The placenta is a complex organ with multiple immune and non-immune cell types that promote fetal tolerance and facilitate the transfer of nutrients and oxygen. The nonhuman primate (NHP) is a key experimental model for studying human pregnancy complications, in part due to similarities in placental structure, which makes it essential to understand how single-cell populations compare across the human and NHP maternal-fetal interface. We constructed a single-cell RNA-Seq (scRNA-Seq) atlas of the placenta from the pigtail macaque ( Macaca nemestrina ) in the third trimester, comprising three different tissues at the maternal-fetal interface: the chorionic villi (placental disc), chorioamniotic membranes, and the maternal decidua. Each tissue was separately dissociated into single cells and processed through the 10X Genomics and Seurat pipeline, followed by aggregation, unsupervised clustering, and cluster annotation. Next, we determined the maternal-fetal origins of cell populations and analyzed single-cell RNA trajectory, Gene Ontology enrichment, and cell-cell communication. Single-cell populations in the pigtail macaque were strikingly similar in their identity and frequency to those found in the human placenta, including cells from trophoblast, stromal cell, immune, and macrophage lineages. An advantage of our approach was the deep sequencing of three tissues at the maternal-fetal interface, which yielded a rich diversity of common and rare single-cell populations. The third-trimester pigtail macaque single-cell atlas enables the identification of cellular subclusters analogous to those in humans and provides a powerful resource for understanding experimental perturbations on the NHP placenta.

IntroductionPregnant individuals are at increased risk of developing severe illness from coronavirus disease 2019 (COVID-19). They also tend to be vaccine-hesitant, which is even more pronounced in certain racial and ethnic minority groups. The study objective was to determine whether social media advertisements promoting COVID-19 vaccination could influence vaccination likelihood among pregnant and recently pregnant participants who self-identified as Black or African American.MethodsParticipants were interviewed individually or in focus groups to explore their attitudes toward vaccination and were asked to rate their likelihood of receiving a COVID-19 vaccine after seeing a panel of advertisements featuring different messengers (e.g., doctors, peers, older adults, and faith leaders) and content types (e.g., social proof, text-heavy, fear-based, and activation). Advertisement ratings were analyzed using linear mixed-effects models to examine the effects of vaccination status, advertisement messenger, and advertisement content type. Interview data were coded and analyzed to identify qualitative themes. Advertisement ratings differed significantly according to vaccine status, religious affiliation, political affiliation, and marital status.ResultsParticipants who were vaccinated, religious, politically conservative, and/or married were more likely to report an increased willingness to receive a COVID-19 vaccine after seeing the social media ads. No specific advertisement messenger or content type was rated as more probable to motivate vaccination. Qualitative analysis identified several themes related to attitudes toward vaccination, including trust in oneself, family, doulas, and faith leaders; mistrust of the government and healthcare system; concerns about medical racism; and personal experiences with individuals recommending or discouraging vaccination.DiscussionAlthough no single advertisement messenger or content type was preferred, personal testimony from a trusted source, such as a faith leader or doula, may support vaccine acceptance among Black pregnant women.

Pregnant individuals have a greater susceptibility to severe disease from the coronavirus 2019 disease (COVID-19). Pregnant people also tend to be vaccine-hesitant, which is even more pronounced in certain racial and ethnic minority groups. The study objective was to determine whether social media ads promoting COVID-19 vaccination could influence vaccination likelihood among pregnant and recently pregnant participants who self-identified as Black or African American. Participants were interviewed individually or in focus groups to explore their attitudes about vaccination and to ask them to rate their COVID-19 vaccination likelihood after seeing a panel of ads featuring different messengers (e.g., doctor, peer, elder, faith leader) and content types (e.g., social proof, text-heavy, fear-based, activation). Ad ratings were analyzed using linear mixed models to examine the effect of vaccination status, ad messenger, and ad content type. Interviews were coded and analyzed for qualitative themes. Ad scores differed significantly by vaccine status, with vaccinated participants rating ads as more likely to inspire vaccine uptake, while unvaccinated participants rated ads negatively. No specific messenger or content type was rated as more probable to motivate vaccination. There was a significant interaction between faith-based messengers and COVID-19 vaccination status, with faith leaders perceived as more favorable by unvaccinated participants (p=0.008). Vaccine-hesitant respondents cited mistrust of healthcare providers and fears of medical racism. Although we did not identify content types that might be helpful in a public health vaccine campaign targeting Black pregnant people, faith leaders may be a trusted messenger for unvaccinated individuals.

