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IntroductionThe prime-boost COVID-19 mRNA vaccination strategy has proven to be effective against severe COVID-19 disease and death. However, concerns have been raised due to decreasing neutralizing antibody levels after COVID-19 vaccination and due to the emergence of new immuno-evasive SARS-CoV-2 variants that may require additional booster vaccinations.MethodsIn this study, we analyzed the humoral and cell-mediated immune responses against the Omicron BA.1 and BA.2 subvariants in Finnish healthcare workers (HCWs) vaccinated with three doses of COVID-19 mRNA vaccines. We used enzyme immunoassay and microneutralization test to analyze the levels of SARS-CoV-2 specific IgG antibodies in the sera of the vaccinees and the in vitro neutralization capacity of the sera. Activation induced marker assay together with flow cytometry and extracellular cytokine analysis was used to determine responses in SARS-CoV-2 spike protein stimulated PBMCs.ResultsHere we show that within the HCWs, the third mRNA vaccine dose recalls both humoral and T cell-mediated immune responses and induces high levels of neutralizing antibodies against Omicron BA.1 and BA.2 variants. Three weeks after the third vaccine dose, SARS-CoV-2 wild type spike protein-specific CD4+ and CD8+ T cells are observed in 82% and 71% of HCWs, respectively, and the T cells cross-recognize both Omicron BA.1 and BA.2 spike peptides. Although the levels of neutralizing antibodies against Omicron BA.1 and BA.2 decline 2.5 to 3.8-fold three months after the third dose, memory CD4+ T cell responses are maintained for at least eight months post the second dose and three months post the third vaccine dose.DiscussionWe show that after the administration of the third mRNA vaccine dose the levels of both humoral and cell-mediated immune responses are effectively activated, and the levels of the spike-specific antibodies are further elevated compared to the levels after the second vaccine dose. Even though at three months after the third vaccine dose antibody levels in sera decrease at a similar rate as after the second vaccine dose, the levels of spike-specific CD4+ and CD8+ T cells remain relatively stable. Additionally, the T cells retain efficiency in cross-recognizing spike protein peptide pools derived from Omicron BA.1 and BA.2 subvariants. Altogether our results suggest durable cellmediated immunity and protection against SARS-CoV-2.

Background Primary diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is based on detection of virus RNA in nasopharyngeal swab samples. In addition, analysis of humoral immunity against SARS-CoV-2 has an important role in viral diagnostics and seroprevalence estimates. Methods We developed and optimized an enzyme immunoassays (EIA) using SARS-CoV-2 nucleoprotein (N), S1 and receptor binding domain (RBD) of the viral spike protein, and N proteins from SARS, Middle East respiratory syndrome (MERS), and 4 low-pathogenic human CoVs. Neutralizing antibody activity was compared with SARS-CoV-2 IgG, IgA, and IgM EIA results. Results The sensitivity of EIA for detecting immune response in COVID-19 patients (n = 101) was 77% in the acute phase and 100% in the convalescent phase of SARS-CoV-2 infection when N and RBD were used as antigens in IgG and IgA specific EIAs. SARS-CoV-2 infection significantly increased humoral immune responses against the 229E and NL63 N proteins. S1 and RBD-based EIA results had a strong correlation with microneutralization test results. Conclusions The data indicate a combination of SARS-CoV-2 S1 or RBD and N proteins and analysis of IgG and IgA immunoglobulin classes in sera provide an excellent basis for specific and sensitive serological diagnostics of COVID-19.

