COMMUNICATIONS ARTICLE B crete OPEN Immune responses to SARS-CoV-2 in three children of parents with symptomatic COVID-19 Shidan Tosif@ '23"4™, Melanie R, Neeland@ '2"4, Philip Sutton@ ', Paul V. Licciardi@ "2, Sohinee Sarkar® ', Kevin J, Selva’, Lien Anh Ha Dol, Celeste Donato', Zheng Quan Toh@ '?, Rachel Higgins”, Carolien Van de Sandt@ *°, Melissa M. Lemke® ®, Christina Y. Lee® °, Suzanne K. Shoffner® ®, Katie L. Flanagan@ 75, Kelly B. Arnold@ ©, Francesca L. Mordant® 4, kim Mulholland!2", Julie Bines'*"2, Kate Dohle?, Daniel G. Pellicci'?, Nigel Curtis!2", Sarah McNab!23, Andrew Steer!2", Richard Saffery® '2, Kanta Subbarao® *, Amy W. Chung® 4, Katherine Kedzierska@ 4, David P. Burgner@ }2™5 & Nigel W. Crawford!2315 ‘Compared to adults, children with severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2) have predominantly mild or asymptomatic infections, but the underlying immunological differences remain unclear. Here, we describe clinical features, virology, longitudinal cellular, and cytokine immune profile, SARS-CoV-2-specitic serology and salivary antibody responses in a {amily of two parents with PCR-confirmed symptomatic SARS-CoV-2 infection and their three children, who tested repeatedly SARS-CoV-2 PCR negative. Cellular immune profiles and cytokine responses of all children are similar to their parents at all timepoints. All family members have salivary anti-SARS-CoV-2 antibodies detected, predominantly igA, that coincide with symptom resolution in 3 of 4 symptomatic members. Plasma from both parents and one child have IgG antibody against the S1 protein and virus-neutralizing activity detected. Using a systems serology approach, we demonstrate higher levels of SARS-CoV-2-spectic antibody features of these family members compared to healthy controls. These data indicate that chiidren can mount an immune response to SARS-CoV-2 without virological confirmation of infection, raising the possibilty that immunity in children can prevent the establishment of ‘SARS-CoV-2 infection. Relying on routine virological and serological testing may not identify ‘exposed children, with implications for epidemiological and clinical studies across the life-span "Department of Paediatris, The University of Melbourne, Meboune, Victoria, Australia. Infection and immursty, Murdoch Children's Research institute, Melbourne, Australia. “Department af Genera! Medicine, The Royal Chidter’s Hospital Melbourne, Victoria, Australia. “Department of Microbiology and Immunology, Peter Doherty nsttute for infection and immunity, The Unversity of Melbourne, Melbourne, Victoria, Austra. ® Deparimento Hematopciess, Sangun Research and Landsener Laboratory, Amsterdam UMC, Unvesty af Amsterdam, Amsterdam, Netherlands, © Department of Biomedical Engineering, University of Michigan, Ml, USA. Department of infectious Diseases, Launceston General Hospital Launceston Tasman, ‘Astala, Schoo of Heath Sciences and Scho! of Medicine University of Tasmania, Launceston, Tasmania, Austalia,° Department of Irmunokony and Pathology, Monash University, Commercial Road, Melbourne, Victoria, Austra, "School of Heath and Biomedical Science, RMIT University, Meboume, Victoria, Australa, Mnfectous Diseases Unit, Department of General Medicine, The Royal Chile's Hospital, Melbourne, Austra Deparment of Gastroenterology, The Royal Childrens Hospital Melbourne, Victora, Australa. "WHO Collaborating Centre for Reference and Resarch on Ivluenza, Melbourne, Australia, “These authors contbuteé equal Shan Tos, Melanie Neeland, "These authors oly supenised this wok: David P. Burner, Nigel W. Craword. emai Shida Tostarchorgau NATURE COWMLNICATIONS| cnz0>Ts703) s/o /101038/st467-02019548 8 wu ature con/natneconmictons 1ARTICLE NATURE COMMUNICATIONS | htips/dotorg/10:1038/241467-020 -19545-8 ‘ date, children represent a small proportion of SARS-CoV- 2 confirmed coronavirus disease (COVID-19) eases!’ ‘Children are predominantly infected from symptomatic household adult contacts. Children have comparatively milder COVID-19 disease and up to one-third are asymptomatic’. The immunological bass for miller pediatric disease is unclear, but may be relevant to other viral pandemics where striking age-related «epidemiological differences were observed. In SARS-CoV-2 infec- tion, reduced respiratory epithelial expression ofthe ACE2 receptor and trained innate immunity in children have been proposed. Investigating immune responses to SARS-CoV-2 across all age groups is key to understanding disease susceptibility, severity determinants, and vaccine candidates. Detailed investigations of immune responses during SARS-CoV-2 infection have been reported in adults, with exposure to SARS-CoV-2 causing specific T cell responses without seroconversion" Data on immune responses in children exposed to SARS-CoV-2 ae limited. Here, we show that three children repeatedly exposed to SA CoV-2 in their household mount cellular and antibody-mediated immune responses similar to their infected parents and specific to SARS-CoV-2, without virological confirmation of infection. Results Patient Characteristics. Two parents (mother 38. years, and father 47 years) residing in Melbourne, Australia, attended a 3- hour wedding inter-state without their children in early March 2020. ‘They returned home 3-days later and developed cough, coryza, and subjective fevers, followed by lethargy and headache fora total of 14 (mother, Al) and 11 days (father, A2) (Fig 1) Seven days after the onset of the parents’ symptoms, child’ one {onale 9 years, C1) developed a mild cough, coryza, sore throat, atxdominal pain, and loose stools, and child 2 (male 7 years, 2) developed mild cough and coryza.Thethied hid (female 5 years, (C3) was asymplomati. Eight days afler the onset of the parents? symptoms, they were noted of an emerging outbreak of SARS- CoV"? traced to the wedding. The parents were SARS-CoV-2 PCR postive on nasopharyngeal (NP) swabs taken the same day. Repeated NP swabs from the children were negative for SARS- CoV-2. Physical distancing precautions were not feasible in the household. Child 3 had particularly clase contact, sloping inthe parents’ bed throughout the period both parents were unwell. All family members recovered fully without requiring medical care SARS-CoV-2 testing. Serial samples, including blood, saliva, NP swabs, feces, and urine, were collected from all family members approximately every 2-3 days (Fig. 1). Dally symptoms were recorded ina standardized diary. Nasopharyngeal swabs from the parents on days 8 and 12 were SARS-CoV-2 PCR positive. All INP, saliva, and stool samples from the children were PCR negative for SARS-CoV-2. Nasopharyngeal swabs from the chil- dren were all positive for shinovirus by a multiplex respiratory viral panel on day 10, Children and parents show an active cellular immune response. ‘We investigated the callular immune response in peripheral blood ‘mononuclear cells (PBMCs) from all family members on days 12, 37, and 88 by flow cytometry. Both parents and children had high proportions of CDS ells