Hormonally inactivated pituitary adenoma: morphological, immunohistochemical and electron-microscopic characteristic

Morphology of hormonally inactivated pituitary adenoma can be different types of tumors. Morphological immunohistochemical and electron-microscopic researches of 23 hormonally inactivated pituitary adenomas has been carried out. We shown that more frequent morphological substrate was gonadotropinoma or "zero-cells" adenoma. According to our results, gonadotropinomas, "zero-cells" adenomas and oncocytomas have similar features and can be put into the same group of tumor Pathomorphologist has to differentiate this group of tumors from others "silent" pituitary adenomas because they have different prognosis for a disease. A research of somatostatin and dopamine receptors expression would be new area for differential diagnosis of these types of adenomas.     

Human corticotroph cell adenomas

Sixty-one pituitary corticotroph adenomas from 47 patients with Cushing's disease, 10 with Nelson's syndrome, and four eucorticoid patients were studied by light microscopy, immunoperoxidase, and electron microscopy. Seventy nine percent of all tumors and 70% of Nelson's cases were microadenomas, sometimes minute. A contiguity between the posterior lobe and the adenoma was seen in ten cases. Spontaneous infarction of the tumor with remission of Cushing's syndrome occurred in one case. Light microscopy revealed that the adenoma cells were basophilic and contained PAS-positive granules also staining with Herlant tetrachrome and lead-hematoxylin. The granules stained positively with antiserum to adrenocorticotrophic hormone (ACTH), beta-lipotropic hormones (beta-LPH) and beta-endorphin. The most characteristic ultrastructural finding was the presence of perinuclear bundles of microfilaments found in all our cases. Oncocytic changes were seen in three tumors. Four silent corticotroph adenomas, two of them originally microadenomas that had enlarged to enclosed adenomas while being treated with bromocriptine for hyperprolactinemia and one a large diffuse invasive tumor, did not differ in their microscopic, immunocytological, or ultrastructural features.     

Molecular cytogenetics of chromosome 11 in pituitary adenomas: a comparison of fluorescence in situ hybridization and DNA ploidy study

Chromosome 11 abnormalities were detected by fluorescence in situ hybridization (FISH) technique and compared with DNA ploidy in 24 surgically removed pituitary adenomas. The tumors were diagnosed and classified by histology, electron microscopy, and pituitary hormone immunocytochemistry. They included 2 densely granulated somatotroph (DG-SM) and 4 sparsely granulated somatotroph (SG-SM) adenomas, 3 SG lactotroph (LT), 2 mixed somatotroph-lactotroph (SM-LT), 4 functioning corticotroph (CRT), 1 silent CRT subtype 1, 1 thyrotroph, 1 mixed thyrotroph-somatotroph, 2 gonadotrophs, and 4 null cell adenomas. FISH analysis with an alpha-satellite DNA probe specific for chromosome 11 showed numerical abnormalities in 16 functioning (94%) and 5 nonfunctioning (71%) adenomas. Ten functioning tumors showed aneuploid histograms, whereas the remaining and all nonfunctioning adenomas were diploid. Aberrant chromosome 11 signals were noted mostly in aneuploid adenomas involving 17% to 100% of their cell population. The severity of chromosome 11 aberrations in adenomas containing extra copies often correlated with a higher DNA index (DI). Monosomy 11 as dominant aberration was noted in a mixed SM-LT and to a lesser degree in 3 CRT adenomas involving 21% to 97% of their cell population. Two of these CRT adenomas were associated with normal DI, whereas the remaining third showed a high DI, indicating increased copy number of chromosomes other than of chromosome 11. In conclusion, chromosome 11 abnormalities are common in all types of pituitary adenomas, occurring more frequently in functioning tumors. Specific numerical abnormalities, such as monosomy and trisomy, tend to be associated with certain adenoma types, whereas tumors with extra chromosome 11 copies often exhibit aneuploid histograms.     

Immunohistological determination of beta-endorphin in chromophobe, clinically hormone-nonproducing hypophyseal adenomas

Pituitary adenomas are usually classified according to the nature of their proper hormonal production. Silent adenomas of the pituitary are tumors without clinical and biochemical evidence of overproduction of any known adenohypophyseal hormones. The proportion of such seemingly nonfunctioning tumors is 20 to 30%. Silent corticotropic adenomas are able to synthesize some normal or abnormal sequences of proopiomelanocortin precursor without any signs of hypercorticism. These tumors were divided into basophilic adenomas with strong periodic acid-Schiff (PAS) positivity and chromophobic adenomas with moderate or no PAS positivity. All of our cases were chromophobic adenomas. Two of the cases were positive for beta-endorphin by immunofluorescence. ACTH immunoreactivity was not present in the cells. Electron microscopic study of the adenoma cells showed small secretory granules with a halo. The diameter of these granules varied from 50 to 250 nm. Automated morphometric and densitometric investigations of silent corticotropic adenomas and adenomas from patients with Cushing's disease gave different karyometric results. The most important practical problem arising from the present investigation was the high frequency of recurrence of silent corticotropic tumors.     

