases. HIV/AIDS,Diabtetes,Cancer,hepatitis 

    ReplyDelete
  7. Good Day everyone, just want to share my experience here so you all should be more careful when contacting any herbalist for solution cos majority of the testimonies are fake and were the ones that fooled me.they all uses the same local code number (+234) cos i have been screwed by some of them.My Husband was suffering from Hsv for years and i went everywhere to get useful information that will help cure or suppress the disease.then i came across testimonies of those that took my hard earn money without any useful information or cure but they keep telling stories.about two months ago, a new staff was employed in my place of work and when He heard about my Husband Disease He was so remorseful and told me about Dr Emmanuel.but i felt it was still one of those Internet guys so i didn't call or text Dr Emmanuel. later that week, He asked me if i`ve heard from Dr Emmanuel and he gave me proof and reasons to try.well, lots of thanks to Dr Emmanuel cos as im writing this message, my Husband is CURED through the herbs that Dr Emmanuel sent to us.so after my experience, i decided to save some people from falling to the hands of SCAMS.thx for your time and if you require the service of Dr Emmanuel, you have to reach him via email: nativehealthclinic@gmail.com  or WhatsApp/Call +2348140073965
    you can also reach me on  +1(862) 260-4433  
    he have cure to all kinds of virus and disease,thank you 

    ReplyDelete
  8. My name is Ryan am from U.S.A. I want to share a testimony of how Dr. DADA  herbal mixture cream saves me from shame and disgrace, my penis was a big problem to me as the size was really so embarrassing,and i was also having weak erection problem. I can't make love to my wife and my penis was just too small a full grown man like me having 4 inches penis and to worsen it i don't last in sex i cant even last two minutes it was really a thing of shame to me. My wife was really tired of me because my sex life was very poor,she never enjoyed sex,i was always thinking and searching for solutions everywhere until when i saw a testimony of how Dr.DADA herbal mixture cream have been helping people regarding their sex life, so i decided to give him a try and to my greatest surprise in less than one week of taking the herbs my penis grow to 8 inches i couldn't believe my eyes and as i speak now my penis is inced 8 es and i do not have week erection again. I can make love to my wife longer in bed. And my marriage is now stable,my wife now enjoy me very well in bed. You can contact him via {drdadaspellhome7@gmail.com) or call him or what-apps him through +2348158568638 .he also specialize on the following things
    BREAST ENLARGEMENT
    BREAST LIFT & REDUCTION
    PENILE/PENIS ENLARGEMENT
    GENERAL BODY SLIMMING
    HIPS & THIGH REDUCTION
    Thanks for the Enlarging my penis sir, you indeed save my marriage...I am really grateful sir,..,,

    ReplyDelete
  9. 5 years ago I had warts, I was treated with some liquid applied to the warts they continued to grow and spread... The next 2 doctors did laser surgery to remove them. 1 year after the surgery, they grew back close to where the 1st ones were' so I was finally told it was hpv. I have had it for very long time, I contract it from my cheated boyfriend and I found out he was also infected and I end up the relationship between us. the warts was so embarrasses because it started spreading all over I have be dealing with this things for very long time the last treatment I take was About 2 years ago I applied natural treatment from Dr onokun herbal cure, a week after applying the treatment all the warts was gone. it's now 2 years and some months I don't have single wart or any symptoms of hpv. wow"" it's great, Dr onokun has finally cured me. Anyone living with hpv contact Dr onokun for natural treatment.
    His email address: dronokunherbalcure@gmail.com  

    ReplyDelete

  10. Grateful to Dr LOSA for all his help for my life, my partner was having Herpes Simplex Virus, and we were planing on getting married on early December, when we decided to get ourselves tested first and we found out that my Girlfriend was HSV 2 Positive, my Girlfriend was so unhappy, because this was a barrier for us to get married, so i went out to search out a remedy for my partner, and it was so surprising how Dr LOSA has been a blessing and healing to the masses, after reading about his work on a blog i immediately emailed him: dr.losaherbalhome@gmail . com and he wrote back to me that my Girlfriend will be OK and that we we get married this same year, i couldn't believe it, after doing all that is needed to be done, i was surprise that after 2weeks of using his medicine my Girlfriend went to the doc and was diagnose negative. For more info contact his
    Whatsapp +2349056464736.OR
    email dr.losaherbalhome@gmail . com and you will get your illness cure and also beware of scams Good Luck
    He cure listed diseases
    CANCER
    HEPATITIS A AND B
    DIABETIC.
    HERPES

    ReplyDelete
  11. Hello everyone, I was infected with the Epstein-Barr virus and I got cured from the Epstein-Barr virus few months ago after i contacted Chief Dr Wealthy. I saw a post on the internet after i have seek healing for several years from different doctors in California. I sent the Doctor a request for help, just a few email i followed his instruction and he sent me the medication. Now i am negative and i referred him to all my friend who had this same sickness and they have gotten their cure too. You can contact him via his email: wealthylovespell@gmail.com or Phone call or whatsAPP: +2348105150446. While i was on his medication i understood that he can also cure HERPES, HIV/AIDS, CANCER, Male/female menopause, Miscarriage, Menstruation problems, PREGNANCY PROBLEM, EPILEPSY, GONORRHEA, LASSA FEVER, OBESITY, KIDNEY FAILURE, HYPERTENSION, FIBROID TUMOR, BRAIN FOG, EPSTEIN-BARR VIRUS and many more.