at day 12 that subsequently decreased (ig 2a), a decline associated with a corresponding increase in the proportion of CD4 T eels in all samples. Strikingly low proportions ‘of monocytes were observed on day 12 in all family members, particularly in C3 (0.12%) relative to her siblings (average 0.5%) and parents average 088%) (Fig, 2a), Monocytes returned to circulating proportions inal family members by day 37 (average 4.1%) and day 88 (average 2.5%). These signatures were also identified by unsu- pervised tistibuted stochastic neighbor embedding (SNE) dimensionality reduction, where (SNE clusters corresponding to €DB "T, CDA, and monocytes in parents and children showed identical sequential changes to those observed by manual gating (ig. 2b). Low proportions of monocytes were observed in all ce- culating subsets with reductions in CD16" subsets most evident (ig. 20. Both parents showed increases in central (Tesy) and effector (Tz) memory CD8'T cells by day 88 (Fig, 2d), and CD8 T cell expression of the exhaustion marker PD increased in ll family ‘members over time (Fig, 2e). CD4 py cells reduced overtime in the parents, and one parent (A2) had a marked dectne in the CD4 effector (Tiea) cell population (Fig, 2). The heterogeneous cellular immune responses observed in all family members at the fist timepoint are consistent with emerging evidence on SARS-CoV2 infection in als, including broad changes in the frequency, phe- notype, and activation status of CD8 and CD4 T cells. Depletion of innate immune cll subs including monocytes (partly ‘CDI6* subsets) has aso been observed in COVID-19! ‘A study comparing the blood and lung profiles of patients with severe COVID-19 revealed that non-classical monocytes prefer entially migrate from the blood into the lungs during disease! ‘We observed further alterations in the myeloid compartment in ‘our whole blood analysis. Low proportions of neutrophils were evident in all family members at day 12, particularly in C3 (6.1%) relative to her siblings (average 104%) and parents (average 15.5%) (Fig. 2). Circulating neutrophils returned to an average of 30.3% in children and 45.4% in parents by day 88, atime point associated with the appearance of low-density immature neutrophils (SSCNCD16"CD14!/-) in PBMCs of all family ‘members (Fig, 2, h). Pre- and immatuse-neutrophils in PBMC fractions have been recently described in SARS-CoV-2 infected adults!”, In our study, parent AT and all children had high proportions of eosinophils at alltime points (Fig. 2), in keeping Fig. 1 Timeline of travel, exposure, symptoms, and selected results. Nasopharyngeal PCR, saliva, an serum antibodies are show fr each parent and child, Key events in the timeline are highlighted according to anumber of days falling the return of parents to the household 2 NATURE COWMUNICATIONS| ngoyTs703)hps/do/101038/st467-020-19545 8 wate con/natnecanmictonsNATURE COMMUNICATIONS | htips//doiorg/101038/541467-02019545-8 ARTICLE Fig. 2 Longitudinal celluiar immune profiling in parents and children. a Major immune coll populations in PBMC at day 12,37, ad 8 in parents (soi line) and children (broken ine) (A (closed cices), A2 (lose squares), C (open ccles), C2 (open squares), C3 (open tangles). B SNE dimensionality reduction of immune cell populations in all PBMC sample across the three-time points. The tSNE pot was generated from a concatenated file containing 300,000 events (20,000 randomly selected lve singe cells per patient pe ime point). € Fraquency of monacye subpopulations in