Validation of new aromatase monoclonal antibodies for immunohistochemistry: progress report
Identifieur interne : 00AF91 ( Main/Exploration ); précédent : 00AF90; suivant : 00AF92Validation of new aromatase monoclonal antibodies for immunohistochemistry: progress report
Auteurs : H. Sasano [Japon] ; D. P. Edwards [États-Unis] ; T. J. Anderson [Royaume-Uni] ; S. G. Silverberg [États-Unis] ; D. B. Evans [Suisse] ; R. J. Santen [États-Unis] ; P. Ramage [Suisse] ; E. R. Simpson [Australie] ; A. S. Bhatnagar [Suisse] ; W. R. Miller [Royaume-Uni]Source :
- Journal of steroid biochemistry and molecular biology [ 0960-0760 ] ; 2003.
Descripteurs français
- Pascal (Inist)
English descriptors
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Abstract
Intratumoral aromatase is a potential therapeutic target for the treatment of postmenopausal estrogen-dependent breast cancers. Therefore, reliable methods should be developed for routine application for the detection of intratumoral aromatase. A multi-center collaborative group has been established to generate and validate new aromatase monoclonal antibodies (MAbs). A recombinant GST-aromatase fusion protein was expressed in baculovirus and the purified protein was used for immunization of mice either as a native or formalin-fixed antigen. Hybridomas were generated using standard techniques and screened biochemically prior to immunohistochemistry (IHC) evaluation in human placenta, ovary and breast cancer tissues. Twenty-three MAbs selected by biochemical assays were further evaluated by IHC of paraffin-embedded tissue sections including normal ovary, and placenta, and a small series of 10 breast carcinomas. Of the 23 MAbs, 2 (clones 677 and F2) were determined to specifically stain cell types known to express aromatase in normal tissues. In breast carcinomas staining of malignant epithelium, adipose tissue, normal/benign and stromal compartments was detected. IHC was performed and independently evaluated by three pathologists (HS, TJA and SGS), each using the same evaluation criteria for staining intensity and proportion of immunopositive cells. With these two MAbs, interpathologist and intralaboratory variations were minimal in comparison with differences which could be detected between tissue specimens and antibodies.
Affiliations:
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Sasano</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Phathology, Tohoku University School of Medicine, 2-1 Seiryou-Machi</s1><s2>Aoba-Ku, Sendai 980-8575</s2><s3>JPN</s3><sZ>1 aut.</sZ></inist:fA14><country>Japon</country><wicri:noRegion>Aoba-Ku, Sendai 980-8575</wicri:noRegion></affiliation></author><author><name sortKey="Edwards, D P" sort="Edwards, D P" uniqKey="Edwards D" first="D. P." last="Edwards">D. P. Edwards</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>University of Colorado Health Science Center</s1><s2>Denver CO</s2><s3>USA</s3><sZ>2 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>University of Colorado Health Science Center</wicri:noRegion></affiliation></author><author><name sortKey="Anderson, T J" sort="Anderson, T J" uniqKey="Anderson T" first="T. J." last="Anderson">T. J. Anderson</name><affiliation wicri:level="4"><inist:fA14 i1="03"><s1>Department of Pathology, University of Edinburgh</s1><s2>Edinburgh, Scotland</s2><s3>GBR</s3><sZ>3 aut.</sZ></inist:fA14><country>Royaume-Uni</country><placeName><settlement type="city">Édimbourg</settlement><region type="country">Écosse</region></placeName><orgName type="university">Université d'Édimbourg</orgName></affiliation></author><author><name sortKey="Silverberg, S G" sort="Silverberg, S G" uniqKey="Silverberg S" first="S. G." last="Silverberg">S. G. Silverberg</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Pathology, University of Maryland School of Medicine</s1><s2>Baltimore, MD</s2><s3>USA</s3><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Baltimore, MD</wicri:noRegion></affiliation></author><author><name sortKey="Evans, D B" sort="Evans, D B" uniqKey="Evans D" first="D. B." last="Evans">D. B. Evans</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Novartis Pharm AG</s1><s2>Basel</s2><s3>CHE</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Suisse</country><wicri:noRegion>Novartis Pharm AG</wicri:noRegion></affiliation></author><author><name sortKey="Santen, R J" sort="Santen, R J" uniqKey="Santen R" first="R. J." last="Santen">R. J. Santen</name><affiliation wicri:level="1"><inist:fA14 i1="06"><s1>Department of Medicine, University of Virginia</s1><s2>Charlottesville, VA</s2><s3>USA</s3><sZ>6 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Charlottesville, VA</wicri:noRegion></affiliation></author><author><name sortKey="Ramage, P" sort="Ramage, P" uniqKey="Ramage P" first="P." last="Ramage">P. Ramage</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Novartis Pharm AG</s1><s2>Basel</s2><s3>CHE</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Suisse</country><wicri:noRegion>Novartis Pharm AG</wicri:noRegion></affiliation></author><author><name sortKey="Simpson, E R" sort="Simpson, E R" uniqKey="Simpson E" first="E. R." last="Simpson">E. R. Simpson</name><affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Prince Henry's Institute of Medical Research</s1><s2>Clayton, Vic.</s2><s3>AUS</s3><sZ>8 aut.