    Contact him today and get your problem solved.

    ReplyDelete
  12. I'm 61 years old, I contracted hpv in 2011' I has be taking lot treatment for it and some months ago the wart stated coming out seriously, I used lot recommendation because there was lot warts around my anus and was so embarrassed. but today I'm totally happy I got the virus eliminated by using natural treatment from Dr Onokun herbal center after his treatment I got cured. all the warts went away' seriously believed Dr Onokun he have the cure for human papillomavirus because he has eliminated hpv been in my body since 2011, Dr Onokun make it possible for me. Here is Dr Onokun email to reach him: Dronokunherbalcure@gmail.com he is welled capable of curing terrible diseases.  

    ReplyDelete
  13. I'm 61 years old, I contracted hpv in 2011' I has be taking lot treatment for it and embarrassedsome months ago the wart stated coming out seriously, I used lot recommendation because there was lot warts around my anus and was so . but today I'm totally happy I got the virus eliminated by using natural treatment from Dr Onokun herbal center after his treatment I got cured. all the warts went away' seriously believed Dr Onokun he have the cure for human papillomavirus because he has eliminated hpv been in my body since 2011, Dr Onokun make it possible for me. Here is Dr Onokun email to reach him: Dronokunherbalcure@gmail.com  he is welled capable of curing terrible diseases.  

    ReplyDelete
  14. HELLO EVERYONE.. FEW MUNINETS TO REDY THIS INFOR ON HERPES CURE 2018..
    2017 MY MOTHER WAS DIAGNOSED OF HERPES/ KNOWN AS GENITAL WARTS ,I SPENT A LOT OF MONEY ON HER MEDICATION TILL A POINT I EVEN LOST HOPE,BECAUSE MY MOTHER WAS GRADUALLY DYING AND LOST HER MEMORY TOO, I WAS SO DESPERATE TO GET MY MOTHER BACK TO NORMAL, ONE DAY MY UNCLE WHO LIVES IN LONDON UNITED KINGDOM TOLD ME ABOUT DR OLIHA ,WHO HELPED HIM GET RID OF HERPES /GENITAL WART WITH HERBAL MEDICINE AND HIS HERBAL SOAP ,I WAS SO SHOCKED WHEN HE TOLD ME ABOUT THIS ,ALTHOUGH I NEVER BELIEVE IN HERB BUT, I KEEP TO BELIEVE BECAUSE MY UNCLE CAN'T TELL ME LIES WHEN IT COMES TO HEALTH CONDITION I CONTACTED DR OLIHA VIA HIS EMAIL; OLIHA.MIRACLEMEDICINE@GMAIL.COM , YOU CAN TALK TO HIM VIA CALL OR WHATSAPP MESSENGER ON +2349038382931 , HE REPLIED AND ASK ME TO SEND MY HOME ADDRESS AND MY MOTHER'S DETAIL AND THEN I PURCHASED THE HERBAL MEDICINE,SENT ME THE HERBAL MEDICINE THROUGH COURIER SERVICE, WHEN I RECEIVED THIS HERBAL MEDICINE USED IT FOR 2 WEEKS, AND 4 DAYS OF USAGE THE WARTS FELL OFF, MY MOTHER I NOW TOTALLY CURED AND MY MOTHER IS LIVING FREE AND HAPPY AGAIN. YOU CAN TALK TO DR VIA HIS MOBILE NUMBER OR WHATS APP HIM ON +2349038382931.ALL THANKS TO DOCTOR DR OLIHA  

    ReplyDelete
  15. I was married at 32 and immediately tried to get pregnant. When I was unable to conceive I had blood tests for fertility and was told that I had an FSH (follicle stimulating hormone) of 54 and would not be able to have children. Even though the doctors knew that I had been diagnosed with Hashimoto’s thyroiditis since age 25, no one bothered to check my thyroid levels. my TSH was measured at .001. My Synthroid dosage was lowered. a friend advise me to contact a spiritualist who help with fertility with his medicine, i collected his contact and explain my situation to him he prepared for me a herbal medicine which i took as describe by him. became pregnant very quickly, I had a successful pregnancy. I have my baby august 2017. to get pregnant at age 35 with my 2nd child in september 2019, thank you sir , this is his email contact if you require his help babaka.wolf@gmail.com or Facebook at priest.babaka














    I was married at 32 and immediately tried to get pregnant. When I was unable to conceive I had blood tests for fertility and was told that I had an FSH (follicle stimulating hormone) of 54 and would not be able to have children. Even though the doctors knew that I had been diagnosed with Hashimoto’s thyroiditis since age 25, no one bothered to check my thyroid levels. my TSH was measured at .001. My Synthroid dosage was lowered. a friend advise me to contact a spiritualist who help with fertility with his medicine, i collected his contact and explain my situation to him he prepared for me a herbal medicine which i took as describe by him. became pregnant very quickly, I had a successful pregnancy. I have my baby august 2017. to get pregnant at age 35 with my 2nd child in september 2019, thank you sir , this is his email contact if you require his help babaka.wolf@gmail.com or Facebook at priest.babaka

    ReplyDelete