Novità nella diagnostica molecolare
dell’allergia alimentare
Magda Lodolini
Laboratorio analisi - Ospedale Maggiore di Bologna
Bologna 27 Febbraio 2015
[email protected]
1
alpha Gal Allergy
Reazione IgE mediata a
Galactose- alpha -1,3-galattosio
Nome sistematico
3- O -α- D -Galactopyranosyl- D -galactopyranose
Altri nomi
Alpha-gal; 3-α- Galactobiose; Gal ( α1- 3) Gal; alfa- D - Gal- (1 → 3) - D - Gal
Numerosi tessuti di tutti i mammiferi inferiori
2
alpha-Gal Allergy
(allergia alla carne rossa)
Quadro clinico
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Reazione allergica ritardata (3-7 ore)
dopo un pasto di carne rossa o frattaglie
Sia la carne cruda che cotta possono causare reazioni
Prurito diffuso, orticaria, angioedema
Manifestazioni gastrointestinali
Difficoltà respiratorie. No asma
Possibile anafilassi
3
alpha-Gal Allergy
Particolarità
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Il primo esempio di food allergy da carboidrati
Reazione clinica ritardata
Non presente asma
Dose dipendente
Spesso sono pazienti non atopici che hanno tollerato la carne
Recede nel tempo se… non esposizione a punture di zecca
4
Zecca
Lone star tick
(Amblyomma americanum)
Zecca europea
(Ixodes ricinus)
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Lone Star Tick
Amblyomma americanum
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Le zecche
Country
Ticks
Food
No. Cases
Australia
I. holocyclus
Kangaroo
400
France
I. ricinus
Pork kidney
40
Sweden
I. ricinus
Red meat
60
Japan
I. sp.
Beef
30
Germany
I. ricinus
Pork kidney
50 + 40
USA
A. americanum
Red meat
1 800
7
•N Engl J Med. 2008 Mar 13; 358(11): 1109–1117.
Cetuximab-Induced Anaphylaxis and IgE Specific for Galactose-α-1,3-Galactose
•doi: 10.1056/NEJMoa074943
•Cetuximab-Induced Anaphylaxis and IgE Specific for
Galactose-α-1,3-Galactose
•Christine H. Chung, M.D., Beloo Mirakhur, M.D., Ph.D., Emily Chan, M.D., Ph.D., Quynh-Thu Le, M.D.,
Jordan Berlin, M.D., Michael Morse, M.D., Barbara A. Murphy, M.D., Shama M. Satinover, M.S., Jacob Hosen,
B.S., David Mauro, M.D., Ph.D., Robbert J. Slebos, Ph.D., Qinwei Zhou, Ph.D., Diane Gold, M.D., Tina Hatley,
M.D., Daniel J. Hicklin, Ph.D., and Thomas A.E. Platts-Mills, M.D., Ph.D.
8
•N Engl J Med. 2008 Mar 13; 358(11): 1109–1117.
Cetuximab-Induced Anaphylaxis and IgE Specific for Galactose-α-1,3-Galactose
•doi: 10.1056/NEJMoa074943
•Cetuximab-Induced Anaphylaxis and IgE Specific for
Galactose-α-1,3-Galactose
•Christine H. Chung, M.D., Beloo Mirakhur, M.D., Ph.D., Emily Chan, M.D., Ph.D., Quynh-Thu Le, M.D.,
Jordan Berlin, M.D., Michael Morse, M.D., Barbara A. Murphy, M.D., Shama M. Satinover, M.S., Jacob Hosen,
B.S., David Mauro, M.D., Ph.D., Robbert J. Slebos, Ph.D., Qinwei Zhou, Ph.D., Diane Gold, M.D., Tina Hatley,
M.D., Daniel J. Hicklin, Ph.D., and Thomas A.E. Platts-Mills, M.D., Ph.D.
•
The high prevalence of hypersensitivity reactions to cetuximab in the Southeast is supported by
our own data from the Tennessee group and in other recent studies.11 The striking difference in
the prevalence of the IgE antibodies against cetuximab provides an explanation for the difference
in rates of clinical hypersensitivity reaction between subjects in Boston or northern California and
those in Tennessee, Arkansas, or North Carolina.6,11,30.31,32 The explanation for the regional
distribution of IgE antibodies against galactose-α-1,3-galactose in the United States is not clear.
•
In conclusion, we have identified a mechanism underlying a hypersensitivity reaction to
cetuximab, preexisting IgE antibodies against an oligosaccharide present on the recombinant
molecule. Our results have implications for evaluating risks associated with antibody-based
therapeutics and for understanding the relevance of IgE antibodies specific for post-translational
modifications of natural and recombinant molecules.
9
J Allergy Clin Immunol. 2011 May;127(5):1286-93.e6.
doi: 10.1016/j.jaci.2011.02.019. Epub 2011 Mar 31.
The relevance of tick bites to the production of IgE
antibodies to the mammalian oligosaccharide
galactose-α- 1 , 3 - g a l a c t o s e .
Commins SP1 , James HR, Kelly LA, Pochan SL, Workman LJ, Perzanowski MS, Kocan KM, Fahy JV, Nganga
LW, Ronmark E, Cooper PJ, Platts-Mills TA.
BACKGROUND:
In 2009, we reported a novel form of delayed anaphylaxis to red meat that is related to serum IgE antibodies to
the oligosaccharide galactose-α- 1,3- galactose (alpha- gal). M ost of these patients had tolerated meat for many
years previously. T he implication is that some exposure in adult life had stimulated the production of these I gE
antibodies.
OBJECTIVES:
We sought to investigate possible causes of this IgE antibody response, focusing on evidence related to tick
bites, which are common in the region where these reactions occur.
