Detecting Allergy by using Latex Allergens: Epidemiological and Clinical Experiences

Adriano Mari M.D.

Gloves: Managing Infection ControlSelecting the right gloves

Rome, Italy – June 24, 2010

Center for Molecular Allergology IDI-IRCCS, Rome, Italy

Allergy Data Laboratories s.c.Latina, Italy

Allergenic Extracts

Improving the Quality of Latex Extract with rHev b 5 Allergen

Hemery ML et al. Allergy 2005;60:131-2

Lundberg M et al. Allergy 2001;56:794-5

Latex + rHev b 5

Latex

Improving the Quality of Extracts with Allergenic Molecules

Mari A, Breiteneder H, Wagner S. J Allergy Clin Immunol 2005;113:S111(N = 28)

Latex Extr. Latex Extr. + rHev b 50.350.7

3.5

17.5

50.0

IgE

kU/l

WHO-IUIS Common name

Hev b 1 Yes Elongation Factor

Hev b 2 Yes Glucanase

Hev b 3 Yes Small Rubber Particle

Hev b 4 Yes Component Microhelix Complex

Hev b 5 Yes

Hev b 6 Yes Hevein

Hev b 7 Yes Patatin

Hev b 8 Yes Profilin

Hev b 9 Yes Enolase

Hev b 10 Yes Mn-SOD

Hev b 11 Yes Hevein-like

Hev b 12 Yes LTP

Hev b 13 Yes Esterase

Hev b Citrate binding Protein No Citrate binding protein

Hev b Hevamine No Hevamine

Hev b IFR No Isoflavone reductase

Hev b Rotamase No Rotamase

Hev b Trx No Thioredoxin

Hev b UDPGP No UDP-glucose Pyrophosphorylases

Mari A. Int.Arch.Allergy Immunol. 2001;125:57-65

33%

44%

5%

18%

MONO OLIGO BORDER PAN

MULTIPLE POLLEN SENSITIZATION

Profilins = PositiveCBPs = Highly PositiveSycamore

Mustard

Oak

Locust t.

Salt Cedar

Lime Rye GrassPellitory

Mugwort

Ragweed

Plantain

Goosefoot

Maple

T. of Heaven

Birch

HazelEucalyptus

Juniper Palm

PoplarOlive

Mari A. Int.Arch.Allergy Immunol. 2001;125:57-65

Elm Pine

Latex

Alle

rgen

ic M

olec

ules

Patient 0

Alle

rgen

ic M

olec

ules

Patient 1

Patient 2

Alle

rgen

ic M

olec

ules

Patient 3

Alle

rgen

ic M

olec

ules

Alle

rgen

ic M

olec

ules

Patient 4

Alle

rgen

ic M

olec

ules

Patient 5

Patient 6

Alle

rgen

ic M

olec

ules

Courtesy of VBC-Genomics, Vienna, Austria

SlideAllergen

IgE (serum)

Anti-IgE Fluorescence

2005

2006

2007

2008

2009

2010

1000

1250

1500

1750

2000Sources

MoleculesJune 2009

Years

Cum

ulat

ive

Num

ber

ReTiMEData Mining

IgE DetectionLab systems

InterAllAllergy e-Record

TD-Synergy

InterAllAllergy e-Record InterAll

Allergy e-Record

ISAC Proteomic Microarray for IgE Detection

Case (I) PR – F 53yoTotal IgE: 50.000 UI/l

Milk (Bos d 4, 5, 6, 8s)

Hen’s Egg (Gal d 1, 2, 4)

Wheat (Tri a 18)

Dog (Can f 1,3)

Plane Tree (Pla a 1, 2)

Cypress (Cup a 1)

Latex (Hev b 6, 11)

5 ; 1

,073

≤ 6-15

; 2,

990

16 -

25 ;

2,58

426

- 35

; 3,

087

36 -

45 ;

3,23

746

- 55

; 1,

711

56 -

65 ;

957

66 ;

769

≥

0

5

10

15

20

*

* ° °

°

°°

Age ranges and subjects-per-group values

Pati

ents

' dis

trib

utio

n (%

)

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Cup a 1

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Der f 2Der p 2

Der p 1Der f 1

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Pru p 3

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Bet v 1

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Mer a 1

Phl p 12Hev b 8

Latex Allergy within a Cohort of Not-at-Risk Subjects with Respiratory

n = 3930

Latex Sensitized = 46 (1.2 %)

Latex Allergy = 35 (0.9 %)

Mari A. et al. Int Arch Allergy Immunol 2007;143:135-43

Mari A. May 2005 Data on file

rBet v 2 nGrPro rPhl p 12 rHev b 80.1

1

10

100

Profilins

IgE

kU/L

n = 31

Comparative Testing for Profilin IgE using Natural or Recombinant Molecules

The ProfilinPollen (1)