BackgroundThe fetal brain undergoes rapid cellular and structural changes in late gestation, when waves of neurogenesis and gliogenesis shape cortical circuitry. The ventricular zone (VZ), subventricular zone (SVZ), periventricular white matter (PVWM), and deep white matter (DWM) are enriched in neuroprogenitor cells, newborn neurons, and interneurons, which are regions challenging to study in the third-trimester human fetal brain. The nonhuman primate (NHP) provides a powerful translational model to overcome this limitation, given its close similarity to human neurodevelopmental trajectories. The study objective was to construct a single-cell RNA-Seq (scRNA-Seq) atlas of the late-gestation fetal brain of the pigtail macaque (Macaca nemestrina), focusing on cells in the periventricular proliferative zone.MethodsA sample of the lateral ventricular wall, subventricular zone, and overlying white/gray matter was dissociated into single cells and processed through 10X Genomics sequencing, SoupX removal of ambient RNA, empty droplet removal, doublet exclusion, and Seurat's pipeline which consists of aggregation, unsupervised clustering, and cluster annotation to create a single-cell RNA-Seq (scRNA-Seq) atlas. We also investigated developmental trajectories using scVelo and CellRank2.ResultsThis analysis captured diverse populations of neuroprogenitors, newborn neurons, developing lineages of excitatory and inhibitory neurons, oligodendrocytes, astrocytes, epithelial and vascular cells.ConclusionsSingle-cell populations from the third-trimester nonhuman primate fetal brain are highly similar to those in the human fetus at the level of major lineages. This late-gestation single-cell atlas of the periventricular proliferative zone provides a unique reference for progenitor, neuronal, glial, vascular, and immune cell states during a critical window of primate neurodevelopment, enabling mechanistic interrogation of how inflammatory, infectious, or hypoxic insults disrupt vulnerable neurogenic niches.

Influenza A virus (IAV) infection during pregnancy is associated with stillbirth and preterm birth, but the degree to which IAV alters placental and fetal immunity is poorly understood. The objective of our study was to determine the immunologic impact of maternal IAV infection on the placenta and fetus in a pigtail macaque (Macaca nemestrina) model. Pregnant pigtail macaques were inoculated with 10 7 plaque forming units (PFU) of IAV [A/California/07/2009 (H1N1)] and underwent necropsy 5 days post-infection (N=11). Results were compared to uninfected historical controls (N=16). IAV inoculation induced maternal pneumonia in all cases. Stillbirth occurred in 18% (2/11) of IAV-infected pregnancies, but not in controls. While vertical transmission was not observed, low-level IAV viral RNA was detected in two placentas. In the placenta, maternal IAV infection was associated with increased IL-1β, IL-18, and IFN-β levels, and an upregulated type I interferon (IFN) transcriptional response. IAV infection was also associated with significantly higher frequencies of intermediate and non-classical monocytes, plasmacytoid dendritic cells, CD4⁺ T cells, and NKT cells in the fetus (lung, lymph node, blood). Although placental immune and transcriptional perturbations were rarely correlated with maternal IAV disease indicators (e.g., maternal lung viral load/IFN-α/IFN-β/IL-6), there were consistent and significant correlations between these metrics and perturbed immune cell populations in the fetus (CD4+ and CD8+ T cells, plasmacytoid dendritic cells, monocyte sub-populations). Maternal IAV infection disrupted both placental and fetal immune environments, but only fetal immune alterations correlated with maternal lung disease severity.