PBMC from parents and chien. d Frequency of CDS T coll nave, efector, and memory subpopulations in PBMC. e Frequency of PD! expressing C8 T cells overtime { Frequency of CDA T cel nave, efector, nd memory subpopulation in PENIC.g tSNE dimensionality reduction of whole blood samples. The ¢SNE plat was generated from a concatenated fle containing 300,000 events (20,000 randomly selected Ive single calls per patient per time point). Coloring epics SSC and CDI6 expression in SNE islands. Granulocyte populations (neutrophils and eosinophils) ave expressed as the proption of leukocytes. IhFrequency of low-density CDI6"SSC neutrophils (CD14 and CD14 in PBMC fraction at day 88.1 Plasma cytokine concentration of tee detectable cytokines, RANTES (blue, MCP-I(pucple), and IL-8 (green in children and parents a day 12 and 37. in SARS-CoV-2 infected patients during tole remains unclear ‘Our analyses highlighted that active cellular immune responses in the family members were not accompanied by a coresponding increase in’ plasma cytokine levels, consistent with mild oF absence of symptoms. We quantified 18 plasma cytokines using a custom multiplex bead array and only IL-8, MCP-1, and CCLS (RANTES) were detectable (Fig. 24), with levels remaining constant over time, excluding C1 and C2 who had a ~2-fold increase in RANTES levels at day 37 (Fig, 2). case of mild adult COVID-19 disease reported an identical plasma. cytokine signature to that observed in our family members'2. RANTES NATURE COWMLNICATIONS| cnz0>Ts703) s/o /101038/st467-02019548 8 wu ature con/natneconmictons aARTICLE NATURE COMMUNICATIONS | htips/dotorg/10:1038/241467-020 -19545-8 Fig. 3 Salivary and plasma antibody responses against SARS-CoV-2 SI protein by ELISA and by microneutralization assay. Ant-SI saivary IgA, 1, and IgM. # Ig ant response that developed concurrent with resolution of symptoms. b Ant plasma IgA, IgG, and IgM. € Neutvalzing antibody activity in plasma, A: mother, A2 father, CI: mae (9 years, C2: male (7 years), C3: female yeas), (P) positive cote hha also recently been shown to be elevated in patients with both mild and severe COVID-19. In critically ill patients, blocking this pathway with a CCR5-targeted monoclonal antibody resulted in the reortion of T cell counts and reduced SARS-CoV-2 viral load SARS-CoV-2 antibodies in the saliva of ll family members. To explore SARS.CoV-2 specific humoral immune responses, we first quantified salivary and plasma antibodies against the SI protein by ELISA. Saliva from all family members tested postive for IgA antibodies against the St protein at all timepoints (Gig. 5), A2 had high levels of salivary ant-S1 IgA at day 12, one Ts703) s/o /101038/st467-02019548 8 wu ature con/natneconmictons 5ARTICLE NATURE COMMUNICATIONS | htips/dotorg/10:1038/241467-020 -19545-8 ‘rely, 96. wel igh binding plats (Thermo Fsber Scenic) were coated wit ‘oc RMD (Sin Boga citd n PRS st gland then incobated a °C ‘remit The flowing dy plates were ached with PBS contain 01% (ee) Tween (PES) and Noche with PHS contig 1% Tween and 10% (w¥) sim mil (285-19) for tro temperate RT), Srl itn (1) of asa samples were prepared in PHS TSM stating at 10 pose cont {omlecent samp and nave conta (pre panei) sample wee st all ‘saps The Hocking ation nas semoved nd 100 lo each sil ton wae de to the pate for 2h at RT. The plats were then washed thre times with 20 per wel of PBST Goat an: human Tg (10000) 9 IM (000) omerash persis (HRD) conjugated secondary antibody (Souther Bete) seas prepared in PRS-TSM, and 