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DQ transgenic mice to ragweed allergen
J ALLERGY CLIN IMMUNOL
Abstracts
S369
VOLUME 104, NUMBER 1, PART 2
opportunity to describe and compare allergic sensitivity to common indoor allergens across the seven sites. ln order to be eligible, skin test reactivity had to be demonstrated to one or more of the following allergens: dust mites (D. pteronyssinus, D. farinae), cockroach, mouse, rat, cat, dog, or mold (Altemaria, Aspergillus, Cladosporium and Penicillum). Of I I I4 asthmatic children who were skin tested, 992 (89%) were skin test positive to one or more indoor allergen and thus were eligible to participate in the study. Nine hundred forty two of these children have completed both baseline clinical and home evaluations and have been enrolled. Skin test reactivity (% positive) is demonstrated in the following rable: BO
BR
CH
DA
NY
SE
TU
TOTAL
DUSTMITE COCKROACH MOLD RAT
59% 68% 43% 40%
64% 81%
51% 61%
38% 61%
84% 80% 73% 16% 22% 30%
55% 79% 41% 22% 33% JO‘%
78% ‘6%
MOUSE CAT
38% 69% 48% IS% 20% 29%
59% 58% 59% 4% 15% 39Qc
62% 69% 50% I96 28% 44%
41% 23%
46% 15% 21% 53%
The most prominent allergens to which the majority of children at all sites reacted were house dust mites and cockroach. Summarizing the data across all sites, it was found that 62% and 69% of the children enrolled reacted to dust mite and cockroach, respectively. Dust mite reactivity was highest in Dallas and Seattle (84% and 78%. respectively) and, at all sites except Seattle, the majority of children were cockroach-allergic. Reactivity to all other allergens depended upon the testing site. The majority of children from Boston demonstrated a positive skin test response to mouse (-5 I %) as did >30% from the Bronx and New York. In contrast, at all other sites, skin test reactivity to mouse occurred in approximately 20% or less of those tested. While mold positivity was greatest in the children from Dallas (73%) and Tucson (59%), 40-45%. of children from all other sites were found to be moldpositive as well. These results extend the findings from the previous National Cooperative Inner City Asthma Study and demonstrate that the majority of inner city children with severe asthma have multiple indoor allergen sensitivities. In addition, despite wide geographic and climatic variation, reactivity to dust mite, cockroach and mold allergens was found to be common among all the children tested.
1083
HLA-DQ/Human roach Allergens
CD4 Restricted Immune Response to Cockin Transgenic Mice B fcrpouchado*. S ChupoWI/*. E Marietta *, C Weilerf. C David* *Department of Immunology, Mayo Clinic, Rochester, MN, USA tDepartment of Internal Medicine, Division of Allergy and Infectious Disease, Mayo Clinic, Rochester, MN, USA We investigated the immune response to the German Cockroach (Blattella germanica). and its major antigen, Blattella germanica group 5(Bla g 5). For this purpose, we used a doubletransgenic, double-knockout mouse expressing human HLADQ8. HLA-DQ6 and CD4 molecules in the absence of mouse class II and mouse CD4. Transgenic mice were primed and challenged with cockroach extract or individual synthetic peptides representing Bla g 5. Strong T cell responses to cockroach extract were detected in both HLA-DQ/hCD4+ transgenic mice. The responses were two times lower in mice expressing HLA-DQ
molecule in the context of mouse CD4. Under similar treatment, no responses were found in the double-knockout Ab’/mCD4’ mice and in mice expressing human CD4 molecule alone. HLADQ/hCD4+ mice produced primarily IL-5, IL-IO, and IL-13. Minimal amounts of IL-4 were detected only in HLA-DQ6/hCD4+ mice. IFN-g production was low in both transgenic mouse, suggesting a predominantly Th2 type response. Cockroach extract immunized HLA DQ8/hCD4+ mice recognized only one of the 20 peptides of Bla g 5 while HLA-DQ6/hCD4+ mice responded primarily to four peptides. Primed with individual peptides, both HLA-DQ/hCD4+ mice responded maximally to peptides IO(residues 91-l IO) and 17(residues 161-180). In addition, HLADQ6/hCD4+ mice responded to peptide 16(residues 15 1- 170). Thus, peptides IO and I7 contained the major T cell HLADQ/hCD4+ epitopes and could be recognized by both HLA-DQ8 and HLA-DQ6 transgenic mice. Transgenic mice represent a new tool for investigating and studying the immune responses to cockroach allergen. Our results suggest that therapeutic strategies aimed at developing antagonist peptides might be a useful treatment (immunotheraphy) for allergic asthma.
