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研究生(外文):Yan-Ming Chen
論文名稱(外文):Identification and analysis of the antigenic and allergenic determinants on the allergen Pen n 18: epitope mapping by rabbit polyclonal antibodies and human IgE.
外文關鍵詞:allergenic determinantantigenic determinantserine proteasehapten
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全世界大約有20%的人口面臨過敏性疾病的威脅。過敏性疾病包括了過敏性鼻炎、關節炎、氣喘及各種過敏性反應等。當人體內具有辨識過敏原特異性的IgE抗體與過敏原結合後,會進一步與其受器發生交聯反應,並導致一些免疫細胞活化及釋放介質引發過敏性反應。黴菌的孢子是空氣中含量最豐富的有機物質,很久之前便發現它是導致呼吸系統性過敏疾病的重要來源之一。一般來說,Aspergillus和Penicillium這兩個屬的黴菌是室內最常見兩種黴菌。直到目前為止,青黴菌屬大約已經有150種以上的種類被發現,根據研究,最常見的5種青黴菌是P. spinulosum、 P. brevicompactum、 P. oxalicum、 P. notatum 和 P. citrunum.
青黴菌屬中的點青黴是一個已知室內型黴菌,會引起支氣管的氣喘疾病。同時皮膚測試中也使用點青黴的粗萃取物來診斷病人是否對黴菌產生過敏性反應。由於近年來重組DNA科技的日新月異,許多過敏原分子特性的分析都可利用重組蛋白進行研究,並加速相關研究的進行。本研究室的學姐利用二維電泳將點青黴的粗萃取物展開後,利用西方墨點法找到了一個分子量38kDa的過敏原,並將其命名為Pen n 18。接下來便利用基因轉殖的技術以大腸桿菌表現出Pen n 18的重組蛋白,並在其N端序列加上六個Histidine。我們利用一根親和力管柱,純化rPen n 18。純化後的重組蛋白rPen n 18使用病人的血清進行西方墨點試驗,發現其仍然具有與IgE抗體結合的能力,證明了此重組蛋白擁有與自然狀態nPen n 18相同的致敏特性,可以做進一步的研究。
為了找出過敏原Pen n 18的抗原及過敏原決定點,我們使用CNBr化學切割及酵素分解的方式將rPen n 18處理成不同大小的片段,並使用高效能液相層析法將這些片段分離並純化,在將其轉印到PVDF纖維膜上以兔子多株抗體及八位病人的血清進行西方墨點試驗。將其中十九段呈現陽性反應的片段以N端蛋白質定序儀分析其N端序列,並利用MALDI-TOF質譜儀分析所有片段的質量數,結果一共得到至少九段含有抗原決定點的區域,以及九段含有過敏原決定點的區域。我們利用電腦模擬Pen n 18分子的三級結構,觀察這些區域的分佈,發現到幾乎所有的抗原或過敏原決定點都包含一些暴露在Pen n 18分子表面的部分。
在本次實驗之中,我們發現重組過敏原rPen n 18能夠表現出與自然狀態Pen n 18相同的IgE抗體結合能力,這也證明使用容易製備及純化的重組蛋白,能夠提供一個明確且便利的方法用來進一步研究過敏原的結構,並用來發展更精確的診斷方式或治療策略。
Almost 20% of the population suffers from Type Ⅰ allergy. The symptoms of Type Ⅰ allergy (allergic rhinitis, conjunctivitis, asthma, and anaphylaxis) result from the cross-link of effector cell-bound IgE antibodies by innocuous allergens. Fungal spores are the most abundant organic particles in the atmosphere and were long recognized as important cause of respiratory allergy. In general, Aspergillus and Penicillium are commonly found indoor. Until now, more than 150 different species have been discovered in the genus Penicillium. The five most frequently found species of Penicillium indoor and outdoor in Topeka KS were P. spinulosum, P. brevicompactum, P. oxalicum, P. notatum, and P. citrunum.
