MILD SARS-COV-2 INFECTIONS AND NEUTRALIZING ANTIBODY TITERS
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Mild SARS-CoV-2 Infections and Neutralizing Antibody Titers Francesco Bonfante, DVM,a,* Paola Costenaro, MD, DTM&H,b,* Anna Cantarutti, PhD,c Costanza Di Chiara, MDb Alessio Bortolami, DVM, PhD,a Maria Raffaella Petrara, PhD,d Francesco Carmona, BSc,e Matteo Pagliari, PhD,a Chiara Cosma, MD,f Sandra Cozzani, MD,b Eva Mazzetto, DVM,a Giovanni Di Salvo, MD,g Liviana Da Dalt, MD,g Paolo Palma, MD,h,i Luisa Barzon, MD,j Giovanni Corrao,k,c Calogero Terregino, MD,a Andrea Padoan, MD,l Mario Plebani, MD,l,f Anita De Rossi, PhD,d,e Daniele Dona, MD, PhD,b Carlo Giaquinto, MDb Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 BACKGROUND: Recent evidence suggests that neutralizing antibodies (nAbs) to severe acute abstract respiratory syndrome coronavirus 2 may persist over time; however, knowledge regarding pediatric subjects is limited. METHODS:A single-center, prospective observational study was conducted on 57 family clusters of coronavirus disease 2019, including children of neonatal and pediatric age attending the University Hospital of Padua (Italy). For each patient, blood samples were collected for both the quantification of nAbs through a plaque reduction neutralizing test and the detection of antinucleocapsid-spike protein immunoglobulin G and/or immunoglobulin M. RESULTS:We analyzed 283 blood samples collected from 152 confirmed coronavirus disease 2019 cases (82 parents and 70 children or older siblings of median age of 8 years, interquartile range: 4–13), presenting asymptomatic or with mildly symptomatic disease. Despite the decrease of immunoglobulin G over time, nAbs were found to persist up to 7 to 8 months in children, whereas adults recorded a modest declining trend. Interestingly, children aged
European countries have been facing naturally acquired nAbs among a Data Collection and Definitions a third wave of the novel cohort of family clusters of COVID- Information collected during the coronavirus disease 2019 (COVID- 19, including adults and children clinic was entered into a Web-based 19) pandemic and the spread of who recovered from asymptomatic database by using the Research several severe acute respiratory or mild symptomatic infections. Electronic Data Capture platform syndrome coronavirus 2 (SARS-CoV- (Vanderbilt University, Tennessee) 2) variants. With the advent of METHODS hosted in the server of the vaccines,1 longitudinal studies of University of Padova. For this both convalescent and vaccinated Study Design and Population study, data were collected patients are of fundamental A single-center, prospective study retrospectively from the existing importance to understand the Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 was conducted on Italian family clinical files and analyzed kinetics of humoral response and clusters of COVID-19 attending the anonymously. Subjects were infer correlates of protection for COVID-19 Family Cluster Follow-up considered patients with confirmed both infection and disease. In this Clinic, at the Department of COVID-19 if they had a record of respect, the titration of neutralizing virological positivity for SARS-CoV- Women’s and Children’s Health of antibodies (nAbs) is key to the University Hospital of Padua 2 by real-time polymerase chain determine the concentration of (Veneto Region, Italy). From March reaction (RT-PCR) according to antibodies preventing cells to be 1, 2020 to September 4, 2020, 57 routine diagnostic molecular infected by SARS-CoV-2.2 families were enrolled meeting the protocols16 and/or resulted following inclusion criteria: (1) positive by either of the 2 Studies including convalescent having children of pediatric age serological tests adopted in this adults reported that humoral (aged 18 years, including older adolescents are key in the confirmed SARS-CoV-2 family siblings and parents, and legally outbreak (Supplemental Fig 6). transmission of infection.15 Little is authorized representatives of Subjects who were asymptomatic known about the kinetics of SARS- subjects aged