Vaccination shows efficacy in protecting from COVID-19, but regime and dosing optimization is still ongoing. Here the authors show that BNT162b2, mRNA-1273, or their combination with ChAdOx1 induces similar antibody responses, and those receiving three doses of BNT162b2 induce neutralizing antibodies against the Omicron variant. Two COVID-19 mRNA (of BNT162b2, mRNA-1273) and two adenovirus vector vaccines (ChAdOx1 and Janssen) are licensed in Europe, but optimization of regime and dosing is still ongoing. Here we show in health care workers (n = 328) that two doses of BNT162b2, mRNA-1273, or a combination of ChAdOx1 adenovirus vector and mRNA vaccines administrated with a long 12-week dose interval induce equally high levels of anti-SARS-CoV-2 spike antibodies and neutralizing antibodies against D614 and Delta variant. By contrast, two doses of BNT162b2 with a short 3-week interval induce 2-3-fold lower titers of neutralizing antibodies than those from the 12-week interval, yet a third BNT162b2 or mRNA-1273 booster dose increases the antibody levels 4-fold compared to the levels after the second dose, as well as induces neutralizing antibody against Omicron BA.1 variant. Our data thus indicates that a third COVID-19 mRNA vaccine may induce cross-protective neutralizing antibodies against multiple variants.

Two COVID-19 mRNA and two adenovirus vector vaccines have been licensed in Europe and various vaccine combinations and dosing strategies have been exploited to maximize the immunity against COVID-19. Here, we show that among health care workers (n=328) two doses of BNT162b2, mRNA-1273, or ChAdOx1 as also a combination of an adenovirus vector and mRNA vaccines induces equally high levels of anti-SARS-CoV-2 spike antibodies and neutralizing antibodies against B.1 and B.1.617.2 when administrated with a long 12-week dose interval. Two doses of BNT162b2 with a short 3-week interval induce 2-3-fold lower titers of neutralizing antibodies compared to the long interval. Third mRNA vaccine dose for the short dose interval group increased the antibody levels 4-fold compared to the levels after the second dose. Importantly, sera from all three-times vaccinated neutralized B.1.1.529 (Omicron). The data indicates that a third COVID-19 mRNA vaccine dose efficiently induces cross-protective neutralizing antibodies against multiple variants.

Zika virus (ZIKV) infections in humans are considered to be mild or subclinical. However, during the recent epidemics in the Pacific Islands and the Americas, the infection was associated with Quillain-Barré syndrome and congenital infections with fetal brain abnormalities, including microcephaly. Thus, more detailed understanding of ZIKV-host cell interactions and regulation of innate immune responses by strains of differential evolutionary origin is required. Here, we characterized the infection and immune responses triggered by two epidemic Asian/American lineage viruses, including an isolate from fetal brains, and a historical, low passage 1947 African lineage virus in human monocyte-derived dendritic cells (DCs) and macrophages. The epidemic Asian/American ZIKV replicated well and induced relatively good antiviral responses in human DCs whereas the African strain replicated less efficiently and induced weaker immune responses. In macrophages both the African and Asian strains showed limited replication and relatively weak cytokine gene expression. Interestingly, in macrophages we observed host protein degradation, especially IRF3 and STAT2, at early phases of infection with both lineage viruses, suggesting an early proteasomal activation in phagocytic cells. Our data indicates that ZIKV evolution has led to significant phenotypic differences in the replication characteristics leading to differential regulation of host innate immune responses.

As SARS-CoV-2 has been circulating for over a year, dozens of vaccine candidates are under development or in clinical use. The BNT162b2 mRNA COVID-19 vaccine induces spike protein-specific neutralizing antibodies associated with protective immunity. The emergence of the B.1.1.7 and B.1.351 variants has raised concerns of reduced vaccine efficacy and increased re-infection rates. Here we show, that after the second dose, the sera of BNT162b2-vaccinated health care workers (n = 180) effectively neutralize the SARS-CoV-2 variant with the D614G substitution and the B.1.1.7 variant, whereas the neutralization of the B.1.351 variant is five-fold reduced. Despite the reduction, 92% of the vaccinees have a neutralization titre of >20 for the B.1.351 variant indicating some protection. The vaccinees’ neutralization titres exceeded those of recovered non-hospitalized COVID-19 patients. Our work provides strong evidence that the second dose of the BNT162b2 vaccine induces efficient cross-neutralization of SARS-CoV-2 variants currently circulating in the world.