P. notatum is a well-know indoor aeroallergen considered as one of the causative agents of extrinsic bronchial asthma and is frequently included in skin test panels for allergic diagnosis. Molecular characterization of allergens by recombinant DNA technology progressed rapidly during the last few years. Pen n 18, a novel allergen from P. notatum was identified by two-dimensional immunoblotting, and followed by the molecular cloning and expression of this allergen as a hexahistidine- tagged fusion protein in Escherichia coli. Further purification by Ni-chelate chromatography led to a homologous protein fraction of rPen n 18. Purified rPen n 18 was tested by immunoblotting with sera from patients with allergy and shown strong reaction to IgE, indicating that the allergenic properties are well conserved in the recombinant-derived protein.
In order to identify the antigenic and allergenic determinants of Pen n 18, rPen n 18 was cleaved by cyanogens bromide, and digested with Lysyl-C endopeptidase, Chymotrypsin and Glu-C endopeptidase to generate various peptide fragments which were then separated by HPLC and electroblotting to PVDF membrane. After immuodetection with rabbit plolyclonal antibodies and sera of eight patients, nineteen positive peptide fragments were analyzed for N-terminal protein sequence and all peptides were submitted for mass analysis. Finally we found at least nine regions contain antigenic determinants and seven regions contain allergenic determinants. Finally, By 3D-structure homology modeling of rPen n 18, we found that almost all positive peptide fragments locate at the exposed regions of the molecule.
In summary, we demonstrated that the recombinant allergen of rPen n 18 exhibited a high potency of reactivity with IgE. The facile production and purification of recombinant mold allergens like rPen n 18 provide a straightforward strategy for a detailed study of epitopes structure as well as the development of accurate diagnosis protocols which may lead to effective therapeutic strategies in the future.
1.Aalberse RC, Gaag R, Leeuwen J. Serologic aspexts of IgG4 antibodies. Prolonged immunization results in an IgG4-restricted response. J. Immunol. (1983)130:722
2.Abad-Zapatero C, Rydel TJ, Erikson J. Revised 2.3 A structure of porcine pepsin: evidence for a flexible subdomain. Proteins Struct. Funct. Genet. (1990) 8: 62-81.
3.Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. (1997) 25: 3389-3402.
4.Ball T, Vrtala S, Sperr WR, Valent P, Susani M, Graft D, Valenta R. Isolation of an immunodominant IgE hapten from an epitope expression cDNA library : dissection of the allergic effector reaction. J. Biol. Chem. (1994) 269: 28323-28328.
5.Barnes PJ. (1992) Asthma in adults, in Current Therapy in allergy, immunology, and rheumatology. 4thed. (Lichtenstein L.M., Fauci A.S. eds.), Mosby, St. Louis, MO.
6.Banani B, Paul Agm Jordan NF, Viswananth PK. Immunological characterizationof Asp f 2, a magor allergen from Aspergillus fumigatus associated with allergic bronchopulmonary aspergillosis. Infect. Immunity (1998) 66: 5175-5182.
7.Berzofsky JA. Intrinsic and extrinsic factors in protein antigenic structure. Sciences (1985) 229:923-940.
8.Betzel C, Belleman M, Pal GP, Bajorath J, Saenger W, Wilson KS. X-ray and model-building studies on the specificity of the active site of proteinase K. Proteins Struct. Funct. Genet. (1989) 4: 157-164.
9.Bloemen PG, van den Tweel MC, Henricks PA. Expression and modulation of adhesion molecules on human bronchial epithelial cells. Am. J. Respir. Cell Mol. Biol. (1993) 9: 586-593.
10.Bode W, Papamokos E, Musil D, Seemuller U, Fritz H. Refined 1.2 A crystal structure of the complex formed between subtilisin Carlsberg and the inhibitor eglin c. Molecular structure of eglin and its detailed interaction with subtilisin. EMBO J., (1986) 5: 813-818.
