Allergy information for: Chestnut (Castanea sativa)

Clinical History

• Number of Studies:6-10
• Number of Patients:21-50
• Symptoms:
  • Fernandez de Corres et al 1993 [327] reported 3 patients with generalized urticaria and/or angioedema; 3 with rhinitis and/or asthma; 1 with anaphylaxis and 2 with mouth itching.
  • Rodriguez et al 1993 [492] reported 2 cases of anaphylaxis after eating chestnuts.
  • Añibarro et al 1993 [252] reported one patient with urticaria and dry cough.
  • Blanco et al 1994 [287] reported 4 patients with systemic anaphylaxis; 4 with angioedema; 1 with rhinoconjunctivitis and 1 with urticaria.
  • Blanco et al 1994 [288] reported 3 patients with systemic anaphylaxis; 4 with angioedema and 1 with asthma.
  • Antico 1996 [263] reported one patient with oral allergy syndrome.
  • Beezhold et al 1996 [281] reported one patient with anaphylaxis.
  • Diaz-Perales et al 2002 [762] reported 8 patients with 4 oral allergy symdrome, 1 angioedema, 1 urticaria and 2 anaphylaxis with chestnut.
  • Garcia-Selles et al. 2002 [594] reported 10/47 peach allergic patients showed symptoms to chestnut with 4 oral allergy symdrome, 3 urticaria and angioedema and 3 anaphylaxis.

Skin Prick Test

• Number of Studies:11-20
• Food/Type of allergen:
  • PBS extract of raw chestnut (Fernandez de Corres et al 1993 [327]).
  • Fresh chestnut (Rodriguez et al 1993 [492]).
  • A chestnut extract; preparation not described (Añibarro et al 1993 [252]).
  • Fresh chestnuts (Blanco et al 1994 [287]; Blanco et al 1994 [288]; Antico 1996 [263]).
  • Dilution 1/10 of a commercial extract (Miles Inc., Elkhart, USA) (Beezhold et al 1996 [281]).
  • PBS extract from peeled chestnut; a chitinase-enriched protein preparation and the purified Cas s 5 allergen (Blanco et al 1999 [289]).
  • Prick-prick with chestnut (Diaz-Perales et al 2002 [762]).
  • Commercial extract (Crespo et al. 2002 [834])
• Protocol: (controls, definition of positive etc)

  Diaz-Perales et al (2002) [762] and Crespo et al. (2002) [834] defined as positive a mean wheal diameter which was 3 mm larger than the saline control after 15 minutes.

• Number of Patients:
  • 8 latex allergic patients (Fernandez de Corres et al 1993 [327]) .
  • 3 latex allergic patients with positive SPT (Rodriguez et al 1993 [492]).
  • 1 patient with latex sensitization (Añibarro et al 1993 [252]).
  • 25 patients with latex allergy (Blanco et al 1994 [287]).
  • 17 patients with immediate avocado hypersensitivity (Blanco et al 1994 [288]).
  • 1 patient with oral allergy syndrome induced by chestnuts (Antico, 1996 [263]).
  • 47 latex allergic patients (Beezhold et al 1996 [281]).
  • 18 patients with latex-fruit allergy (Blanco et al 1999 [289]).
  • 47 of peach allergics, 20 artimisia pollen allergic patients and 12 controls (Garcia-Selles et al. 2002 [594])
  • 8 latex allergic patients with a history of symptoms to chestnut, avocado or banana (Diaz-Perales et al 2002 [762]).
  • 65 fruit allergic patients (Crespo et al. 2002 [834])
• Summary of Results:
  • 6/8 latex allergic patients showed positive SPT to chestnut (Fernandez de Corres et al 1993 [327]) .
  • Three latex allergic patients gave positive SPT with chestnut (Rodriguez et al 1993 [492]).
  • One patient showed positive SPT to latex and chestnut (Añibarro et al 1993 [252]).
  • 9/25 patients with latex allergy showed chestnut hypersensitivity with positive SPT (Blanco et al 1994 [287]).
  • 10/17 patients with immediate avocado hypersensitivity showed positive SPT to chestnut and 8/17 of these had clinical manifestations (Blanco et al 1994 [288]).
  • A patient with oral allergy syndrome induced by chestnuts had a negative SPT to chestnut (Antico, 1996 [263]).
  • 13/47 latex allergic patients gave positive SPT to chestnut (Beezhold et al 1996 [281]).
  • The purified allergen Cas s 5 (called Cas s 1 in the article) elicited positive SPT in 13 of 18 (72%) patients with latex-fruit allergy. By contrast its class II chitinase equivalent did not show positive SPT responses (Blanco et al 1999 [289]).
  • 11/47 (23%) of peach allergics and 3/20 (15%) of Artimisia pollen allergic patients gave positive SPT to chestnut (Garcia-Selles et al. 2002 [594]). Positive SPT to chestnut only occurred in patients with a positive SPT to Artemisia.
  • Mean wheal diameters of 6-9mm for the 8 patients (Diaz-Perales et al 2002 [762]).
  • 22/65 gave either a positive SPT or CAP-RAST test (Crespo et al. 2002 [834])

