Adaptive immune responses to SARS-CoV-2 persist in the pharyngeal lymphoid tissue of children.

Most studies of adaptive immunity to SARS-CoV-2 infection focus on peripheral blood, which may not fully reflect immune responses at the site of infection. Using samples from 110 children undergoing tonsillectomy and adenoidectomy during the COVID-19 pandemic, we identified 24 samples with evidence of previous SARS-CoV-2 infection, including neutralizing antibodies in serum and SARS-CoV-2-specific germinal center and memory B cells in the tonsils and adenoids. Single-cell B cell receptor (BCR) sequencing indicated virus-specific BCRs were class-switched and somatically hypermutated, with overlapping clones in the two tissues. Expanded T cell clonotypes were found in tonsils, adenoids and blood post-COVID-19, some with CDR3 sequences identical to previously reported SARS-CoV-2-reactive T cell receptors (TCRs). Pharyngeal tissues from COVID-19-convalescent children showed persistent expansion of germinal center and antiviral lymphocyte populations associated with interferon (IFN)-γ-type responses, particularly in the adenoids, and viral RNA in both tissues. Our results provide evidence for persistent tissue-specific immunity to SARS-CoV-2 in the upper respiratory tract of children after infection.

Association of anosmia and neutralizing antibody production in adolescents with SARS-CoV-2.

BACKGROUND: To monitor olfactory/gustatory dysfunction and its relationship to SARS-CoV-2 IgG antibody responses in an adolescent population. METHODS: Adolescents with changes in olfactory/gustatory functions were enrolled in a 15-month study. The patients were evaluated with 1) SNOT-22, 2) pediatric smell wheel, and 3) SARS-CoV-2 antibody testing. The relationship between these scores and length of anosmia, and the amount of SARS-CoV-2 IgG antibodies were assessed. A brain MRI was performed in cases of persistent special sensory symptoms. RESULTS: Eighteen patients were identified with smell and/or taste complaints. Most of the patients were female (67%) and median age was 15 years (range 11-17). Twelve patients had prior SARS-CoV-2 PCR testing, with only five patients with a positive result. The median SNOT-22 score was 16 (range 0-52) and the median smell wheel score was 6.5 (range 1-11). Patients with taste difficulty were more likely to have a score less than eight. 78% of the patients tested positive for antibodies and there was a strong negative correlation between smell wheel score and antibody level (Spearman, ρ = -0.798, p = 0.002). Five patients underwent MRI scan, and all resulted as normal olfactory bulb structures. 66% received nasal corticosteroids. 11 patients presented in follow up. CONCLUSIONS: Adolescents presenting to a pediatric ENT clinic during the SARS-CoV-2 pandemic were likely to have prolonged (>6 weeks) symptoms of SARS-CoV-2. The majority do not report positive PCR testing result but do report systemic symptoms including anosmia. This suggests that anosmia may be both a late and prolonged symptom of SARS-CoV-2.

Examining multi-level immune response to determine prevalence of COVID-19 in pediatric tonsillectomy.

OBJECTIVE: To determine the prevalence of COVID-19 in a cohort of children undergoing tonsillectomy through assessment of B cell immune responses to SARS-CoV-2 in both peripheral blood and tonsil tissue. METHODS: In this cohort study at a tertiary pediatric hospital (Children's National Hospital) in Washington, DC, we recruited 100 children undergoing tonsillectomy from late September 2020 to January 2021. Serum, peripheral blood cells, and tonsil tissue were collected and examined for immune reactivity to SARS-CoV-2. Parent-reported clinical histories were compared to antibody and B-cell responses. RESULTS: Among 100 children undergoing tonsillectomy, 19% had evidence of immune responses to SARS-CoV-2 (CoV2+), indicating prior COVID-19. In all seropositive participants, we detected SARS-CoV-2 specific B cells in both peripheral blood mononuclear cells and tonsils, providing evidence for tissue-specific immunity in these children. Of the 19, 63% reported no known history of COVID-19, and an additional 3 were asymptomatic or unaware of an acute infection when detected on pre-surgery screen. Hispanic children represented 74% of CoV2+ subjects compared to 37% of the full cohort. 100% of CoV2+ children lived in a zip code with poverty level >10%. CONCLUSIONS: Nearly one-fifth of children undergoing tonsillectomy at an urban U.S. hospital had evidence of prior COVID-19 during the early pandemic, with the majority unaware of prior infection. Our results underscore the ethnic and socio-economic disparities of COVID-19. We found concordant evidence of humoral immune responses in children in both blood and tonsil tissue, providing evidence of local immune responses in the upper respiratory tract. LEVEL OF EVIDENCE: 3 Laryngoscope, 2022.