Allergenicity to worldwide invasive grass Cortaderia selloana as environmental risk to public health

1
Vol.:(0123456789)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s
Alle genici y o wo ldwide
in asi e g ass Co ade ia selloana
as en i onmen al isk o public
heal h
Fe nando Rod íguez1*, Manuel Lomba de o‑Vega2,7*, Lucía San Juan3,
Le icia de las Vecillas1, So ía Alonso4, E a Mo chón1, Diego Liendo5, Ma a U anga6 &
Albe o Ganda illas3*
Alle gies o g ass pollen a ec s abou 20% o he popula ion wo ldwide. In he las ew decades, he
Sou h Ame ican g ass Co ade ia selloana (CS, Pampas g ass) has expanded wo ldwide in a a ie y
o coun ies including he USA, Aus alia and Wes e n Eu ope. In many o hese loca ions, CS has
s ikingly sp ead and has now been classi ied an in asi e species. Many pe nicious consequences o
CS ha e been epo ed o local biodi e si y, landscape and s uc u es. Howe e , he e ec on human
heal h has no been s udied. To in es iga e his issue, we ha e chosen a Eu opean egion on he
no he n cos o Spain whe e CS sp ead is o e whelming, Can ab ia. We ob ained CS pollen ex ac
and analysed he alle genic eac ion o 98 pa ien s ha we e alle gic o pollen o local g asses. We
de e mined he skin eac ion and he p esence o speci ic IgE an ibodies (sIgE) o CS o o a ypical
au och honous g ass, Phleum p a ense. We also compa ed he seasonal symp oms wi h epo ed
g ass pollen coun s in he a ea. The esul s s ongly sugges ha CS can cause espi a o y alle gies a
a simila ex en o he local g asses. Gi en ha CS pollinises la e han he local g asses, his would
ex end he pe iod o g ass alle gies in he egion o abou h ee mon hs e e y yea , as s a ed by mos
o he pa ien s. This is he i s s udy epo ed on he e ec s o he s iking expansion o CS on human
heal h. Conside ing he s ong impac ha espi a o y alle gies ha e on he popula ion, ou esul s
sugges ha CS can cu en ly cons i u e a ele an en i onmen al heal h issue.
G ass pollen is one o he main causes o espi a o y alle gies wo ldwide and he i s cause in No h Ame ica and
Eu ope, wi h es ima ed 20% o he popula ion a ec ed1. Co ade ia selloana (CS) is a g ass o he Poaceae amily,
o he Dan honioideae sub amily, commonly known as Pampas g ass and na i e o Sou h Ame ica. Howe e , in
he las ew decades CS was in oduced in a wide di e si y o coun ies wo ldwide including he USA, Aus alia
and Wes e n Eu ope2. In hese loca ions, CS has s ikingly sp ead, and i is classi ied as an in asi e species.
Wi hin Eu ope, F ance, G ea B i ain, Po ugal and Spain a e s ongly colonised. The Uni ed S a es Depa men
o Ag icul u e, in a epo o 2014, s a ed: ’Co ade ia selloana ob ained a ela i ely high impac po en ial isk
sco e because i impac s na u al, an h opogenic, and p oduc ion sys ems’3. Fo his eason, i has been o bid-
den o comme cialise, plan o main ain in a a ie y o coun ies. One o such coun ies is Spain, whe e CS has
in ensi ely sp ead along he no he n cos including he egions o Galicia, As u ias, Can ab ia and he Basque
Coun y4,5–7. Fi s epo men ioning Co ade ia in Spain a e om 1953 in Can ab ia8.
The alle gic incidence o CS is unknown. CS has been e e ed o as a dange o au och honous species,
s ongly a ec ing biodi e si y and landscape. Mo eo e , i is spo adically men ioned in some enues and dis-
cussion g oups as a dange o humans, because o ma e ial machine y damage and heal h, such as cu s due o
he sha p na u e o i s lea es, o alle gic eac ions in con ac wi h he skin9–11. Howe e , despi e he s iking
OPEN
1Alle gy Se ice, Hospi al Uni e si a io Ma qués de Valdecilla (HUM), Valdecilla 25, 39008 San ande , Spain. 2CMC
R&D Depa men , ALK-Abelló S.A., Miguel Fle a 19, 28037 Mad id, Spain. 3Ins i u e o Resea ch Ma qués de
Valdecilla, A e He e a O ia SN, 39011 San ande , Spain. 4Alle gy Se ice, Sie allana Hospi al, To ela ega,
Spain. 5Depa men o Plan Biology and Ecology, Uni e si y o he Basque Coun y (UPV/EHU), Bilbao,
Spain. 6Asociación Can ab ia Sin Plume os, Liaño, Spain. 7INSERM, Occi anie, Mon pellie , F ance. *email:
[email p o ec ed]; Manuel.Lomba de oV[email p o ec ed]; [email p o ec ed]
2
Vol:.(1234567890)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
expansion o he g ass in egions whe e i is no au och honous, he e a e no s udies on he impac on human
heal h so a epo ed wo ldwide.
CS has s ongly colonised ex ensi e a eas o Can ab ia, a ypical No he n Spanish egion (Fig.1A,B) o abou
500,000 inhabi an s, mos signi ican ly du ing 1990–2008, a pe iod o in ensi e oad and house building6,12. CS
has mainly in aded he coas bu i also has eached he inland moun ains (Fig.1B–F). The plan has sp ead
by human ac i i ies. I is used in mo o ways o e ain he oad slope soil and as a na u al ba ie 13,14 and i is
anspo ed wi h cons uc ion agg ega es and g a el om s one qua ies. The e o e, i is consis en ly ound nex
o oads, new buildings o small pa hs co e ed wi h g a el and is abundan a ound s one qua ies (Fig.1C–F).