IgE assay (by RAST, CAP etc)

• Number of Studies:0
• Food/Type of allergen:

  Diaz-Perales et al. (1998) [315] and Blanco et al. (1999) [289] made a crude extract by grounding mature nuts in a mortar with cold acetone and then defatting three times with the same solvent (1:5 w/v) for 1 hour at 4° C. The dried residues was extracted with 0.1 M sodium phosphate buffer (pH 7.0) in 0.15 M NaCl (1:5 w/v) for 1 hour at 4° C. The supernatant was centrifuged, dialyzed against water and freeze-dried.

  Commercial extracts were used in the CAP assay by Crespo et al. (2002) [834].

• IgE protocol:CAP-FEIA
• Number of Patients:

  Brehler et al (1997) [292] used sera from 136 patients with immediate hypersensitivity to latex.

  Diaz-Perales et al. (1998) [315] used sera from 4 patients (testing 20) with immediate hypersensitivity to chestnut and avocado according to the clinical history, positive skin prick test responses with the corresponding food extract and specific IgE levels to chestnut or avocado higher than 3.5 kU/L.

  Blanco et al. (1999) [289] used sera from 18 patients (selected from 60) with immediate hypersensitivity to chestnut and avocado according to the clinical history and positive skin prick test responses with the corresponding food extract.

  Diaz-Perales et al (1999) [44] used sera from 8 patients with immediate hypersensitivity to banana, chestnut, and/or avocado, according to the clinical history, positive skin prick test responses with the corresponding food extracts and specific IgE levels higher than 3.5 kU/L to at least 2 of the 3 foods. As control, sera was used from 5 patients with latex allergy but without accompanying food allergy.

  Diaz-Perales et al (2002) [762] used sera from 8 latex allergic patients with a history of symptoms to chestnut, avocado or banana, together with 8 atopic controls (several patients are common between the above studies).

  Diaz-Perales et al (2000) [43] used a pool of six sera from patients allergic to apple and peach.

  Crespo et al. (2002) [834] tested sera from 65 fruit allergic patients.

• Summary of Results:

  Brehler et al (1997) [292] found specific IgE antibodies against chestnut allergens in 47/136 patient's sera.

  Blanco et al. (1999) [289] found that IgE from sera of 10/18 patients bound to chestnut extract (0.4-23.6 kU/l). This was lower than for latex and slightly lower than avocado where 16/18 sera were CAP positive.

  Diaz-Perales et al (2002) [762] found IgE of 1.1-23.6 kU/l for chestnut (higher for latex). The sera was used to investigate the recombinant allergen Cas s 5.

  Crespo et al. (2002) [834] reported that 22/65 patients gave either a positive SPT or CAP-RAST test to chestnut.

Immunoblotting

• Immunoblotting separation:

  Diaz-Perales et al. (1998) [315], Blanco et al. (1999) [289], Diaz-Perales et al (1999) [44] and Diaz-Perales et al (2002) [762] used 1D SDS-PAGE (15% polyacrylamide) without treating the samples with heating or reducing agents.

