Path. Res. Pract. 187,539-540 (1991)
Further Immunohistochemical Study of Crooke's Hyalin Y. Uei, M. Kanzaki 1 and T. Yabana2 Cytology Section, Clinical Laboratory, National Cancer Center Hospital; lClinical Laboratory, Kamo Hospital; 2Clinical Laboratory, Yamada Red Cross Hospital, Japan
SUMMARY
For analysis of the cytokeratin (CK) of Crooke's cells, 28 post-mortem pituitary glands with unequivocal Crooke's hyaline change were investigated immunohistochemically using monoclonal antibodies for CK subfamilies. Crooke's hyalin was positive for CK 8 {molecular weight 52.5 kilodalton (KD)} and 18 (45 KD) but negative for skin-, corneaand esophageal-typed CKs.
Introduction Previously we investigated immunohistochemically hyaline changes of Crooke's cells using a monoclonal antibody for each intermediate filament and concluded that Crooke's hyalin was composed of abnormally accumulated CK4. The antibody for CK (KL 1) used was that for a broad range of molecular weights 5 • The present study was carried out to further analyze the CK using anticytokeratin monoclonal antibodies of more restricted specificity.
Material and Methods The materials and methods employed were similar to those described by the authors elsewhere4, except that horizontal serial sections of each pituitary gland were used instead of mirror image sections. The monoclonal antibodies for CK subfamilies used are listed in Table 1. Trypsin digestion was performed by reaction with 0.1 % trypsin in 0.1 % calcium chloride solution (pH 7.8) at for 30 minutes.
3rc
Results Crooke's hyalin was positive for CK 8 (52.5 KD) and 18 (45 KD) but negative for CK 1 (68 KD), 5 (58 KD), 10 (56.5 KD), 11 (56 KD), 3 (64 KD), 13 (51 KD), 19 (40 KD) and 7 (54 KD). An antibody for CK 14 (50 KD) (CBL 197) failed to react even with a positive control. © 1991 by Gustav Fischer Verlag, Stuttgart
Fig. 1. Serial sections of Crooke's cells. - A: ACTH stain (x 400). - B: Cytokeratin stain (MA-902) (x 400). 0344-0338/91/0187-0539$3.50/0
540 . Y. Uei, M. Kanzaki and T. Yabana Table 1. Monoclonal antibodies for cytokeratins Cat. No. (Source)
Basic (Type II) No.
Cytokeratin subfamily Acidic (Type I) Mol.wt (KD) Mol.wt (KD) No.
MA-904 (ENZO) MA-903 (ENZO) AE 5 (lCN) AE 8 (lCN) MA-902 (ENZO) Ks 19.1 (lCN) C-6417 (Sigma) C-8541 (Sigma) CBL 197 (CBL)
1 1,5 3
68 68,58 64
8
52.5
7
10,11
56.5,56
13
51
19
40
18 14
45 50
54
Markers for Skin-type Skin-type Cornea-type Esophageal-type Simple epithelia Simple epithelia Simple epithelia Simple epithelia Keratinocytes
Trypsin digestion
+
+ +
+ +
abnormal accumulation of a cellular structural protein4 • The monoclonal antibody (KL 1) used in this study was that generated against a broad range of CKs recognizing epithelial cells from the upper layer of the epidermis, gastrointestinal and respiratory mucosae and from other epithelia including the thyroid and thymus s . It is of interest to determine what subunit molecular weight CKs Crooke's hyalin is composed of. Nineteen different CK polypeptides have been identified with molecular weights ranging from 40 to 68 KD1. Recently monoclonal anticytokeratin antibodies of more restricted specificity have become available. Using these antibodies, it was confirmed that the CK of Crooke's hyalin consisted of CKs 8 and 18. There have only been few reports on Crooke's hyaline change from the immunohistochemical viewpoint 2, 3. According to Neumann et aI., Crooke's hyalin was positive for CK with each of the five antisera they used 3, but their results disagree slightly with ours concerning the exact subfamilies involved. References
Fig. 2. Serial sections of Crooke's cells. - A: ACTH stain (x 400). - B: Cytokeratin stain (C-8541) (x 400).
Discussion Our previous study identified the Crooke's hyalin found in ACTH cells as CK, which therefore represents an
1 Cooper 0, Schermer A, Sun T-T (1985) Biology of disease. Classification of human epithelia and . their neoplasms using monoclonal antibodies to keratin: Strategies, applications and limitations. Lab Invest 52: 243-256 2 Halliday WC, Asa SL, Kovacs K, Scheithauer BW (1987) Immunohistochemical study of intermediate filaments in the human pituitary. Lab Invest 56: 29 A 3 Neumann PE, Horoupian DS, Goldman JE, Hess ME (1984) Cytoplasmic filaments of Crooke's hyaline change belong to the cytokeratin class. An immunocytochemical and ultrastructural study. Am J Pathol116: 214-222 4 · Uei Y, Kanzaki M, Ohta T. Immunohistochemical study of Crooke's hyaline change. Endocr Pathol: In press 5 Viac J, Reano A, Brochier J, Staquet M-J, Thivolet J (1983 ) Reactivity pattern of a monoclonal antikeratin antibody (KL 1). J Invest Dermatolog 81: 351-354
Received December 15, 1990 . Accepted January 16, 1991
Key words: Crooke's cell- Cytokeratin Yoshio Uei, M.D., Head, Cytology Section, Clinical Laboratory, National Cancer Center Hospital, 1-1 Tsukiji 5 chome, Chuo-ku, Tokyo 104, Japan
Further Immunohistochemical Study of Crooke's Hyalin Y. Uei, M. Kanzaki 1 and T. Yabana2 Cytology Section, Clinical Laboratory, National Cancer Center Hospital; lClinical Laboratory, Kamo Hospital; 2Clinical Laboratory, Yamada Red Cross Hospital, Japan
SUMMARY
For analysis of the cytokeratin (CK) of Crooke's cells, 28 post-mortem pituitary glands with unequivocal Crooke's hyaline change were investigated immunohistochemically using monoclonal antibodies for CK subfamilies. Crooke's hyalin was positive for CK 8 {molecular weight 52.5 kilodalton (KD)} and 18 (45 KD) but negative for skin-, corneaand esophageal-typed CKs.