Plans o limi ing and e adica ing he g ow h o his in asi e plan ha e been deba ed in he local pa liamen
due o p essu e o ecologis o ganisa ions al hough only limi ed p og ammes we e implemen ed. Cu en ly,
Figu e1. Co ade ia selloana (CS) has s ongly in aded no he n Spain. (A) Cu en sp ead o CS wo ldwide
(yellow/o ange spo s). F om GBIF.o g, GBIF Home Page. A ailable om: h ps:// www. gbi . o g/ speci es/ 27045
19. (B) Le : loca ion o Can ab ia egion in Spain (le ). Righ : cu en sp ead o CS in he egion is s iking, no
only on he coas bu also inland (blue line). Sou ce: LIFE S op Co ade ia, h p:// s opc o ad e ia. o g/ langu age/
en/ ea ly ‐wa ni ngne wo k/. (C–F) Rep esen a i e pho og aphs o he o e whelming p esence o CS in Can ab ia
egion, on he no he n coas o Spain. CS has sp ead nea he coas , nex o mo o ways and new house
buildings (C) bu is also no o ious inland, nex o newly cons uc ed a eas (D) and e en in disc e e loca ions a
he moun ains, whe e g a el has been used on small pa hs (E). S one qua ies whe e he g a el is anspo ed
om, a e equen ly su ounded by CS (F).
3
Vol.:(0123456789)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
he Eu opean Union is unding a egional ne wo k o igh ing he inland expansion o he g ass and di using
he apidly inc easing p oblem among he Eu opean socie y (3.5 million eu os o 2018–202215). Howe e , he
p esence o CS on he no he n cos o Spain is s ill o e whelming.
Can ab ia is a good pa adigma ic e i o y o in es iga e he alle genic e ec s o CS on he human popula ion.
In Can ab ia, au och honous g ass pollens peak om Ap il o July1,16,17, when hey cause a concomi an peak o
hay e e . I is es ima ed ha he pe cen o he popula ion su e ing om g ass-associa ed hay e e in Can ab ia
is abou 19% o pa ien s diagnosed o hinoconjunc i i is and 14% o as hma ic pa ien s18. In con as o he
au och honous g asses, CS in he No h o Spain lou ishes om mid Augus o Oc obe 5. G ass pollens o he
Pooideae sub amily, he main g asses ound in empe a e clima es o he No h Hemisphe e, con ain p o eins
wi h simila i ies in hei an igens19,20. We ques ioned whe he pa ien s alle gic o he au och honous g ass pol-
len (Phleum p a ense, Phl, as ep esen a i e species) migh also be alle gic o CS pollen. Alle genic molecules
o g oups 1 and 5 (Phl p1 and Phl p5) a e main an igens inducing alle gies due o hei high capaci y o bind o
immunoglobulin IgE o he human immune sys em. To in es iga e his issue, we analysed he skin eac ion o
Phl and CS ex ac s o 98 pa ien s o Can ab ia ha we e alle gic o local g ass pollen. In addi ion, we de e mined
he p esence o speci ic IgE an ibodies (sIgE) o Phl and CS pollen ex ac s and o he single alle gens Phl p 1,
Phl p 5, Phl p 7 and Phl p 12 in blood se um. We also compa ed he seasonal symp oms wi h epo ed g ass
pollen coun s. The esul s e y s ongly sugges ha CS is a signi ican cause o espi a o y alle gies, a a simila
ex en as he local g ass. This migh hus ex end he pe iod o espi a o y alle gies in he egion o mo e han
h ee mon hs e e y yea . This is he i s s udy epo ed on he e ec s o he s iking expansion o CS on human
heal h and i has implica ions in all he egions o he wo ld whe e CS has become a widesp ead in asi e g ass.
Conside ing he implica ions ha espi a o y alle gies ha e on heal h, no only by he di ec e ec s bu also
by allowing oppo unis in ec ions, ou esul s sugges ha CS can cons i u e a signi ican public heal h issue.
This isk mus be added o he ecological impac , in o de o encou age e o s o e adica ing CS om in aded,
non-au och honous egions.
Ma e ials and me hods
Se ing. This s udy was conduc ed in Can ab ia, a egion o he No h coas o Spain.
Design and pa ien s. A c oss-sec ional s udy wi h p ospec i e da a collec ion was pe o med a he
Alle gy Se ices o he Ma qués de Valdecilla Uni e si y Hospi al in San ande and he Sie allana Hospi al in
To ela ega (Can ab ia, Spain).
98 pa ien s diagnosed o hinoconjunc i i is, as hma o bo h, caused by sensi iza ion o g ass pollen, we e
included in a sequen ial way om Oc obe 2015 o Ma ch 2016.
W i en in o med consen was ob ained om all pa ien s be o e en e ing he s udy. The s udy me he p in-
ciples o he 1975 Helsinki decla a ion and was e iewed and app o ed by he local Resea ch Commi ee o
Can ab ia (CEIC e e ence numbe 2015.207).
A se um sample was ob ained om each pa ien and s o ed a –20°C un il used.
Pollen ex ac p epa a ion. All me hods we e pe o med in acco dance wi h he ele an guidelines and
egula ions.