(>15 years). These age classes samples, stratified by age classes validation of immunoassays for were deemed instrumental for a and by time between serological SARS-CoV-2, we calculated measures translation of results into the sampling and baseline, categorizing of diagnostic accuracy of the CLIA context of school-targeted subjects into 3 intervals, namely 1 assay. vaccination and sero-surveillance to 2, 3 to 6 and 7 to 8 months. The campaigns. 1-way analysis of variance and the Analyses were performed by using independent samples t test were the Statistical Analysis System Serological Assays performed, when appropriate. software (version 9.4; SAS Institute, Plasma was stored at 80 C before Associations between antibody Inc, Cary, NC). Statistical testing for the quantification of titers, baseline intervals and age, significance was set at the .05 level. nAbs through a high-throughput were assessed with linear All P values were 2-sided. Graphs Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 method for plaque reduction regression models. Strength of were made by using GraphPad neutralizing test (PRNT).18 Another associations between variables was Prism version 9 (GraphPad aliquot was analyzed with the assessed by Pearson correlation Software, Inc, La Jolla, CA). chemiluminescence immunoassay coefficient by using the logarithm (CLIA) MAGLUMI 2019-nCoV (base 10) of the antibody titers, RESULTS Immunoglobulin M (IgM) and given data skew. From March 1, 2020, to December Immunoglobulin G (IgG).18 Further 3, 2020, we prospectively evaluated Use of the robust variance details on the 2 assays are 57 family clusters of COVID-19 estimator to account for reported in the Supplemental (Supplemental Fig 5). A serological correlations within patients with Information. assessment was performed at least multiple blood samplings did not once on 209 recruited subjects. SARS-CoV-2 Viral Load Measurement change the CIs considerably in the Subjects who had previously tested unadjusted analyses, so correlation A selection of NPSs of enrolled positive for SARS-CoV-2 by real- structures were omitted from all subjects that had been originally time RT-PCR (111 of 209) were analyses. Among a subcohort of screened at the Padova University considered to have confirmed subjects that agreed to be sampled Hospital were made available for COVID-19, together with individuals again after enrollment, a quantification of the viral load. who had no record of virological dependent t test for subject-paired Copies of SARS-CoV-2 were positivity but showed evidence of samples was used to compare the quantified by a homemade multiplex seropositivity by either of the 2 GMT and 95% CI. quantitative assay on the basis of a serological tests adopted in this 1-step digital droplet polymerase To test the robustness of our data study (44 of 209). Descriptive chain reaction (ddPCR).19 Results sets against selection bias, we analysis and additional information were expressed as SARS-CoV-2 conducted a x2 test and verified on baseline identification are copies per 5 ml. Further details are the homogeneity within each age provided as Supplemental reported in the Supplemental class and time window of (1) the Information (Supplemental Table 2, Information. temporal distribution of serological Supplemental Fig 6). Three out of samplings (P 5 .4363) and (2) the 73 children were excluded from the Statistical Analyses analyses (see Supplemental Fig 5). proportion of cases identified by Descriptive statistics were used virological or serological methods In