</sZ></inist:fA14><country>Australie</country><wicri:noRegion>Prince Henry's Institute of Medical Research</wicri:noRegion></affiliation></author><author><name sortKey="Bhatnagar, A S" sort="Bhatnagar, A S" uniqKey="Bhatnagar A" first="A. S." last="Bhatnagar">A. S. Bhatnagar</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Novartis Pharm AG</s1><s2>Basel</s2><s3>CHE</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>Suisse</country><wicri:noRegion>Novartis Pharm AG</wicri:noRegion></affiliation></author><author><name sortKey="Miller, W R" sort="Miller, W R" uniqKey="Miller W" first="W. R." last="Miller">W. R. Miller</name><affiliation wicri:level="4"><inist:fA14 i1="08"><s1>Department of Experimental Oncology, University of Edinburgh</s1><s2>Edinburgh, Scotland</s2><s3>GBR</s3><sZ>10 aut.</sZ></inist:fA14><country>Royaume-Uni</country><placeName><settlement type="city">Édimbourg</settlement><region type="country">Écosse</region></placeName><orgName type="university">Université d'Édimbourg</orgName></affiliation></author></analytic><series><title level="j" type="main">Journal of steroid biochemistry and molecular biology</title><title level="j" type="abbreviated">J. steroid biochem. mol. biol.</title><idno type="ISSN">0960-0760</idno><imprint><date when="2003">2003</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of steroid biochemistry and molecular biology</title><title level="j" type="abbreviated">J. steroid biochem. mol. biol.</title><idno type="ISSN">0960-0760</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Breast</term><term>Estrogen synthase</term><term>Immunohistochemistry</term><term>Malignant tumor</term><term>Monoclonal antibody</term><term>Progress report</term><term>Validation</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Validation</term><term>Estrogen synthase</term><term>Anticorps monoclonal</term><term>Immunohistochimie</term><term>Rapport activité</term><term>Sein</term><term>Tumeur maligne</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Intratumoral aromatase is a potential therapeutic target for the treatment of postmenopausal estrogen-dependent breast cancers. Therefore, reliable methods should be developed for routine application for the detection of intratumoral aromatase. A multi-center collaborative group has been established to generate and validate new aromatase monoclonal antibodies (MAbs). A recombinant GST-aromatase fusion protein was expressed in baculovirus and the purified protein was used for immunization of mice either as a native or formalin-fixed antigen. Hybridomas were generated using standard techniques and screened biochemically prior to immunohistochemistry (IHC) evaluation in human placenta, ovary and breast cancer tissues. Twenty-three MAbs selected by biochemical assays were further evaluated by IHC of paraffin-embedded tissue sections including normal ovary, and placenta, and a small series of 10 breast carcinomas. Of the 23 MAbs, 2 (clones 677 and F2) were determined to specifically stain cell types known to express aromatase in normal tissues. In breast carcinomas staining of malignant epithelium, adipose tissue, normal/benign and stromal compartments was detected. IHC was performed and independently evaluated by three pathologists (HS, TJA and SGS), each using the same evaluation criteria for staining intensity and proportion of immunopositive cells. With these two MAbs, interpathologist and intralaboratory variations were minimal in comparison with differences which could be detected between tissue specimens and antibodies.</div></front></TEI><affiliations><list><country><li>Australie</li><li>Japon</li><li>Royaume-Uni</li><li>Suisse</li><li>États-Unis</li></country><region><li>Écosse</li></region><settlement><li>Édimbourg</li></settlement><orgName><li>Université d'Édimbourg</li></orgName></list><tree><country name="Japon"><noRegion><name sortKey="Sasano, H" sort="Sasano, H" uniqKey="Sasano H" first="H." last="Sasano">H. Sasano</name></noRegion></country><country name="États-Unis"><noRegion><name sortKey="Edwards, D P" sort="Edwards, D P" uniqKey="Edwards D" first="D. P." last="Edwards">D. P. Edwards</name></noRegion><name sortKey="Santen, R J" sort="Santen, R J" uniqKey="Santen R" first="R. J." last="Santen">R. J. Santen</name><name sortKey="Silverberg, S G" sort="Silverberg, S G" uniqKey="Silverberg S" first="S. G." last="Silverberg">S. G. Silverberg</name></country><country name="Royaume-Uni"><region name="Écosse"><name sortKey="Anderson, T J" sort="Anderson, T J" uniqKey="Anderson T" first="T. J." last="Anderson">T. J. Anderson</name></region><name sortKey="Miller, W R" sort="Miller, W R" uniqKey="Miller W" first="W. R." last="Miller">W. R. Miller</name></country><country name="Suisse"><noRegion><name sortKey="Evans, D B" sort="Evans, D B" uniqKey="Evans D" first="D. B." last="Evans">D. B. Evans</name></noRegion><name sortKey="Bhatnagar, A S" sort="Bhatnagar, A S" uniqKey="Bhatnagar A" first="A. S." last="Bhatnagar">A. S. Bhatnagar</name><name sortKey="Ramage, P" sort="Ramage, P" uniqKey="Ramage P" first="P." last="Ramage">P. Ramage</name></country><country name="Australie"><noRegion><name sortKey="Simpson, E R" sort="Simpson, E R" uniqKey="Simpson E" first="E. R." last="Simpson">E. R. Simpson</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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