Introduction Previously we investigated immunohistochemically hyaline changes of Crooke's cells using a monoclonal antibody for each intermediate filament and concluded that Crooke's hyalin was composed of abnormally accumulated CK4. The antibody for CK (KL 1) used was that for a broad range of molecular weights 5 • The present study was carried out to further analyze the CK using anticytokeratin monoclonal antibodies of more restricted specificity.
Material and Methods The materials and methods employed were similar to those described by the authors elsewhere4, except that horizontal serial sections of each pituitary gland were used instead of mirror image sections. The monoclonal antibodies for CK subfamilies used are listed in Table 1. Trypsin digestion was performed by reaction with 0.1 % trypsin in 0.1 % calcium chloride solution (pH 7.8) at for 30 minutes.
3rc
Results Crooke's hyalin was positive for CK 8 (52.5 KD) and 18 (45 KD) but negative for CK 1 (68 KD), 5 (58 KD), 10 (56.5 KD), 11 (56 KD), 3 (64 KD), 13 (51 KD), 19 (40 KD) and 7 (54 KD). An antibody for CK 14 (50 KD) (CBL 197) failed to react even with a positive control. © 1991 by Gustav Fischer Verlag, Stuttgart
Fig. 1. Serial sections of Crooke's cells. - A: ACTH stain (x 400). - B: Cytokeratin stain (MA-902) (x 400). 0344-0338/91/0187-0539$3.50/0
540 . Y. Uei, M. Kanzaki and T. Yabana Table 1. Monoclonal antibodies for cytokeratins Cat. No. (Source)
Basic (Type II) No.
Cytokeratin subfamily Acidic (Type I) Mol.wt (KD) Mol.wt (KD) No.
MA-904 (ENZO) MA-903 (ENZO) AE 5 (lCN) AE 8 (lCN) MA-902 (ENZO) Ks 19.1 (lCN) C-6417 (Sigma) C-8541 (Sigma) CBL 197 (CBL)
1 1,5 3
68 68,58 64
8
52.5
7
10,11
56.5,56
13
51
19
40
18 14
45 50
54
Markers for Skin-type Skin-type Cornea-type Esophageal-type Simple epithelia Simple epithelia Simple epithelia Simple epithelia Keratinocytes
Trypsin digestion
+
+ +
+ +
abnormal accumulation of a cellular structural protein4 • The monoclonal antibody (KL 1) used in this study was that generated against a broad range of CKs recognizing epithelial cells from the upper layer of the epidermis, gastrointestinal and respiratory mucosae and from other epithelia including the thyroid and thymus s . It is of interest to determine what subunit molecular weight CKs Crooke's hyalin is composed of. Nineteen different CK polypeptides have been identified with molecular weights ranging from 40 to 68 KD1. Recently monoclonal anticytokeratin antibodies of more restricted specificity have become available. Using these antibodies, it was confirmed that the CK of Crooke's hyalin consisted of CKs 8 and 18. There have only been few reports on Crooke's hyaline change from the immunohistochemical viewpoint 2, 3. According to Neumann et aI., Crooke's hyalin was positive for CK with each of the five antisera they used 3, but their results disagree slightly with ours concerning the exact subfamilies involved. References
Fig. 2. Serial sections of Crooke's cells. - A: ACTH stain (x 400). - B: Cytokeratin stain (C-8541) (x 400).
Discussion Our previous study identified the Crooke's hyalin found in ACTH cells as CK, which therefore represents an
1 Cooper 0, Schermer A, Sun T-T (1985) Biology of disease. Classification of human epithelia and . their neoplasms using monoclonal antibodies to keratin: Strategies, applications and limitations. Lab Invest 52: 243-256 2 Halliday WC, Asa SL, Kovacs K, Scheithauer BW (1987) Immunohistochemical study of intermediate filaments in the human pituitary. Lab Invest 56: 29 A 3 Neumann PE, Horoupian DS, Goldman JE, Hess ME (1984) Cytoplasmic filaments of Crooke's hyaline change belong to the cytokeratin class. An immunocytochemical and ultrastructural study. Am J Pathol116: 214-222 4 · Uei Y, Kanzaki M, Ohta T. Immunohistochemical study of Crooke's hyaline change. Endocr Pathol: In press 5 Viac J, Reano A, Brochier J, Staquet M-J, Thivolet J (1983 ) Reactivity pattern of a monoclonal antikeratin antibody (KL 1). J Invest Dermatolog 81: 351-354
Received December 15, 1990 . Accepted January 16, 1991
Key words: Crooke's cell- Cytokeratin Yoshio Uei, M.D., Head, Cytology Section, Clinical Laboratory, National Cancer Center Hospital, 1-1 Tsukiji 5 chome, Chuo-ku, Tokyo 104, Japan