The fetal origins of neuropsychiatric disorders are poorly understood but have been linked to viral or inflammatory injury of the developing brain. The fetal white matter is particularly susceptible to injury as myelination, axonal growth, and deep white matter tracts become established. We have used the pigtail macaque (Macaca nemestrina) to study the maternal and fetal effects of influenza A virus (FLUAV) and Zika virus (ZIKV) infection during pregnancy, in cohorts with different time intervals between inoculation and delivery. We observed a striking histopathological alteration in a subset of astrocytes which contained granular cytoplasmic inclusions (“inclusion cells”, ICs) within a specific region of the deep cerebral white matter in the fetal brains from specific FLUAV and ZIKV cohorts. Immunohistochemical and ultrastructural characteristics of ICs indicated that they are astrocytes (GFAP+) undergoing autophagocytosis (p62+) with activated lysosomes (LAMP1+, LAMP2+) and reactive changes in neighboring microglia. There was also a positive correlation between the number of ICs and LAMP1 or LAMP2 immunoreactivity in the fetal brain (LAMP1: rho 0.66; LAMP2: rho 0.54, p < 0.001 for both). Interestingly, ICs were significantly more prevalent in the 5-day FLUAV cohort and the 21-day intermediate ZIKV cohort than in controls (p < 0.005 and p = 0.04, respectively), but this relationship was not apparent in the ZIKV cohort with a shorter (2–3 days) or longer (months) time course. Virologic and immunologic assays indicated that the appearance of these cells was not linked with fetal brain infection. ICs were not observed in a macaque model of perinatal hypoxic ischemic encephalopathy. These alterations in fetal white matter are pathologically abnormal and may represent a transient neuropathologic finding that signifies a subtle brain injury in the fetus after maternal viral infection.

BackgroundPreterm birth is a leading cause of neonatal mortality, which is often complicated by intrauterine infection and inflammation. We have established a nonhuman primate model of Group B Streptococcus (GBS, Streptococcus agalactiae) infection-associated preterm birth. Immune checkpoints are modulators of the immune response by activating or suppressing leukocyte function and are understudied in preterm birth. The objective of this study was to spatially profile changes in immune protein expression at the maternal-fetal interface during a GBS infection with a focus on immune checkpoints.MethodsTwelve nonhuman primates (pigtail macaques, Macaca nemestrina) received a choriodecidual inoculation of either: 1) 1-5 X 108 colony forming units (CFU) of hyperhemolytic/hypervirulent GBS (GBSΔcovR, N=4); 2) an isogenic/nonpigmented strain (GBS ΔcovRΔcylE, N=4); or, 3) saline (N=4). A Cesarean section was performed at preterm labor or 3 days after GBS infection or 7 days after saline inoculation. Nanostring GeoMx® Digital Spatial Profiling technology was used to segment protein expression within the amnion, chorion, and maternal decidua at the inoculation site using an immuno-oncology panel targeting 56 immunoproteins enriched in stimulatory and inhibitory immune checkpoint proteins or their protein ligands. Statistical analysis included R studio, Kruskal-Wallis, Pearson and Spearman tests.ResultsBoth inhibitory and stimulatory immune checkpoint proteins were significantly upregulated within the chorioamniotic membranes and decidua (VISTA, LAG3, PD-1, CD40, GITR), as well as their ligands (PD-L1, PD-L2, CD40L; all p<0.05). Immunostaining for VISTA revealed positive (VISTA+) cells, predominantly in the chorion and decidua. There were strong correlations between VISTA and amniotic fluid concentrations of IL-1β, IL-6, IL-8, and TNF-α (all p<0.05), as well as maternal placental histopathology scores (p<0.05).ConclusionDifferential regulation of multiple immune checkpoint proteins in the decidua at the site of a GBS infection indicates a major perturbation in immunologic homeostasis that could benefit the host by restricting immune-driven pathologies or the pathogen by limiting immune surveillance. Protein expression of VISTA, an inhibitory immune checkpoint, was upregulated in the chorion and decidua after GBS infection. Investigating the impact of innate immune cell expression of inhibitory immune checkpoints may reveal new insights into placental host-pathogen interactions at the maternal-fetal interface.