total, 152 confirmed COVID-19 for comparing the distribution of (P 5 .6568). Moreover, we cases were studied: 70 children or sex, age, disease-related conducted a x2 test to verify older siblings and 82 parents with symptoms, and pediatric among subjects who contributed median ages of 8 (interquartile comorbidities between patients with either 1, 2, or 3 samples the range [IQR], 4–13) and 42 years infected with COVID-19 and homogeneity of sex (P 5 .6082), (IQR, 34–46), respectively. Of 152 uninfected patients. age (P 5 .0973), family position cases, 38, 97, and 17 were sampled (P 5 .3971) and severity of once, twice and 3 times, The humoral response was symptoms (P 5 .6947). respectively. assessed by comparing the geometric mean titer (GMT) and The diagnostic sensitivity of the Analyzing all 283 blood samples the 95% confidence interval (CI) of CLIA and PRNT assays were collected from confirmed COVID-19 IgM, IgG, and PRNT50 values in the assessed on subjects with a positive cases, we observed that nAbs overall data set, including both NPS result. Considering the PRNT persisted in the population, independent and subject-paired assay as reference method for the (Fig 1A) recording a modest PEDIATRICS Volume 148, number 3, September 2021 3
Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 FIGURE 1 Stability of SARS-CoV-2 nAb titers over time. A, PRNT50 titers from 283 serum samples collected at a median time of 132 days (IQR, 79–187) from infection onset, overall and stratified by 3 age classes, including children aged
Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 FIGURE 3 Performance of SARS-CoV-2 CLIA IgG and PRNT titers over time. A, Decreasing levels of SARS-CoV-2 CLIA IgG levels observed for all classes of age (aged
Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 FIGURE 4 nAb titers according to COVID-19 disease severity. A, Clinical presentation of COVID-19 in children aged
TABLE 1 Subject-Paired Serological Data of 76 Subjects Who Were Sampled Twice at Periods of 72 Days (SD ±22) and 169 Days (SD ±26) From Baseline and Data From 50 Subjects, for Whom Paired Samples Were Available at 99 Days (SD ±35) and 234 Days (SD ±10) From Baseline A B a First Sample Second Sample (5–6 mo) P First Sample Latest Sample (7–9 mo) Pa Aged
clusters of COVID-19. In our immunodominant proteins, blunting decrease between 3 and 6 months cohort, antibodies neutralizing the neutralizing potential.26 from infection for mild cases. In SARS-CoV-2 virus persisted over a Recently, this mechanism has been addition, Chia et al9 identified 5 period of 2 to 8 months from explored for influenza, proving that profiles of antibody responses and infection, recording only a modest children aged
19,32 analyses confirmed our shedding and transmission. It is conclusions against the exclusion of the utmost importance to identify ABBREVIATIONS of few cases. age- and time-matched correlates of CI: confidence interval protection to finally translate CLIA: chemiluminescence In the absence of correlates of serological data into useful elements immunoassay protection for nAbs acquired after for the design of vaccines and COVID-19: coronavirus disease infection, it is not advisable to immunization campaigns for SARS- 2019 translate our data into predictions CoV-2. ddPCR: digital droplet of a superior immunity of children polymerase chain to reinfection. According to clinical ACKNOWLEDGMENTS reaction studies and experimental animal We are grateful to Dr Franco Pisetta GMT: geometric mean titer Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 work, superior nAbs for SARS-CoV-2 and to all the family pediatricians of IgG: immunoglobulin G might translate into protection from Veneto Region for collaborating in IgM: immunoglobulin M COVID-19 disease and higher viral this study. Moreover, we are grate- IQR: interquartile range clearance in the upper respiratory ful to all families that participated nAb: neutralizing antibody tract, leading to a reduction in to the study. NPS: nasal-pharyngeal swab PRNT: plaque reduction neutralizing test RT-PCR: real-time polymerase chain reaction SARS-CoV-2: severe acute respiratory syndrome coronavirus 2 PEDIATRICS (ISSN Numbers: Print, 0031-4005; Online, 1098-4275). contributed to the writing and supervised the project; Drs Palma, Da Dalt, Di Salvo, and Terregino supervised the project; and all authors approved the final manuscript as submitted. DOI: https://doi.org/10.1542/peds.2021-052173 Accepted for publication Jun 14, 2021 Address correspondence to Francesco Bonfante, DVM, Istituto Zooprofilattico Sperimentale delle Venezie, viale dell’Universita 10, 35020 Legnaro, Padua, Italy. E-mail: FBonfante@izsvenezie.it PEDIATRICS (ISSN Numbers: Print, 0031-4005; Online, 1098-4275). Copyright © 2021 by the American Academy of Pediatrics FINANCIAL DISCLOSURE: The authors have indicated they have no financial relationships relevant to the article to disclose. FUNDING: Partially supported by the ORCHESTRA project, Horizon 2020 research and innovation program under grant agreement 101016167 to Dr Giaquinto. In addition, this work has received funding from the EU Horizon 2020 (RECOVER) under grant agreement 101003589 to Dr Giaquinto, and Fondazione Cassa di Risparmio di Padova e Rovigo, Progetti di Ricerca COVID-19 (Dr De Rossi participant). POTENTIAL CONFLICT OF INTEREST: The authors have indicated they have no potential conflicts of interest to disclose. COMPANION PAPER: A companion to this article can be found online at www.pediatrics.org/cgi/doi/10.1542/peds.2021-052173. REFERENCES 19: the dose matters. EClinicalMedicine. 4. Seow J, Graham C, Merrick B, et al. Lon- 1. Hodgson SH, Mansatta K, Mallett G, Harris 2020;26:100545 gitudinal evaluation and decline of anti- V, Emary KRW, Pollard AJ. What defines an 3. Moshe M, Brown JC, Flower B, et al. body responses in SARS-CoV-2 infection efficacious COVID-19 vaccine? A review of Declining prevalence of antibody positiv- [published online ahead of print July the challenges assessing the clinical effi- ity to SARS-CoV-2: a community study of 11, 2020]. medRxiv. doi:10.1101/ cacy of vaccines against SARS-CoV-2. Lan- 365,000 adults [published online ahead 2020.07.09.20148429 cet Infect Dis. 2021;21(2):e26–e35 of print October 27, 2020]. medRxiv. 5. Ibarrondo FJ, Fulcher JA, Goodman- 2. Verkerke HP, Maier CL. Towards charac- doi:https://doi.org/10.1101/2020.10.26. Meza D, et al. Rapid decay of anti-SARS- terized convalescent plasma for COVID- 20219725 CoV-2 antibodies in persons with mild PEDIATRICS Volume 148, number 3, September 2021 9
covid-19. N Engl J Med. 2020; in Wuhan: a retrospective observational infection in hospitalized pediatric and 383(11):1085–1087 study. Lancet Infect Dis. 2021;21(5): adult patients. Sci Transl Med. 6. Ripperger TJ, Uhrlaub JL, Watanabe M, 617–628 2020;12(564):eabd5487 et al. Orthogonal SARS-CoV-2 serological 16. Lavezzo E, Franchin E, Ciavarella C, et al; 25. Weisberg SP, Connors TJ, Zhu Y, et al. assays enable surveillance of low-preva- Imperial College COVID-19 Response Distinct antibody responses to SARS- lence communities and reveal durable Team. Suppression of a SARS-CoV-2 out- CoV-2 in children and adults across the humoral immunity. Immunity. break in the Italian municipality of Vo’. COVID-19 clinical spectrum. Nat Immu- 2020;53(5):925–933.e4 Nature. 2020;584(7821):425–429 nol. 2021;22(1):25–31 7. Wajnberg A, Amanat F, Firpo A, et al. 17. World Health Organization. Clinical 26. Kim JH, Skountzou I, Compans R, Jacob Robust neutralizing antibodies to Management of COVID-19: Interim J. Original antigenic sin responses to SARS-CoV-2 infection persist for influenza viruses. J Immunol. Guidance, May 27, 2020. Geneva, Downloaded from http://publications.aap.org/pediatrics/article-pdf/148/3/e2021052173/1227777/peds_2021052173.pdf by guest on 11 January 2022 months. Science. 