Yellow jacket venom allergens, hyaluronidase and phospholipase: Sequence similarity and antigenic cross-reactivity with their hornet and wasp homologs.

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Yellow jacket venom allergens, hyaluronidase and phospholipase: Sequence similarity and antigenic cross-reactivity with their hornet and wasp homologs and possible implications for clinical allergy Te Piao King, PhD, Gang Lu, MD, Manuel Gonzalez, PhD, Naifeng Qian, MS, Lyudmila Soldatova, PhD Journal of Allergy and Clinical Immunology Volume 98, Issue 3, Pages 588-600 (September 1996) DOI: 10.1016/S0091-6749(96)70093-3 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Amino acid sequences of two yellow jacket and one hornet phospholipase: Ves v 1, Ves m 1, and Dol m 1. Underlined regions of Ves v 1 represent sequences established by Edman degradation. Residues of Ves m 1 or Dol m 1 that differ from those of Ves v 1 are indicated by dots. Data for Dol m 1 and Ves m 1 are from Soldatova et al.10 and Hoffman,11 respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid phospholipases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for reduced nVes v 1 and for fragment M of nDol m 1, 102-221, respectively. Lanes 1 and 2, nVes v 1 and rVes v 1; lanes 3 and 4, nDol m 1 and rDol m 1; lanes 5 and 6, nPol a 1 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid phospholipases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for reduced nVes v 1 and for fragment M of nDol m 1, 102-221, respectively. Lanes 1 and 2, nVes v 1 and rVes v 1; lanes 3 and 4, nDol m 1 and rDol m 1; lanes 5 and 6, nPol a 1 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid phospholipases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for reduced nVes v 1 and for fragment M of nDol m 1, 102-221, respectively. Lanes 1 and 2, nVes v 1 and rVes v 1; lanes 3 and 4, nDol m 1 and rDol m 1; lanes 5 and 6, nPol a 1 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Amino acid sequences of yellow jacket, hornet, and honeybee hyaluronidases: Ves v 2, Dol m 2, and Api m 2. Data for Dol m 1 and Api m 1 are from Lu et al.9 and Gmachl and Kreil,22 respectively. Underlined regions of Ves v 2 were determined by protein sequencing. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid hyaluronidases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for rVes v 2 and rDol m 2, respectively. Lanes 1 and 2, nVes v 2 and rVes v 2; lanes 3 and 4, nDol m 2 and rDol m 2; lanes 5 and 6, nPol a 2 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid hyaluronidases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for rVes v 2 and rDol m 2, respectively. Lanes 1 and 2, nVes v 2 and rVes v 2; lanes 3 and 4, nDol m 2 and rDol m 2; lanes 5 and 6, nPol a 2 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 SDS gel electrophoresis and immunoblot of reduced natural and recombinant vespid hyaluronidases. A, Dye-stained gel. B and C, Immunoblots probed with mouse sera specific for rVes v 2 and rDol m 2, respectively. Lanes 1 and 2, nVes v 2 and rVes v 2; lanes 3 and 4, nDol m 2 and rDol m 2; lanes 5 and 6, nPol a 2 and standards, respectively. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 5 Proliferation assay with spleen cells from mice immunized with reduced vespid phospholipases. Natural Dol m 1–, nVes v 1–, and nPol a 1–specific spleen cells were used in A and B, C and D, and E and F, respectively. Stimulating antigens are nDol m 1 (•), nVes v 1 (■), nPol a 1 (▴), and their respective reduced derivatives are represented by open symbols. Background proliferations were 4100, 8200, and 8300 cpm in these respective graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 6 Proliferation assay with spleen cells from mice immunized with recombinant hornet and yellow jacket hyaluronidases. Recombinant Dol m 2– and rVes v 2–specific spleen cells were used in A and B and C and D, respectively. Stimulating antigens are nDol m 2 and rDol m 2 (• and ○, respectively), nVes v 2 and rVes v 2 (■ and 2, respectively) and nPol a 2 (▴). Background proliferations were 7500 ± 500 cpm in these graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 6 Proliferation assay with spleen cells from mice immunized with recombinant hornet and yellow jacket hyaluronidases. Recombinant Dol m 2– and rVes v 2–specific spleen cells were used in A and B and C and D, respectively. Stimulating antigens are nDol m 2 and rDol m 2 (• and ○, respectively), nVes v 2 and rVes v 2 (■ and 2, respectively) and nPol a 2 (▴). Background proliferations were 7500 ± 500 cpm in these graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 6 Proliferation assay with spleen cells from mice immunized with recombinant hornet and yellow jacket hyaluronidases. Recombinant Dol m 2– and rVes v 2–specific spleen cells were used in A and B and C and D, respectively. Stimulating antigens are nDol m 2 and rDol m 2 (• and ○, respectively), nVes v 2 and rVes v 2 (■ and 2, respectively) and nPol a 2 (▴). Background proliferations were 7500 ± 500 cpm in these graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 6 Proliferation assay with spleen cells from mice immunized with recombinant hornet and yellow jacket hyaluronidases. Recombinant Dol m 2– and rVes v 2–specific spleen cells were used in A and B and C and D, respectively. Stimulating antigens are nDol m 2 and rDol m 2 (• and ○, respectively), nVes v 2 and rVes v 2 (■ and 2, respectively) and nPol a 2 (▴). Background proliferations were 7500 ± 500 cpm in these graphs. Journal of Allergy and Clinical Immunology 1996 98, 588-600DOI: (10.1016/S0091-6749(96)70093-3) Copyright © 1996 Mosby, Inc. Terms and Conditions