Aln g 2Amb a 8Ant o 12Art v 4*Ave s 12Bet v 2*Bra n (#)Car b (#)Cas s (#)Che a 2*Cor a 2*Cyn d 12*Fra e 2

Pollen (2)

Hel a 2*Hum j (#)Lil l (#)Lol p 12Mer a 1*Ole e 2*Ory s 12Par j 3*Phl p 12*Pho d 2*Phr a 12Pla a (#)Poa p 12

Food (2)

Cuc s (#)Cum c (#)Dau c 4*Foe v (#)Gly m 3*Lit c 4Lyc e 1*Mal d 4Mus xp 1*Pru av 4*Pru du 2Pru p 4*Pyr c 4*….

Food (1)

Act c (#)Ana c 1*Api g 4*Ara h 5*Aspa o (#)Cap a 2*Cit la (#)Cit s 4*Cor s (#)Cro s (#)Cuc m 2*Cuc ma (#)Cuc p (#)

Pollen (3)

Que a (#)Ric c (#)Sec c 12Tri a 12Zea m 12Zyg f (#)Hev b 8*Ara t 8Fra a (#)Jug r (#)Nic t (#)Pap s (#)Pers a (#)….

Mari A. Int Arch Allergy Immunol 2002;129:286-95

Latex SPT-ve

CCDs -ve CCDs +ve0

25

50

75

100

p<0.0001

(n = 210) (n = 281)

Late

x Ig

E (%

)

Latex SPT-ve (n = 491)[r = 0.57; p < 0.0001]

0.1 1 10 1000.1

1

10

100

Latex IgE (kUA/L)

CCDs

- Ig

E (k

UA/

L)

Comparative Testing for Latex in CCD-IgE+ Subjects

Comparative Testing for Latex Extracts and Recombinant Allergenic Molecules

Mari A. et al. Int Arch Allergy Immunol 2007;143:135-43

Latex Allergy (n = 19) CCD-IgE Sera (n = 5)

Latex Extract

SPT

Latex Extract

IgE

rHev b Allergens

IgE

Latex Extract

SPT

Latex Extract

IgE

rHev b Allergens

IgE

Hev b 2

Sequence Homology IgE Co-recognition

Sequence Homology and IgE Co-recognitionVisualized by the Allergome O-ring

Allergenic Molecule-based IgE Prevalence (Allergens = 75; n = 23,077;

n+ at least 1 allergen = 16,408;71.1%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Hev b 8

Hev b 6Hev b 5

Allergenic Molecule-based IgE PrevalenceMono-sensitised patients

(Allergens = 75; n+ to 1 allergen = 2,565; 15.63%)

0 5 10 15 20 25 30 35 40 45 50

Prevalence (%)

Scala E et al Clin Exp Allergy 2010;40:911-21)

Cup a 1

Pru p 3Hev b 6 (n=17)

Scala E et al Clin Exp Allergy 2010;40:911-21)

5≤ 6-15

16 -

2526

- 35

36 -

4546

- 55

56 -

65 66≥

0

10

20

30

40

50

60 Milk

Egg

Mites

Cat

Cypress

Grasses

Parietaria

PR-10

Profilin

LTP

TropomyosinAge groups

Prev

alen

ce (

%)

Allergenic Molecule-based IgE PrevalenceAge distribution – Signature allergens

(Allergens = 75; n+ = 16,408)

Conclusions

• Allergenic molecules, replacing the extracts, are driving the current allergy diagnosis

• Microtechnology is a powerful approach allowing CRD, or, better, the Molecule-based Allergy Diagnosis, including Latex allergy

• Allergenic molecules as markers give us a perfect view on patients’ risk for allergic reactions

• The overall latex allergy phenomenon should be deeply re-evaluated using modern approches

Gabriele CarabellaVincenzo Lavinio

Allergome June 2010

www.allergome.org

Giorgio Perotti Alessandro BrunettiSara NutiSilvia Monti

adriano.mari@allergome.org

Center for Molecular Allergology

Istituto Dermopatico dell’ImmacolataIstituto Ricovero e Cura a Carattere Scientifico

IDI-IRCCS – Rome, Italy

Claudia Alessandri, MDMaria Livia Bernardi, MDRosetta Ferrara, MDDonato Quaratino, MDAlessandra Zaffiro, MDDanila Zennaro, MD

Marina Liso, BDPaola Palazzo, BD

Debora Pomponi, BDIvana Giangrieco, Ch

Lisa Tuppo, BDGiuseppe Costanzi, Tc

Chiara Rafaiani, Tc

Chiara Rasi, BDMario Santoro, MS

a.mari@panservice.it

Experimental Allergy UnitEnrico Scala, MD