Avsikten med denna studie har varit att analysera Sveriges 19 lärosätens kursplaner för ämnet svenska i årskurs 4–6 samt jämföra dessa med lgr11s läroplan. Studien syftar till att belysa hur ämnet svenska framställs i kursplaner på svenska lärarutbildningar med inriktning mot årskurs 4–6 i grundskolan, samt till att analysera förhållandet mellan dessa kursplaner och kursplanen i svenska (Lgr11) med avseende på det ämnesinnehåll som beskrivs. De frågeställningar som besvaras i studien är följande: 1. Hur är svenskkurserna på landets lärarutbildningar organiserade med avseende på struktur, innehåll och poäng, samt vilka olika typer av läroplaner går det att urskilja i de olika kursplanerna? 2. Vilka motsvarigheter till det centrala innehållet för årskurs 4–6 finns i lärosätenas kursplaner i ämnet svenska? Förekommer andra innehållspunkter eller saknas punkter som finns i Lgr11? 3. Var ligger ämnets tyngdpunkter och vilka ämneskonceptioner går att identifiera i de olika kursplanerna? Läroplansteori är en central utgångspunkt för detta arbete. Den handlar om hur innehåll, mål och metodik implementeras i läroplaner för att forma elever efter samhällets grundtankar, bland annat om vad som är av vikt i utbildningen. Det går att urskilja olika typer av läroplaner som alla lägger olika vikt på de delar som ska tillsammans utgöra en fullständig plan för elevers undervisning. De läroplanstyper som har använts i analysen för att besvara fråga 1 är: innehållsbaserad, processfokuserad, målstyrd och resultatfokuserad. Dessa fyra jämförs med de 19 lärosätenas kursplaner för att få fram vilken typ som varje kursplan efterliknar mest. Begreppet ämneskonceptioner används för att förklara hur ämnet svenska kan uppfattas på olika sätt. Det finns främst fyra olika synsätt på hur man kan tolka ämnet och dess mening i utbildning. Svenska som ett färdighetsämne innebär att eleverna skall lära sig den formella tekniken inom delområden för ämnet. Svenska som ett litteraturhistoriskt bildningsämne innebär att kulturarvet ses som centralt för ämnet och den tredje ämneskonceptionen är svenska som ett erfarenhetspedagogiskt ämne, vilket innebär att man utgår från elevernas förutsättningar och erfarenheter när man designar undervisningen. Svenska som ett demokratiämne innebär att svenskämnet ses som ett demokratifostrande ämne som skall bidra till att alla elever kan delta i samhället på lika villkor. För att besvara första delen av fråga 1 användes en tematiskt innehållsbaserad textanalys för att skilja på de strukturer som de 19 kursplanerna innefattar samt ta reda på hur många poäng kurserna ger och vilka avsnitt de består av. För att besvara fråga två och tre utfördes en etnografisk innehållsanalys. Resultatet av den första frågeställningen, hur ser strukturen ut och vilka typer går att urskilja, pekar mot att kursplanerna skiljer sig från varandra, där de flesta lägger större vikt på mål än vad de gör på bedömning. Vilka läroplanstyper som främst går att sammankoppla till kursplanerna är den målfokuserade, efter det den processcentrerade och de läroplaner som framkommer minst ur analysen är den resultatcentrerade samt innehållscentrerade. Ur analysen som utfördes för att besvara de två sista frågeställningar framkommer det att de centrala delar som går att finna i läroplanen även tas upp i kursplanerna, men med vissa undantag där avsnitt inte är framskrivna av ett lärosäte, som annars används i Lgr11. Majoriteten av kursplanerna har spår av alla ämneskonceptioner, men synen att ämnet svenska är ett erfarenhetsdemokratiskt ämne och svenska som ett demokratiämne är främst representerade och svenska som bildningsämne som är den minst vanliga. Resultaten gällande de vanligaste kursplanerna på grundlärarutbildningen 4–6, går att koppla samman med vad Benita Berg (2014) i sin studie redovisar, vilket är att ett av de viktigaste ändamål med lgr11 är att eleverna uppnår målen och därför är de ofta explicit uttalade. Sedan hur eleverna gör det i förhållande till bestämda metoder och explicit innehåll är inte lika tydliga (Berg, 2014). Däremot så är innehållet i lgr11 mer bestämt än i majoriteten av kursplanerna. Vad som främst framkommer utav denna studie är att lgr11 bidrar till att lärare har en bred grund att stå på då det kommer till hur den planerar och utför undervisning. Det finns däremot mycket fritt handlingsutrymme som läraren får, målen är ofta konkreta men innehållet och metoderna är ofta öppna för egen tolkning. Detta är även något som går att finna i de 19 kursplanerna men då ofta med ännu färre strikta ramar.