11.Bush RK, Yunginger JW. Standaridization of fungal allergens. Clinical reviews in allergy. (1987) 5(1): 3-21.
12.Cairns JA, Walls AF. Mast cell tryptase is a mitogen for epithelial cells. Stimulation of IL-8 production and intercellular adhesion molecule-Ⅰ expression. J. Immunol. (1996) 156: 275-283.
13.Chang TW. The pharmacological basis of anti-IgE therapy. Nature Biotech. (2000) 18: 157- 162.
14.Chow LP, Su NY, Yu CJ, Chiang BL, Shen HD. Identification and expression of Pen c 2, a novel allergen from Pencicillium citrinum. Biochem. J. (1999) 341:51-59.
15.Christian B, Gour PP, Wolfram S. Three-dimensional structure of proteinase K at 0.15-nM resolution. Eur. J. Biochem. (1988) 178: 155-171.
16.Cooke RA, Berngard JH, Hebald S, Stull A. Serological evidence of immunity with co —existing sensitization in hay fever type of human allergy. J. Exp. Med. (1935) 62: 733-750.
17.Cromwell O, Hamid Q, Corrigan CJ, Barkans I, Meng Q, Collins PD, Kay AB. Expression and generation of interleukin-8, IL-6 and granulocytemacrophage colony-stimulating factor by bronchial epithelial cells and enhancement by IL-1 beta anad tumour necrosis factor-alpha. Immunology (1992) 77: 330-337.
18.Dudler T, Machado DC, Kolbe L, Annand RR, Rhodes N, Gelb MH, Koelsch E, Suter M, Helm BA.A link between catalytic activity, IgE-independent mast cell activation, and allergenicity of bee venom phospholipase A2. J. Immunol. (1995) 155: 2605-2613.
19.Dreskin SC, Metzger H. The high-affinity receptor for immunoglobulin E. Prospects for the therapy of immediate hypersensitivity reactions. JAMA. (1988) 260(9):1265-8.
20.Eijsink VGH, Vriend G, van de Burg B, Venema G, Stulp BK. Contribution of the C-terminal amino acid to the stability of Bacillus subtilis neutral protease. Protein Engng. (1990) 4: 99-104.
21.Fedorov AA. Ball T. Valenta R. Almo SC. X-ray crystal structures of birch pollen profilin and Phl p 2. Int. Arch. Allergy Immunol. (1997) 113: 109-113.
22.Ferreira FD, Mayer P, Sperr WR, Valent P, Seiberler S, Ebner C, Liehl E, Scheiner O, Kraft D, Valenta R. Induction of IgE antibodies with predefined specificity in rhesus monkeys with recombinant birch pollen allergens, Bet v1 and Bet v 2. J. Allergy Clin. Immunol. (1996) 97: 95-103.
23.Fullmer CS. Identification of cysteine-containing peptides in protein digests by high-performance liquid chromatography. Anal. Biochem. (1984) 142: 336-339.
24.Gajhede M, Osmark P, Poulsen FM, Ipsen G, Larsen JN, Joost van Beerven RJ, Schou C, Lowenstein H. Spangfort MD. X-ray and NMR structure of Bet v1, the origin of birch pollen allergy. Nat. Struct. Biol. (1996) 3: 1040-1045.
25.Garcia ZE, Rothenberg ME, Ownbey RT, Celestin J, Leder P, Lusher AD. Human eotaxin is a specific chemoattractant for eosinophil cells and provides a new mechanism to explain tissue eosinophilia. Nature Med. (1996) 2: 449-56.
26.Geysen HM, Rodda SJ, Mason TJ, Tribbick G, Schoofs PG. Strategies for epitope analysis using peptide synthesis. J. Immunol. Methods. (1987) 102: 259-274.
27.Greer J. Comparative model-building of the mammalian serine proteases. J. Mol. Biol, (1981) 153: 1027-1042.
