Stromal cells in cerebellar haemangioblastomas: an immunocytochemical study

The nature of the stromal cells in formalin-fixed paraffin-embedded material from 23 cerebellar haemangioblastomas was investigated using antisera to intermediate filaments (glial fibrillary acidic protein, vimentin and desmin), histiocytic markers (alpha 1-antitrypsin, alpha 1-antichymotrypsin and lysozyme), glycolytic enzymes (alpha and gamma enolase and aldolase C4) and the endothelial markers, factor VIII related antigen and Ulex europaeus I lectin. Most stromal cells stained positively for vimentin and the glycolytic enzymes. Occasional process-bearing cells within the stroma stained strongly for glial fibrillary acidic protein, alpha 1-antitrypsin and alpha 1-antichymotrypsin. No stromal cell staining for desmin, lysozyme or the endothelial markers was observed, although the latter stained the vascular endothelium within all neoplasms. The findings do not support previous suggestions of an endothelial or histiocytic origin for the stromal cells. They appear to be a heterogeneous population including entrapped reactive astrocytes and locally-derived non-angiogenic cells of neuroectodermal (pial) origin.     

Primary retroperitoneal dysgerminoma presenting as an adrenal tumor: a case report and literature review

Primary extragonadal germ cell tumors are rare and mostly occur in young men with predominance of nonseminomatous histology. We report an undescribed case of primary retroperitoneal dysgerminoma presenting as an adrenal tumor in a 17-year-old girl. Surgery was performed on a 10 × 9.5 cm sized adrenal gland tumor and the resected tumor showed unequivocal histological features of dysgerminoma. The diagnosis was confirmed by the tumor's germ cell immunophenotype. Postoperative ultrasonography, CT and PET over a 6-month period revealed no evidence of ovarian lesion. The patient is stable, but with a suspicious residual tumor after adjuvant chemotherapy.     

Progression of cutaneous squamous cell carcinoma in immunosuppressed patients is associated with reduced CD123+ and FOXP3+ cells in the perineoplastic inflammatory infiltrate

Aims: Squamous cell carcinoma of the skin (SCC) increases dramatically in organ transplant recipients (OTRs). The aim was to determine whether qualitative and quantitative differences in perineoplastic inflammation in OTRs contribute to the increased carcinogenesis. Methods and results: We studied the perineoplastic inflammatory infiltrate in SCC, assessing depth, density and phenotype (CD3, 4, 8, FOXP3, CD123 and STAT1) by immunohistochemistry in paired biopsy specimens of intraepithelial and invasive SCC in immunocompetent patients and OTRs. Considerable inflammation was observed in all intraepithelial SCC (inflammatory infiltrate depth 2.80 ± 2.21 mm immunocompetent patients, 2.15 ± 2.95 mm OTRs). Inflammation was more pronounced in invasive SCC of immunocompetent patients (4.60 ± 4.67 mm) and OTRs (3.30 ± 5.90 mm) (P < 0.005). The density of perineoplastic inflammatory infiltrates increased from intraepithelial to invasive SCC (P = 0.005). OTRs showed a lower density of perineoplastic inflammatory infiltrate (P = 0.041). OTRs also showed reduced CD3+ T‐lymphocyte and CD8+ cytotoxic T‐lymphocyte proportions in intraepithelial SCC (P = 0.025 and 0.027, respectively). FOXP3+ regulatory T‐lymphocyte proportions in OTRs’ invasive SCC were markedly diminished (P = 0.048). CD123+ plasmacytoid dendritic cells increased in the progression from intraepithelial to invasive SCC in immunocompetent patients (P = 0.040). CD123+ cells were reduced in all SCC of OTRs (P = 0.036). Conclusions: Perineoplastic inflammation in intraepithelial SCC is pronounced both in immunocompetent patients and OTRs. Inflammation increases further in invasive SCC. OTRs show reduced proportions of FOXP3+ regulatory T cells and CD123+ plasmacytoid dendritic cells. This distinct inflammatory infiltrate may result in increased cutaneous carcinogenesis and more aggressive behaviour of SCC in OTRs.     