0 jf this secondary atibesy was aed each, srl oe For IgA of bate (SI) as sed in PBST and {Med wo each wl for 1, flowed by the don of Suepavidn-HAP to each tre for 20min Pes were washed wth PAST flamed by dated water and [Soy of 337 55"tcrmeybereidne (TMS, Sera Car) subsite solton was ‘ded fr 9 in. The reaction was topped by headin of OL 1M. Shonphorc ci ad opal denis rune wing a microplate ede (io ‘eA a 450 am (60 rm eetence ite doin ters wee eluted folowing tackgiound soneton of the respcine egal conte ea nea ay. Salva $1 protein ELISA Silva pooled unde the tongue was done int a0 mL. fue and stored at 80°C antl anal mano Msasorp 96 wel ELISA plates (Tero Fer Seti) were cold overnight at °C with 2 recom ant SARS-CoV 22019-nCaV St protein (Sino Beal ted in PHS. Wels sree Nace eth 10% sim mk i PAST (P85 + 01% Tees 20) t room {empsatre for Ih, Told srl tions of save samples in PBST mere {canted tothe FLISA plats induct) and inated a oom emperstare for Th Saliva fom an asypeonars india confirmed ga For SAIS (Cov 2b lineal testing was use as negve carl, Sala from a conan {divide nected wih SARS-COV-2 war ed ae 1 putve conta ‘Antibody Binding was detected with built otman fg 110 Sgn ‘lei and ig (110,00, Asay Mate or Thor at rom temper, hen Sweptviin HAP (000 Lif echnolgts) in PEST for 4 mina oo ey Ferre Cores deeoped wih TMU sation Sigma-Aldrich) and Os with the econ topped wang ?M #80, Absorbance at 150m was red o8 =| Imcroplt reser Examples of titatione ae shown in Supplementary Fe. 2. OD {aloe wh neste control salva were stated Km te te same teach “luton, then endpoint ters alate. Microneutalisation assay SARS Ca¥ 2 valae CoV/Austela/V1CD1/2020"" psd in Vero cals a stored at 0°. Sv fefold dons of et natvted plasma wee incubated with 100 TID of SARS-CoV-2 for Iran rsd vt infectivity was assed in quart walls of Vero cls vil copa fect was rend on dey 8. The ‘Seutling sober ealunad wing te Red/Marac cod, Systems serology. Hay prtipons, Age matched chien underging ele ‘We tonal ge 5-9) were erated athe Lauscoion Genera Hosa (asian) and oper filing the ceri for oso they were ‘onniered erably, showing no gn of imane eompromine Healy dle donors (ge 36-88) wee terse va the Unies of Meboure. Al aly donors were rected prior SARS.C3N-2 pandeme Heparin! Mond seceded for 10min 300g ta collet pls which was enema 20°C ‘a reuird ‘Coupling of arated beds A custo CoV maples ay mas designed”, ith SARS‘CoV'2 Spe 1 (Sino Biologia, SARS-CoV"? Spike 2 SARS-CoV Spc I (ACKO items USA), and HCY (29, NS, OCS) spe (Si roe) ab well ap SARS-CoV’? RBD (produced under HHSN27220140004C, ‘nd blind trough BEI Resources, NIAID, NIH USA), SARS Cov RBD (git ‘Eom Dale Godfrey and buh SARS-CoV-2 and HKUL Teme Spe (gf fom ‘Adam What). Teanos toxoid (Signa Aldrich) and inflacva hemagyuiin (GnICA2A Sino Bilal were ako add to the sy ax ptive conto “Antigens wee coaenly coupled to magnet cabonate beads (ho ad) using 2 ‘wo sep carbide rection and locks with. BSA Before Beng ‘ouspendel pd stored in PAS 005% som azide for we. Lines beaded miter asa. The types aod subse of putogen spac antibodies preven! in ote plasma wore uss wing he above ‘ule asp Bly 204 of working bead mast (000 beads per bead ‘egion) and 20 of dite pasa inal ein 1100) were aed yer well and ‘neinted owen st °C ona shaker Pathogen specie anos were dtc hing 14 tent detectors One step dtection wan done wing phyeoeythnn| (Eh conjugate mouse ant-haman pag Ig 4, IgA? Sothern Bech "ppl, 25 lie), where detectors wee added to the beads, washed then rea bythe MagPan