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IgE and IgG4 immvnoprinting patterns of patients sensitive to Dactylis glomerata pollen allergens are restricted by HLA class II genes
158
Abstracts
IV:S. Genetics and Specific Immune Response in Allergy and Asthma 0858
0857 TCR-a RESTRICTION OF IMMUNOGLOBULIN E RESPONSES TO SPECIFIC ANTIGEN Moffatt Miriam, Schou Carsten, Faux Jennie, Comelis Francois, Young Robert, Cookson William Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK
EVIDENCE FDR THE ROLE OF HLA-D ALLELES IN THE SUSCEPTIBILITY OF TYPE I ALLERGYTO HEVEIN OF HEVEA LATEX Rihs Hans-Peter. Chen Zhiping, Cremer Reinhold. Allmers Henning. Baur Xaver. BGFA Bochum, Kinderkrankenhaus Koln, Germany.
The particular allergens that atopic individuals react to is of clinical importance. House Dust Mite (HDM) allergy has been shown to be associated with an increased risk ofasthma. Important candidate loci for controlling the specificity of the 19E response include the HLA Class II genes and the genes of the T Cell Receptor (TCR). Previously we have shown strong genetic linkage between the TCRalo locus on chromosome 14, and 19E responses to a number of purified allergens. Genetic linkage was demonstrated in two independent sets of subjects, one British and one Australian, lotalling 823 subjects. These individuals, plus an additional set of 435 Australian subjects, were Iyped for HLA-DRB and for a biallelic polymorphism in the V «8.1 gene. In the first Australian data set, a significant association between allele I of Va8.1 and increased 19E responses to HDM (p = 0.01) and Derp II (p = 0.006) was seen. Confirmation of the association for HDM was seen in the second set of Australian subjects (p = 0.047) giving a combined result ofp = 0.001. Derp I and Derp II data for the second set was not available. Multiple regression analysis with 19E titre to Der p II as Ihe dependent variable and Va8 I and HLA-DR types as independent variables, showed ORB 1*02 also 10 be positively associated with 19E titres 10 Der p II (p = 0 004) in Australian data set I. It would appear therefore that interacting HLA-DR and TCRa restriction of 19E responses 10 particular antigens occurs.
Latex ellergy has become a serious problem for health cere workers (HCWs) and spina bifida (SB) patients due to the increased use of gloves and medical devices made of latex. The aim of this study was to elucidate the role of HLA-D alleles in latex allergy and in the specific immune response to the 4.7 kDa Hevein from Hevea brasiliensis. We investigated 216 latex-exposed subjects (111 HCWs. 105 SB patients) and so non-allergic blood donors (controls). 130 subjects (79 HCWs, 51 SB patients) showed positive anti latex-lgE values with the Pharmacia-CAP system (>0.35 kU!I). 30 randomly selected latex-lgE positive subjects (20 HCWs and 10 SB patients) were tested for their Hevein-specific IgE-concentrations by EAST, 18 were positive (specific IgE> 0.35 kU/I), 12 were negative. Statistical analysis (Fisher's exact test, 2-sided) of the generic HLA-DRBI allele distri bution revealed a significantly elevated ORB1 '04 frequency (67%) in Hevein-positive subjects when compared with Heveinnegative subjects (0%, p = 0.0003, pc =0.008) and with healthy controls (24%, p = 0.00 14, pc = 0.0361. Further, the frequency of ORB1 '04 was also elevated in the group of latex-lgE positive HCWs (44%) when compared with controls (24%, p=0.0089, pc e n.s.l. The data available so far indicate evidence for an association between the specific immune response to Hevein and HLA-DRB1 '04 in HCWs.