28.Gross E. The cyanogen bromide reaction. Method Enzymol. (1967) 11: 238.
29.Gros P, Betzel C, Dauter Z, Wilson KS, Hol WG.Molecular dynamics refinement of a thermitase-eglin-c complex at 1.98 A resolution and comparison of two crystal forms that differ in calcium content. J. Mol. Biol. (1989) 210(2): 347-67.
30.Guex N, Peitsch MC. SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis (1997) 18: 2714-2723.
31.Hakkaart GA, Aalberse RC, van Ree R. Epitope mapping of the house-dust-mite allergen Der p 2 by means of site-directed mutagenesis. Allergy. (1998) 53: 165-172.
32.Hawkes R, Niday E, and Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal. Biochem. (1982) 119: 142-147.
33.Hirono S, Akagawa H, Mitsui Y, Iitaka Y. Crystal structure at 2.6 A resolution of the complex of subtilisin BPN' with streptomyces subtilisin inhibitor. J. Mol. Biol, (1984) 178: 389-413.
34.Hopp TP, Woods KR. A computer program for predicting protein antigenic determinants. Mol. Immunol. (1983) 20(4): 483-489.
35.Horner WE, Helbling A, Salvaggio JE, Lehrer SB. Fungal allergens. Clin.Microbiol.Rev (1995) 8:161-179.
36.Juniper CP, How MJ, Goodwin BFJ, Kinshott AK. Bacillus subtilis enzymes:A 7-years clinical, epidemiological and immunological study of an industrial allergen. J. Soc. Occup. Med. (1997) 27: 3-12
37.Kay AB. (1997) Allergy and Allergic Diseases. Oxford; Blackwell Science.
38.Kelley LA, MacCallum RM, Sternberg MJE. Enhanced Genome Annotation using Structural Profiles in the Program 3D-PSSM. J. Mol. Biol. (2000) 299(2): 499-520.
39.Laffer S, Vrtala S, Duchene M, van Ree R, Kraft D, Scheiner O. IgE-binding capacity of recombinant timothy grass(Phleum pratense) pollen allergens. J. Allergy Clin. Immunol. (1994) 94: 88-94.
40.Laver WG, Air GM Websteer GR, Smith-gill SJ. Epitopes on protein antigens: misconceptions and realities. Cell (1990) 61:553-556.
41.Lebecque S, Dotecek C, Laffer S, Visco V, Denepoux S, Pin JJ, Guret C, Boltz-Nitulescu G, Weyer A, Valenta R. Immunologic characterization of monoclonal antibodies that modulate human IgE binding to the mahor birch pollen allergen Bet v1. J. Allergy. Clin. Immunol. (1997) 99: 374-384.
42.Leibers V, Sander I, Kampen VV, Raulf-Heisoth M, Rozynek P, Baur X. Overview on denominated allergens. Clin. Exp. Allergy. (1996) 26: 494-516
43.Licorish K, Novey HS, Kozak P, Fairshter RD, Wilson AF. Role of Alternaria and Penicillium spores in the pathogenesis of asthma. J. Allergy. Clin. Immunol. (1985) 76: 819-825.
44.Loveless MH. Immunological studies of pollinosis. I. The presence of two antibodies related to the same pollen-antigen in serum of treated hay fever patients. J. Immunol. (1940) 38: 25-50.
45.Machado DC, Horton D, Harrop R,Peachell PT, Helm BA. Potential allergens stimulate the release of mediators of the allergic response from cells of mast cell lineage in the absence of sensitization with antigen-specific IgE. Eur. J. Immunol. (1996) 26: 2972-2980.
46.Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membrane. J. Biol. Chem. (1987) 262: 10035- 10038.
47.Menz G, Dolecek C, Schőnheit-Kenn U, Ferreira F, Moser M, Schneider T. Serological and skin-test diagnosis of birch pollen allergy with recombinant Bet v Ⅰ, the major birch pollen allergen. Clin. Exp. Allergy (1996) 26: 50-60.