Clq pati (MP Biomedical USA) was ft bitinyeted (Them aber Sentie, USA), tea tetramer with Septvain RPE GAPE: Thermo her Scent) before mers or tetameric Clq-PE were eg wed in one-step ection Tor the detection of Fey binding 0 tp detection was done by Bt ding soluble coin Fey ders higher aay plymarphisns Fey +13, lowr ality ppmorphlams Bylla RIS, Fey, higher any ppmorpbisns FyBl-ViS8 lower tiny polymorphisms Fella £138 Sql 25 lel git om Brace Wines and Mark Hoga to the Bed, veshing flowed bythe addon of SAPE. Lew for Ip two step detection ss dae using ntfsd moe at aman It (mab M2: Mab Techy Spi, 25 lel llowed by SAPE. Assays were repeated in dplste Data ne procsing for Sens Serology Anas, In the multivariate als potve contol antigen (Tetanae apd HI) were rene. Al vit dye Wve used fo each india: Dats was ight-shied and then log ransfonned (Gogto(e +1). Right siting was performed on each etre (detector atiaen ed tha cots cept oes today, by abling te mio vl for {hat etre toll sapien that str, Fr al tart anal te at ‘were eeatcentere end varias sled foreach etre ung the acre once. In Maa, ture Slaton To determine the minimal ct of stres(sgnair) nes to dai the aos cata tree tp procs was se aed Fs the ha were randomly sampled without replacenet fo generate 200 subset ll Slasser were resampled tthe ne of Ue small law fr clei utc, ‘which cotced fr any potent fects of ds sa nals ding {hguliation, Hai Ne tealarieation was then ap to each af be 2000 ‘sampled subsets ost atures most acted wih cor clssiiations. The ie Net hyperparaeterp, wae set ove el weight betwen the U1 ‘oem and? horn snail withthe pe fonction fr the et able shrinkage ad sletion (LASSO) and ilgerepreston, respectively which allows for beter aati of colinear dats which maybe limited fn LASSO ‘egreson'. The fequency at which ech fsture war seated across the 2000 iterations was wed to determine the sgatres y sing a sequent tp forward Algorithm that tray added single etre nto a PLSDA model tring wih {te eat that ha the highest reuenc of selection othe Howes requeny of Seton. Mode prediction performance was ase each step an eat by Tov cos vation Gascon ero The model wth he lest, lastion er within a D4 ference beeen the minima cain sro wat sclced a hemi sate oly one feature ma sled the tex bet et of etre wa chosen, const etre see were allege, ‘he th mallet oh gest to etre was chosen based on interpreaiity "PLSDA, Paral Least Sars Diinant Anal (PLSDA) performed a [Bgemsectors PLS totbox in Mas, ward in conncton with Hate Ne ‘dribd dove to dently and wacslne sare tht diapiah cohorts This Superised method assigns a Tuding to cach eure within a gien spate, and ‘pier the lina combination of ann (Ise vara) tht be separator the categorical groups A festa with sigh lading mgt inate peter Imporanc or sparating the groups rom ane anae. ach sample then ed pled ing te nda response earners expensed troup the intent arabes (LV9. The sores and lading can then be cross-eeenced 0 ‘dtcrnine which etre ronda in asosaton with which ati groups (ostvely landed feature are higher ie posiiely scoring groups ae). A mls te created with 10 fold ers-radaton, where Realy 10% ofthe dataset uta the at set andthe et ed Wain the mod Mol performances ‘measured tough clan ero (average torn the teaig Se) a well as ‘ont alain errr (average erin the ttn) with ales eae 20 Beng, Ihst All models were ohonogondizsd 1 ene cer visualization of ress Tiara! Catenin Cabo clacton catering mas uid fo the Healthy Househld Coe and sed on th fest sled sgatres dkseribed above sing