Julio C Valencia1, Wendy K Steagall1, Yi Zhang1, Patricia Fetsch2, Andrea Abati2, Katsuya Tsukada1, Eric Billings3, Vincent J Hearing4, Zu-Xi Yu5, Gustavo Pacheco-Rodriguez1, Joel Moss6. 1. Cardiovascular and Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health. 2. Cytopathology Section, National Institutes of Health, Bethesda, MD. 3. Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health. 4. Pigment Cell Biology Section, National Cancer Institute, National Institutes of Health, Bethesda, MD. 5. Pathology Core, National Heart, Lung, and Blood Institute, National Institutes of Health. 6. Cardiovascular and Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health. Electronic address: mossj@nhlbi.nih.gov.
Abstract
BACKGROUND: Lymphangioleiomyomatosis (LAM) is characterized by the proliferation in the lung, axial lymphatics (eg, lymphangioleiomyomas), and kidney (eg, angiomyolipomas) of abnormal smooth muscle-like LAM cells, which express melanoma antigens such as Pmel17/gp100 and have dysfunctional tumor suppressor tuberous sclerosis complex (TSC) genes TSC2 or TSC1. Histopathologic diagnosis of LAM in lung specimens is based on identification of the Pmel17 protein with the monoclonal antibody HMB-45. METHODS: We compared the sensitivity of HMB-45 to that of antipeptide antibody αPEP13h, which reacts with a C-terminal peptide of Pmel17. LAM lung nodules were laser-capture microdissected to identify proteins by Western blotting. RESULTS: HMB-45 recognized approximately 25% of LAM cells within the LAM lung nodules, whereas αPEP13h identified > 82% of LAM cells within these structures in approximately 90% of patients. Whereas HMB-45 reacted with epithelioid but not with spindle-shaped LAM cells, αPEP13h identified both spindle-shaped and epithelioid LAM cells, providing greater sensitivity for detection of all types of LAM cells. HMB-45 recognized Pmel17 in premelanosomal organelles; αPEP13h recognized proteins in the cytoplasm as well as in premelanosomal organelles. Both antibodies recognized a Pmel17 variant of approximately 50 kDa. CONCLUSIONS: Based on its sensitivity and specificity, αPEP13h may be useful in the diagnosis of LAM and more sensitive than HMB-45.
BACKGROUND:Lymphangioleiomyomatosis (LAM) is characterized by the proliferation in the lung, axial lymphatics (eg, lymphangioleiomyomas), and kidney (eg, angiomyolipomas) of abnormal smooth muscle-like LAM cells, which express melanoma antigens such as Pmel17/gp100 and have dysfunctional tumor suppressor tuberous sclerosis complex (TSC) genes TSC2 or TSC1. Histopathologic diagnosis of LAM in lung specimens is based on identification of the Pmel17 protein with the monoclonal antibody HMB-45. METHODS: We compared the sensitivity of HMB-45 to that of antipeptide antibody αPEP13h, which reacts with a C-terminal peptide of Pmel17. LAM lung nodules were laser-capture microdissected to identify proteins by Western blotting. RESULTS:HMB-45 recognized approximately 25% of LAM cells within the LAM lung nodules, whereas αPEP13h identified > 82% of LAM cells within these structures in approximately 90% of patients. Whereas HMB-45 reacted with epithelioid but not with spindle-shaped LAM cells, αPEP13h identified both spindle-shaped and epithelioid LAM cells, providing greater sensitivity for detection of all types of LAM cells. HMB-45 recognized Pmel17 in premelanosomal organelles; αPEP13h recognized proteins in the cytoplasm as well as in premelanosomal organelles. Both antibodies recognized a Pmel17 variant of approximately 50 kDa. CONCLUSIONS: Based on its sensitivity and specificity, αPEP13h may be useful in the diagnosis of LAM and more sensitive than HMB-45.
Authors: Elena A Goncharova; Dmitry A Goncharov; Andrew Eszterhas; Deborah S Hunter; Marilyn K Glassberg; Raymond S Yeung; Cheryl L Walker; Daniel Noonan; David J Kwiatkowski; Margaret M Chou; Reynold A Panettieri; Vera P Krymskaya Journal: J Biol Chem Date: 2002-06-03 Impact factor: 5.157
Authors: T Kobayashi; K Urabe; S J Orlow; K Higashi; G Imokawa; B S Kwon; B Potterf; V J Hearing Journal: J Biol Chem Date: 1994-11-18 Impact factor: 5.157
Authors: Connie G Glasgow; Gustavo Pacheco-Rodriguez; Wendy K Steagall; Mary E Haughey; Patricia A Julien-Williams; Mario P Stylianou; Bernadette R Gochuico; Joel Moss Journal: Chest Date: 2017-05-30 Impact factor: 9.410