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Time-resolved Fluorescence in Immunocytochemical Detection of Prostate-specific Antigen in Prostatic Tissue Sections

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Abstract

Chelates with fluorescent lanthanides such as europium and terbium are widely used in immunofluorometric assays, e.g. for the measurement of different molecular forms of prostate-specific antigen (PSA) in serum for detection and monitoring of prostate cancer. These chelates have also been introduced as non-radioactive labels in immunocytochemistry and in situ hybridization. In the present study, sections of non-malignant prostate were investigated using monoclonal IgGs against PSA. Detection of specific immunostaining employing time-resolved fluorescence with europium-labeled streptavidin was compared with conventional detection by streptavidin conjugated to horse-radish peroxidase. The high PSA concentration in the tissue produced high intensity, specific time-resolved fluorescence signals in the epithelial cells of the prostate gland without disturbance from non-specific tissue autofluorescense. This allowed short exposure times to be used which resulted in insignificant photobleaching. Two of the three europium-chelates evaluated yielded high signal intensities. Counterstaining was found to be optimal with Gill No. 1-Haematoxylin solution and Merckoglas™ was the best mounting medium for the europium chelates tested. In conclusion, time-resolved fluorescence imaging is an attractive alternative to conventional detection of streptavidin conjugated to horse-radish peroxidase, as it provides linear, high intensity, specific signals subsequent to the decay of non-specific tissue autofluorescence.

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References cited

  • Abrahamsson P-A, Lilja H, Falkmer S, Wadström LB (1988) Immunohistochemical distribution of the three predominant secretory proteins in the parenchyma of hyperplastic and neoplastic prostate glands. The Prostate 12: 39-46.

    Google Scholar 

  • Ahiyama K, Nakamura T, Iwamura S, Hara M(1987) The chymotrypsinlike activity of the prostate-specific antigen, g-semino protein. FEBS Lett 225: 168-172.

    Google Scholar 

  • Beverloo HB, Van Schadewijk A, Van Gelderen Boele S, Tanke HJ (1990) Inorganic phosphors as new luminescent labels for immunocytochemistry and time-resolved microscopy. Cytometry 11: 784-792.

    Google Scholar 

  • Beverloo HB, Van Schadewijk A, Zijlmans HJ, Verwoerd NP, Bonnett J, Vrolijk H, Tanke HJ (1993) A comparison of the detection sensitivity of lymphocyte membrane antigens using fluorescein and phosphor immunoconjugates. J Histochem Cytochem 41: 719-725.

    Google Scholar 

  • Bjartell A, Björk T, Matikainen M-T, Abrahamsson P-A, Di Santágnese A, Lilja H (1993) Production of alpha-1-antichymotrypsin by PSA-containing cells of human prostate epithelium. Urology 42: 502-510.

    Google Scholar 

  • Björk T, Bjartell A, Abrahamsson P-A, Hulkko S, Di Santágnese A, Lilja H (1994). Alpha-1-antichymotrypsin production in PSA-containing cells is common in prostate cancer but rare in benign prostatic hyperplasia. Urology 3: 427-434.

    Google Scholar 

  • Brawer MK, Lange PH (1989) Prostate-specific antigen in management of prostatic carcinoma. Urology (Suppl 5) 33: 11-16.

    Google Scholar 

  • Brawer MK (1991) Prostate specific antigen. Acta Onkologica 30: 161-168.

    Google Scholar 

  • Catalona W, Smith D, Ratliff TL, Dodds KM, Coplen DE, Yuan JJJ, Petros JA and Andriole GL (1991) Measurements of prostate-specific antigen in serum as a screening test for prostate cancer. N Engl J Med 324: 1156-1161.

    Google Scholar 

  • Clements JA (1994) The human kallikrein family: a diversity of expression and function. Mol Cell Endocrinol 99: C1-C6.

    Google Scholar 

  • de Haas RR, Verwoerd NP, Van Der Corput MP, Van Gijlswijk RP, Siitari-H, Tanke HJ (1996) The use of peroxidase-mediated deposition of biotin-tyramide in combination with time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization. J Histochem Cytochem 44: 1091-1099.

    Google Scholar 

  • Fazekas De St Groth S, Scheidegger D (1980) Production of Monoclonal Antibodies: Strategy and Tactics. J Immunol Methods 35: 1-21.

    Google Scholar 

  • Frazier HA, Humphrey PA, Burchette JL, Paulson DF (1992) Immunoreactive prostate-specific antigen in male peri-urethral glands. J Urol 147: 246-248.

    Google Scholar 

  • Hemmilä I, Dakabu S, Mukkala V-M, Siitari H, Lövgren T (1984) Europium as label in time-resolved immunoflouorometric assays. Analyt Biochem 137: 335-343.

    Google Scholar 

  • Hemmilä IA (1991) Applications of fluorescence in immunoassays. New York, John Wiley & Sons.

    Google Scholar 

  • Hsu S, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in Immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29: 577-580.

    Google Scholar 

  • Lilja H (1985) A kallikrein-like serine protease in prostatic fluid cleaves the predominant seminal vesicle protein. J Clin Invest 76: 1899-1903.

    Google Scholar 

  • Lilja H, Abrahamsson P-A (1988) Three predominant proteins secreted from the human prostate gland. Prostate 12: 29-38.