1084 Immune Transgenic Eric
Response Mice
Marietta*.
of HLA-DQ, to Ragweed Koji
lijimaf,
HLA-DR, and HLA-DR/DQ Allergen Sveflana Chapoval*, Michele
Smart*,
Chella
David*
*Department of Immunology, Mayo Clinic tAllergic Diseases Research Laboratory, Mayo Clinic Transgenic mice expressing human DQ6, DQS, DR2. DR3, and DR3/DQ6 genes in Class II knock-out (AbO) mice were generated. The in vitro and in vivo responses of transgenic mice and their negative littermates to short ragweed allergen were examined. Mice were immunized with whole allergenic extract subcutaneously and lymph node cells were challenged in vitro with either extract or individual peptides representing antigen 5 from short ragweed. For the in vivo studies, transgenic mice were actively immunized and later challenged intranasally with SRW allergenic extract. We have found that DQ and DR mice differ in the set of epitopes recognized in vitro and in some characteristics of the in vivo response to SRW (levels of eosinophilia, cytokines, airway hyperreactivity). Lately we have found that SRW-sensitized HLA-DQ6 transgenic mice develop substantial BAL eosinophilia in response to i.n. challenge with low concentration of naturally processed single peptide from antigen 5. Recently, we have generated and characterized models of SRW -induced astbma using transgenic mice with human HLA-DQ (DQ6 or DQS), human CD4 (hCD4) or both on a genetic background that lacks MHC II and mouse CD4. The HLA-DQ/hCD4 transgenic mice developed a pulmonary eosinophilia and associated lung tissue damage, an increase in total protein, eosinophil peroxidase, IL-5 and IL-I 3 production in bronchoalveolar lavage. This was absent in double knockout mice and mice expressing HLA-DQ8 or hCD4 alone. Thus, specific HLA Class I1 molecules are critical to SRWinduced asthma. Our experimental uansgenic animal models of asthma are useful for elucidating the multiple mechanisms that contribute to disease and can be invaluable to study the efficacy of potential therapies. These findings also provide the rationale for developing strategies to target HLA II molecules to effectively control allergic asthmatic conditions.
Abstracts
S369
VOLUME 104, NUMBER 1, PART 2
opportunity to describe and compare allergic sensitivity to common indoor allergens across the seven sites. ln order to be eligible, skin test reactivity had to be demonstrated to one or more of the following allergens: dust mites (D. pteronyssinus, D. farinae), cockroach, mouse, rat, cat, dog, or mold (Altemaria, Aspergillus, Cladosporium and Penicillum). Of I I I4 asthmatic children who were skin tested, 992 (89%) were skin test positive to one or more indoor allergen and thus were eligible to participate in the study. Nine hundred forty two of these children have completed both baseline clinical and home evaluations and have been enrolled. Skin test reactivity (% positive) is demonstrated in the following rable: BO
BR
CH
DA
NY
SE
TU
TOTAL
DUSTMITE COCKROACH MOLD RAT
59% 68% 43% 40%
64% 81%
51% 61%
38% 61%
84% 80% 73% 16% 22% 30%
55% 79% 41% 22% 33% JO‘%
78% ‘6%
MOUSE CAT
38% 69% 48% IS% 20% 29%
59% 58% 59% 4% 15% 39Qc
62% 69% 50% I96 28% 44%
41% 23%
46% 15% 21% 53%
The most prominent allergens to which the majority of children at all sites reacted were house dust mites and cockroach. Summarizing the data across all sites, it was found that 62% and 69% of the children enrolled reacted to dust mite and cockroach, respectively. Dust mite reactivity was highest in Dallas and Seattle (84% and 78%. respectively) and, at all sites except Seattle, the majority of children were cockroach-allergic. Reactivity to all other allergens depended upon the testing site. The majority of children from Boston demonstrated a positive skin test response to mouse (-5 I %) as did >30% from the Bronx and New York. In contrast, at all other sites, skin test reactivity to mouse occurred in approximately 20% or less of those tested. While mold positivity was greatest in the children from Dallas (73%) and Tucson (59%), 40-45%. of children from all other sites were found to be moldpositive as well. These results extend the findings from the previous National Cooperative Inner City Asthma Study and demonstrate that the majority of inner city children with severe asthma have multiple indoor allergen sensitivities. In addition, despite wide geographic and climatic variation, reactivity to dust mite, cockroach and mold allergens was found to be common among all the children tested.