Detailed analysis of the T-cell lymphocytic infiltrate in penile lichen sclerosus: an immunohistochemical and molecular investigation

AIM: To determine the extent of clonal outgrowth in the lymphocytic tissue infiltrate of lichen sclerosus (LS). The presence of T cells with a monoclonally rearranged T-cell receptor gamma-gene (TCRgamma) has been described in up to 50% of biopsies of vulvar and penile LS. MATERIAL AND RESULTS: We analysed 33 foreskin specimens with LS for the presence of clonal T cells by conventional polymerase chain reaction (PCR) analysis and with TCRgamma-PCR-based fluorescent fragment analysis. Eighteen of 33 patients revealed a band indicating a monoclonally rearranged TCRgamma on conventional PCR analysis. Subsequent TCRgamma-PCR-based fluorescent fragment analysis identified 8/18 patients with monoclonal T-cell DNA ranging from 1.4% to 23.1% of total T-cell DNA analysed and a size range from 56 to 72 base pairs. Four of 18 patients had an oligoclonal and 6/18 patients revealed a polyclonal banding pattern. The lymphocytic infiltrate contained low numbers of gammadelta T cells and cytotoxic T cells in comparable numbers to the low percentage of clonal TCRgamma DNA. CONCLUSIONS: The low percentage of clonal TCRgamma DNA argues against a systemic neoplastic disease, but rather for a local immune disorder. The target antigen of the clonal outgrowth is unknown, but an exaggerated antigen-dependent proliferation of T cells due to chronic local antigen exposure, probably an infectious antigen, is the most likely explanation.     

Coexistence of gonadoblastoma and dysgerminoma in a dysgenetic gonad on touch preparation: a case report

Swyer syndrome is known as pure gonadal dysgenesis. Individuals with Swyer syndrome are phenotypically female with unambiguously female genital appearance at birth, and normal Mullerian structures. The condition usually first becomes apparent in adolescence with delayed puberty and amenorrhea due to the fact that the gonads have no hormonal or reproductive potential. These individuals are characterized by 46XY karyotype, primary amenorrhea, tall stature, female external genitalia and normal but hypoestrogenised vagina and cervix. A high incidence of gonadoblastoma and germ cell malignancies has been reported in dysgenetic gonads, and therefore, the current practice is to proceed to a gonadectomy once the diagnosis is made. Herein, we report a case of gonadoblastoma and dysgerminoma diagnosed on touch preparation in a dysgenetic gonad of a 16-year-old patient with Swyer syndrome.     

Mantle cell lymphoma: A cytopathological and immunocytochemical study

Mantle cell lymphoma (MCL) is a rare type of non-Hodgkin's lymphoma which is thought to derive from the cuff of the lymphoid follicle. The histopathologic and immunophenotypic features of MCL are well described. The literature contains few cytopathologic studies of collected cases of MCL. Review of files from the National Institutes of Health from 1989 through June 1993 revealed a total of 24 positive specimens from eight patients with a diagnosis of MCL. The specimens consisted of ten pleural effusions, ten cerebrospinal fluids (csf), and four fine-needle aspirations. CSF involvement was noted in 3 of 8 (37.5%) patients and was associated with disease progression. The cardinal morphologic features on air-dried, Diff-Quik-stained material are a monotonous population of relatively small atypical lymphoid cells with enlarged, frequently grooved nuclei, coarse chromatin and small nucleoli, scant cytoplasm, and an absence of large or “transformed” lymphoid cells. Immunocytochemistry is characterized by expression of one or more pan-B-cell markers, immunoglobulin light-chain restriction, and positivity for the pan-T-cell antigen CD5. When these morphologic and immunocytochemical characteristics are present, the specific diagnosis of MCL can be suggested on cytologic specimens. Diagn Cytopathol 1996;14:32–37. © 1996 Wiley-Liss, Inc.     

Qligodendrogliomas: a clinical, histological, immunocytochemical and lectin-binding study

We have studied 27 oligodendrogliomas with a panel of antibodies (vimentin, GFAP, S-100 protein, myelin basic protein, CAM 5.2) and of lectins (WGA, Con A, PNA, RCA, DBA, SBA) to different glycoproteins. There were 16 well-differentiated tumours, including one gliofibrillary and 11 anaplastic oligodendrogliomas, three of which were gliofibrillary. Four cases showed positivity for vimentin, three of which were anaplastic tumours. Fifteen cases were positive for S-100 protein (nine well-differentiated and six anaplastic tumours) and 13 contained GFAP-positive cells (three well-differentiated and 10 anaplastic tumours). WGA binding was positive in 75% of well-differentiated and 63% of anaplastic oligodendrogliomas, the corresponding figures were 50% and 45% for PNA, 37% and 81% for Con-A and 25% and 54% for RCA. On the basis of the results with lectin binding, we believe that there are changes in the spectrum of tumour cell-associated lectin-like proteins during malignant transformation. Our observations also suggest that the pattern of lectin expression can undergo substantial changes in the course of differentiation.     