unsupervised average linkage hercieal cstering of 2 ‘Sore dat Eucdden datancs ya wed a the datance mete ‘Sofie PISDA modes wer competed using the Eger PLS tolbox a Mat Hierarchical Clsering was completed using MATLAB 20175 (avers, Natick, MA). PLSDA scorer nd ladinge pote wer lt in Pi verion #0, Flow cytometry of PBMC and whole blood lon was slain EDTA tues from each partiput at day 1,37, and 86. nme lowing clin 10D lot whol ood was aquotd fr fw cytometry ana The remaining EDTA, Fao samples were posse int pasos and PBMC For fw eto analysis ef wile Hood samples, whole cod was Hed wh 1m ed al Isis tur fr 10min at room temperature. Cells were washed with ml PBS and ntrued at 350 for Sia. Fllowing two ore washes, ell were es feed PS for wabity staining using near ined vay dye acorn to ‘anulsctrer nractons For faye cyeomety analysis ah elted PMI, ‘all were wath in mil PS prior lysing ung BVO vit dy ‘cording to manuatre’ instructions For batt whole Blood and PRMC sa fle the vay dye reaton was topped by the akin of FACS blr (2% Frat incited FCS in 2m PBS FDTA) and es wee cnt at 380 for Sminute Cols were then reunpended in human FC bck according to mana facture’ isto for Sin a room temperature. The wae ad or PRMC amuody cocks Gapplementary Tale 1) made wp a 2 concentration were {Lcd wa the cele and ete for 20min oe olin sen cele ssre washed with 2m FACS bute and centfiged at 350g for min. Calls sre then resnpended in 2% PPA fora 20min fation one, washed. 2nd ‘suspended sl FACS ber fo argon wig the BD L3R X29 Fortes 6 NATURE COWMUNICATIONS| ngoyTs703)hps/do/101038/st467-020-19545 8 wate con/natnecanmictonsNATURE COMMUNICATIONS | htips//doiorg/101038/541467-02019545-8 ARTICLE oral dow cyte experinents, compensation was performed athe ne of snplescqoiston sig compensation beads. Sopplementry Fig 1 epi the ‘amu gating sto or PHM and wale bond samples Ross wee alee (paul aig and SNE anya) using How Veron 16 salwar The SNE plats mee generated from a concatenated fle containing 00 eens (2000 randomly sac ive sige alls pr patient pr te in) Manual gated results af presented es proportion f Ine cll ora Proprtion of parent gate (or PBMC) o a proportion a leakocyes (or whole ‘ond. Data were pled In Prism version 80. Plasma cytokines Plsia was ted 1:2 and for assent of etkins Shing the human slblepotin comet: head sry erat (HD Races) cording to manicures itrctons.Cjlomste Bead aay data were Sire on 3 BD LSR Il X20 Foren snd analy sing the FCAP Array Sifu The filling 18 cyekines were gant Hele If. 6, Ne ‘TNFa, MIP-lay MCP Ie I- RANTES 1290 101-211-519, iL- 15, TPy and 117A. ll ones expe fr I 8. MCP. Ie pd RANTES fll tnlow the nit of detextion of the saya both atone and were ede Gem fate analy Res are pot np and plete sing Pee tics. Human experimental work was conduct scconding to the Desaation of "Helinki principles ad according to the Austalian Notional Health ad Medical ‘Revwarch Counc Code of Practice, All donor ott eal gran pore sete informed consent The sty we approved bythe Human Reseach cs {Commit (HEC) of the University of Nelorne (hk ID #14435894. ‘2081, #167306, 2056689, #1995465) for healthy als, Tasmanian Heal {nd Modal HIRE (007479 for ely child danas Forte fm cte a, this ject reeived eiclupproval fom The Royal Chen's Howie Mel toute Hunan Research Bs Comite (HREC): HRECIS@@6RCHM.2018, Reporting summary Fit nfcnaton on ech dsp lilo he Nae esearch epoeing Sumary Ink to ths are Data availability [Alaa ppg nding of stay rv Source aa ae provid th thie paper he fom th athors pon reqs Received: 21 July 2020; Accepted: 20 October 2020; Published online: 1 November 2020 References 1» The epdemilogial chance ofan outrak of 2019 noel cronavirs| lneanes(COVID TS) China. 