Co ade ia selloana (CS) pollen was ob ained comme cially (Ibe -Polen, Jaén, Spain) and hen ex ac ed a a
1:10 (w/ ) a io in PBS pH 6.5 wi h magne ic s i ing o 90min. a 5°C. The soluble ac ion was sepa a ed by
cen i uga ion. A e dialysis agains PBS, he ex ac was il e ed h ough 0, 22µm il e s. P o ein con en was
de e mined by B ad o d me hod (BioRad, He cules, CA, USA). Two di e en ba ches we e ob ained (07 and
09) wi h consis en esul s.
Pa o he ex ac was adjus ed o 0.25mg p o ein/ml and o mula ed in PBS wi h 50% glyce ol, phenol 0.51%
(SPT bu e ). The emaining ex ac was s o ed in aliquo s a −20°C.
Phleum p a ense (Phl) pollen ex ac was made as desc ibed o CS. The o igin o he pollen in his case was
ALK Sou ce Ma e ials, Pos Falls, Idaho, USA.
The p o ein p o iles o he CS o he Phl ex ac s we e de e mined by polyac ylamide elec opho esis in he
p esence o sodium dodecyl sulpha e (SDS-PAGE) unde educing condi ions (In i ogen-No ex icine gels
10–20% ac ylamide, Fishe Scien i ic, SL, Mad id Spain).
Skin p ick es . Pa ien s we e skin p ick es ed (SPT) wi h a comme cial ex ac (ALK-Abelló, S.A. Mad id,
Spain) o Phl and he CS ex ac . His amine dihyd ochlo ide solu ion (10mg/ml) and SPT bu e we e used as
posi i e and nega i e con ol(no eac ion), espec i ely.
The SPT wheal a eas we e measu ed by planime y. A cu -o a ea o 7 mm2 (abou 3mm a e age diame e )o
highe was conside ed a posi i e es esul (his amine).
The CS ex ac was es ed in 10 con ol subjec s, ha we e no sensi ised o g ass pollen, wi h nega i e
esul (no eac ion).
IgE assays. Se um samples we e es ed o IgE an ibodies agains Phleum p a ense (Phl) pollen ex ac and
he alle gens Phl p 1, Phl p 5, Phl p 7 (polcalcin) and Phl p 12 (p o ilin) (ImmunoCap FEIA, The mo Fishe
Scien i ic, Ba celona, Spain).
In addi ion, speci ic IgE agains Phl and CS pollen ex ac s was de e mined by RAST (Radio Alle go So ben
Tes ). Pape discs we e ac i a ed wi h CNB and sensi ised wi h he pollen ex ac s as desc ibed by Ceska e al.21.
Phl and CS discs we e incuba ed o e nigh wi h 50 µL o he pa ien ’s se um and a e washing (0.1% Tween-20
in PBS), wi h app oxima ely 100,000cpm o he iodine 125–labeled an i-IgE mAb HE-2 o 3h as desc ibed22.
4
Vol:.(1234567890)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
Finally, he discs we e washed, and hei adioac i i y was de e mined in a gamma coun e . sIgE alues in kilou-
ni s pe li e we e de e mined by in e pola ing in a s anda d cu e buil up wi h Lolium pe enne—sensi ised discs
and 4 dilu ions o a se um pool om pa ien s wi h g ass alle gy, which was p e iously calib a ed in a bi a y kU/l.
A cu -o alue o 0.35 kU/l was conside ed posi i e o bo h ImmunoCap and RAST. The e was a e y sig-
ni ican co ela ion be ween he sIgE agains Phl de e mined by bo h me hods ( Spea man = 0.8874, p < 0.0001).
RAST inhibi ion assay. Pape discs we e sensi ised as abo e in he IgE assays sec ion and hen incuba ed
wi h 50 µL o a se um pool om all pa ien s combined. 50 µL o (inhibi o y) CS ex ac solu ion (in se ial dilu-
ions) we e added on o he pape discs and incuba ed o e nigh a oom empe a u e. All o he incuba ions we e
pe o med as indica ed abo e in he IgE assays sec ion. The % o inhibi ion was de e mined o each ex ac
dilu ion by adioac i e coun s (cpm) and calcula ed by means o he ollowing equa ion:
Cpmx co esponds o he mean adioac i i y o he discs incuba ed wi h inhibi o a a gi en X dilu ion.
cpm100% co esponds o he blank con ol samples o he assay (no se um pool added). cpm0% co esponds o
he signal ob ained wi h no inhibi o ex ac added.
Resul s
To in es iga e whe he pa ien s alle gic o he local pollen eac o CS pollen, we chose a coho o 98 pa ien s
om Can ab ia. Table1 shows he demog aphic and clinical cha ac e is ics o he pa ien s. All o hem we e
diagnosed wi h hini is du ing he sp ing season and g ass pollen sensi isa ion. In addi ion o nasal symp oms,
98% had also associa ed conjunc i i is, 31.6% su e ed om as hma and 8.2% om u ica ia. Only 12.2% had
ood alle gies and 2 ou o 98 d ug alle gies. 53.06% o he pa ien s unde wen g ass pollen immuno he apy.
76.5% o he pa ien s e e ed li ing in a eas wi h high p esence o CS. 78.6% o pa ien s p esen ed a wo sening
o hei pollen alle gic symp oms om Augus o No embe (“delayed eac i a ion”). In addi ion, 56.12% o he
coho we e polysensi ised including o he pollens such as Plan ago spp. (18/98), ees (9/98), Pa ie a ia spp.
(6/98), animal dande (11/98) o house dus mi es (38/98).