Effects of endotoxin-treatment on inflammatory cell infiltrates in murine Meth A sarcoma

The effect of intravenously injected endotoxin on inflammatory cells within solid Meth A tumours was studied and central hyperaemia, necrosis and early collapse were observed macroscopically at 4, 24 and 48 h, respectively. The effects were studied in semithin sections and cytocentrifuge preparations of the tumours. The inflammatory cell reaction evoked by the tumours in untreated animals was relatively slight. It was located predominantly around the lateral margins of the tumours and only a few inflammatory cells were found inside the tumour. Prominent effects of endotoxin included a transient increase of mononuclear inflammatory cells in the centre of the tumour by 4 h and a reduction of the influx of lymphocytes, observed in and around the margin of control tumours, by 48 h. Mast cells formed an important part of the inflammatory cell infiltrate, but no distinct changes in number and appearance were observed with time or following treatment. Total host cell numbers within tumours did not increase significantly upon endotoxin-treatment. Results suggest that a direct cytotoxic action of host cells cannot account for the extensive tumour damage observed. Rather, endotoxin-induced regression seems to be related to decreased lymphocyte numbers.     

The distribution of renin-containing cells in kidneys with renal artery stenosis—an immunocytochemical study

In 10 kidneys removed for hypertension due to renal artery stenosis, the histological appearances varied from negligible ischaemic damage to end-stage ischaemic atrophy. We stained the renin-containing cells in tissue sections using an antiserum to pure human renin and an immunoperoxidase technique. In all kidneys there was hyperplasia of the renin-containing cells both in juxtaglomerular apparatuses (JGAs) and in arteries outside the JGA, where these cells extended proximally as far as the Interlobular arteries. We mapped the distribution of renin-containing cells and found them in all zones of the renal cortex; in three kidneys they were predominantly in the superficial cortex; in four they were distributed more evenly throughout the width of the cortex; but in three kidneys the normal gradient was reversed, with most of the cells being in the juxtamedullary cortex and, in two of the cases, little or no stainable renin in the superficial cortical JGAs. We suggest that these abnormalities in the distribution of renin-containing cells could affect both the pattern of intrarenal blood flow and the site in the kidney at which secreted renin enters the blood.     

Quantitative immunocytochemical study of islet cell populations in diabetic calmodulin-transgenic mice

The present study describes the changes in the endocrine pancreas of severely diabetic calmodulin-transgenic mice using light microscopic immunocyto-chemical and morphometric techniques. A marked reduction in the number and volume of islets, together with distortion of their normal architecture, was found in diabetic mice. In addition, the volume density of both endocrine tissue and B-cells was decreased. An irregular distribution of non-B-cells was also observed in diabetic animals. The volume density and the percentage of A-cells appeared increased. However, when quantified per area unit, the number of all the islet cell types diminished, although only the decrease in B-cell number was statistically significant. The decrease in B-cell mass might account for the diabetic state developed in this animal model.     

The microscopic features of inflammatory abdominal aortic aneurysms: discriminant analysis

Up to 15% of abdominal aortic aneurysms are designated as inflammatory. They are characterized by marked fibrous thickening of the aneurysmal wall, with the fibrosis extending into the adjacent retroperitoneum. Thirty-five abdominal aortic aneurysms were studied, 15 inflammatory and 20 atherosclerotic. Of the inflammatory group, 10 were symptomatic and five asymptomatic. For each resection specimen, 59 microscopic features (variables) were scored semi-quantitatively. Discriminant function analysis showed that endarteritis obliterans, fibrosis around nerves or ganglia at the outer margin of mural fibrosis, and the thickness of the combined fibrotic media and adventitia gave a satisfactory high discrimination between atherosclerotic and inflammatory aneurysms. When these three variables are used together, a histological diagnosis of inflammatory aneurysm can be made with an expected accuracy in excess of 80%.     