1083
HLA-DQ/Human roach Allergens
CD4 Restricted Immune Response to Cockin Transgenic Mice B fcrpouchado*. S ChupoWI/*. E Marietta *, C Weilerf. C David* *Department of Immunology, Mayo Clinic, Rochester, MN, USA tDepartment of Internal Medicine, Division of Allergy and Infectious Disease, Mayo Clinic, Rochester, MN, USA We investigated the immune response to the German Cockroach (Blattella germanica). and its major antigen, Blattella germanica group 5(Bla g 5). For this purpose, we used a doubletransgenic, double-knockout mouse expressing human HLADQ8. HLA-DQ6 and CD4 molecules in the absence of mouse class II and mouse CD4. Transgenic mice were primed and challenged with cockroach extract or individual synthetic peptides representing Bla g 5. Strong T cell responses to cockroach extract were detected in both HLA-DQ/hCD4+ transgenic mice. The responses were two times lower in mice expressing HLA-DQ
molecule in the context of mouse CD4. Under similar treatment, no responses were found in the double-knockout Ab’/mCD4’ mice and in mice expressing human CD4 molecule alone. HLADQ/hCD4+ mice produced primarily IL-5, IL-IO, and IL-13. Minimal amounts of IL-4 were detected only in HLA-DQ6/hCD4+ mice. IFN-g production was low in both transgenic mouse, suggesting a predominantly Th2 type response. Cockroach extract immunized HLA DQ8/hCD4+ mice recognized only one of the 20 peptides of Bla g 5 while HLA-DQ6/hCD4+ mice responded primarily to four peptides. Primed with individual peptides, both HLA-DQ/hCD4+ mice responded maximally to peptides IO(residues 91-l IO) and 17(residues 161-180). In addition, HLADQ6/hCD4+ mice responded to peptide 16(residues 15 1- 170). Thus, peptides IO and I7 contained the major T cell HLADQ/hCD4+ epitopes and could be recognized by both HLA-DQ8 and HLA-DQ6 transgenic mice. Transgenic mice represent a new tool for investigating and studying the immune responses to cockroach allergen. Our results suggest that therapeutic strategies aimed at developing antagonist peptides might be a useful treatment (immunotheraphy) for allergic asthma.
1084 Immune Transgenic Eric
Response Mice
Marietta*.
of HLA-DQ, to Ragweed Koji
lijimaf,
HLA-DR, and HLA-DR/DQ Allergen Sveflana Chapoval*, Michele
Smart*,
Chella
David*
*Department of Immunology, Mayo Clinic tAllergic Diseases Research Laboratory, Mayo Clinic Transgenic mice expressing human DQ6, DQS, DR2. DR3, and DR3/DQ6 genes in Class II knock-out (AbO) mice were generated. The in vitro and in vivo responses of transgenic mice and their negative littermates to short ragweed allergen were examined. Mice were immunized with whole allergenic extract subcutaneously and lymph node cells were challenged in vitro with either extract or individual peptides representing antigen 5 from short ragweed. For the in vivo studies, transgenic mice were actively immunized and later challenged intranasally with SRW allergenic extract. We have found that DQ and DR mice differ in the set of epitopes recognized in vitro and in some characteristics of the in vivo response to SRW (levels of eosinophilia, cytokines, airway hyperreactivity). Lately we have found that SRW-sensitized HLA-DQ6 transgenic mice develop substantial BAL eosinophilia in response to i.n. challenge with low concentration of naturally processed single peptide from antigen 5. Recently, we have generated and characterized models of SRW -induced astbma using transgenic mice with human HLA-DQ (DQ6 or DQS), human CD4 (hCD4) or both on a genetic background that lacks MHC II and mouse CD4. The HLA-DQ/hCD4 transgenic mice developed a pulmonary eosinophilia and associated lung tissue damage, an increase in total protein, eosinophil peroxidase, IL-5 and IL-I 3 production in bronchoalveolar lavage. This was absent in double knockout mice and mice expressing HLA-DQ8 or hCD4 alone. Thus, specific HLA Class I1 molecules are critical to SRWinduced asthma. Our experimental uansgenic animal models of asthma are useful for elucidating the multiple mechanisms that contribute to disease and can be invaluable to study the efficacy of potential therapies. These findings also provide the rationale for developing strategies to target HLA II molecules to effectively control allergic asthmatic conditions.