48.Morgenstern. DIALIGN 2: improvement of the segment-to-segment approach to multiple sequence alignment. Bioinformatics. (1999) 15: 211-218.
49.Moser M, Crameri R, Menz G, Schineider T, Dudler T, and Virchoe C. Cloning andexpression of recombinant Aspergillus fumigatus 1/a with IgE binding and typeⅠ skin test activity. J. Immunol. (1992) 149: 454-460.
50.Muilenberg M, Burge H, Sweet T, Solomon W. Penicillium Species in and out of doors in Topeka KS. J. Allergy. Clin. Immunol. (1990) 85(2): 247.
51.Müller GA, Smith AM, Williams DC Jr, Hakkaart GA, Aalerse RC, Chapman MD, Rule GS, Benhamin DC. Expression and secondary structure determination by NMR methods of the mahor house dust mite allergen Der p2. J. Biol. Chem. (1997) 272: 26893-26898.
52.Nishiyama C, Hatanaka H, Ichikawa S, Fukada M, Akagawa-Chihara M, Yuuki T, Yokota T, Inagaki F, Okumura Y. Analysis of human IgE epitope of Der f 2 with anti-Der f 2 mouse monoclonal antibodies. Mol. Immunol. (1999) 36: 53-60.
53.Norman PS. Modern concepts of immunotherapy. Curr. Opin. Immunol. (1993) 5: 968-973.
54.Ong EK, Knox RB, Singh MB. Mapping of the antigenic and allergenic epitopes of Lol p VB using gene fragmentation. Mol. Immunol. (1995) 32: 295-302.
55.Pauli G, Oster JP, Deviller P, Heiss S, Bessot JC, Susani M. Skin testing with recombinant allergens rBet v 1 and birch profiling, rBet v 2: diagnostic value for birch pollen and associated allergies. J. Allergy. Clin. Immunol. (1996) 97: 1-10.
56.Peitsch MC. ProMod and Swiss-Model: Internet-based tools for automated comparative protein modelling. Biochem. Soc. Trans. (1996) 24: 274-279.
57.Pitt JI. (1988) A laboratory guide to common Penicillin species. Commonwealth Scientific and Industrial Research Organization, North Ryde, N.S.W. Australia.
58.Reichard U, Buttner S, Hiffert H, Staib F, Ruchel R. Purification and characterization of an extracellular serine proteinase from Apergillus fumigatus and its detection in tissue. J. Med. Microbiol. (1990) 33: 243-251.
59.Robinson BW, Venaille TJ, Mendis AHW, McAleer R. Allergens as proteases: An Aspergillus fumigatus proteinase directly induces human epithelial celll detachment. J. Allergy. Clin. Immunol. (1990) 86: 726-731
60.Ruef C, Jefferson DM, Schlegel HS, Suter S. Regulation of cytokine secretion by cystic fibrosis airway epithelial cells. Eur. Resp. J. (1993) 6: 1429-36
61.Schramm G, Bufe A, Petersen A, Haas H, Schlaak M, Becker WM. Mapping of IgE-binding epitopes on the recombinant major group I allergen of velvet grass pollen, rHol l 1. J. Allergy Clin. Immunol. (1997) 99: 781-787.
62.Segal DM, Taurog JD, Metzger H. Dimeric immunoglobulin E serves as a unit signal for mast cell degranulation. Proc. Natl. Acad. Sci. USA (1977) 74: 2993-2997.
63.Seiberler S, Scheiner O, Kraft D, Lonsdale D, Valenta R. Characterization of a birch pollen allergen, Bet v Ⅲ, representing a novel class of Ca2+ binding proteins: Specific expression in mature pollen and dependence of patients IgE-binding on protein bound CA2+. EMBO J. (1994) 13: 3481-3486.
64.Selo I, Clement G, Bernard H, Chatel JM, Creminon C, Peltre G, Wal JM. Allergy to bovine β-lactoglobulin: specificity of human IgE to tryptic peptides. Clin. Exp. Allergy. (1999) 29: 1055-1063.