Spindle cell tumours of the skin of debatable origin. An immunocytochemical study

Twelve cases of malignant spindle-cell and sarcomatoid tumours of the skin of debatable nature were studied by immunocytochemical methods, using four antisera which might help contribute to resolution of the problems. The initial diagnosis made on structural grounds was confirmed by immunocytochemistry in six of eight cases in which a specific diagnosis had been made (one melanoma, three squamous carcinomas and two atypical fibroxanthomas). One case, initially regarded as AFX was reclassified as a squamous carcinoma, while a further case of possible AFX could not be confirmed by immunocytochemical study. Of the four cases in which structural examination was inconclusive, two were identified as squamous carcinomas and one as a melanoma by virtue of tumour markers. The fourth case was an intriguing actin-rich tumour of uncertain nature. Immunocytochemistry, despite certain limitations, has a valuable role to play in the analysis of the problematic spindle-cell malignant and pseudomalignant tumours of the skin.     

Inflammatory pseudotumor of lymph node and spleen: An entity biologically distinct from inflammatory myofibroblastic tumor

Inflammatory pseudotumors (IPTs) of the lymph node and spleen are an uncommon, benign cause of lymphadenopathy and/or splenomegaly that often bear striking clinicopathologic similarities to the inflammatory myofibroblastic tumors (IMTs) found in soft tissues. These tumors have classically been grouped together under the umbrella category of "inflammatory pseudotumor." Recent evidence shows that IMTs are in fact neoplastic processes that often harbor balanced chromosomal translocations involving the ALK kinase gene. These translocations result in expression of ALK kinase in IMTs as assessed by immunohistochemical studies. However, the relationship between IMT and IPT of the lymph node and spleen is uncertain. To determine if ALK tyrosine kinase expression is also present in IPT, 13 cases of IPT (9 involving lymph nodes, 4 splenic lesions) were examined for the presence of ALK tyrosine kinase by immunohistochemical staining on paraffin-embedded tissue. In addition, in situ hybridization studies for Epstein-Barr virus--encoded RNAs (EBER) and immunoperoxidase studies for human herpesvirus-8 (HHV8)--specific proteins were performed. All cases had clinical, morphologic, and immunophenotypic findings typical of IPT and had varying proportions of fibroblastic and inflammatory components. Age ranged from 11 to 75 (median, 40) years; 8 subjects were male, and 5 were female. None of the cases (0 of 13) had positive staining for ALK kinase or HHV8, and in 1 a lymph node (1 of 13) was focally positive for EBV (EBER) by in situ hybridization. The absence of ALK kinase as detected by immunohistochemical studies in IPT of the lymph node and spleen suggests that this entity is biologically distinct from the histologically similar IMT.     

Folic acid in the monkey brain: an immunocytochemical study

The present report describes the first visualization of folic acid-immunoreactive fibers in the mammalian central nervous system using a highly specific antiserum directed against this vitamin. The distribution of folic acid-immunoreactive structures was studied in the brainstem and thalamus of the monkey using an indirect immunoperoxidase technique. We observed fibers containing folic acid, but no folic acid-immunoreactive cell bodies were found. In the brainstem, no immunoreactive structures were visualized in the medulla oblongata, pons, or in the medial-caudal mesencephalon, since at this location immunoreactive fibers containing folic acid were only found at the rostral level in the dorsolateral mesencephalon (in the mesencephalic-diencephalic junction). In the thalamus, the distribution of folic acid-immunoreactive structures was more widespread. Thus, we found immunoreactive fibers in the midline, in nuclei close to the midline (dorsomedial nucleus, centrum medianum/parafascicular complex), in the ventral region of the thalamus (ventral posteroinferior nucleus, ventral posteromedial nucleus), in the ventrolateral thalamus (medial geniculate nucleus, lateral geniculate nucleus, inferior pulvinar nucleus) and in the dorsolateral thalamus (lateral posterior nucleus, pulvinar nucleus). The highest density of fibers containing folic acid was observed in the dorsolateral mesencephalon and in the pulvinar nucleus. The distribution of folic acid-immunoreactive structures in the monkey brain suggests that this vitamin could be involved in several mechanisms, such as visual, auditory, motor and somatosensorial functions.     