  Diaz-Perales et al. (2000) [43] separated samples by 1D SDS-PAGE on aSDS-PAGE was carried out on 4% to 20% precast polyacrylamide gels (Bio-Rad) without treating the samples with heating or reducing agents.

• Immunoblotting detection method:

  Diaz-Perales et al. (1998) [315], Blanco et al. (1999) [289], Diaz-Perales et al (1999) [44] and Diaz-Perales et al (2002) [762] soaked the gels for 15 minutes in buffer and electrotransferrred to PVDF membranes for 60 minutes at 80V. Membranes were washed, blocked and incubated with sera (1:3 dilution), then alkaline phosphatase conjugated monoclonal anti-human IgE was added. Allergens were revealed with 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium.

  Diaz-Perales et al. (2000) [43] electrotransferred proteins onto nitrocellulose membranes. After blocking with 5% w/v BSA in phosphate buffered saline, PBS, for 2 hours at room temperature, the blots were incubated overnight with each individual serum (1:10 dilution in 0.5% BSA, 0.05% v/v Tween-20 in PBS). After washing 4 times with 0.1% v/v Tween-20 in PBS, membranes were overlaid with mouse anti-human IgE mAb HE-2 ascitic fluid (1:3000 dilution) for 2 hours and then washed and incubated with a rabbit anti-mouse IgE peroxidase-conjugated antibody (DAKO A/S, Denmark) for 1 hour. Detection of IgE-binding components was achieved by enhanced chemiluminescence.

• Immunoblotting results:

  Diaz-Perales et al. (1998) [315] reported that a single band of 32 kDa was recognized in both chestnut and avocado extracts by IgE from either a pool of the four sera or each individual serum. Purified purified chestnut and avocado class I chitinases which were also recognized and the class II chitinases were not. Preincubation with the purified class I chitinases could fully inhibit binding of IgE from the pooled sera to chestnut extracts. The N-terminal sequences of the class I chitinases were determined.

  Blanco et al. (1999) [289] found that IgE from pooled sera of patients bound to a single band of 32 kDa in both the crude chestnut and avocado extracts. This band coincided with one from the purified chestnut and avocado class I chitinases which were also recognized, although the class II chitinases were not.

  Diaz-Perales et al (1999) [44] found that sera from patients with latex allergy alone did not recognise allergens in chestnut extracts. Sera from patients with latex/fruit allergy recognised allergens from chestnut, cherimoya, passion fruit, kiwi, latex, papaya, mango, potato, tomato and wheat. All except the potato band were also recognised by anti-class I chitinase IgG.

  Diaz-Perales et al (2002) [762] found bands at 31 kDa with both native and recombinant Cas s 5 which could be inhibited by the Cas s 5 preparations or by latex hevein, Hev b 6.

  Diaz-Perales et al. (2000) [43] reported that the serum pool reactive to apple and peach LTPs, also contains specific IgE to the expected low molecular weight bands of the mugwort and chestnut extracts, as well as to the corresponding isolated LTPs. The chestnut polypeptide shows an even stronger reaction than the apple and peach allergens.

Oral provocation

• Number of Studies:1-5
• Food used and oral provocation vehicle:The food was masked in a mix of orange and pineapple juice, sugar, wheat meal and liquid colouring (Crespo et al. 2002 [834]).
• Blind:Yes
• Number of Patients:Crespo et al. (2002) [834] challenged 18 patients with chestnut who had either a positive SPT or CAP-RAST positive sera.
• Dose response:No information
• Symptoms:Crespo et al. (2002) [834] report 2/18 positive challenges with chestnut but no symptoms were described. 4 additional patients with a convincing history of anaphylaxis were not challenged.