0859
0860
HLA-D GENES AND THE IgE IMMUNE RESPONSIVENESS TO A RECOMBINANT MITE ALLERGEN
IgE AND IgG4 IMMVNOPRINTING PATTERNS OF PATIENTS SENSITIVE TO Dactylis glomerata POLLEN ALLERGENS ARE RESTRICTED BY HLA CLASS II GENES
B. Martinez, S. Jimenez, GB.Ferrara", L. Caraballo InstituteofImmunological Research, University of Cartagena, Colombia 'Institutefor AdvancedBiotechnology, Geneva, Italy Genes on the HLA complex may be controlling the specificity of the 19E responsiveness in allergic diseases. In this work we studied the distribution of HLA-DRB I and TAP alleles (pCRlSSO typed) in 95 asthmatic patients and 89 non-allergic controls of the same ethnic (mulatto) group. Both groups were selected on the basis of their 19E responsiveness to a purified recombinant allergen (BIM), obtained from a cDNA library of the mite Blomia tropicalis(Bt) and representing a major allergen of this mite. 19E binding of sera to BtM was determined by plaque immunoassay. 45% of patients with 19E antibodies to BtM were HLA-DRB 1*03 (no particular subtype involved), while only 7% ofBtM-lgE-negative controls had this allele (p~O.OI). The frequencies of TAP alleles in patients and controls were: TAPI'010180.2% and 72%; TAPI*02011: 37.5% and 48.4%; TAPI*0301: 93% and 7.2%; TAP I '0401 135% and 8.2% TAPZ-AlE: 50% and 63%; TAP2-B/G: 62% and 68%; TAP2-C/D:28% and 23%; TAP2-H: 8.4% and 3.5%. Our results suggest that HLA-DRBI'03, is related with the 19E responsiveness to a major Bt allergen. No association was found belween this response and TAP alleles. Supported by a grant ofCOLCIENCIAS.
0861
Inacio Filipc", Peres Maria Joao *, Correia Sara', Desvaux FrancoisXavier#, Peltre Gabriel#, *Unidade Irnuno-Alergologia, Hospital Seuibal, Settibal , Portugal ; #Unlle d'lmmuno-Allergie, Institut Pasteur, Paris, France. A group of 50 patients allergic to Dactylisglomerata (Dac g) pollen allergens living in the same geographic area for more than 5 years was studied for the specificity of their IgE and IgG4 antibodies to the pollen. Two blood samples were collected 3 to 4 years apart. A soluble extract of Dac g pollen was separated by isoelectric focusing and blotted onto rutrocellulose membranes. Specific IgE and IgG4 response to pollen was evaluated by probing nitro-cellulose strips with patient sera and antihuman IgE and [gG4 as secondary antibodies. The patients were divided inlo two groups: group I recognised always the same small number of allergens and group 1I recognized a more complex allergen pattern that may vary. Both groups were typed for lILA-DPBI, DQAI, DQBI and DRB I by PCR-RFLP. The group of patients whith no cbanged pattern of allergens recognised had a strong association with HLA DRB1*0301 and ORB I*040I.
0862
ro
ASSOCIATION OF HLA CLASS 1I HAPLOTYPES WITH RESPONSE TO INHALANT ALLERGENS.
O'Amato Mauro, Picardi Adele, di Pietro Angelo, Malricardi Paolo, Testa Benedetto, ArianoRenate,Maggi Enrico,PlebaniAlessandro, Sac
FX.Desvaux I", Elnacro '". J.Gayraud '", F Marrache 0', V. Lepage'" M.N.Loste'" D.Charron "'and G.Peltre 'I>,
HLA-DRBI' ALLELES AS GENETIC RISK FACTORS IN ALLERGY PARIETARIA. A MULTICENTERL1NKAGE STUDY
Istitulo di Biologia Cellulare, CNR. Roma; Istituto di Genetica e Biofisica, CNR. Napoli; OASRS, Pratica di mare; Ospedale Ge.... e Maria, Napoli; PolicIiDioo di Careggi, Firenze; CIiDica PedialricaOspedali Civili, BIescia,U Facolta di Medicina, Napoli;Ospedale Giovanni da Procida, Salerno. In a previous epidemiological survey we have shown thai allergyto Parielaria and antibody (ab) response to the major allergen Par 0 I are positively associated with HI.A-DRBI' 1101104. In the presentfamily study we asked the questionsa) whether selected HLA-DRB 1* alleles were genetically lioked with allergy to Parietaria and lor with thepresenceof ab 10 Par oland b) whether the genetic risk representedby predisposing allels was comparable between monosensitized and polysensitized highly atopicpatients. Sixtynuclearfamilieswith two Parietariaallergicsibsand at least one non allergic parent were recruited in 6 ClinicalCenters. Skin prick tests (S1) were perfonoedand clinical history was recorded for all subjects. lILA·ORBI· alleles were typed osing the beteroduplex methodology. Total serum 19Elevel and Par 0 I specificab levels (lgG and 19E)were determined(pRISTand DARIA.respectively). In a preliminaryanalysisallele sharing betweensib pairs from informativeparents wasdetormined. Sharingof one and two alleleswere 90 and 80% and 75 and 60% in pain; concordant for ST positivity to Parietaria and for ab response to Par 0 I, respectively, whereas in absenceof linkagethe expectedvalues are SO and 25%.