Abstract Introduction The onset of puberty brings changes in sleep patterns in neurotypical school-aged children, shifting circadian timing and decreasing total sleep time. Sleep problems affect over 80% of school-aged autistic children. The predominant autism sleep phenotype involves longer sleep onset latency, night wakings, and early morning waking—defining features of insomnia. Research on the relationship of puberty to sleep problems, sleep duration, and sleep onset timing in autistic children is limited. Methods 232 school-aged children (age 6-13 years; Male n=142, Female n=90) participated in a sleep study through the longitudinal, multisite Infant Brain Imaging Study (IBIS) network. Participants were ascertained at 6-months-old for having an older autistic sibling (high-likelihood; HL) or no family history of autism (lower-likelihood; LL). The HL group included 47 diagnosed with autism (HL+), 104 without autism (HL-), and an LL comparison group of 81. The participants with actigraphy were categorized as Pre-puberty (n=89) or In-puberty (n=38) based on Tanner Staging. Measures include sleep problems (Children's Sleep Habits Questionnaire, CSHQ), sleep duration and sleep onset time (via 10 days actigraphy and sleep diary). Results Sleep problems were significantly increased in the HL+ group compared to HL- and LL groups (CSHQ; p< 0.01; χ2=23.736). Sleep onset times were later for the In-Puberty versus Pre-Puberty HL- group by 52.2 minutes (Kruskal-Wallis; p=0.01), and later, but not statistically significant, for the HL+ group by 43.2 minutes and LL group by 6.6 minutes. Sleep duration was lower, but not statistically significant, for the In-Puberty versus Pre-Puberty HL+ group by 36.6 minutes and HL- group by 37.2 minutes, and higher for the LL group by 3 minutes (ns). Conclusion Overall, parent-reported sleep problems were increased in school-age HL autistic children compared with HL- and LL children. Objective measures revealed In-Puberty HL+ and HL- had notably later sleep onset times and decreased sleep duration compared with Pre-Puberty. However, only HL- sleep onset time was significantly later. This pattern suggests sleep characteristics could be associated with familial autism and puberty. Future studies with a larger sample are needed to further evaluate the effect of puberty on sleep in autism. Support (if any) R01 HD101578

The coronavirus disease 2019 (COVID-19) pandemic exposed the vulnerability of pregnant women to excess morbidity and mortality, as well as the disproportionate disease burden in certain racial, ethnic, and sociodemographic groups. Vaccine hesitancy represents a major threat to public health, and crafting messages that reach vulnerable groups and address their intersectionality remains a weakness for pandemic preparedness. We sought to investigate factors that influenced vaccine acceptance and social media ad response in a mixed-methods study of Spanish-speaking women living in the rural Western United States who were pregnant or recently pregnant between November 2022 and June 2023. Direct interviews were translated, transcribed, and coded, while the ad ratings were analyzed using linear mixed models. Participants most favorably rated ads that featured doctors and text-heavy content describing benefits of vaccination. Qualitative data illustrated how information from trusted medical providers along with generational and cultural history of vaccine acceptance positively impacted perspectives on vaccination. Immigration status had varying influences on vaccination perspectives. Future vaccination campaigns targeting Spanish-speaking pregnant individuals in rural communities should use medical providers as ad messengers and dispel fears that vaccine acceptance may lead to problems with immigration status.

Auditory processing differences, including hyper- or hyposensitivity to sound, aversions to sound, and difficulty listening under noisy, real-world conditions, are commonly reported in autistic individuals. However, the developmental course and functional impact of these auditory processing differences are unclear. In this study, we investigate the prevalence, developmental trajectory, and functional impact of auditory processing differences in autistic children throughout childhood using a longitudinal study design. Auditory processing differences were measured using the Short Sensory Profile, a caregiver questionnaire, in addition to adaptive behaviors and disruptive/concerning behaviors at 3, 6, and 9 years of age. Our results showed that auditory processing differences were reported in greater than 70% of the autistic children in our sample at all three timepoints, maintained a high prevalence through 9 years of age, and were associated with increased disruptive/concerning behaviors and difficulty with adaptive behaviors. Furthermore, in our sample of children, auditory processing differences at age 3 years predicted disruptive/concerning behaviors and difficulty with adaptive behaviors at age 9 years. These findings warrant further investigations of the potential benefit of incorporating measures of auditory processing during routine clinical evaluations as well as interventions targeting auditory processing differences in autistic children.

This mixed-method study investigated vaccine hesitancy among pregnant women living in rural western United States and their response to social media ads promoting COVID-19 vaccine uptake. Thirty pregnant or recently pregnant participants who live in rural zip codes in Washington, Oregon, California, and Idaho were interviewed between November 2022 and March 2023. Interviews were transcribed and coded, while the ad ratings were analyzed using linear mixed models. The study identified five main themes related to vaccine uptake, including perceived risk of COVID, sources of health information, vaccine hesitancy, and relationships with care providers. Participants rated ads most highly that used peer-based messengers and negative outcome-based content. Ads with faith-based and elder messengers were rated significantly lower than peer messengers (p = 0.04 and 0.001, respectively). An activation message was also rated significantly less favorably than negative outcome-based content (p = 0.001). Participants preferred evidence-based information and the ability to conduct their own research on vaccine safety and efficacy rather than being told to get vaccinated. Primary concerns of vaccine-hesitant respondents included the short amount of time the vaccine had been available and perceived lack of research on its safety during pregnancy. Our findings suggests that tailored messaging using peer-based messengers and negative outcome-based content can positively impact vaccine uptake among pregnant women living in rural areas of the Western United States.

severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19 with a spectrum of disease outcomes in pregnancy and rare congenital infections. Viral tropism for cells in placental tissues is suggested to be low, but SARS-CoV-2 prevalence in placental tissues from women with COVID-19 is unknown. The placenta can produce a robust antiviral innate immune response, though it is unknown whether SARS-CoV-2 induces placental innate immunity and pathologic changes. Though SARS-CoV-2 detection in placental tissues is thought to be infrequent, immune factors associated with viral persistence are unknown. Placental tissues were collected from pregnant women, who were either healthy (N=20; 10 labored, 10 unlabored) or had active (N=53) or resolved COVID-19 disease (N=90) at delivery. Resolved COVID-19 disease was defined as delivery >10 days from symptom onset or diagnosis. We extracted RNA from chorionic villi (CV) and chorioamniotic membranes (CAM) for quantitative PCR for SARS-CoV-2 and a panel of interferon (IFN; ifnb), IFN stimulated genes (ISG; mxa, ifit1) and cytokines (il6). Placental pathology was evaluated in samples where SARS-CoV-2 RNA was detected. Statistical analysis included Student’s t-test and Fisher’s exact test. SARS-CoV-2 viral RNA (vRNA) was detected in placental tissues from 3.5% (5/143) of women with a history of COVID-19 (2 active, 3 resolved; 3 CAM, 3 CV) and 0% (0/20) in healthy controls. Median viral load was 1.0 x 107 copies/mg (range: 4 x 105 – 1 x 1011 copies/mg). An extremely high viral load (1.2 x 1011 copies/mg) was detected in CV 6 days after COVID-19 diagnosis in a patient with a placental basal infarct. The longest interval between diagnosis and SARS-CoV-2 vRNA detection in the placenta (CAM) was 17 weeks after COVID-19 diagnosis. In the CAM, innate immune gene expression was significantly higher in patients with either active or resolved COVID-19 versus controls (active: ifit1, mxa, il6; resolved: ifnb, ifit1, mxa, il6; all p<0.05). In CV, a more limited panel of innate immune genes was significantly elevated compared to controls (active: ifnb; resolved: ifnb and il6; all p<0.05). Notably, the case with detectable SARS-CoV-2 vRNA 17 weeks after COVID-19 diagnosis had very low il6 gene expression in the CV compared to controls. Our data suggest SARS-CoV-2 vRNA can rarely persist in the placenta up to 4 months after diagnosis. High viral loads may correlate with placental pathology. The persistence of the innate immune response, most prominent in CAM, suggests the placental antiviral response remains active for weeks to months after COVID-19. Further study is necessary to determine the impact of the placental innate immune response on viral clearance and whether a sustained elevation in innate immunity may impact fetal health.

Group B streptococci (GBS) are bacteria that can cause preterm birth and invasive neonatal disease. Heterogeneous expression of virulence factors enables GBS to exist as both commensal bacteria and to become highly invasive. A molecular epidemiological study comparing GBS bacterial traits, genotype and host characteristics may indicate whether it is possible to predict the risk of perinatal invasive GBS disease and more accurately target intrapartum antibiotic prophylaxis. A total of 229 invasive GBS isolates from Swedish pregnant women or neonates were assessed for virulence and phenotypic traits: hemolysis zone, hemolytic pigment (Granada agar), Streptococcus B Carrot Broth (SBCB) assay, CAMP factor, and hyaluronidase activity. Genes regulating hemolytic pigment synthesis ( covR/covS , abx1 , stk1 , stp1 ) were sequenced. Of the virulence factors and phenotypes assessed, a Granada pigment or SBCB score ≥ 2 captured more than 90% of EOD isolates with excellent inter-rater reliability. High enzyme activity of hyaluronidase was observed in 16% (36/229) of the invasive GBS isolates and notably, in one case of stillbirth. Hyaluronidase activity was also significantly higher in GBS isolates obtained from pregnant/postpartum individuals versus the stillbirth or neonatal invasive isolates (p < 0.001). Sequencing analysis found that abx1 (g.T106I), stk1 (g.T211N), stp1 (g.K469R) and covS (g.V343M) variants were present significantly more often in the higher (Granada pigment score ≥ 2) versus lower pigmented isolates (p < 0.001, each variant). Among the 203 higher Granada pigment scoring isolates, 22 (10.8%) isolates had 3 of the four sequence variants and 10 (4.9%) had 2 of the four sequence variants. Although heterogeneity in GBS virulence factor expression was observed, the vast majority were more highly pigmented and contained several common sequence variants in genes regulating pigment synthesis. High activity of hyaluronidase may increase risk for stillbirth and invasive disease in pregnant or postpartum individuals. Our findings suggest that testing for GBS pigmentation and hyaluronidase may, albeit imperfectly, identify pregnant people at risk for invasive disease and represent a step towards a personalized medical approach for the administration of intrapartum antibiotic prophylaxis.