CS pollen ex ac is no comme cially a ailable o un skin p ick es s o sIgE de e mina ion. The e o e, we
isola ed and p epa ed a CS pollen ex ac by a s anda d ex ac ion p o ocol used o pollens (see Ma e ials and
Me hods). The yield p o ein/pollen was abou 50mg/g, a ypical concen a ion ob ained o o he g ass pollens
(ou own unpublished da a). G ass-speci ic ELISA assays showed ha he CS ex ac did no con ain g oup 5
an igen, as expec ed o a non-Pooideae sub amily g ass (< 0.3µg g oup 5/mL23). The p o ile o he p o ein ex ac
by SDS-PAGE shows a g oup o 25–37 kD bands wi h he mobili y o he g ass g oup 1 alle gens and i migh
co espond o he homologous CS g oup 1 (a ow, Fig.2A;19,24).
Isola ed CS pollen ex ac was used on cu aneous es s on he pa ien coho , in pa allel wi h Phl pollen
ex ac , as a ep esen a i e o he local au och honous g ass pollens. All 98 pa ien s ga e a posi i e esponse by
skin p ick es o Phl pollen ex ac and 89% o he pa ien s we e also posi i e o CS pollen ex ac (Table1).
Mo eo e , he e was a signi ican co ela ion be ween he a ea o he papule o Phl and o CS ( Pea son = 0.2558,
p = 0.01; Fig.2B). As a con ol, 10 pa ien s nega i e o skin eac ion o Phl we e ound also nega i e o CS
ex ac . These esul s show a s ong coincidence in he cu aneous eac ion o CS and o he local g ass. To u he
s udy he in e species c oss eac ion o he pa ien se a, we un by RAST ( adio alle go so ben es ) inhibi ion
assays. As shown in Fig.2C, Phl ex ac signi ican ly compe ed wi h CS ex ac o bind he se um sIgE om he
pa ien s.
Supplemen a y TableI displays he esul s o sIgE masu emen . We de e mined sIgE o Phl and o he alle -
gens Phl p 1, Phl p 5, Phl p 7 and Phl p 12 by ImmunoCap (The mo Fishe ) and o CS by RAST. All pa ien s had
se um sIgE o Phl by bo h ImmunoCap and RAST, in ag eemen wi h he skin p ick es esul s. We de e mined
he co ela ion be ween bo h echniques in de ec ing he sIgE o Ph. The ela ion was Spea man = 0.8874,
p < 0.0001. Values ob ained by RAST we e below hose ob ained by ImmunoCAP ( ac o = 0.36) and he linea
ange o RAST (0.17–27) was sho e han o ImmunoCAP (0.35–100). Ne e heless, he co ela ion be ween
bo h echniques was good, indica ing ha he sIgE da a ob ained by ImmunoCap can be compa ed wi h he
sIgE da a ob ained by RAST (Supplemen a y Fig.1). All pa ien s bu se en con ained sIgE speci ic o CS ex ac .
In e es ingly, wi hin he se en pa ien s wi h a nega i e sIgE es o CS, 5 displayed a nega i e skin esponse o CS
and he o he 2 displayed a weal smalle han 14 mm2. The e o e, he e was a s ong co ela ion be ween he skin
esponse and he sIgE o CS in se um (Fishe ’s exac es , p < 0.0001; Supplemen a y TableII).
We measu ed he p esence o sIgE o he indi idual alle gens Phl p 1, Phl p 5, Phl p 7 and Phl p 12 in he se a
om he pa ien s (Supplemen a y TableI). Fo he pan-alle gens Phl p 7 (polcalcin) and Phl p 12 (p o ilin), only
27 pa ien s (27.5%) had sIgE o any o hem. Consequen ly, he pa ien sensi isa ion o hese alle gens canno
explain he high c oss-sensi isa ion o Phl and CS in his g oup o pa ien s. The p e alence o sIgE o Phl p 1 was
e y high (98%) and only wo pa ien s (# 45 and 83) we e nega i e o IgE o Phl p 1. Consis en ly, hese pa ien s
also displayed a nega i e skin esponse o CS ex ac . The p e alence o Phl p 5 was lowe bu s ill impo an
(72%). Twen y-se en pa ien s o he coho displayed no IgE o Phl p 5 in se um. Howe e , o hese, only i e
pa ien s we e nega i e o skin esponse o he CS ex ac . The e was a signi ican linea eg ession be ween he
sIgE o he whole Phl ex ac and he sIgE o Phl p 1 (Fig.3A) o Phl p 5 (Fig.3B). F om he slope o he eg ession