(I) : Immuno-Allergie, Institut Pasteur, Paris, France. (2) : Hasp. Setubal, Lab. Imuno-Alergologia, Sett1bal, Portugal. (3) : ESFHLA & Allergy Network, France (4): Institut Biomedical des Cordelicrs, Paris France.
HLA haplotypes, IJ(E and IgG response were studied in 33 nuclear families with at least 2 affected sibs and no more than one allergic parent. All family members were assessed for the presence of atopic allergy by clinical history and skin prick tests with ten common inhalant allergens (ALK, Danemark). Clinical history using a questionnaire (asthma, rhinitis, conjunctivitis, atopic dermatitis, food and drug reactions) was analysed. HLA-DNA typing was performed by PCR-RFLP (according to lILA and Allergy workshop guidelines), and immunoprinting for IgE and IgG were made for grasses and birch pollen. Relation between HLA genotyping on one hand, and molecular and clinical phenotyping on the other hand will be discussed.
Abstracts
IV:S. Genetics and Specific Immune Response in Allergy and Asthma 0858
0857 TCR-a RESTRICTION OF IMMUNOGLOBULIN E RESPONSES TO SPECIFIC ANTIGEN Moffatt Miriam, Schou Carsten, Faux Jennie, Comelis Francois, Young Robert, Cookson William Nuffield Department of Clinical Medicine, University of Oxford, Oxford, UK
EVIDENCE FDR THE ROLE OF HLA-D ALLELES IN THE SUSCEPTIBILITY OF TYPE I ALLERGYTO HEVEIN OF HEVEA LATEX Rihs Hans-Peter. Chen Zhiping, Cremer Reinhold. Allmers Henning. Baur Xaver. BGFA Bochum, Kinderkrankenhaus Koln, Germany.
The particular allergens that atopic individuals react to is of clinical importance. House Dust Mite (HDM) allergy has been shown to be associated with an increased risk ofasthma. Important candidate loci for controlling the specificity of the 19E response include the HLA Class II genes and the genes of the T Cell Receptor (TCR). Previously we have shown strong genetic linkage between the TCRalo locus on chromosome 14, and 19E responses to a number of purified allergens. Genetic linkage was demonstrated in two independent sets of subjects, one British and one Australian, lotalling 823 subjects. These individuals, plus an additional set of 435 Australian subjects, were Iyped for HLA-DRB and for a biallelic polymorphism in the V «8.1 gene. In the first Australian data set, a significant association between allele I of Va8.1 and increased 19E responses to HDM (p = 0.01) and Derp II (p = 0.006) was seen. Confirmation of the association for HDM was seen in the second set of Australian subjects (p = 0.047) giving a combined result ofp = 0.001. Derp I and Derp II data for the second set was not available. Multiple regression analysis with 19E titre to Der p II as Ihe dependent variable and Va8 I and HLA-DR types as independent variables, showed ORB 1*02 also 10 be positively associated with 19E titres 10 Der p II (p = 0 004) in Australian data set I. It would appear therefore that interacting HLA-DR and TCRa restriction of 19E responses 10 particular antigens occurs.