Streptococcus agalactiae, or group B streptococcus (GBS), is an important pathogen for pregnant women and neonates, which can cause preterm birth, stillbirth, maternal sepsis and neonatal invasive disease. GBS are highly heterogeneous in their virulence factor expression and thus can exist as commensal bacteria or become highly invasive. In the U.S., universal screening for GBS is performed at 37-38 weeks gestation and women testing positive are treated with intrapartum antibiotic prophylaxis to prevent neonatal invasive disease. However, these measures do not prevent all GBS disease. If identification of specific GBS virulence factors correlates with invasive disease, their inhibition can prevent GBS perinatal disease and potentially reduce the use of antepartum antibiotics. The study objective was to determine if key virulence factors [e.g., hemolytic pigment (beta-hemolysin), CAMP factor, hyaluronidase] correlate with maternal and neonatal GBS invasive disease.

The influenza A virus (IAV) 2009 H1N1 pandemic was associated with an increased risk of maternal mortality, preterm birth, and stillbirth. The underlying mechanism for severe maternal lung disease and stillbirth is incompletely understood, but IAV infection is known to activate innate immunity triggering the release of cytokines. Elucidating the impact of progesterone (P4), a key hormone elevated in pregnancy, on the innate immune and inflammatory response to IAV infection is a critical step in understanding the pathogenesis of adverse maternal-fetal outcomes. IAV H1N1 pdm/09 was used to infect cell lines Calu-3 (lung adenoma) and ACH-3P (extravillous trophoblast) with or without P4 (100 nM) at multiplicity of infections (MOI) 0, 0.5, and 3. Cells were harvested at 24 and 48 hours post infection (hpi) and analyzed for cytopathic effects (CPE), replicating virus (TCID50), cytotoxicity (Lactate Dehydrogenase (LDH) assay), and NLRP3 inflammasome activation (caspase-1 activity, fluorometric assay). Activation of antiviral innate immunity was quantified (RT-qPCR, Luminex) by measuring biomarker gene and protein expression of innate immune activation (IFIT1, IFNB), inflammation (IL6), interferon signaling (MXA), chemokines (IL-8, IL-10). Both Calu-3 and ACH-3P were highly permissible to IAV infection at each timepoint as demonstrated by CPE and recovery of replicating virus. In Calu-3, progesterone treatment was associated with a significant increase in cytotoxicity, increased gene expression of IL6, and increased protein expression of IFN-β, IL-6, and IL-18. Conversely, in ACH-3P, progesterone treatment was associated with significantly suppressed cytotoxicity, decreased gene expression of IFNB, IL6 and IL1B, and increased protein expression of IFN-β and IL-6. In both cell lines, caspase-1 activity was significantly decreased after progesterone treatment, indicating NLRP3 inflammasome activation was not underlying the higher cell death in Calu-3. In summary, these data provide evidence that progesterone plays a dual role by ameliorating viral infection in the placenta but exacerbating influenza A virus-associated injury in the lung through nongenomic modulation of the innate immune response.