IgE cross-reactivity and Polysensitisation

Other Clinical information

Reviews (0)

References (18)

• Diaz-Perales A, Sanchez-Monge R, Blanco C, Lombardero M, Carillo T, Salcedo G.
  What is the role of the hevein-like domain of fruit class I chitinases in their allergenic capacity?
  Clin Exp Allergy. 32(3): 448-54.. 2002
  PUBMEDID: 11940077
• García Sellés FJ, Diaz-Perales A, Sanchez-Monge R, Alcantara M, Lombardero M, Barber D, Salcedo G, Fernandez-Rivas M
  Patterns of reactivity to lipid transfer proteins of plant foods and Artemisia pollen: an in vivo study
  Int Arch Allergy Immunol. 128(2):115-122. 2002
  PUBMEDID: 12065911
• Beezhold D.H., Sussman G. L., Liss G.M. and Chang N.S.
  Latex allergy can induce clinical reactions to specific foods.
  Clin. Exp. Allergy 26: 416-422.. 1996
  PUBMEDID: 8732238
• Blanco C., Diaz-Perales A., Collada C., Sánchez-Monge R., Aragoncillo C., Castillo R., Ortega O., Alvarez M., Carrillo T. and Salcedo G.
  Class I chitinases as potential panallergens involved in the latex-fruit syndrome.
  J Allergy Clin Immunol. 103(3 Pt 1):507-513. 1999
  PUBMEDID: 10069887
• Blanco C., Carrillo T., Castillo R., Quiralte J. and Cuevas M.
  Latex allergy: Clinical features and cross-reactivity with fruits.
  Ann. Allergy 73(4): 309-314.. 1994
  PUBMEDID: 7943998
• Blanco C., Carrillo T., Castillo R., Quiralte J. and Cuevas M.
  Avocado hypersensitivity.
  Allergy 49: 454-459.. 1994
  PUBMEDID: 8074265
• Añibarro B., Garcia-Ara M.C. and Pascual C.
  Associated sensitization to latex and chestnut.
  Allergy 48: 130-131. 1993
  PUBMEDID: 8457033
• Rodriguez M, Vega F, Garcia MT, Panizo C, Laffond E, Montalvo A, Cuevas M
  Hypersensitivity to latex , chestnut , and banana.
  Ann. Allergy 70: 31-34.. 1993
  PUBMEDID: 7678723
• Fernandez de Corres L, Moneo I, Munoz D, Bernaola G, Fernandez E, Audicana M, Urrutia I
  Sensitization from chestnuts and bananas in patients with urticaria and anaphylasis from contact with latex.
  Ann. Allergy 70:35-39.. 1993
  PUBMEDID: 7678724
• Kos T., Hoffmann-Sommergruber K., Ferreira F., Hirschwehr R., Ahorn H., Horak F., Jager S., Sperr W., Kraft D., Scheiner O.
  Purification, characterization and N-terminal amino acid sequence of a new major allergen from European chestnut pollen--Cas s 1.
  Biochem. Biophys. Res. Commun. 196:1086-1092.. 1993
  PUBMEDID: 7504464
• Hirschwehr R, Jager S, Horak F, Ferreira F, Valenta R, Ebner C, Kraft D, Scheiner O.
  Allergens from birch pollen and pollen of the European chestnut share common epitopes.
  Clin Exp Allergy 23(9):755-761. 1993
  PUBMEDID: 10779306
• Brehler R, Theissen U, Mohr C, Luger T
  "Latex-fruit syndrome": frequency of cross-reacting IgE antibodies.
  Allergy 52: 404-410.. 1997
  PUBMEDID: 9188921
• Antico A.
  Oral allergy syndrome induced by chestnut (Castanea sativa).
  Ann. Allergy 76: 37-40.. 1996
  PUBMEDID: 8564626
• Crespo JF, Rodriguez J, James JM, Daroca P, Reano M, Vives R
  Reactivity to potential cross-reactive foods in fruit-allergic patients: implications for prescribing food avoidance
  Allergy. 57(10):946-9. 2002
  PUBMEDID: 12269944
• Diaz-Perales A, Collada C, Blanco C, Sanchez-Monge R, Carrillo T, Aragoncillo C, Salcedo G
  Class I chitinases with hevein-like domain, but not class II enzymes, are relevant chestnut and avocado allergens.
  J. Allergy Clin. Immunol 102: 127-133. 1998
  PUBMEDID: 9679856
• Mari A, Wallner M, Ferreira F.
  Fagales pollen sensitization in a birch-free area: a respiratory cohort survey using Fagales pollen extracts and birch recombinant allergens (rBet v 1, rBet v 2, rBet v 4).
  Clin Exp Allergy 33(10):1419-1428.. 2003
  PUBMEDID: 14519150
• Diaz-Perales, A, Collada C, Blanco C, Sanchez-Monge R, Carrillo T, Aragoncillo C. and Salcedo G
  Cross -reactions in the latex -fruit syndrome: A relevant role of chitinases but not of complex asparagine-linked glycans.
  J.Allergy Clin.Immunol. 104:681-687.. 1999
  PUBMEDID: 10482846
• Diaz-Perales A, Lombardero M, Sanchez-Monge R, Garcia-Selles FJ, Pernas M, Fernandez-Rivas M, Barber D, Salcedo G.
  Lipid-transfer proteins as potential plant panallergens: cross-reactivity among proteins of Artemisia pollen, Castanea nut and Rosaceae, with different IgE-binding capacities.
  Clin Exp Allergy 2000, 30: 1403-1410.. 2000
  PUBMEDID: 10998016

