ABSTRACT
BACKGROUND: Novel rabbit monoclonal antibodies (RabMab) for estrogen (ER), progesterone (PR) receptors and HER2 evaluation by immunohistochemistry have recently been commercially released. We compared the RabMab anti-ER, anti-PR and anti-HER2 to mouse monoclonal antibodies (Mab) using tissue microarrays (TMA) of breast carcinomas. METHODS: Two TMA containing breast carcinomas were built. Sections were immunostained using anti-ER and anti-PR, Mab and RabMab. The sections stained for ER and PR were evaluated considering positive those tumors in which more than 1 percent of the tumor cell nuclei stained moderate or strong. For HER2, the immunostained sections were evaluated using the ASCO/CAP guidelines for HER2. Chromogenic in situ hybridization (CISH) was used as the gold standard for HER2 evaluation. CISH was evaluated using the Zymed HER2 CISH interpretation guidelines. RESULTS: RabMab against ER have similar staining patterns compared to the 6F11 (Mab), but stronger than 1D5 (Mab) from three different suppliers. The RabMab against PR provide stronger and sharper immunohistochemical signals compared to Mab. The detection of HER2 protein overexpression was more prevalent with the polyclonal antibodies and RabMab than with the Mab. These were more specific than the RabMab, which were more sensitive when compared to CISH. CONCLUSION: The novel RabMab against ER and PR showed higher intensity of staining than the Mab. The RabMab against HER2 is more sensitive than Mab, however, Mab presented more specificity than RabMab when compared to CISH for HER2 evaluation of breast carcinomas.
OBJETIVOS: Novos anticorpos monoclonais de coelho (RabMab) para a avaliação imuno-histoquímica de receptores de estrógeno (RE), progesterona (RP) e HER2 foram lançados comercialmente. Comparamos os RabMab anti-RE, anti-RP e anti-HER2 com os anticorpos monoclonais de camundongo (Mab) utilizando tissue microarrays (TMA) de carcinomas de mama. MÉTODOS: Foram construídos dois TMAs de carcinomas de mama. As secções foram marcadas usando anti-RE, anti-RP e anti-HER2, Mab e RabMab através de imuno-histoquímica. As secções marcadas para RE e RP foram avaliadas considerando positivos aqueles tumores nos quais mais de 1 por cento dos núcleos coraram moderadamente ou forte. Para HER2, as secções foram avaliadas utilizando as recomendações da ASCO/CAP para HER2. Hibridização in situ cromogênica (CISH) foi usada como padrão-ouro para avaliação de HER2. CISH foi avaliado utilizando as recomendações da Zymed. RESULTADOS: Os RabMab anti-RE apresentam intensidade de coloração semelhante ao 6F11 (Mab), porém maior que o 1D5 (Mab) proveniente de três diferentes fabricantes. Os RabMab anti-RP apresentaram sinal imunoistoquímico mais forte e delimitado comparado aos Mab. A detecção da superexpressão da proteína HER2 foi mais prevalente entre os anticorpos policlonais e RabMab, que se mostraram mais sensíveis quando comparados com o CISH. CONCLUSÃO: Os novos RabMab anti-RE e RP proporcionaram maior intensidade de coloração que os Mab. O RabMab anti-HER2 apresentou maior sensibilidade que os Mab, porém os Mab apresentaram maior especificidade quando comparados com o CISH para a avaliação de HER2 em carcinomas de mama.