Latex ellergy has become a serious problem for health cere workers (HCWs) and spina bifida (SB) patients due to the increased use of gloves and medical devices made of latex. The aim of this study was to elucidate the role of HLA-D alleles in latex allergy and in the specific immune response to the 4.7 kDa Hevein from Hevea brasiliensis. We investigated 216 latex-exposed subjects (111 HCWs. 105 SB patients) and so non-allergic blood donors (controls). 130 subjects (79 HCWs, 51 SB patients) showed positive anti latex-lgE values with the Pharmacia-CAP system (>0.35 kU!I). 30 randomly selected latex-lgE positive subjects (20 HCWs and 10 SB patients) were tested for their Hevein-specific IgE-concentrations by EAST, 18 were positive (specific IgE> 0.35 kU/I), 12 were negative. Statistical analysis (Fisher's exact test, 2-sided) of the generic HLA-DRBI allele distri bution revealed a significantly elevated ORB1 '04 frequency (67%) in Hevein-positive subjects when compared with Heveinnegative subjects (0%, p = 0.0003, pc =0.008) and with healthy controls (24%, p = 0.00 14, pc = 0.0361. Further, the frequency of ORB1 '04 was also elevated in the group of latex-lgE positive HCWs (44%) when compared with controls (24%, p=0.0089, pc e n.s.l. The data available so far indicate evidence for an association between the specific immune response to Hevein and HLA-DRB1 '04 in HCWs.
0859
0860
HLA-D GENES AND THE IgE IMMUNE RESPONSIVENESS TO A RECOMBINANT MITE ALLERGEN
IgE AND IgG4 IMMVNOPRINTING PATTERNS OF PATIENTS SENSITIVE TO Dactylis glomerata POLLEN ALLERGENS ARE RESTRICTED BY HLA CLASS II GENES
B. Martinez, S. Jimenez, GB.Ferrara", L. Caraballo InstituteofImmunological Research, University of Cartagena, Colombia 'Institutefor AdvancedBiotechnology, Geneva, Italy Genes on the HLA complex may be controlling the specificity of the 19E responsiveness in allergic diseases. In this work we studied the distribution of HLA-DRB I and TAP alleles (pCRlSSO typed) in 95 asthmatic patients and 89 non-allergic controls of the same ethnic (mulatto) group. Both groups were selected on the basis of their 19E responsiveness to a purified recombinant allergen (BIM), obtained from a cDNA library of the mite Blomia tropicalis(Bt) and representing a major allergen of this mite. 19E binding of sera to BtM was determined by plaque immunoassay. 45% of patients with 19E antibodies to BtM were HLA-DRB 1*03 (no particular subtype involved), while only 7% ofBtM-lgE-negative controls had this allele (p~O.OI). The frequencies of TAP alleles in patients and controls were: TAPI'010180.2% and 72%; TAPI*02011: 37.5% and 48.4%; TAPI*0301: 93% and 7.2%; TAP I '0401 135% and 8.2% TAPZ-AlE: 50% and 63%; TAP2-B/G: 62% and 68%; TAP2-C/D:28% and 23%; TAP2-H: 8.4% and 3.5%. Our results suggest that HLA-DRBI'03, is related with the 19E responsiveness to a major Bt allergen. No association was found belween this response and TAP alleles. Supported by a grant ofCOLCIENCIAS.