Biochemical Information for Cas s 1

• Allergen Name:Cas s 1
• Alternatve Allergen Names:
• Allergen Designation:Major
• Protein Family:Bet v 1 family Pfam PF00407
• Sequence Known?:N-terminal fragment only
• Allergen accession No.s:Q9S8Q4:Swissprot:http://ca.expasy.org/cgi-bin/niceprot.pl?Q9S8Q4
• 3D Structure Accession No.:Not determined
• Calculated Masses:N/A
• Experimental Masses:22 kDa
• Oligomeric Masses:Monomer
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Most allergens of the Bet v 1 family are thermolabile allergens.
• Nature of main cross-reacting proteins:Cas s 1 is a member of the Bet v 1 family. IgE binding to Cas s 1 could be completely inhibited by pre-incubating sera with purified recombinant Bet v I (Hirschwehr et al. 1993 [71]).
• Allergen properties & biological function:Cas s 1 is a member of the PR10 family with a possible defence function against plant pathogens.
• Allergen purification:Purified from an aqueous extract of chestnut pollen by affinity chromatography using an immobilized anti-Bet v 1 antibody (BIP1), eluting with 0.1 M glycine/HCl at pH 2.5, followed by anion-exchange chromatography using a DEAE column (DEAE-5WP, TOSO-HAAS, Japan) equilibrated with 20 mM Tris/HCl at pH 8.3. Elution used a gradient of 0-0.5 M NaCl.
• Other biochemical information:

  Kos et al. (1993) [99] report the N-terminal sequence GVFTHESQETSVIPPARLFKAFVL.
  
  

References (2)

• Kos T., Hoffmann-Sommergruber K., Ferreira F., Hirschwehr R., Ahorn H., Horak F., Jager S., Sperr W., Kraft D., Scheiner O.
  Purification, characterization and N-terminal amino acid sequence of a new major allergen from European chestnut pollen--Cas s 1.
  Biochem. Biophys. Res. Commun. 196:1086-1092.. 1993
  PUBMEDID: 7504464
• Hirschwehr R, Jager S, Horak F, Ferreira F, Valenta R, Ebner C, Kraft D, Scheiner O.
  Allergens from birch pollen and pollen of the European chestnut share common epitopes.
  Clin Exp Allergy 23(9):755-761. 1993
  PUBMEDID: 10779306

Biochemical Information for Cas s 5

• Allergen Name:Cas s 5
• Alternatve Allergen Names:
• Allergen Designation:Major
• Protein Family:Two domains, chitin binding - PF00187 and chitinase I - PF00182.
• Sequence Known?:Yes
• Allergen accession No.s:

  U48687, Swissprot: http://ca.expasy.org/cgi-bin/niceprot.pl?Q42428

• 3D Structure Accession No.:Not determined
• Calculated Masses:33946 Da with signal
  31978 Da (19-316)
• Experimental Masses:31 kDa
• Oligomeric Masses:monomer
• Allergen epitopes:Diaz-Perales et al (2002) [762] show that specific IgE bound only to recombinant Cas s 5 and not to the recombinant catalytic domain.
• Allergen stability:
  Process, chemical, enzymatic:

  Sánchez-Monge et al (2000) [1076] show that green bean class I chitinase is denatured by boiling and does not bind anti-hevein IgE. Diaz-Perales et al (2003) [904] show that the avocado allergen (Prs a 1) is rapidly degraded by pepsin but that the fragments retain IgE binding potency. These results may suggest that Cas s 5 would be destroyed by boiling but might survive digestion.