line, we can conclude ha e e y alle gen accoun s o abou 50% o he o al IgE esponse o he whole ex ac ,
being he IgE- esponse o Phl p 1 sligh ly highe . The eac ion o Phl p 1 plus Phl p 5 is simila o he eac ion o
whole Phl ex ac (Fig.3C), s ongly sugges ing ha g oups 1 and 5 a e he main alle gens o Phl and hey accoun
o mos o he IgE o he whole Phl ex ac . The e is a signi ican co ela ion be ween hesIgE o CS ex ac and
hesIgE o Phl whole ex ac , oPhl p1 o oPhl p5 (Table2). The co ela ion is s onge o he whole ex ac o
100
×

1−

cpmx −cpm100%

/

cpm0% −cpm100%

5
Vol.:(0123456789)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
Pa ien Age (yea s) SexaYea s li ing in
Can ab ia Exposu ebClinical symp omscMon hs wi h symp oms O he sensi isa ionsd
Cu aneous eac ione
C. selloana P. p a ense
E0416001 50 M 10 (*) RC May–Oc HDM/plan ago 47 59
E0416002 38 F 38 (*) RCA Ma –Oc –40 24
E0416003 23 M 23 (–) RCAU Ma –Sep HDM/dog 30 21
E0416004 44 10 (*) RCA Ap –Sep HDM 47 45
E0416005 46 M 46 (*) RC Ma –Oc HDM/pla anus 10 22
E0416006 49 M 20 (*) RC Jun–Oc – 67 34
E0416007 41 F 41 (–) RCA Ma –Aug Dog 32 113
E0416008 27 F 27 (*) RC May–Aug HDM 31 65
E0416009 27 F 27 (*) RC Ma –Oc –49 76
E0416010 55 M 55 (*) RC Ap /Oc – 30 54
E0416011 50 F 20 (*) RC Jul–Oc – 35 38
E0416012 36 M 36 (*) RC May–Oc – 16 33
E0416013 20 M 20 (*) RC Ap –Aug HDM 39 94
E0416014 42 M 37 (*) RC Ma –Sep – 19 92
E0416015 39 M 12 (*) RCA May–Oc HDM 18 25
E0416016 45 M 45 (*) RC May–Sep – 21 20
E0416017 52 M 31 (*) RCA Ma –Jul –63 92
E0416018 45 M 40 (*) RC May–Sep – 42 48
E0416019 34 M 34 (*) RC May–Oc – 67 78
E0416020 30 M 25 (*) RC Feb–No Ca 29 48
E0416021 44 M 21 (*) RCA May–Se – 111 162
E0416022 38 M 38 (*) RC Ap –Aug HDM 737
E0416023 43 M 7 (–) RC Feb–No Plan ago 14 23
E0416024 50 M 50 (*) RC May–Se – 6(N) 39
E0416025 33 F 33 (*) RC Ap –Oc –49 28
E0416026 29 F 29 (*) RC Ap –Se – 37 77
E0416027 48 M 48 (*) RC May–Oc HDM 68 35
E0416028 41 M 4 (*) RC Ap –Jun – 86 48
E0416029 42 M 14 (*) RCU Ap –Sep HDM 21 48
E0416030 29 M 29 (*) RC May–Sep HDM 48 50
E0416031 42 M 42 (*) RCA Ma –Aug HDM 36 48
E0416032 48 M 15 (–) RC Ma –Aug 0 49 69
E0416033 25 M 25 (–) RC May–Sep HDM 34 169
E0416034 39 F 15 (*) RC Ap –Jul –6(N) 83
E0416035 53 F 17 (*) RC Ap –No –27 38
E0416036 48 M 6 (*) RC Ap –Oc HDM 61 33
E0416037 63 F 63 (*) RC Ap –Jul –27 33
E0416038 58 M 58 (*) RC May–Aug – 23 22
E0416039 39 M 39 (*) RC May–Aug HDM 50 114
E0416040 40 F 40 (*) RC May–Oc HDM 18 37
E0416041 31 F 31 (*) RC May–Aug – 1(N) 26
E0416042 29 F 29 (*) RCAU May–Jul – 13 31
E0416043 32 F 18 (*) RC Jul–Sep HDM 24 171
E0416044 42 F 8 (*) RCA Ap –Aug Pa ie a ia 17 22
E0416045 42 M 2 (*) RC Jul–Sep – 1(N) 37
E0416046 22 F 22 (*) RCA Ma –Aug HDM/pa ie a ia 79 64
E0416047 34 M 34 (*) RCA May–Oc HDM/ca 1(N) 18
E0416048 39 M 39 (*) RCA May–Jul – 41 42
E0416049 41 M 15 (*) RC Ap –Oc HDM 57 44
E0416050 28 F 27 (*) RC Ap –No HDM 15 23
E0416051 30 M 30 (*) RC Ma –May HDM/plan ago/cup esa-
ceous/pa ie a ia 32 128
E0416052 63 M 63 (*) RC May–Oc HDM 40 37
E0416053 22 M 20 (*) RC Ap –Aug Dog 21 75
E0416054 32 F 32 (*) RC May–Oc HDM 22 67
Con inued

6
Vol:.(1234567890)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
Table 1. Demog aphic and clinical da a and SPT esul s. SPT Skin p ick es . a F emale, M male. b Exposu e
(*) means ha he pa ien li es in an a ea in which C. selloana plan s ha e been iden i ied. c A a hma, C
conjunc i i is, R hini is, U u ica ia. d HDM House Dus Mi es. e Wheal a ea (mm2). Nega i e eac ion o C.
selloana is highligh ed in i alic numbe s (N). Numbe s in bold indica e posi i e eac ion (>6).
