In 2020 Lao PDR had low reported COVID-19 cases but it was unclear whether this masked silent transmission. A seroprevalence study was done August - September 2020 to determine SARS-CoV-2 exposure. Participants were from the general community (n=2433) or healthcare workers (n=666) in five provinces and bat/wildlife contacts (n=74) were from Vientiane province. ELISAs detected anti- SARS-CoV-2 Nucleoprotein (N; n=3173 tested) and Spike (S; n=1417 tested) antibodies. Double-positive samples were checked by IgM/IgG rapid tests. Controls were confirmed COVID-19 cases (n=15) and pre-COVID-19 samples (n=265). Seroprevalence for the general community was weighted to account for complex survey sample design, age and sex. In pre-COVID-19 samples, 5·3%, [95% CI=3·1-8·7%] were anti-N antibody single-positive and 1·1% [0·3-3·5%] were anti-S antibody single positive. None were double positive. Anti-N and anti-S antibodies were detected in 5·2% [4·2-6·5%] and 2·1% [1·1-3·9%] of the general community, 2·0% [1·1-3·3%] and 1·4% [0·5-3·7%] of healthcare workers and 20·3% [12·6-31·0%] and 6·8% [2·8-15·3%] of bat/wildlife contacts. 0·1% [0·02-0·3%] were double positive for anti-N and anti-S antibodies (rapid test negative). We find no evidence for significant SARS-CoV-2 circulation in Lao PDR before September 2020. This likely results from early decisive measures taken by the government, social behavior, and low population density. High anti-N /low anti-S seroprevalence in bat/wildlife contacts may indicate exposure to cross-reactive animal coronaviruses with threat of emerging novel viruses. Agence Française de Développement. Additional; Institut Pasteur du Laos, Institute Pasteur, Paris and Luxembourg Ministry of Foreign and European Affairs (“PaReCIDS II”).

Autoantibodies characteristic for anti‐phospholipid syndrome (APS) and systemic lupus erythematosus (SLE) are anti‐β₂‐glycoprotein I (β₂GPI) antibodies and anti‐DNA antibodies, respectively, and almost half of APS cases occur in SLE. Anti‐β₂GPI antibodies are recognized to play a pivotal role in inducing a prothrombotic state, but the precise mechanism has not been fully elucidated. In a widely accepted view, binding of anti‐β₂GPI antibodies to cell surface β₂GPI in monocytes and endothelial cells triggers the Toll‐like receptor 4‐myeloid differentiation primary response 88 (TLR)‐4‐MyD88) signaling pathway which leads to activation of p38 mitogen‐activated protein kinase (MAPK), mitogen‐activated protein kinase kinase 1/extracellular signal‐regulated kinases (MEK‐1/ERK) and/or nuclear factor kappa B (NF‐κB) and expression of tissue factor (TF). However, resting cells do not express substantial amounts of TLR‐4. Previously, we generated a mouse monoclonal anti‐β₂GPI antibody WB‐6 and showed that it induced a prothrombotic state – including TF expression on circulating monocytes – in normal mice. In the current study, we aimed to clarify the mechanism of interaction between WB‐6 and resting monocytes, and found that WB‐6 exhibits binding activity to DNA and enters living monocytes or a monocytic cell line and, to a lesser extent, vascular endothelial cells. Treatment of the cells with DNase I reduced the internalization, suggesting the involvement of cell surface DNA in this phenomenon. Monocytes harboring internalized WB‐6 expressed TF and tumor necrosis factor (TNF)‐α which, in turn, stimulated endothelial cells to express intercellular adhesion molecule 1 (ICAM‐I) and vascular cell adhesion molecule 1 (VCAM‐I). These results suggest the possibility that a subset of anti‐β₂GPI antibodies with dual reactivity to DNA possesses ability to stimulate DNA sensors in the cytoplasm, in addition to the cell surface receptor‐mediated pathways, leading to produce proinflammatory and prothrombotic states.

Introduction: Measles is an endemic but largely neglected disease in Lao People’s Democratic Republic. New-borns are protected by maternal antibodies, but antibody waning before measles vaccination at 9 months of age leaves infants susceptible to infection. In this study, the susceptibility window of infants was determined to generate scientific evidence to assess the national measles immunization strategy. Methods: Between 2015 and 2016, demographic data, medical history, and blood samples were collected from 508 mother-child pairs at the provincial hospital in Vientiane. The samples were screened with a commercial kit detecting anti-measles IgG antibodies. Results: The large majority (95.7%) of the mothers were seropositive for anti-measles IgG and antibody titers of the mothers and infants were highly correlated (p < 0.01). While at birth 97.7% of the infants were seropositive, seropositivity rates decreased to 74.2% two months later to reach only 28.2% four months after birth (p < 0.01). Just before the first dose of the measles-rubella vaccine, scheduled at 9 months of age, was actually given, less than 14% of the infants were seropositive. Conclusion: This alarmingly wide susceptibility gap due to rapid maternal antibody decay leaves infants at risk of measles infection and serious disease complications. A high herd immunity is crucial to protect young infants and can be achieved through improved routine vaccination coverage and (expanded age group) supplementary immunization activities.