    Google Scholar 

  • Lilja H, Christensson A, Dahlén U, Matikainen M-T, Nilsson O, Pettersson K Lövgren T (1991) Prostate-specific antigen in human serum occurs predominantly in complex with alpha-1-antichymotrypsin. Clin chem 37: 1618-1625.

    Google Scholar 

  • Lövgren T, Hemmiläi, Pettersson K, Halonen P (1985) Timeresolved flurometry in immunoassays. In (Collins W. ed.) Alternative Immunoassays, JohnWiley & Sons Ltd. Chichester, UK, pp. 203-217 (Review).

    Google Scholar 

  • Mukkala V-M, Helenius M, Hemmilä I, Kankare J, Takalo H (1993) Development of luminescent europium (III) and terbium (III) chelates of 2,2: 6, 2-terpyridine derivatives for protein labeling. Helvetica Chimica Acta 76: 1361-1378.

    Google Scholar 

  • Oesterling JE (1991) Prostate specific antigen: A critical assessment on the most useful tumor marker for adenocarcinoma of the prostate. J Urol 145: 907-923.

    Google Scholar 

  • Papsidero LD, Wang MC, Valenzuela LA, Murphy GP, Chu TM (1980) A prostate antigen in sera of prostatic cancer patients. Cancer Res 40: 2428-2432.

    Google Scholar 

  • Pettersson K, Piironen T, Seppälä M, Liukkonen L, Christensson A, Matikainen M-T, Suonpää M, Lövgren T, Lilja H (1995) Free and complexed Prostate-specific antigen (PSA): In vitro stability, epitope map, and development of immunofluoreometric assays for specific and sensitive detection of free PSA and PSA-alpha-1-antichymotrypsin complex. Clin Chem 41: 1480-1488.

    Google Scholar 

  • Seveus L,VäisäläM, Syrjänen S, SandbergM,Kuusisto A, Harjo R, Salo J, Hemmilä I, Kojola H, Soini E (1992) Time-resolved fluorescence imaging of europium chelate label in immunohistochemistry and in situ hybridization. Cytometry 13: 329-338.

    Google Scholar 

  • Shi Z-R, Itzkowitz SH, Kim YS (1988) A comparison of three Immunoperoxidase techniques for antigen detection in colorectal carcinoma tissues. J Histochem Cytochem 36: 317-322.

    Google Scholar 

  • Soini EJ, Lövgren T (1987) Time-resolved fluorescence of lanthanide probes and applications in biotechnology. CRC Crit Rev Anal Chem 18: 105-154.

    Google Scholar 

  • Soini EJ, Pelliniemi LJ, Hemmilä IA, Mukkala V-M, Kankare JJ, Fröjdman K (1988) Lanthanide chelates as new fluorochrome labels for cytochemistry. J Histochem Cytochem 36: 1449-1451.

    Google Scholar 

  • Stamey TA, Yang N, Hay AR, McNeil JE, Freika FS, Redwine E (1987) Prostate-specific antigen as a marker for adenocarcinoma of the prostate. N Engl J Med 317: 909-916.

    Google Scholar 

  • Takalo H, Mukkala V-M, Mikola H, Liitti P, Hemmilä I (1994) Synthesis of europium (III) chelates suitable for labeling of bioactive molecules. Bioconjugate Chem 5: 278-282.

    Google Scholar 

  • Takalo H, Hänninen E, Kankare J (1993) Luminescence of europium (III) chelates of substituted 4-phenylethynylpyridines as ligands. Helv Chim Ascta 76: 877-883.

    Google Scholar 

  • Wang MC, Valenzuela CA, Murphy GP, Chu TM (1979) Purification of a human prostate-specific antigen. Invest Urol 17: 159-163.

    Google Scholar 

  • van Gijlswijk RP, Zijlmans HJ, Wiegant J, Bobrow MN, Erickson TJ, Adler KE, Tanke HJ, Raap AK (1997) Fluorochrome-labeled tyramides: use in immunocytochemistry and fluorescence in situ hybridization. J Histochem Cytochem 45: 375-382.

    Google Scholar 

  • Verwoerd NP, Hennink EJ, Bonnet J, Van Der Geest CR, Tanke HJ (1994) Use of ferro-electric liquid crystal shutters for time-resolved fluorescence microscopy. Cytometry 16: 113-117.

    Google Scholar 

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Authors and Affiliations

  1. Department of Urology, Lund University, University Hospital, Malmö, Sweden

    Anders Bjartell

  2. Department of Biotechnology, University of Turku, Finland

    Kim Pettersson & Elise Nilsson

  3. Department of Biotechnology, University of Turku, Finland

    Timo Lövgren & Hans Lilja

  4. Department of Biotechnology, University of Turku, Finland

    Sari Laine

Authors
  1. Anders Bjartell
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  2. Sari Laine
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  3. Kim Pettersson
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  4. Elise Nilsson
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  5. Timo Lövgren
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  6. Hans Lilja
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Bjartell, A., Laine, S., Pettersson, K. et al. Time-resolved Fluorescence in Immunocytochemical Detection of Prostate-specific Antigen in Prostatic Tissue Sections. Histochem J 31, 45–52 (1999). https://doi.org/10.1023/A:1003504115690

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  • DOI: https://doi.org/10.1023/A:1003504115690

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