A Silent Follicle-Stimulating Hormone-Producing Pituitary Adenoma in a Teenage Male

An 18-year-old male was referred to Toranomon Hospital seeking reoperation for recurrent clinically nonfunctioning pituitary adenoma. A pituitary macroadenoma was first suspected at age 15 due to intractable headaches. Endocrine data were unremarkable except slightly elevated serum follicle-stimulating hormone (FSH). Transsphenoidal surgery done at another hospital achieved partial tumor removal but the remaining tumor regrew 2 years after surgery. The recurrent tumor was completely and selectively removed on repeat surgery at Toranomon Hospital. Pathological examination confirmed a silent FSH-producing pituitary adenoma. Forty-five patients less than 20 years old underwent transsphenoidal surgery for pituitary adenoma at Toranomon Hospital between 1993 and 2010. Of the 45 patients, 36 (80.0%) had clinically functioning adenomas and the other 9 (20.0%) had clinically non-functioning adenomas. No patients, other than the present case, had a silent gonadotroph adenoma. In contrast, among 579 patients over 20 years old undergoing surgery for nonfunctioning pituitary adenomas between 2006 and 2010 at Toranomon Hospital, 304 (52.3%) had silent gonadotroph adenomas. Gonadotroph adenomas are more common with aging: for example, 37 (61.7%) of 60 patients more than 70 years old at the time of operation had gonadotroph adenomas. In conclusion, gonadotroph adenomas, especially silent gonadotroph adenomas, are extremely rare in childhood and adolescence.     

Pituitary adenomas. An immunohistochemical study of hormone production and chromogranin localization.

Tumors from 42 surgically resected pituitaries and from 13 autopsy cases were studied immunohistochemically with polyclonal antisera to 7 anterior pituitary hormones and with a newly developed monoclonal antibody directed against human chromogranin for evaluation of the distribution of chromogranin in normal and neoplastic pituitaries. In addition, a prospective study was done for assessment of the prevalence, morphology, and endocrine cell types of pituitary tumors in 100 autopsy subjects. When these 55 pituitary adenomas were examined with monoclonal antibody (LK2H10) directed against human chromogranin, selective staining of normal adenohypophyseal cell types and pituitary tumors was observed. Most null-cell adenomas (12/14) were positive for chromogranin, whereas all prolactin (PRL)-producing adenomas (19/19) were negative. Growth hormone (GH) adenomas were focally positive (9/9). All oncocytomas (2/2), 1 thyrotropin (TSH) adenoma, and a follicle-stimulating hormone/luteinizing hormone adenoma were positive for chromogranin. One or more adenomas were present in 14% of the autopsy cases. The tumors occurred most frequently in patients in the fifth through the seventh decades of life. Immunohistochemical staining of 13 adenomas revealed 1 TSH, 1 ACTH, and 4 PRL-producing tumors, whereas 7 other tumors, which were null-cell or undifferentiated adenomas, failed to stain for any of the seven principle pituitary hormones. These results indicate that antibody LK2H10 to human chromogranin is useful in the immunohistochemical characterization of pituitary adenomas. Incidental pituitary microadenomas from autopsy-derived pituitaries most commonly produce PRL, or they belong to the null-cell or undifferentiated tumor group.     