Xaysomboun province has some of the lowest health indicators in Lao People’s Democratic Republic (PDR). This cross-sectional study aimed to determine the vaccination, susceptibility and exposure status of the population to hepatitis B virus (HBV), measles, rubella, and tetanus. Participants aged 5 years and older were randomly selected from four districts. From each enrolled participant, demographic data and 5 mL of blood sample were taken. HBV surface antigen (HBsAg) and antibodies against HBV, measles, rubella, and tetanus were detected by ELISA. A total of 363 participants (age 5 to 80 years) were included. HBV exposure, as determined by anti-HBV core (anti-HBc) antibodies, was 56.2% overall, and was significantly higher among those aged ≥21 years (78.1%). HBsAg was detected in 9.4% overall and increased to 20% in ages 31–40 years. Only 13.8% of participants had serology indicative of vaccination (anti-HBs positive, anti-HBc negative). Seroprotection against measles was 74.6% overall but only 41.7% in children aged 5–10 years. Anti-rubella IgG was 94.2% overall and high in all age groups. Tetanus seroprevalence was only 47.4% overall but significantly higher in females aged 31–40 (75.6%). We suggest strengthening of routine and booster HBV, measles, and tetanus vaccine coverage in Xaysomboun province.

Background: Despite the high prevalence of chronic hepatitis B virus (HBV) infection in adults in Lao People’s Democratic Republic (Lao PDR), Lao health care workers (HCWs) have previously been shown to have low levels of protection against infection. Furthermore, the prevalence of hepatitis D virus (HDV), which increases disease severity in individuals infected with HBV, is not known in Lao PDR. Objective: This study aimed to estimate the exposure and seroprotection against HBV, as well as exposure to HDV, in Lao HCWs from 5 provinces. Methods: In 2020, a total of 666 HCWs aged 20 to 65 years from 5 provinces of Lao PDR were recruited, and their sera were tested by enzyme-linked immunosorbent assay to determine their HBV and HDV coinfection status. Results: HBV exposure, as indicated by the presence of anti–hepatitis B core antibodies, was 40.1% (267/666) overall and significantly higher for HCWs from Oudomxay province (21/31, 67.7%; adjusted odds ratio 3.69, 95% CI 1.68‐8.12; P=.001). The prevalence of hepatitis B surface antigen was 5.4% (36/666) overall and increased with age, from 3.6% (9/248) in those aged ≤30 years to 6.8% (8/118) in those aged ≥50 years. Only 28.7% (191/666) of participants had serological indication of immunization. We could find no evidence for HDV exposure in this study. Conclusions: The study found intermediate hepatitis B surface antigen prevalence among HCWs in Lao PDR, with no evidence of HDV coinfection. Notably, a significant proportion of HCWs remains susceptible to HBV, indicating a substantial gap in seroprotection against the disease.

The epidemiology of typhoid fever in Lao People`s Democratic Republic is poorly defined. Estimating the burden of typhoid fever in endemic countries is complex due to the cost and limitations of population-based surveillance; serological approaches may be a more cost-effective alternative. ELISAs were performed on 937 serum samples (317 children and 620 adults) from across Lao PDR to measure IgG antibody titers against Vi polysaccharide and the experimental protein antigens, CdtB and HlyE. We measured the significance of the differences between antibody titers in adults and children and fitted models to assess the relationship between age and antibody titers. The median IgG titres of both anti-HylE and CdtB were significantly higher in children compared to adults (anti-HylE; 351.7 ELISA Units (EU) vs 198.1 EU, respectively;p<0.0001 and anti-CdtB; 52.6 vs 12.9 EU;p<0.0001). Conversely, the median anti-Vi IgG titer was significantly higher in adults than children (11.3 vs 3.0 U/ml;p<0.0001). A non-linear trend line fitted to the anti-CdtB and anti-HlyE IgG data identified a peak in antibody concentration in children <5 years of age. We identified elevated titers of anti-HlyE and anti-CdtB IgG in the serum of children residing in Lao PDR in comparison to adults. These antigens are associated with seroconversion after typhoid fever and may be a superior measure of disease burden than anti-Vi IgG. This approach is scalable and may be developed to assess the burden of typhoid fever in countries where the disease may be endemic, and evidence is required for the introduction of typhoid vaccines.