• Nature of main cross-reacting proteins:There are cross reactions with other class I plant endochitinases of fruits like banana, avocado and kiwi fruit. It is a panallergen involved in the fruit-latex syndrome.
• Allergen properties & biological function:Cas s 5 is a class I endochitinase with a possible defence function against plant pathogens.
• Allergen purification:

  Collada et al (1992) [306] purified basic endochitinases from Castanea sativa cotyledons and Allona et al. (1996) [6] expressed a chitinase (Cas s 5) in a bacterial system.

  Diaz-Perales et al (1998) [315] started from a PBS extract of chestnut which was salted-out with 80% saturated (NH₄)₂SO₄. The pellets were collected by centrifugation, dissolved in PBS buffer, and then the pH was brought to 3.5 by addition of 0.1 mol/L HCl. After centrifugation, the supernatants were dialyzed against distilled H₂O and freeze-dried. 30 mg of protein was dissolved in 20 mmol/L Tris-HCl (pH 8.0) and loaded onto a column (45 × 2.5 cm) of regenerated chitin, which was equilibrated with the same buffer. Elution was carried out with 300 ml (3 hours) of 20 mmol/L sodium acetate (pH 5.5) followed by 400 ml (4 hours) of 20 mmol/L acetic acid (pH 3.2) and chitinases containing fractions were pooled, concentrated, and desalted. These were subjected to cation-exchange chromatography with a Mono S HR 5/5 column (Pharmacia). The column was equilibrated with 10 mmol/L sodium acetate (pH 5.3) (buffer A), and eluted with a salt gradient to 0.5 mol/L NaCl in the same acetate buffer (buffer B). The two-step linear gradient was 0% to 50% of buffer B in 40 minutes and 50% to 100% of buffer B in 20 minutes (1 ml/min). SDS-PAGE and immunodetection was used.

  Diaz-Perales et al (2002) [762] expressed both recombinant Cas s 5 and Cas s 5 with its hevein domain deleted in Pichia. These were purified by gel filtration on Superdex 75 and cation exchange on Mono-S (both columns from Amersham Pharmacia) to give single bands on SDS-Page (Coomassie Blue stained) at the calculated masses.

• Other biochemical information:Both a Bet v 1 homologue and a class I endochitinase have been called Cas s 1 in the literature. The class I endochitinase was renamed as Cas s 5.

References (7)

• Diaz-Perales A, Sanchez-Monge R, Blanco C, Lombardero M, Carillo T, Salcedo G.
  What is the role of the hevein-like domain of fruit class I chitinases in their allergenic capacity?
  Clin Exp Allergy. 32(3): 448-54.. 2002
  PUBMEDID: 11940077
• Diaz-Perales A, Collada C, Blanco C, Sanchez-Monge R, Carrillo T, Aragoncillo C, Salcedo G
  Class I chitinases with hevein-like domain, but not class II enzymes, are relevant chestnut and avocado allergens.
  J. Allergy Clin. Immunol 102: 127-133. 1998
  PUBMEDID: 9679856
• Moneo I., LLamazares A., Curiel G. and Martinez J.
  Characterization of latex and chestnut antigens by immunoblotting.
  Ann. Allergy 75: 1-5.. 1995
  PUBMEDID: 7583867
• Allona I., Collada C., Casado R., Paz-Ares J. and Aragoncillo C.
  Bacterial expression of an active class Ib chitinase from Castanea sativa cotyledons.
  Plant Mol. Biol. 32: 1171-1176.. 1996
  PUBMEDID: 9002617
• Collada C., Casado R., Fraile A. and Aragoncillo C.
  Basic endochitinases are major proteins in Castanea sativa cotyledons.
  Plant Physiol 100: 778-783.. 1992
  PUBMEDID:
• Sanchez-Monge R, Blanco C, Perales AD, Collada C, Carrillo T, Aragoncillo C, Salcedo G
  Class I chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating
  J Allergy Clin Immunol. 106(1):190-5.. 2000
  PUBMEDID: 10887324
• Diaz-Perales A, Blanco C, Sanchez-Monge R, Varela J, Carrillo T, Salcedo G
  Analysis of avocado allergen (Prs a 1) IgE-binding peptides generated by simulated gastric fluid digestion
  J Allergy Clin Immunol. 112(5):1002-7. 2003
  PUBMEDID: 14610495