METHODS:
Serum assays were carried out with biotinylated proteins and extracts bound to a streptavidin ImmunoCAP.
RESULTS:
Prospective studies on IgE antibodies in 3 subjects after tick bites showed an increase in levels of IgE to alphagal of 20- fold or greater. Other evidence included (1) a strong correlation between histories of tick bites and
levels of IgE to alpha-gal (χ (2) = 26.8, P < .001), (2) evidence that these I gE antibodies are common in areas
w here the tick A mblyomma americanum is common, and (3 ) a signif icant correlation between I gE antibodies to
alpha- gal and I gE antibodies to proteins derived from A americanum (r( s) = 0.75, P < .001).
CONCLUSION:
10
Pediatrics
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Tabella 1
Pediatrics vol. 131, n. 5-2013
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Fig 2
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Pediatrics vol. 131, n. 5-2013
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Fig 3
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Diagnosi differenziale
EBM nella diagnostica molecolare
• LATTE
dd clinica > 5 aa, allergia latte di NUOVA insorgenza:
dd molecolare Bos d 4 Bos d 5 Bos d 8
•
CARNE rara sensibilizzazione a epitopi specifici della carne.
spesso sostenuta da cross reattività con SA e actine.
•
CAT-PORK syndrome
Sostenuta da SA gatto (Fel d2)
Reazione clinica rapida dopo ingestione di carne maiale
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EBM nella diagnostica molecolare
GATTO /
ALFA GAL
IgE vs E2
No sintomi
Raro Fel d 1
Freq Fel d5
Commins SP Current Allergy Asthma Resp.
2013,13:72-77
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EBM nella diagnostica molecolare
Dosaggio IgE vs alpha Gal
utile strumento diagnostico e preventivo nei casi di :
• Reazione clinica dopo ingestione di carne
• Reazione intensa dopo puntura di zecca (misura preventiva)
• Tests sierologici positivi per uno o più estratti
manzo, maiale, agnello (cross reattività?)
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L’allergene che non c’è
MAIN VEGETABLE FOOD ALLERGENS
CUPIN
SUPERFAMILY
VICILINS
Ara h 1/Jug r 2
Ses i 3/Ana o 1
Cor a 11
LEGUMINS
PROLAMIN
SUPERFAMILY
2S ALBUMINS
Ber e 1/Ses i 1-2
Ana o 3/Jug r 1
Sin a 1/Bra j 1
Ara h 2-6-7/Ric c 1-3
nsLTP
Ara h 3-4/Cor a 9
Ana o 2/Jug r 4
GERMIN-LIKE
PROTEINS
Cit s 1 (orange)
Chili Pepper
(mugwort-celery
spice syndrome)
Pru p 3/Cor a 8
Mal d 3/Jug r 3
-Amilase Inhib.
Rice
Prolamins of
Cereals
PROTEINS of the
DEFENSE SYSTEM
PROTEINS of the
DEFENSE SYSTEM (II)
PATHOGENESIS
RELATED Pr.
STRUCTURAL AND
METABOLIC proteins
THIOL
PROTEASES
PR-1 (Cuc m 3)
PR-2 (1,3-glucanase)
Papain-like
cystein proteases
Hev b 2/Banana
Potato/tomato/etc.
PR-1 (Cuc m 3)
PR-3 (Class 1 chitinase)
Actinidin (Act c 1)
Ficin,bromelain,papain
Gli m Bd (soy)
Subtilisin-like
serin proteases
Hev b 6.02/Pers a 1
Cas s 5/Banana/etc.
PR-4 (Win-like proteins)
Bra r 2
PR-5 (TLP)
Pru av 2 /Mal d 2
Cap a 1/Act c 2
PR-9 (Peroxydases)
Tri a 19/Sec c 20
Tri a Bd (wheat)
PR-10 (Intracellular PRPs)
Mal d 1/Api g 1/etc
PR-14 (LTP)
Cuc m 1 (Melon)
PROTEASE
INHIBITORS
Type Kunitz (Soy)
 amylase inhibitors
(cereals)
STORAGE
proteins
Patatin (Sola t 1)
ENZYMES
Isoflav. reduct. like (Bet v 6)
(Pyr c 5)(Orange)
Cyclophilin (carrot)
Fructofurosanidases
Lyc e 2 (tomato)
Oxydases
Api g 5 (celery)
Liases
Alliin liase (garlic)
1-ascorbate
peroxidase (bell pepper)
PROFILINS
Api g 4//Mal d 4
Ara h 5/Cuc m 2
OLEOSINS
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Diagnostica molecolare frutta secca
• PR10 Bet v1 like
• PROFILLINE
• LTP
• STORAGE PR.
proteine labili; SOA
Cross reattive, labili, SOA
proteine stabili, spesso anche
severe reazioni
proteine stabili. Severe reazioni
• OLEOSINE ?????
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OLEOSINE
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Allergeni recentemente scoperti
Proteine idrofobiche sulla superficie dei corpi oleosi
Ruolo strutturale P.M. 16-24 KDa
Presenti in sesamo, nocciola, arachide, semi soia
Cross reattività
Sintomi spesso severi
SPT e CAP spesso negativi
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OLEOSINE
Allergene
Fonte allergenica
Funzione biologica Esposizione
Ara h 10
Arachide
oleosina
ingestione
Ara h 11
Arachide
oleosina
ingestione
Cor a 12
Nocciola
oleosina
ingestione
Cor a 13
Nocciola
oleosina
ingestione
Ses i 4
Sesamo
oleosina
ingestione
Ses i 5
Sesamo
oleosina
ingestione
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