0861
Inacio Filipc", Peres Maria Joao *, Correia Sara', Desvaux FrancoisXavier#, Peltre Gabriel#, *Unidade Irnuno-Alergologia, Hospital Seuibal, Settibal , Portugal ; #Unlle d'lmmuno-Allergie, Institut Pasteur, Paris, France. A group of 50 patients allergic to Dactylisglomerata (Dac g) pollen allergens living in the same geographic area for more than 5 years was studied for the specificity of their IgE and IgG4 antibodies to the pollen. Two blood samples were collected 3 to 4 years apart. A soluble extract of Dac g pollen was separated by isoelectric focusing and blotted onto rutrocellulose membranes. Specific IgE and IgG4 response to pollen was evaluated by probing nitro-cellulose strips with patient sera and antihuman IgE and [gG4 as secondary antibodies. The patients were divided inlo two groups: group I recognised always the same small number of allergens and group 1I recognized a more complex allergen pattern that may vary. Both groups were typed for lILA-DPBI, DQAI, DQBI and DRB I by PCR-RFLP. The group of patients whith no cbanged pattern of allergens recognised had a strong association with HLA DRB1*0301 and ORB I*040I.
0862
ro
ASSOCIATION OF HLA CLASS 1I HAPLOTYPES WITH RESPONSE TO INHALANT ALLERGENS.
O'Amato Mauro, Picardi Adele, di Pietro Angelo, Malricardi Paolo, Testa Benedetto, ArianoRenate,Maggi Enrico,PlebaniAlessandro, Sac
FX.Desvaux I", Elnacro '". J.Gayraud '", F Marrache 0', V. Lepage'" M.N.Loste'" D.Charron "'and G.Peltre 'I>,
HLA-DRBI' ALLELES AS GENETIC RISK FACTORS IN ALLERGY PARIETARIA. A MULTICENTERL1NKAGE STUDY
Istitulo di Biologia Cellulare, CNR. Roma; Istituto di Genetica e Biofisica, CNR. Napoli; OASRS, Pratica di mare; Ospedale Ge.... e Maria, Napoli; PolicIiDioo di Careggi, Firenze; CIiDica PedialricaOspedali Civili, BIescia,U Facolta di Medicina, Napoli;Ospedale Giovanni da Procida, Salerno. In a previous epidemiological survey we have shown thai allergyto Parielaria and antibody (ab) response to the major allergen Par 0 I are positively associated with HI.A-DRBI' 1101104. In the presentfamily study we asked the questionsa) whether selected HLA-DRB 1* alleles were genetically lioked with allergy to Parietaria and lor with thepresenceof ab 10 Par oland b) whether the genetic risk representedby predisposing allels was comparable between monosensitized and polysensitized highly atopicpatients. Sixtynuclearfamilieswith two Parietariaallergicsibsand at least one non allergic parent were recruited in 6 ClinicalCenters. Skin prick tests (S1) were perfonoedand clinical history was recorded for all subjects. lILA·ORBI· alleles were typed osing the beteroduplex methodology. Total serum 19Elevel and Par 0 I specificab levels (lgG and 19E)were determined(pRISTand DARIA.respectively). In a preliminaryanalysisallele sharing betweensib pairs from informativeparents wasdetormined. Sharingof one and two alleleswere 90 and 80% and 75 and 60% in pain; concordant for ST positivity to Parietaria and for ab response to Par 0 I, respectively, whereas in absenceof linkagethe expectedvalues are SO and 25%.
(I) : Immuno-Allergie, Institut Pasteur, Paris, France. (2) : Hasp. Setubal, Lab. Imuno-Alergologia, Sett1bal, Portugal. (3) : ESFHLA & Allergy Network, France (4): Institut Biomedical des Cordelicrs, Paris France.
HLA haplotypes, IJ(E and IgG response were studied in 33 nuclear families with at least 2 affected sibs and no more than one allergic parent. All family members were assessed for the presence of atopic allergy by clinical history and skin prick tests with ten common inhalant allergens (ALK, Danemark). Clinical history using a questionnaire (asthma, rhinitis, conjunctivitis, atopic dermatitis, food and drug reactions) was analysed. HLA-DNA typing was performed by PCR-RFLP (according to lILA and Allergy workshop guidelines), and immunoprinting for IgE and IgG were made for grasses and birch pollen. Relation between HLA genotyping on one hand, and molecular and clinical phenotyping on the other hand will be discussed.