The pathology of nonfunctional pituitary adenomas

The nonfunctional adenomas also known as undifferentiated, null cells, or nonsecretory adenomas are endocrinologically silent neoplasms of the anterior lobe of the pituitary gland. They constitute 50% of all pituitary adenomas in the present report. Most tumors are large, often growing beyond the confines of the sella turcica and characterized anatomically by displacement and compression of the adjacent sellar structures including the optic chiasm, hypothalamus, and third ventricle. Others may be truly invasive when they locally or diffusely infiltrate the adjacent structures. They may grow through the dura and the bone into the cranial cavity and/or the sphenoidal sinus and the nasopharynx or infiltrate one or both cavernous sinuses. Seventy-six adenomas were divided into 48 nononcocytic adenomas and 28 oncocytic adenomas or oncocytomas occurring in older patients. By light microscopy using conventional histologic stains, the majority of nononcocytic tumors were chromophobic. The oncocytomas were slightly acidophilic with a large granular cytoplasm. With the peroxidase-anti-peroxidase (PAP) method with anti-sera for the anterior pituitary hormones, the adenoma cells show no immunostaining. Ultrastructurally, the nononcocytic tumors possess a modest number of small secretory granules but no specific ultrastructural features. The oncocytic tumors are characterized by an excessive number of mitochondria in their cytoplasm. With the PAP method using cytochrome C oxidase antiserum, specific identification of mitochondria can be made. As the nonfunctional adenomas are devoid of anterior pituitary hormones they have been called "null cell" adenomas. However, most of the null-cell adenomas are positive for chromogranin. The diagnosis of nonfunctional adenoma can be suspected clinically when in the absence of signs of hypersecretion of pituitary hormones, there is radiologic evidence of ballooning of the sella and bulging of its diaphragm. An invasive adenoma can be detected by radiologic signs of destruction of the sella turcica and its boundaries or by neurosurgical intervention when there are findings of invasion of the dura, the parasellar, and suprasellar structures.     

Pathobiologic study of pituitary tumors: report of 62 cases with a review of the recent literature

Advances in radioimmunoassay procedures, immunocytochemistry, neuroradiologic imaging, and the surgical and medical treatment of pituitary adenomas have led to reappraisal of their classification as well as refinements in the diagnostic approaches used by pathologists. Sixty-two pituitary adenomas are described, and recent advances in this field are reviewed. Most of the patients were adults, but one of the adrenocorticotropic hormone (ACTH)-producing adenomas occurred in an 11-month-old infant. Endocrine-inactive tumors (43.5 per cent) were less common than hormone-producing tumors (56.5 per cent). Local invasion was most common in the former group, followed by ACTH-producing and other hormone-producing tumors. Ultrastructural features correlated with hormonal levels in the growth hormone(GH)-secreting tumors but not in the prolactin(LTH)- or ACTH-producing tumors. The formation of 7-nm filaments in the cytoplasm of tumor cells, corresponding to Crooke's hyaline change on light microscopy, was characteristic of ACTH-producing tumors. Ultrastructural changes in the ACTH granules suggested that the filaments may be derived from the feedback action of cortisol. Prior to surgery, a Rathke's cleft cyst and a chordoma were mistaken for endocrine-inactive pituitary adenomas. In two additional cases ectopic ACTH-producing tumors of lung clinically mimicked pituitary adenoma. Conversely, one pituitary adenoma mimicked sphenoid wing meningioma. Clinical, hormonal, and radiologic data and immunocytochemical and electron microscopic studies are needed for accurate pathologic interpretation and classification of pituitary adenomas.     

Silent corticotroph adenomas: further clinical and pathological observations

Adrenocorticotroph cell pituitary adenomas immunoreactive for adrenocorticotropic hormone (ACTH) but unassociated with preoperative signs of hypercortisolism constitute between 6% and 43% of all ACTH adenomas. Few large series have been published. At our referral center for pituitary diseases, we have encountered 12 patients with silent ACTH adenomas, none of whom exhibited definite clinical features of hypercortisolism preoperatively. Two patients presented with apoplexy, and in 2 patients preoperative neuroimaging studies mimicked craniopharyngioma. Pathological examination revealed 8 adenomas with variably basophilic cytoplasm (type I, including 1 each with coarse basophilic granules and Crooke's hyaline change) and 4 with predominantly chromophobic cytoplasm (type II). Diffuse versus patchy (30% to 50% of cells) immunostaining best distinguished these 2 types; calcitonin staining was focal or negative in both. Two patients had unexpected postoperative courses consistent with acute cortisol insufficiency; 1 patient suffered from a severe flu-like illness, and the other had dizziness and was found to have a serum cortisol level of < 1.0 microg/dL. Both patients improved after cortisol replacement followed by a slow taper. Another patient developed 2 separate pituitary adenomas, a silent ACTH adenoma followed by a pure prolactinoma resected months later. Clonality studies demonstrated that the 2 tumors had arisen from different clonal populations. These cases offer additional insights into clinical, neuroimaging, histological, and biological features of silent ACTH adenomas. Because 2 of these patients seemed to require postoperative cortisol supplementation that otherwise would not have been given, clinicians should be notified about ACTH immunostaining in adenomas from patients without preoperative diagnoses of Cushing's disease, to optimize postoperative care.     