2020;370(6521): 2009;183(5):3294–3301 Switzerland: World Health Organiza- 1227–1230 tion; 2020 27. Meade P, Kuan G, Strohmeier S, et al. 8. Isho B, Abe KT, Zuo M, et al. Persistence Influenza virus infection induces a nar- 18. Padoan A, Bonfante F, Pagliari M, et al. of serum and saliva antibody responses row antibody response in children but Analytical and clinical performances of to SARS-CoV-2 spike antigens in COVID-19 a broad recall response in adults. five immunoassays for the detection of patients. Sci Immunol. 2020;5(52):1–21 MBio. 2020;11(1):e03243-19 SARS-CoV-2 antibodies in comparison 9. Chia WN, Zhu F, Ong SWX, et al. Dynam- with neutralization activity. EBioMedi- 28. Selva KJ, van de Sandt CE, Lemke MM, ics of SARS-CoV-2 neutralising antibody cine. 2020;62:103101 et al. Systems serology detects func- responses and duration of immunity: a tionally distinct coronavirus antibody 19. Cotugno N, Ruggiero A, Bonfante F, et longitudinal study. Lancet Microbe. features in children and elderly. Nat al; CACTUS Study Team. Virological and 2021;2(6):e240–e249 Commun. 2021;12(1):2037 immunological features of SARS-CoV-2- 10. Lau EHY, Tsang OTY, Hui DSC, et al. Neu- infected children who develop neutraliz- 29. Westerhuis BM, Aguilar-Bretones M, tralizing antibody titres in SARS-CoV-2 ing antibodies. Cell Rep. Raadsen MP, et al. Severe COVID-19 infections. Nat Commun. 2021;12(1):63 2021;34(11):108852 patients display a back boost of sea- 11. Edridge AWD, Kaczorowska J, Hoste 20. Garcia-Beltran WF, Lam EC, Astudillo MG, sonal coronavirus-specific antibod- ACR, et al. Seasonal coronavirus protec- et al. COVID-19-neutralizing antibodies ies [published online ahead of print tive immunity is short-lasting. Nat Med. predict disease severity and survival. October 12, 2020]. medRxiv. 2020;26(11):1691–1693 Cell. 2021;184(2):476–488.e11 doi:https://doi.org/2020.10.10. 20210070 12. Liu W, Fontanet A, Zhang PH, et al. Two- 21. Ripperger TJ, Uhrlaub JL, Watanabe M, year prospective study of the humoral et al. Detection, prevalence, and dura- 30. Bolotin S, Tran V, Osman S, et al. SARS- immune response of patients with tion of humoral responses to SARS-CoV- CoV-2 seroprevalence survey estimates severe acute respiratory syndrome. 2 under conditions of limited popula- are affected by anti-nucleocapsid anti- J Infect Dis. 2006;193(6):792–795 tion exposure [published online ahead body decline. J Infect Dis. of print August 15, 2020]. medRxiv. 2021;223(8):1334–1338 13. Dong Y, Mo X, Hu Y, et al. Epidemiology doi:10.1101/2020.08.14.20174490 31. Long QX, Tang XJ, Shi QL, et al. Clinical of COVID-19 among children in China. and immunological assessment of 22. Gudbjartsson DF, Norddahl GL, Melsted Pediatrics. 2020;145(6):e20200702 asymptomatic SARS-CoV-2 infections. P, et al. Humoral immune response to SARS-CoV-2 in Iceland. N Engl J Med. Nat Med. 2020;26(8):1200–1204 14. Di Nardo M, van Leeuwen G, Loreti A, et al. A literature review of 2019 novel 2020;383(18):1724–1734 32. Corbett KS, Flynn B, Foulds KE, et al. coronavirus (SARS-CoV2) infection in 23. Yang HS, Costa V, Racine-Brzostek SE, et Evaluation of the mRNA-1273 vaccine neonates and children. Pediatr Res. al. Association of age with SARS-CoV-2 against SARS-CoV-2 in nonhuman pri- 2021;89(5):1101–1108 antibody response. JAMA Netw Open. mates. N Engl J Med. 2021;4(3):e214302 2020;383(16):1544–1555 15. Li F, Li YY, Liu MJ, et al. Household transmission of SARS-CoV-2 and risk 24. Pierce CA, Preston-Hurlburt P, Dai Y, et factors for susceptibility and infectivity al. Immune responses to SARS-CoV-2 10 BONFANTE et al
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