Ovarian mixed germ cell tumor composed of dysgerminoma, endodermal sinus tumor, choriocarcinoma and mature teratoma in a 44-year-old woman: Case report and literature review

A case of ovarian mixed germ cell tumor In a 44-year-old woman was examined. The tumor was well circumscribed, measured 15 times 11 times 10cm and appeared solid and partly cystic on the cut surface. Light microscopic examinations revealed that the tumor was composed of four different neoplastic germ cell elements, Intermingled with each other. They are: (i) choriocarcinoma, immunohistochemically positive for human placental lactogen (hPL) and human chorionic gonadotropin (hCG); (ii) dysgerminoma, positive for placental alkaline phosphatase; (iii) endodermal sinus tumor positive for α-fetoproteln (AFP); and (lv) mature teratoma. Among these histological types, dysgermlnoma occupied more than 50% of the neoplasm. The patient was diagnosed as a stage la ovarian mixed germ cell tumor and was subsequently treated with chemotherapy. A second-look laparotomy after completion of chemotherapy revealed no residual tumors in the abdomen and the patient Is alive and well 15 months after operation. This Is the fourth reported case of ovarian mixed germ cell tumor arising In patients over 40 years old.     

Vulvar inflammatory dermatoses for the non-dermatopathologist: an approach for the practising surgical pathologist

Recognition of tissue patterns of injury will help the non-dermatopathologist when confronted with biopsies of vulvar inflammatory dermatoses. This paper defines and discusses the most common inflammatory patterns including: spongiosis, psoriasiform, interface and vesiculobullous, providing a brief overview of the clinical, histologic and differential diagnoses of the more common lesions to affect the vulva. Also included are entities with specific predilection for the female gynaecologic tract including Zoon’s vulvitis and Behcet’s disease. A comprehensive review of this topic is beyond the scope of this mini-symposium. However we hope to provide an approach to the diagnosis of vulvar inflammatory dermatitis which will be of practical use to the surgical pathologist.     

Whipple''s disease: a histological, immunocytochemical and electronmicroscopic study of the immune response in the small intestinal mucosa

Whipple's disease is a multisystem disorder with protean manifestations and with poorly understood aetiopathogenesis. It is unclear how the immune system reacts, whether it functions normally or not, whether it protects the organism or is defeated in one way or another by the 'Whipple bacillus'. The purpose of our study was to assess humoral and cellular immunity at the level of the intestinal mucosa. This histochemical, immunocytochemical and electronmicroscopic study, based on 16 cases, has shown that the changes in components of the mucosal immune system in Whipple's disease are quite different from normal. The phagocytic capacity of the macrophages, assessed microscopically, is abnormal, the number of intra-epithelial lymphocytes is increased, the CD 4/CD 8 cell ratio is decreased and the IgM positive cells in the lamina propria outnumber the IgA positive cells. These changes may be inter-dependent.     

Cytologic appearance of myospherulosis of the breast diagnosed by fine-needle aspirates: a clinical, cytological and immunocytochemical study of 23 cases

The objective of this study was to introduce the clinical and cytological aspects of myospherulosis. A total of 5,174 consecutive breast fine needle aspiration (FNA) cytology cases were reviewed, among which 23 cases of myospherulosis of the breast were found, all in female patients. The main findings of myospherulosis, best seen with the Papanicolaou stain, consisted in the observation of spherules that were homogeneously smooth or contained one or more internal dense bodies. Routine Papanicolaou-stained slides with or without Romanowsky staining were analyzed. Immunocytochemistry was conducted for carbonic anhydrase 1 (CA1), glycophorin C, KP1, and PGM1. The patients' ages ranged from 41 to 79 years (mean age: 56 years). Of the 23 patients, 21 had a previous history of breast surgery. Cytologically malignant or suspicious diagnoses were made in four of the 23 cases. The size of parent bodies varied from 18.2 to 151 μm (mean, 52 μm). The size of spherules ranged from 2.1 to 16.4 μm (mean, 6.6 μm). Immunocytochemistry showed that the myospherules reacted with anti-CA1 and anti-glycophorin C antibodies. Most breast myospheruloses occur in patients with a history of breast surgery. Immunocytochemistry for CA1 and glycophorin C can enhance the diagnosis of myospherulosis.