Contribution of immunohistochemistry, electron microscopy, and cell culture to the characterization of nonfunctioning pituitary adenomas: A study of 40 cases

We studied 40 endocrinologically inactive pituitary adenomas by immunohistochemistry, electron microscopy, and cell culture in order to determine the incidence of gonadotropic adenomas and to classify nonfunctioning adenomas. Immunohistochemical studies using a large panel of monoclonal and polyclonal antibodies identified the following nonfunctioning adenomas: 20 gonadotropic adenomas, four silent corticotropic adenomas, one plurihormonal adenoma, and 15 nonsecreting adenomas. Among nonsecreting adenomas, ultrastructural study of 13 cases identified seven null cell adenomas and six oncocytomas. Silent corticotropic adenomas were classified into subtypes I, II, and III according to Kovacs and Horvath. Most often, gonadotropic adenomas displayed a varying number of oncocytic cells, characteristic secretory granules, and a prominent Golgi apparatus. Postembedding immunoelectron microscopy was performed on eight gonadotropic or nonsecreting adenomas, but this technique did not provide any additional information. Six gonadotropic adenomas and 10 so-called nonsecreting adenomas were studied in primary cell cultures. The six gonadotropic adenomas and seven of the 10 nonsecreting adenomas released gonadotropins in the culture medium. The use of in vitro results as a supplementary diagnostic criterion allowed classification of the 40 nonfunctioning adenomas as follows: 27 gonadotropic adenomas, four silent corticotropic adenomas, one plurihormonal adenoma, and eight nonsecreting adenomas. These results demonstrate a high proportion of gonadotropic adenomas among nonfunctioning adenomas (67.5%) and the usefulness of several techniques in characterizing this type of pituitary adenoma.     

Pituitary adenomas and granular cell tumors. Incidence, cell type, and location of tumor in 100 pituitary glands at autopsy.

Incidentally detected pituitary adenomas were investigated in 100 pituitary glands at autopsy to determine the number, cell type, and location of tumors, and the presence of coexisting granular cell tumors in the neurohypophysis. Pituitary glands were sagittally sectioned at 1.5-mm intervals in toto and embedded in 1 cassette to orient location of each tumor. Twenty-four pituitary glands harbored adenomas, most smaller than 3 mm and the largest 6 x 5 x 4 mm. Two pituitary glands contained double adenomas of immunocytochemically different cell types. Of the 26 adenomas, 10 had lactotrophs, 2 had mixed lactotrophs-somatotrophs, 1 had mixed lactotrophs-luteinizing hormone cells, and 12 were nonfunctioning. One adenoma with adenocorticotropic hormone cells was also detected. Thus 25 of 26 (96%) adenomas were either lactotrophic or nonfunctioning; this percentage is much higher than that of surgically resected tumors. Twenty-two tumors were contiguous with or adjacent to the capsule from which the adenomas originated. Nine granular cell tumors were noted in the neurohypophysis; 3 coexisted with pituitary adenomas. Fourteen additional cases revealed small granular cell nests. Thus the incidental finding of nonfunctioning pituitary adenomas is relatively common in adults (24% of cases in this study), and the coexistence of pituitary adenomas and granular cell tumors may suggest a possible histogenic connection between anterior and posterior pituitary tumorigenesis.     

Incidence,Pathology, and Recurrence of Pituitary Adenomas: Study of 647 Unselected Surgical Cases

The incidence of various types of unselected pituitary adenomas based on correlation of pathologic and clinical data was assessed. We investigated 647 cases of unselected pituitary adenomas, which were surgically removed between 1980 and 1993. All cases were examined by immunohistochemistry and electron microscopy. The mean age of patients was 44.0 years with 40.0 years for women (55.2%) and 49.1 years for men (44.8%). Age distribution indicated a remarkable sex difference: 52.4% of women and 26.8% of men were between 21 and 40 years at the time of surgery. Based on immunohistochemistry and electron microscopy, prolactin (PRL) cell adenomas represented 26.3% of tumors, growth hormone (GH) cell adenomas 12.5%, adrenocorticotrophic hormone (ACTH) cell adenomas 12.4%, oncocytomas 12.4%, and gonadotroph cell adenomas 9.4%. Seventy-three percent of the prolactinomas occurred in women and 73.8% of the oncocytomas were found in men. The incidence of pediatrics pituitary adenomas was 4.6%. All 647 cases were followed up; the mean follow-up period was 96.6 months. In 40 patients (6.2%), the adenoma recurred. Recurrence was common in functioning ACTH cell adenomas (8 cases: 9.5%) followed by silent adenomas (7 cases: 25.9%). Recurrence was noted after 2–96 months (average 28.7 months) following surgery. The shortest remission period was found in a patient with oncocytoma followed by a patient with prolactinoma.     