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Se Inmate crg10 1128" Sclimmunolabirt1# (202) 22, Xu, Rta sil: pte diagnostic ue and wansmison of 218-200¥. “ntemaiona our of Oral Sete 12 btpo. or 0 1036168 020 ‘ona (2000, 25, Robbin D. Fc a. Convergent antibody responses o SARS-Co¥-2 in omalevcent indian, Nate S84, 437-412 (202). 24, Coney D, Donte CM, Rocio ara, Kron, C.D. Nove G1OP 11d) rvs train mortem tron, Australia Ene Df Di 19, 13241327 2013) 25, Gham J ta proved molec diagnos of COVID-19 by the nove, igh senve and specie COVID-19-RURpHal eine eee (eanscipton PCR ay valde in vito and wih lie pecimen. 1 li ‘Mier 5%, hiss eoory0.121}CM.00310-20 2020), 26 Amanat Fel A serology to detect SARS-CoV-2 seroconversion in Inumans. medi the reprint rer for heh sens pean 1 1101/202008.72008795 (200). 27, Galt a lation and ap sharing of the 2019 nove oroavias (Galis;Cov-2) frm the at putin apse wt COVID-I9 Austria. ‘Mod. At 212, 459-452 (3020, 28, Hoy, KV. a Prophylas With «Middle East Repraory Syndrome Coronavirus (ERS Ca¥) specie human monoclonal antibody proses "Abdi fm MERS-CoV infection. J. Ic Dis 213, 1557-1561 2010, 29, Sabra etal. Pir infection and pusve afr of rewtaegatody prevent elcaton of severe au espratorysyndtome eoronavins inthe esprtry tract of mie J Vio 78 3572-3577 200). ‘Sle, K} ta Dinettes seolgyfetores in chien, dry ond {GOViD patents. made the propria seer for helen, 220.2005 20112009859, ep devon 101101202008. 2058459 eon, 231. Gann, BM. tA Ral fr Fe anton in theapeatcnonelonal ntdy. ‘mediated prteton api cl ins Cell Host Mice 24, 221-233 235, cars 52 Hu Zou 1. H, Regulation and vale selection va the dst Wt. Sa. So 8 67, 301-320 205). 38, Necland, MR eal. Mass ctomeury revel cellular fingerprint assoc ih IgE pean tolerance and lrg in ery i Na: Commun. 191 e000, ‘Acknowledgments Van lad Why i gry git for proud sing th eon! Feiner MLN. suppose by = Mel bourne Chiles leCouse Felli, BL i supored by NHMRC Carer NATURE COWMLNICATIONS| cnz0>Ts703) s/o /101038/st467-02019548 8 wu ature con/natneconmictons 7ARTICLE NATURE COMMUNICATIONS | htips//dotorg/10 1038/241467-020-19545-8 Developmen Few (1146190), 25 suport by aS Foundation Fela. ‘hs work ma sppoted by lk Ma Taunton to KIC AWC. apd KS. the 2 ik ‘oundaon oH the Clio Craig Foundation KLF abd KK, NHMRC Laden Invnigatr Grat o KA (117871), NMC lvesigator Gra to KS. (177179 [NHMRC Pram Gren © KX (107198), Resch Grats Cun fhe Heng Kong Special Admins Rein. 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Cormespondence and equ fr mater shold he reed 1 8 ‘Peer reve Information Nate Conmankaton thanks Annable De St Mask, Duane Wevemann andthe eer, annymouy, eviews) fc hcg tbe Reprints and permison information is ail a pinnae compins Publier note Springer alae eine ns wih sep a jas hin sb maps and ttt ton, I) Oren Acs This tle esl under «Crave Cameos Ae | oe eestor blger neal eget ban and readin i any mela forma ot oa ou ge ape ‘ret othe en sae) andthe wore, provi # kw the Cave Commons ikem and ode hangs wee ae The ngs or ote i pay miter a his ‘sliced nr ative Common ene ee nd ere Teme and your seal we aot permed by xauorysqpltin or ect the fred ua wl sd stain permion dey fn the copii let Ts ‘ew copy ths hsv icescommonsonenety/ 4 1 the Autos 2020 8 NATURE COWMUNICATIONS| ngoyTs703)hps/do/101038/st467-020-19545 8 wate con/natnecanmictons