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Identification and evaluation of a major cytotoxin of A. fumigatus

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Abstract

Aspergillus fumigatus is a highly pathogenic fungus causing a wide spectrum of diseases in immunocompromised as well as immunocompetent hosts. The present work was undertaken to evaluate the cytotoxic nature of fractionated antigens of A. fumigatus against the mammalian cell lines (J774, RAW, CHO and L929). An enriched protein antigenic fraction of A. fumigatus was subjected to con A Sepharose and phenyl Sepharose chromatography. Antigenic fractions, ConAub (conA unbound) and PSC III (fraction III of phenyl Sepharose column) containing low mw antigens showed higher cytotoxicity as compared to other antigenic fractions. PSC III was further purified on HPLC resulting in an 18 kDa homogeneous protein. The purified protein showed high ELISA absorbance values for specific IgG and IgE antibodies in sera of ABPA patients. Monoclonal antibody raised against Asp fl, a major allergen/antigen of A. fumigatus recognised the purified 18 kDa by ELISA and western blot. The 18 kDa allergen/antigen or Asp fl showed similar toxicity towards all the four cell lines (macrophage and fibroblast) with an IC50 of 75 ng/ml or 4.16 nM. Reduction in toxicity of 18 kDa at low temperatures and potentiation in presence of ammonium chloride and monensin indicates mechanism of internalisation of 18 kDa in eukaryotic cells is similar to α-sarcin. The present work shows that the 18 kDa allergen/antigen (Asp fl) is a major cytotoxin secreted by A. fumigatus which may play multiple roles in the pathogenesis of Aspergillosis through allergenicity, antigenicity and cytotoxicity. (Mol Cell Biochem 167: 89-97, 1997)

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References

  1. Bardana EJ Jr.: The clinical spectrum of aspergillosis. Part 2: classification of saprophytic, allergic and invasive variants of Aspergillus fumigatus. CRC Crit Rev Clin Lab Sci 13: 85, 1980

    Google Scholar 

  2. Kurup VP, Kumar A: Immunodiagnosis of aspergillosis. Clin Microbiol Rev 4: 439, 1991

    Google Scholar 

  3. Kolattukudy PE, Lee JD, Rogers LM, Zimmerman P, Ceselski S, Fox B, Stein B, Copelan EA: Evidence for possible involvement of an elastinolytic serine protease in aspergillosis. Infec Immun 6: 2357, 1993

    Google Scholar 

  4. Olson BJ, Jennings JC, Roga V, Junek AJ, Shuurmans DM: Alpha-sarcin a new antitumor agent. II. Fermentation and antitumor spectrum. Appl Microbiol 13: 314, 1965

    Google Scholar 

  5. Conde FP, Orlandi R, Canevari S, Mezzanzanica D, Ripamonti M, Munoz SM, Jorge P, Colnaghi MI: The Aspergillus toxin restrictocin is a suitable cytotoxin agent for generation of immunoconjugates with monoclonal antibodies directed against human carcinoma cells. Eur J Biochem 178: 795, 1989

    Google Scholar 

  6. Fernandez-Luna JL, Lopez-Otin C, Soriano F, Mendez E: Complete aminoacid sequence of the Aspergillus cytotoxin mitogillin. Biochemistry 24: 861, 1985

    Google Scholar 

  7. Arruda LK, Platts-Mills TAE, Fox JW, Chapman MD: Aspergillus fumigatus allergen 1, a major IgE binding protein, is a member of the mitogillin family of cytotoxins. J Exp Med 172: 1529–1532, 1992

    Google Scholar 

  8. Banerjee B, Chetty A, Joshi AP, Sarma PU: Identification and partial characterisation of diagnostically relevant antigens of A. fumigatus. Asian Pac J Allergy and Immunology 18: 13–18, 1990

    Google Scholar 

  9. Banerjee B, Madan T, Sharma GL, Prasad HK, Nath I, Sarma PU: Characterisation of 45 kd glycoprotein antigen of A. fumigatus. Serodiag and Immunotherap 7, 1996, In press