Aim: Claimed intake of alcohol after a traffic incident, called the hip-flask defence, can be objectively assessed by different methods. One of them is the use of two consecutive ethanol concentrations in urine and the ratio between ethanol concentrations in urine and blood. Another one is the concentrations of ethyl glucuronide (EtG) and ethyl sulphate (EtS) in blood and their ratio to ethanol. The experimental basis for both these models is from single dose studies only. The aim of this study was therefore to describe the kinetics of ethanol, EtG and EtS after ingestion of two repeated doses of ethanol and to investigate the usefulness of the different models for the assessment of the hip-flask defence. Methods: Thirty-five subjects ingested a first dose of 0.51 g of ethanol per kilo body weight, and two hours later a second dose (the hip-flask drink) of 0.25, 0.51 or 0.85 g of ethanol per kilo body weight. Ten urine and 17 blood samples were collected and analysed for ethanol, EtG and EtS using fully validated methods. It was investigated if all subjects fulfilled the criteria for recent drinking, according to the two different models, when using the samples collected 180-240 minutes after start of first dose drinking. According to the first model, increase in urinary ethanol concentrations and a ratio UAC/BAC below 1.3 indicated recent drinking. According to the second model, increase in blood EtG concentrations and a ratio ethanol (g/kg)/EtG (mg/L) above 1 indicated recent drinking. Results: All subjects in the high dose group fulfilled all criteria for recent drinking. One subject in the medium dose group and nine subjects in the low dose group failed to show increasing UAC and/or a UAC/ BAC ratio below 1.3. One subject in the low dose group failed to show increasing concentrations of blood EtG, but all subjects showed a ratio ethanol/EtG above 1. Conclusions: The present study showed, by the use of experimental data, that both two models used to investigate the hip-flask defence can be used, but only when the hip-flask dose is sufficiently high. (C) 2020 The Author(s). Published by Elsevier B.V.

Ebolaviruses (EBOV) cause severe disease with a recent outbreak in West Africa in 2014-2015 leading to more than 28 000 cases and 11 300 fatalities. This emphasizes the urgent need for better knowledge on these highly pathogenic RNA viruses. Host innate immune responses play a key role in restricting the spread of a viral disease. In this study we systematically analyzed the effects of cloned EBOV genes on the main host immune response to RNA viruses: the activation of RIG-I pathway and type I and III interferon (IFN) gene expression. EBOV VP24, in addition of inhibiting IFN-induced antiviral responses, was found to efficiently inhibit type III IFN-λ1 gene expression. This inhibition was found to occur downstream of IRF3 activation and to be dependent on VP24 importin binding residues. These results emphasize the importance of VP24 in EBOV infection cycle, making VP24 as an excellent target for drug development.

The Zika virus (ZIKV) is a member of the Flaviviridae family and an important human pathogen. Most pathogenic viruses encode proteins that interfere with the activation of host innate immune responses. Like other flaviviruses, ZIKV interferes with the expression of interferon (IFN) genes and inhibits IFN-induced antiviral responses. ZIKV infects through epithelial barriers where IFN-&lambda, 1 is an important antiviral molecule. In this study, we analyzed the effects of ZIKV proteins on the activation of IFN-&lambda, 1 promoter. All ZIKV proteins were cloned and transiently expressed. ZIKV NS5, but no other ZIKV protein, was able to interfere with the RIG-I signaling pathway. This inhibition took place upstream of interferon regulatory factor 3 (IRF3) resulting in reduced phosphorylation of IRF3 and reduced activation of IFN-&lambda, 1 promoter. Furthermore, we showed that ZIKV NS5 interacts with the protein kinase IKK&epsilon, which is likely critical to the observed inhibition of phosphorylation of IRF3.