65.Sheffer AL. Allergen-specific immunotherapy: a role for T-cell anergy. Ann. Allergy (1993) 71: 327-329.
66.Shen HD, Lin WL, Liaw SF, Tam MF, Han SH. Characterization of the 33-kilodalton major allergen of Penicillium citrinum by using MoAbs and N-terminal amino acid sequencing. Clin. Exp. Allergy (1997) 27: 79-86.
67.Shen HD, Tam MF, Chou H, Han SH. The importance of serine proteinases as aeroallergens associated with asthma. Int. Arch. Allergy Immunol. (1999) 119: 259-264.
68.Shin DS, Compadre CM, Malekt SJ, Kopper RA, Sampson H, Huang SK, Burks AW, Bannon GA. Biochemical and Structural Analysis of the IgE Binding Sites on Ara h1, an Abundant and Highly Allergenic Peanut Protein. J. Bio. Chem. (1998) 273: 13753-13759.
69.Siezen RJ, Vos WM, Leunissen JAM, Dijkstra BW. Homology modeling and protein engineering strategy of subtilases, the family of subtilisin-like serine proteinases.Protein enginnering. (1991) 4: 719-737.
70.Sithigorngul P, Stretton AO, Cowden C. A versatile dot-ELISA method with femtomole sensitivity for detecting small peptides. J. Immunol. Methods. (1991) 141: 23-32.
71.Smith PK, Krohn RI, Germanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC. Measurement of protein using bicinchoninic acid. Anal. Biochem. (1985) 150: 76-85.
72.Stanley JS, King N, Burks AW, Huang SK,Sampson H,Cockrell G, Helm RM, West CM, Bannon GA. Identification and mutational analysis of the immunodominant IgE binding epitopes of the major peanut allergen Ara h2. Arch. Biochem. Biophys. (1997) 342: 244-253.
73.Stellato C, Beck LA, Corgone CA. Expression of the chemokine RANTES by a human bronchial cell line. Modulation by cytokines and glucocorticoids. Immunol. (1995) 155: 410-8.
74.Stellato C, Collins P, Ponath PD. Production of the novel C-C chemokine MCP-4 byairway cell and comparison of its biological activity to other CC chemokines. J Clin Invest (1997) 99: 926-36.
75.Stewart GA, Thompson PJ, McWilliam AS. Biochemical properties of aeroallergens: Contributory factors in allergic sensitization? Prediatr. Allergy Immunol (1993) 4: 163-172.
76.Stewart GA, Thompson PJ. The biochemistry of common aeroallergens. Clin. Exp. Allergy (1996) 26: 1020-44.
77.Stone KL, LoProsti MB, Crawford JM, DeAngelis R, Williams KR. (1989) Enzyme digestion of proteins and HPLC peptide isolation in “A practical guide to protein and peptide purification for microsequencing”, (Matsudaira PT. eds), Academy Press, CA.
78.Su NY, Yu CJ, Shen HD, Pan FM, Chow LP. Pen c 1, a novel enzyme allergen protein from Penicillin citrium. Purification, characterization, cloning and expression. Eur. J. Biochem. (1999) 261: 115-23.
79.Sutcliffe MJ, Haneef I, Camey D, Blundell TL. Knowledge based modelling of homologous proteins, Part I: Three-dimensional frameworks derived from the simultaneous superposition of multiple structures. Protein Engng, (1987) 1: 377-384.
80.Sutton BJ, Gould HJ. The human IgE network. Nature (1993) 366: 421-28.
81.Taipale J, Lohi J, Saarinen I, Kovanen PT, Keski OJ. Human mast cell chymase and leukocyte elastase release latent transforming growth factor-beta 1 from the extracellular matrix of cultured human epithelial and endothelial cells. J. Biol. Chem. (1995) 270: 4689-96.