BACKGROUND:COVID-19 is caused by the SARS-CoV-2 virus and is associated with critical illness requiring hospitalization, maternal mortality, stillbirth, and preterm birth. SARS-CoV-2 has been shown to induce placental pathology. However, substantial gaps exist in our understanding of the pathophysiology of COVID-19 disease in pregnancy and the long-term impact of SARS-CoV-2 on the placenta and fetus. To what extent a SARS-CoV-2 infection of the placenta alters the placental antiviral innate immune response is not well understood. A dysregulated innate immune response in the setting of maternal COVID-19 disease may increase the risk of inflammatory tissue injury or placental compromise and may contribute to deleterious pregnancy outcomes. OBJECTIVE:We sought to determine the impact of a maternal SARS-CoV-2 infection on placental immune response by evaluating gene expression of a panel of 6 antiviral innate immune mediators that act as biomarkers of the antiviral and interferon cytokine response. Our hypothesis was that a SARS-CoV-2 infection during pregnancy would result in an up-regulated placental antiviral innate immune response. STUDY DESIGN:We performed a case-control study on placental tissues (chorionic villous tissues and chorioamniotic membrane) collected from pregnant patients with (N=140) and without (N=24) COVID-19 disease. We performed real-time quantitative polymerase chain reaction and immunohistochemistry, and the placental histopathology was evaluated. Clinical data were abstracted. Fisher exact test, Pearson correlations, and linear regression models were used to examine proportions and continuous data between patients with active (<10 days since diagnosis) vs recovered COVID-19 (>10 days since diagnosis) at the time of delivery. Secondary regression models adjusted for labor status as a covariate and evaluated potential correlation between placental innate immune gene expression and other variables. RESULTS:SARS-CoV-2 viral RNA was detected in placental tissues from 5 women with COVID-19 and from no controls (0/24, 0%). Only 1 of 5 cases with detectable SARS-CoV-2 viral RNA in placental tissues was confirmed to express SARS-CoV-2 nucleocapsid and spike proteins in syncytiotrophoblast cells. We detected a considerably lower gene expression of 5 critical innate immune mediators (IFNB, IFIT1, MXA, IL6, IL1B) in the chorionic villi and chorioamniotic membranes from women with active or recovered COVID-19 than controls, which remained significant after adjustment for labor status. There were minimal correlations between placental gene expression and other studied variables including gestational age at diagnosis, time interval between COVID-19 diagnosis and delivery, prepregnancy body mass index, COVID-19 disease severity, or placental pathology. CONCLUSION:A maternal SARS-CoV-2 infection was associated with an impaired placental innate immune response in chorionic villous tissues and chorioamniotic membranes that was not correlated with gestational age at COVID-19 diagnosis, time interval from COVID-19 diagnosis to delivery, maternal obesity, disease severity, or placental pathology.

Objective: This randomized, multisite, intent-to-treat study tested the effects of 2 levels of treatment intensity (number of hours) and 2 treatment styles on the progress of young children with autism spectrum disorder (ASD). We predicted that initial severity of developmental delay or autism symptoms would moderate the effects of intensity and style on progress in 4 domains: autism symptom severity, expressive communication, receptive language, and nonverbal ability. Method: A total of 87 children with ASD, mean age 23.4 months, were assigned to 1 of 2 intervention styles (naturalistic developmental/behavioral or discrete trial teaching), each delivered for either 15 or 25 hours per week of 1:1 intervention for 12 months by trained research staff. All caregivers received coaching twice monthly. Children were assessed at 4 timepoints. Examiners and coders were naive to treatment assignment. Results: Neither style nor intensity had main effects on the 4 outcome variables. In terms of moderating the effects of initial severity of developmental delay and of autism symptom severity, neither moderated the effects of treatment style on progress in any of the 4 domains. In terms of treatment intensity, initial severity moderated effect of treatment intensity on only 1 domain, namely, change in autism symptom severity; in a secondary analysis, this effect was found in only 1 site. Conclusion: Neither treatment style nor intensity had overall effects on child outcomes in the 4 domains examined. Initial severity did not predict better response to 1 intervention style than to another. We found very limited evidence that initial severity predicted better response to 25 vs 15 hours per week of intervention in the domains studied. Clinical trial registration information: Intervention Effects of Intensity and Delivery Style for Toddlers With Autism: https://clinicaltrials.gov/; NCT02272192

Group B streptococci (GBS) are bacteria that commonly reside in the female lower genital tract as asymptomatic members of the microbiota. However, during pregnancy, GBS can infect tissues at the maternal-fetal interface, leading to preterm birth, stillbirth, or fetal injury.

We found low prevalence of SARS-CoV-2 (2.7% [5/188]) among pregnant and postpartum patients with universal testing. Prevalence among symptomatic patients was similar under initial targeted screening (22.2% [4/18]) and universal approaches (19.1% [8/42]). Among 170 asymptomatic patients, 2 were positive or inconclusive, respectively; repeat testing at 24 hours was negative.