Biochemical Information for Cas s 8

• Allergen Name:Cas s 8
• Alternatve Allergen Names:Non-specific lipid transfer protein, nsLTP
• Allergen Designation:None
• Protein Family:non-specific lipid transfer proteins (LTPs) Pfam PF00234
• Sequence Known?:

  N-terminal only

• Allergen accession No.s:

  N/A

• 3D Structure Accession No.:Not determined
• Calculated Masses:N/A
• Experimental Masses:9725 Da by MALDI mass spectra
  13 kDa by SDS-PAGE
• Oligomeric Masses:Monomer
• Allergen epitopes:Not known
• Allergen stability:
  Process, chemical, enzymatic:Cas s 8 is a non-specific LTP and these are generally thermostable.
• Nature of main cross-reacting proteins:There is IgE cross-reactivity with other nsLTPs especially that from peach. None of the patients studied by García Sellés et al. (2002) [594] were SPT positive to chestnut only and only 5/47 were positive for a single food (all peach). 10/47 patients were SPT positive to peach, apple, Artemisa and chestnut which was the second largest group.
• Allergen properties & biological function:Cas s 8 is a non-specific lipid transfer protein (ns LTP). There is an expandable cavity between the four alpha-helices which can bind one or two lipids. nsLTPs have also been reported to act as plant defense proteins against bacterial and fungal infections and form the PR14 family of pathogenesis related proteins.
• Allergen purification:Chestnut LTP was isolated by Diaz-Perales et al. (2000) [43] from ground hulled chestnuts, which were defatted with acetone (1:5 w/v; 1 h; 4 °C), and then with ethanol:ether (1:3 v/v; 1 h; 4 °C). The dried material was then extracted with PBS buffer (0.1 M sodium phosphate, pH 7.0, 0.15 M NaCl; 1:5 w/v; 1 h; 4 °C). The extract was gel-filtrated on a Sephacryl S200 column (2.5 X 85 cm; Pharmacia Biotech, Uppsala, Sweden), using 0.1m ammonium acetate as elution buffer (1mL/min). Fractions enriched in chestnut LTP were pooled, freeze-dried, and then separated by RP-HPLC on a preparative Vydac-C4 column (22 X 250 mm; particle size 10 µm), eluting with a three-step linear gradient of acetonitrile in 0.1% trifluoroacetic acid (10% during 45 min; 10% to 35% in 140 min; 35% to 50% in 100 min and 50% to 85% in 15 min; 1 mL/min).
• Other biochemical information:Diaz-Perales et al. (2000) [43] report the 36 amino acid N-terminal sequence SITCTQVSKSLMPCLTYLKSNGGSPPPGTCCQGVKL.

References (2)

• García Sellés FJ, Diaz-Perales A, Sanchez-Monge R, Alcantara M, Lombardero M, Barber D, Salcedo G, Fernandez-Rivas M
  Patterns of reactivity to lipid transfer proteins of plant foods and Artemisia pollen: an in vivo study
  Int Arch Allergy Immunol. 128(2):115-122. 2002
  PUBMEDID: 12065911
• Diaz-Perales A, Lombardero M, Sanchez-Monge R, Garcia-Selles FJ, Pernas M, Fernandez-Rivas M, Barber D, Salcedo G.
  Lipid-transfer proteins as potential plant panallergens: cross-reactivity among proteins of Artemisia pollen, Castanea nut and Rosaceae, with different IgE-binding capacities.
  Clin Exp Allergy 2000, 30: 1403-1410.. 2000
  PUBMEDID: 10998016