'Acidophilic' pituitary tumors: a reappraisal with differential staining and immunocytochemical techniques.

The cells of pituitary adenomas classified as acidophilic on PAS-light green-orange G staining could be further differentiated with the Brookes technique: they stained nonspecifically bluish-gray, orange with orange G, or red with carmoisine. On immunostaining for growth hormone and prolactin, the gray cells were either negative or reactive for prolactin, the orange cells contained growth hormone, and the red cells contained prolactin. Of 28 tumors, eight showed no immunostaining, 11 stained only for prolactin, three stained only for growth hormone, five contained mostly growth hormone cells and some prolactin cells, and one contained predominantly prolactin cells but also numerous growth hormone cells. Immunoreactive growth hormone granules in adenoma cells were usually arranged randomly; prolactin granules were often concentrated along one nuclear pole. This study emphasizes the tinctorial and immunocytochemical heterogeneity of "acidophilic" adenomas.     

Immunohistological studies on clinically silent pituitary adenomas

Clinically inactive adenomas represent 30.7% of all pituitary tumors in our surgical collection of 616 cases. Ninety-six tumors were studied immunohistologically with many antibodies for their hormone content. Morphological classification of these adenomas reveals oncocytic adenomas in 42 cases (44%), small cell chromophobe adenomas in 33 (34%), large cell chromophobe adenomas in 14 cases (15%), undifferentiated mucoid cell adenomas in 4 cases (4%) and undifferentiated acidophil adenomas in 3 cases (3%). Immunohistological studies performed with the six pituitary hormones GH, prolactin, ACTH, TSH, LH and FSH and additionally with α-subunit demonstrated nearly all possible combinations of hormones in adenoma cells. The most frequently occurring (29%) was LH (in 3% of adenomas alone); α-subunit followed in frequency (24%), with FSH present in 21%. Combinations of 2 hormones were detected in 16%, of 3 in 13% and of more than 3 hormones in 2%. All 6 hormones and α-subunit were negative in 48% of adenomas. It must be concluded 1) that many clinically silent adenomas are LH- or FSH- or α:-subunit-positive and therefore probably originate from gonadotropic cells, and 2) that clinically silent adenomas of acidophil cell type or mucoid cell type are rare. Although many of these adenomas apparently do not secrete the hormone which they immunohistologically contain, determining the plasma levels of the gonadotropins, and especially the α-subunit in clinical studies, may obtain a suitable and helpful clinical marker in the diagnosis of “endocrine inactive” adenomas, and especially of their recurrences.     

垂体生长激素细胞腺瘤的电镜及免疫电镜观察

24 cases of growth hormone(GH)-producing pituitary adenomas were studied with electron microscopy and immunoelectron microscopy by protein A-gold complex, 6 cases were identified as densely granulated GH adenoma and 15 cases as sparsely granulated GH adenoma, among which 4 cases were proved by immunoelectron microscopy to be containing granules with prolactin(PRL) activity simultaneously. Intracytoplasmic fibrous bodies were often seen in the sparsely granulated cells anyhow, not all those cells with fibrous bodies possess the secretory granules with GH activity, and fibrous bodies were also detected in some PRL cells of certain mixed type adenoma. This suggests that fibrous bodies might not be the specific morphological feature of pituitary growth hormone cell adenomas.     

Localization of insulin-like growth factor-II mRNA in human pituitary adenomas

Insulin-like growth factors (IGFs) have been reported to promote cell proliferation in many tumours, but their contribution to pituitary adenoma development and growth has not been characterized. We report the presence of insulin-like growth factor II (IGF-II) mRNA in pituitary adenomas using in situ hybridization (ISH). The intensity of IGF-II hybridization signal was correlated with adenoma type, and the presence of Ki-67. Among the 109 adenomas examined, 55 (50.4%) were positive for IGF-II mRNA. All acidophil stem cell, functioning corticotrophic and plurihormonal adenomas contained the message; a high incidence of signal was found among sparsely (7/8) and densely (4/6) granulated growth hormone (GH) cell adenomas, mixed GH cell–prolactin (PRL) cell adenomas (6/7), thyrotrophic (4/6) and null-cell (6/7) adenomas. Less frequently, IGF-II mRNA was localized in mammosomatotrophic, silent subtype 3, gonadotrophic, and oncocytic adenomas, whereas all sparsely granulated PRL cell adenomas and silent corticotrophic adenomas of subtypes 1 and 2 were negative. The MIB-1 labelling index was significantly higher in adenomas with a moderate to intense IGF-II signal than in adenomas with weak or no signal. The results suggest that IGF-II, when highly expressed, may have a role in pituitary adenoma proliferation.