Subject(s)
Animals , Female , Humans , Mice , Rabbits , Antibodies, Monoclonal , Breast Neoplasms/chemistry , Carcinoma, Ductal, Breast/chemistry , /analysis , Receptors, Estrogen/analysis , Receptors, Progesterone/analysis , Microarray Analysis/methods , /immunology , Receptors, Estrogen/immunology , Receptors, Progesterone/immunology , Staining and Labeling , Statistics, NonparametricABSTRACT
Los receptores esteroidales sexuales del tracto genital de la hembra tienen importancia ya que a través de ellos actúan las hormonas responsables de su desarrollo y de sus cambios morfofuncionales. Dado que en su mecanismo de control uno de los factores a tenerse en cuenta son las posibles diferencias entre las distintas especies, los objetivos de nuestro estudio fueron comparar en dos especies distintas la ovina (O) (n=6) y la canina (P) (n=6) la presencia de receptores de estrógenos (RE), progesterona (RP) y de CBG en ovario, tuba uterina y útero de hembras prepúberes (O:n=3; P:n=·3) y adultas (O:n=3; P:n=3). Se realizaron tinción con H-E e inmunocitoquimica según técnica de Stenberger. Los resultados revelaron: en ambas especies en animales durante el ciclo estral inmunorreactividad (IR) positiva marcada para estrógenos en útero, oviducto y ovario siendo comparativamente los RE más numerosos en oviducto de perra que en oveja. En ambos animales en anestro, la IR para RE fue negativa o leve en útero, no evidenciándose su presencia en otros órganos. Los RP variaron según el estadio del ciclo, no mostrando diferencias destacables entre las especies. En las hembras prepúberes de oveja fueron detectados RE Y RP, mientras que en perras éstos no fueron evidenciados. La CBG mostró IR positiva en el tracto de ambos animales durante el ciclo y fue negativa en prepúberes para ambas especies. Se concluye que existen diferencias, especialmente en hembras prepúberes, donde la presencia de RE y RP es detectable en ovejas no así en perras, siendo además, la presencia de RE comparativamente más marcada en tuba uterina de perra en ciclo que en oveja en el mismo estadio.
The sexual steroid receptors of the genital tract of the female has importance since on them the hormones responsible for their development and morphofuncional changes act. Due that in their control mechanism one of the factors to be kept in mind is the possible differences among the diverse species. The objective of our study was to compare in two different species sheep (O) ( n=6) and canine (P) ( n=6) the presence of receptors of estrogens (RE), progesterone (RP) and the presence of CBG in ovary, oviduct and uterus of female prepúberes (O:n=3; Pn=·3) and mature: (O:n=3; Pn=·3) We use H-E tint and and inmunocito chemistry according to technique of Stenberger. The results revealed: in both species of animals during the estral cycle: Inmunorreactivity (RI) positive marked for estrogens in uterus, oviduct and ovary being comparatively much more numerous RE in dog oviduct that in sheep. In both animals in anestro RE was negative or light in uterus, without evidenced of their presence in other organs. The RP varied according to the stage of the cycle not showing remarkables differences among the species. In the prepuber female sheep RE and RP were detected while in dogs they were not evidenced. The CBG showed positivity in the tract of both animals during the cycle and it was negative in both prepuber species We concludes that differences exist especially in prepuber female where the presence of RE and RP are detectable in sheep and no detectable in dogs, being further the comparatively presence of RE in dog oviduct in cycle more marked that in sheep in the same stage.
Subject(s)
Dogs , Receptors, Estrogen/agonists , Receptors, Estrogen/immunology , Receptors, Progesterone/agonists , Receptors, Progesterone/immunology , Dogs , Immunohistochemistry , Immunohistochemistry/veterinary , Reproduction/immunology , SheepABSTRACT
To determine the progesterone receptor status in thyroid gland. This study was based on immunohistochemical staining of formalin fixed paraffin embedded tissues, for progesterone receptors, in 50 previously diagnosed cases of various thyroid lesions and surrounding normal thyroid tissue. Out of 50 cases, 8 were nodular goiter, 9 cases of adenoma, 19 papillary carcinoma, 10 follicular carcinoma and four cases were of medullary carcinoma. Surrounding normal tissue was available in 4 non-neoplastic and 21 neoplastic lesions. Overall male patients comprised 20% [10 cases] and females 80% [40 cases]. Although a wide range of lesions in both the sexes including wide age range were available, none of our cases were positively stained for progesterone receptors. In contrary to earlier reports by immunohistochemical method using monoclonal mouse anti-PR antibody clone PgR 636, on formalin-fixed paraffin embedded thyroid tissues, the progesterone receptors were not detectable in our human samples. The effect of progesterone on thyroid gland may be an indirect one