82.Tanabe S, Tesaki S, Yanagihara Y. Mita H, Takahashi K, Arai S, Watanabe M. Inhibition of basophil histamine release by a haptenic peptide miture prepared by chymotryptic bhdrolysis of wheat fluor. Biochem. Biophys. Res. Commun. (1996) 223: 492-495.
83.Thompson RA, Kumararatne DS. Hyper-IgE syndrome and H2-receptor blockade. Lancet. (1989) 2: 630.
84.Twardosz A, Hayek B, Seiberler S, Vangelista L, Elfman L, Gronlund H, Kraft D, Valenta R. Molecular characterization, expression in Escherichia coli, and epitope analysis of a two EF-hand calcium-binding birch pollen allergen, Bet v 4. Biochem. Biophy. Res. Communi. (1997) 239: 197-204.
85.Valenta R, Flicker S, Eibensteiner PB, Steinberger P, Laffer S, Dolecek C, Kraft D. Recombinant allergen-specific antibody fragments: tools for diagnosis, prevention and therapy of type I allergy. Biol. Chem. (1997) 378: 745-749.
86.Valenta R, Sperr WR, Ferreira F, Valent P, Sillaber C, Tejkl M, Duchene M, Ebner C, Lechner K, Kraft D, Scheiner O. Induction of specific histamine release from basophils with purified natural and recombinant birch pollen allergens. J. Allergy Clin. Immunol. (1993) 91: 88-97
87.Valenta R, Vrtala S, Ebner C, Kraft D, Scheiner O. Diagnosis of grass pollen allergy with recombinant timothy grass(Phleum pratense) pollen allergens. Int. Arch. Allergy Immunol. (1992) 97: 287-94.
88.Van-Neerven RJ, van de Pol MM, Wierenga EZ, Aalberse RCm Jansen HM, Kapsenberg ML. Peptide specificity and HLA restriction do not dicate lymphokine production by allergen-specific T Lymphocute clones. Immunology (1994) 82: 351-536.
89.Vignola AM, Campbell AM, Chanez P. HLA-DR and ICAM-1 expression on bronchial epithelial cells in asthma and chronic bronchitis. Am. Rev. Respir. Dis. (1993) 148: 689-94.
90.Vrtala S, Ball T, Spitzauer S, Pandjaitan B, Suphioglu C, Knox B, Sperr WR, Valent P, Kraft D, Valenta R. Immunization with purified natural and recombinant allergens induces mouse IgG1 antibodies that recognize similar epitopes as human IgE and inhibit the human IgE-allergen interaction and allergen-induced basophil degranulation. J. Immunol. (1998) 160: 6137-6144.
91.Vrtala S, Hirtenlehner K, Vangelista L, Pastore A, Eichler HG, Sperr WR, Valent P, Ebner C, Kraft D, Valenta R. Conversion of the major birch pollen allergen, Bet v 1, into two nonanaphylactic T cell epitope-containing fragments: candidates for a novel form of specific immunotherapy. J. Clin. Invest. (1997) 99: 1673-1681.
92.Vrtala S, Mayer P, Ferreira F, Susani M, Sehon AH, Kraft D, Valenta R. Induction of IgE antibodies in mice and rhesus monkeys with recombinant birch pollen allergens: different allergenicity of Bet v1 and Bet v 2. J. Allergy Clin. Immunol. (1996) 98: 913-921.
93.Wei DL, Chen JH, Jong SC, and Shen HD. Indoor airborne Penicillium species in Taiwan. Current Microbiology, (1993) 26: 137-140.
94.Yasueda H, Mita H, Akiyama K, Shida T, Ando T, Sugiyama S, Yamakawa H. Allergens from Dermatophogoides mites with chymptrypic activity. Clin. Exp. Allergy (1993) 12: 384-390.
95.吳家興等(1998)台灣北部國中學生氣喘盛行率調查. 中華衛誌 17: 214-225.
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