  10. Bir N: Immunochemical and biochemical characterisation of antigens of A. fumigatus. Thesis, 1996

  11. Laemmli UK: Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680, 1970

    Google Scholar 

  12. Towbin H, Stachalin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gel to nitrocellulose sheets, procedure and some applications. Proc Natl Acad Sci 76: 4350, 1979

    Google Scholar 

  13. Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55, 1983

    Google Scholar 

  14. Latge JP, Moutaouakil M, Debeaupuis JP, Bouchara JP, Haynes K, Prevost MC: The 18 kilodalton antigen secreted by Aspergillus fumigatus. Infect Immun 59: 2586, 1991

    Google Scholar 

  15. Smith JM, Tang CM, Noorden SV, Holden DW: Virulence of Aspergillus fumigatus double mutants lacking restrictocin and an alkaline protease in a low dose model of invasive aspergillosis. Infect Immun 62: 5247, 1994

    Google Scholar 

  16. Gasset M, Onaderra M, delPozo AM, Schiavo G, Laynez J, Usobiaga P, Gavilanes JG: Effect of antitumor protein α-sarcin on the thermotropic behaviour of acid phospholipid vesicles. Biochim Biophys Acta 1068: 9, 1991

    Google Scholar 

  17. Deurs BV, Petersen OW, Olsnes S, Sandvig K: The ways of endocytosis. International Review of Cytology 117: 131, 1989

    Google Scholar 

  18. Youle RJ, Newton D, Wu Y, Gadina M, Rybak SM: Cytotoxic ribonucleases and chimeras in cancer therapy. Crit Rev Therap Drug Carrier Sys 10: 1, 1993

    Google Scholar 

  19. Arruda LK, Mann BJ, Chapman MD: Selective expression of a major allergen and cytotoxin Asp fl in A. fumigatus. J Immunol 149: 3354–3359, 1992

    Google Scholar 

  20. Endo Y, Huber PW, Wool IG: The ribonuclease activity of the cytotoxin alpha sarcin. J Biol Chem 258: 2662, 1982

    Google Scholar 

  21. Orlandi R, Canevari S, Conde FP, Leoni F, Mezzanzenica D, Colnaghi MI: Immunoconjugate generation between the ribosome inactivating protein restrictocin and antihuman breast carcinoma MAB. Cancer Immunol Immunother 26: 114, 1988

    Google Scholar 

  22. Pai LH, Pastan l: Immunotoxins and recombinant toxins for cancer treatment. Important advances in oncology. In: Vincent T DeVita, Samuel Hellman and Steven A Rosenberg (eds). JB Lippincott Company, Philadelphia 1994, p 3

    Google Scholar 

  23. Vitetta ES, Thorpe PE, Uhr JW: Immunotoxins: magic bullets or misguided missiles? TiPS 14: 148, 1993

    Google Scholar 

  24. Jin FS, Youle RJ, Johnson VG, Shiloach J, Fass R, Londo DL, Bridges SH: Suppression of the immune response to immunotoxins with anti-CD4 monoclonal antibodies. J Immunol 146: 1806, 1991

    Google Scholar 

  25. Roga V, Hedeman LP, Olson BH: Evaluation of mitogillin (NSC-69529) in the treatment of naturally occurring canine neoplasms. Cancer Chemother Rep Part 1 55: 101, 1971

    Google Scholar 

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  1. Centre for Biochemical Technology, Mall Road, Delhi-, 110007, India

    Taruna Madan, Naveen Arora & P. Usha Sarma

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  1. Taruna Madan
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  2. Naveen Arora
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Madan, T., Arora, N. & Sarma, P.U. Identification and evaluation of a major cytotoxin of A. fumigatus. Mol Cell Biochem 167, 89–97 (1997). https://doi.org/10.1023/A:1006823706119

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