Pa ien Age (yea s) SexaYea s li ing in
Can ab ia Exposu ebClinical symp omscMon hs wi h symp oms O he sensi isa ionsd
Cu aneous eac ione
C. selloana P. p a ense
E0416055 41 F 7 (*) RCA May–Sep HDM/pa ie a ia/
plan ago 15 41
E0416056 44 F 44 (*) RC Ap –No HDM/ho se/dog/ca 34 79
E0416057 23 F 23 (–) RCA May–Sep HDM 13 49
E0416058 41 F 41 (*) RC Ap –Sep – 38 36
E0416059 31 F 31 (*) RCAU Feb–No Ca /dog/plan ago/HDM 815
E0416060 41 M 36 (*) RC May–Sep – 37 45
E0416061 29 F 29 (*) RC Ap –Sep – 29 68
E0416062 44 M 43 (–) RCA May–Jul Ca 535
E0416063 50 F 50 (–) RC Jun–No HDM 11 21
E0416064 26 F 1.5 (*) RC Ma –Oc –17 46
E0416065 69 M 69 (–) RCA May–No – 5(N) 66
E0416066 39 F 31 (*) RCA Ap –Aug –27 47
E0416067 40 M 40 (*) RCA May–No Plan ago 10 14
E0416068 26 F 26 (*) RC May–Sep Plan ago 34 39
E0416069 67 M 67 (–) RC May–Sep – 12 42
E0416070 70 F 70 (*) RC May–No HDM 48 46
E0416071 32 F 32 (*) RC Ap –Oc HDM 20 20
E0416072 30 F 30 (*) RC Ap –Jul 130 34
E0416073 18 F 18 (–) R May–Jun HDM/plan ago 13 26
E0416074 50 M 24 (*) RC May–Oc – 54 53
E0416075 35 M 35 (–) RC Ap –Aug –75 77
E0416076 23 M 23 (*) RCA Ap –Aug HDM/plan ago 71 96
E0416077 38 F 38 (*) RCA May–Oc HDM 38 22
E0416078 34 F 34 (–) RC Ap –Oc Pa ie a ia 37 57
E0416079 23 M 23 (*) RC Ap –Oc HDM 1(N) 24
E0416080 36 M 36 (*) RCA Ap –Sep – 37 36
E0416081 32 F 32 (*) RCA Ap –Jul HDM/pa ie a ia 28 78
E0416082 36 F 9 (–) RC May–Jun – 18 28
E0416083 31 M 31 (–) RC Ap –Jul –329
E0416084 23 M 23 (*) RCA May–Jul – 27 56
E0416085 39 F 39 (*) RC May–Aug HDM 67 55
E0416086 29 F 10 (–) RCAU May–Sep Pla anus/cup esaceous/
plan ago 11 39
E0416087 18 F 12 (*) RCA Ap –Sep Plan ago 16 96
E0416088 46 M 15 (–) RCA Ma –Sep HDM/ca /dog/ho se/
pla anus/cup esaceous/
plan ago 32 32
E0416089 30 F 30 (–) RCAU Ma –Jul Pla anus/plan ago 33 138
E0416090 23 M 23 (–) RCU Ma –Oc –18 52
E0416091 39 F 39 (*) RCA Ma –No HDM/plan ago 17 80
E0416092 20 M 20 (*) RCU Ap –Jul –107 65
E0416093 63 F 63 (–) RC Ap –Aug –20 23
E0416094 23 F 23 (–) RC Ma –Sep Plan ago/pla anus 529
E0416095 44 F 26 (–) RC Ap –Jul Pla anus/plan ago/
cup esaceous 34 39
E0416096 43 F 43 (–) R May–Jun – 1(N) 89
E0416097 37 F 37 (*) RC May–Oc – 80 27
E0416098 30 M 30 (*) RCA Ma –Jul Dog/cup esaceous/
plan ago 20 56
7
Vol.:(0123456789)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
o Phl p 1 ( = 0.75) han o Phl p 5 ( = 0.55). These esul s sugges ha he common eac ion obse ed in he
pa ien s o pollen ex ac o Phl and CS migh eside in he an igenic G oup 1 ha is ubiqui ous in all g asses25.
We analysed he measu ed g ass pollen concen a ion along he yea in he egion. The egional agency
Heal h Depa men o he nea by Basque Coun y de ec ed a sp ing main peak o g ass pollen a ound May
and a second, Augus - o-Oc obe peak, in he ai o Bilbao, a ci y 50km o Can ab ia wi h a simila clima e
and densi y o CS. I is in e es ing ha mos pa ien s (78.6%) in he s udy men ioned a second alle gic eac ion
a ound Sep embe –Oc obe (Table1, Supplemen a y Fig.2). This indica es a imely co ela ion be ween g ass
pollen and he e e ed alle gic symp oms by he pa ien s.
Discussion
We could no ind in he li e a u eany epo on he impac o CS on human heal h. This is somehow su p is-
ing and highligh s he need o s udies on he issue, conside ing he widesp ead p esence o his in asi e plan
wo ldwide1–5. Conce ns abou he consequences o CS expansion a e e iden among p o essionals ega ding he
impac o CS in ecology, indus y o heal h9–12. Ou s udy add esses o he i s ime he po en ial alle genic
e ec s o CS pollen. Gi en he wide impac o g ass alle gy in he popula ion, his cons i u es a public heal h issue.
We he e p esen se e al lines o e idence s ongly sugges ing ha pa ien s alle gic o pollen o no he n Span-
ish au och honous g asses, such as Phl, a e also alle gic o pollen o CS: (i) 89% o he pa ien s alle gic o Phl
we e sensi ised o CS, as e iden bo h by skin eac ion and by sIgE in se um; (ii) he imely coincidence along
he yea o alle gy symp oms epo ed by pa ien s, g ass pollen coun s and lou ishing o CS; (iii) he p esence
in CS o a p o ein band wi h a mobili y compa ible wi h g ass alle genic g oup 1 and he s ong p e alence o
his g oup in he sIgE o Phl.
