ABSTRACT
Background: TLR7/8 are immune receptors expressed in megakaryocytes which detects single-stranded RNA viruses such as SARS-CoV- 2. There is increasing evidence that in addition to raised platelet counts, severe infection with SARS-CoV- 2 increases the risk of venous, arterial and microvascular coagulation. Aim(s): To determine if ssRNA viruses are capable of increasing thrombopoeisis through direct interaction with megakaryocytes. Method(s): Cells were incubated with and without Gardiquimod (GDQ), a specific agonist of TLR7/8 in cord blood derived (CBMKs) and mouse bone marrow derived megakaryocytes (mMKs). TLR7/8-/- iPSC derived megakaryocytes (iPSC-MKs) were produced using CRISPR Cas9 editing of iPSCs and forward programming using an doxycyclin inducible cassette. GFP labelled SARS-CoV- 2 virus was incubated with the TLR7/8-/- iPSC- MKs and wild-type iPSC-MKs. Result(s): Incubation with GDQ increased platelet production in CBMKs and mMKs, and increased platelet function. Increased platelet counts were seen in mice treated with GDQ, and mice infected with influenza. Incubation with GDQ induced increased expression of IL1beta in the parental iPSC-MKs, however in the TLR7-/- and TLR7/8-/- MKs, no increased expression was observed. There was a significant increase in platelet production from the parental iPSC-MKs in response to incubation with GDQ, which was not seen in the TLR7-/- and TLR7/8-/- MKs. Incubation of the GFP-labelled SARS-CoV- 2 virus with wild-type MKs did not lead to a significant increase in fluorescence. Only very low level viral sequences were found in the cells post-incubation demonstrating that penetration within the MKs is unlikely to be of significance. Studies are ongoing to ascertain whether SARS-CoV- 2 induces outside in signalling leading to changes in transcription within the MKs (such as elevation of IL1beta). Conclusion(s): TLR7/8 agonists, including ssRNA genome viruses, increase platelet production and functionality from megakaryocytes. SARS-CoV- 2 does not appear to penetrate and significantly replicate within MKs, however incubation with megakaryocytes did show elevated expression of IL1beta. (Figure Presented).
ABSTRACT
In the context of high rates of domestic violence and abuse (DVA) during the pandemic, specialist DVA services have been required to adapt rapidly to continue to deliver essential support to women and children in both refuges and the community. This study examines service users’ experiences and views of DVA service provision under COVID-19 and discusses implications for future practice. Data are drawn from a wider evaluation of DVA services in five sites in England. Fifty-seven semi-structured interviews and five focus groups were conducted with 70 female survivors and seven children accessing DVA services during the pandemic. Analysis identified key themes in respect of the influence of COVID-19 on the experience of service delivery. COVID-19 restrictions had both positive and negative implications for service users. Remote support reduced face-to-face contact with services, but consistent communication counteracted isolation. Digital practices offered effective means of providing individual and group support, but there were concerns that not all children were able to access online support. Digital support offered convenience and control for survivors but could lack privacy and opportunities for relationship-building. The pivot to remote delivery suggests directions where DVA services can expand the range and nature of future service provision. © Centre for Gender and Violence Research.