050100 15
0200
0
50
100
150
SPT Phleum
(mm
2
)
SPT Co ade ia
(mm
2
)
BA
C
CS PhL
12M
250
150
100
75
50
37
25
20
15
10
43 kD
G1[ ]G1,5
Figu e2. Co ade ia selloana (CS) pollen sha es an igens and inmmunogenici y wi h au hoc onous g ass
Phleum p a ense (Phl). (A) SDS‐PAGE p o ile o CS (lanes 1, 2) o Phl pollen ex ac (lanes 3,4) ex ac . Lane
1 and 2 co esponds o 20µl and 40µl o CS pollen ex ac , espec i ely (see also Supplemen a y Fig.3)
ep esen a i e o wo independen ba ches. Lane 3 and 4 co espond o 20µl om wo di e en ba ches o Phl
pollen ex ac . M: he molecula weigh ma ke s. B acke s indica e he posi ion o he alle genic g oups (G)
acco ding o he documen ed appa en molecula weigh s. (B) Co ela ion be ween SPT esul o PhL and
CS Pea son : 0.2558; R2: 0.06543; p alue : 0.0110 ( wo‐ ailed). (C) IgE C oss‐ eac i i y o CS and Phl pollen
an igens as measu ed by adioalle goso ben (RAST) inhibi ion assays. No e ha i he e we e no c oss‐ eac ion
he Phl plo should be la o ce o (b oken line).
8
Vol:.(1234567890)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
020406080100
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 1, kU/L
A
02040608
01
00
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 5, kU/L
B
020406080
100
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 1 + 5, kU/L
C
Figu e3. Linea eg ession o sIgE o Phleum p a ense whole ex ac (ImmunoCap) e sus sIgE o Phl p 1 (A),
sIgE o Phl p 5 (B) and sIgE o Phl p 1 + Phl p 5 (C).
Table 2. Co ela ion be ween sIgE o C. selloana and o Phl p 1 sIgE, o Phl p 5 sIgE and o P. p a ense sIgE.
Phl p: Phleum an igen g oup.
sIgECS (Ku/L) s. sIgE Phl p 1 (kU/L) sIgECS (Ku/L) s. sIgE Phl p 5 (kU/L) sIgECS (Ku/L) s. sIgE Phl (kU/L)
ImmunoCap
Spea man 0.755 0.552 0.7476
95% con idence in e al 0.6513 o 0.8311 0.3920 o 0.6795 0.6400 o 0.8264
P ( wo‐ ailed) < 0.0001 < 0.0001 < 0.0001
Signi ican ? (alpha = 0.05) Yes Yes Yes
Numbe o XY pai s 98 98 96
9
Vol.:(0123456789)
Scien i ic Repo s | (2021) 11:24426 | h ps://doi.o g/10.1038/s41598-021-03581-5
www.na u e.com/scien i ic epo s/
The high c oss-sensi isa ion o Phl and CS pollen in his coho o pa ien s is no explained by a possible
eac ion o pan-alle gens, such as p o ilin (Phl p 12) and polcalcin (Phl p 7), since only 27.5% o he pa ien s
con ained se um sIgE agains hem. G oup 1 is a majo g ass alle gen ubiqui ous in all g asses in con as o
g oup 5 which is absen in non-Pooideae g asses as is he case o CS. The da a p esen ed in his s udy s ongly
sugges s ha g ass g oup 1 migh be he culp i o he obse ed c oss-sensi isa ion be ween au och honous
g asses (in his s udy, Phl) and CS.
Au och honous g asses in No he n Spain lou ish om Ap il o July1,16,17,26, while CS lou ishes om Augus
o Oc obe 5,7. F om a clinical poin o iew, mos pa ien s (78.6%) e e ed a la e alle gic symp oms eac i a ion
a ound Sep embe –Oc obe coinciden wi h a second, Augus - o-Oc obe , peak o g ass pollen coun s in he
ai . A p esen , he e a e no comme cial ex ac s o CS o immuno he apy. Howe e , he o e all imp o emen
o symp oms usually epo ed by alle gic pa ien s ha we e ea ed wi h con en ional g ass immuno he apy,
du ing bo h pollina ion peaks, sugges s ha hey migh ha e been p o ec ed also o CS pollen. This in addi ion
holds clinical in e es o hose egions whe e CS is au och honous and possibly alle genic.
The implica ions o he esul s in o public heal h- ela ed issues a e many and di e se. Fi s , he esul s encou -
age he in e na ional communi y o un alle genic es s o CS and o biochemically cha ac e ise he eac ion
o CS. Second, he esul s sugges ha CS migh leng hen he g ass alle gy season in e i o ies whe e CS has
expanded, by causing a second la e peak, addi ional o he peak due o he au och honous g asses. To no e,
comme cially a ailable g ass immuno he apy migh be bene icial o pa ien s alle gic o CS wo ldwide. Thi d,
gi en ha CS is banned in many coun ies and s a es, since i is conside ed anin asi e species3, a demons a ed
impac on human heal h would encou age policy make s o un p og ammes o e adica ing his plan in non-
au och honous a eas. The esul s p o ide an example o he global e ec s ha alien in asi e species can ha e
on human heal h.
Recei ed: 5 Augus 2021; Accep ed: 25 No embe 2021
Re e ences
1. Ga cía-Mozo, H. Poaceae pollen as he leading ae oalle gen wo ldwide: A e iew. Alle gy Eu . J. Alle gy Clin. Immunol. 72(12),
1849–1858 (2017).
2. Lamb inos, J. G. The expansion his o y o a sexual and asexual species o Co ade ia in Cali o nia, USA. J. Ecol. 89(1), 88–98
(2001).
3. Uni ed S a e Depa men o Ag icul u e. Weed Risk Assessmen o Co ade ia selloana. (2014).
4. Global Biodi e si y In o ma ion Facili y. Co ade ia selloana Asch. & G aebn. Syn. Mi eleu . Fl. 2(1): 325 1900. h ps:// www. gbi .
o g/ speci es/ 27045 23. Accessed 2 Ma 2021.
5. Pa do-P imoy, D. & Fagúndez, J. Assessmen o he dis ibu ion and ecen sp ead o he in asi e g ass Co ade ia selloana in
Indus ial Si es in Galicia, NW Spain. Flo a Mo phol. Dis ib. Func . Ecol. Plan s. 259, 151465. h ps:// doi. o g/ 10. 1016/j. lo a. 2019.
151465 (2019).
6. Campos, J. A., He e a, M., Biu un, I. & Loidi, J. The ole o alien plan s in he na u al coas al ege a ion in cen al-no he n
Spain. Biodi e s. Conse . 13(12), 2275–2293 (2004).
7. He e a, M. & Campos, J. The eed o he pampas (Co ade ia Selloana) Bizkaia: P ac ical guide o i s con ol. Ins . Es ud. Te i .
Bizcaia Dipu . Fo al. Bizk. 1, 30 (2006).
8. Guinea E. Geog a ía Bo ánica de San ande . [Bo anical Geog aphy o San ande ]. Publicaciones de la Exma. Dipu ación P o incial
de San ande (1953).
9. Queensland Go e nmen . Co ade ia selloana. h ps:// keyse e . lucid cen al. o g/ weeds/ da a/ media/ H ml/ co a de ia_ sello ana.
h m. Accessed 2 Ma 2021.
10. Co ade ia selloana isk assessmen . h p:// www. hea . o g/ pie / w a/ aus alia/ cosel- w a. h m. Accessed 2 Ma 2021.
11. Co ade ia Species. Sil e Pampas G ass, U uguayan Pampas G ass Co ade ia selloana. h ps:// da es ga den. com/ guides/ p / go/
1291/. Accessed 2 Ma 2021.
12. Ea ly Wa ning Ne wo k—LIFE S op Co ade ia—Medidas u gen es de lucha con a el plume o de la Pampa. [U gen measu es
o comba he Pampa]. h p:// s opc o ad e ia. o g/ langu age/ en/ ea ly- wa ni ng- ne wo k/. Accessed 2 Ma 2021.
13. Ruiz de la To e, J. e al. Ca álogo de especies ege ales a u iliza en plan aciones de ca e e as [Ca alog o plan species o be used
in oads]. Minis e io de Ob as Públicas y U banismo SGT, edi o . Vol. P ime a. EPES lndusl ias G á icasi (2019).
14. Plan aciones|Minis e io de T anspo es, Mo ilidad y Agenda U bana [Plan a ions|Minis y o T anspo , Mobili y and U ban
Agenda]. h ps:// www. mi ma. gob. es/ ca e e as/ no ma i a- ecni ca/ 14- plan acion es. Accessed 22 Oc 2021.
15. Backg ound and objec i es: LIFE S op Co ade ia: Medidas u gen es de lucha con a el plume o de la Pampa. [U gen measu es
o comba he Pampas g ass]. h p:// s opc o ad e ia. o g/ langu age/ en/ backg ound- and- objec i es/. Accessed 2 Ma 2021.
16. Subiza, J. G amíneas : Ae obiología y polinosis en España. [G asses: Ae obiology and pollinosis in Spain]. Ale gol Inmunol Clin.
18, 7–23 (2003).
17. Ja o, V., Rod íguez-Rajo, F. J., Seijo, M. C. & Ai a, M. J. Poaceae pollen in Galicia (N.W. Spain): Cha ac e isa ion and ecen ends
in a mosphe ic pollen season. In . J. Biome eo ol. 53(4), 333–344 (2009).
18. Ale gológica-2005. Fac o es epidemiológicos, clínicos y socioeconómicos de las en e medades alé gicas en España. [Epidemiologi-
cal, clinical and socioeconomic ac o s o alle gic diseases in Spain]. (SEAIC, 2006).
19. Ande sson, K. & Lidholm, J. Cha ac e is ics and immunobiology o g ass pollen alle gens. In . A ch. Alle gy Immunol. 130(2),
87–107 (2003).
20. Bu e, A., Sch amm, G., Keown, M. B., Schlaak, M. & Becke , W. M. Majo alle gen Phl p Vb in imo hy g ass is a no el pollen
RNase. FEBS Le . 363(1–2), 6–12 (1995).
21. Ceska, M., E iksson, R. & Va ga, J. M. Radioimmunoso ben assay o alle gens. J. Alle gy Clin. Immunol. 49(1), 1–9 (1971).
22. Sánchez-Mad id, F., Mo ago, G., Co bi, A. L. & Ca ei a, J. Monoclonal an ibodies o h ee dis inc epi opes on human IgE: Thei
use o de e mina ion o alle gen-speci ic IgE. J. Immunol. Me hods. 73(2), 367–378 (1984).
23. Ramí ez, J. e al. G oup 5 de e mina ion in Pooideae g ass pollen ex ac s by monoclonal an ibody-based ELISA: Co ela ion wi h
biologic ac i i y. Alle gy Eu . J. Alle gy Clin. Immunol. 52(8), 806–813 (1997).
24. WHO/IUIS Alle gen Nomencla u e Sub-Commi ee. WHO/IUIS Alle gen Nomencla u e Home Page. h p:// www. alle gen. o g/.
Accessed 2 Ma 2021.