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PubMedDate not supplied

Role of immunocytochemistry in diagnosis of prostatic neoplasia by fine needle aspiration biopsy.

The efficacy of immunocytochemical staining for prostate-specific antigen (PSA) and prostate-specific acid phosphatase (PSAP) was studied in aspiration biopsy specimens from 19 patients. Eighteen patients had prostatic carcinoma and one had hyperplasia of prostate. Specimens were obtained from both the primary tumors and metastatic sites. Immunoperoxidase staining was performed on alcohol-fixed cytology smears (some prepared up to 9 years previously) using appropriate antisera followed by an avidin-biotinylated horseradish peroxidase complex. Results were scored according to the percentage and intensity of positively stained malignant cells. Corresponding histologic specimens were stained and scored in a similar fashion. Correlations were made between the staining characteristics of the tumor markers and grade of tumor, using the University of Texas M.D. Anderson Hospital classification of prostate carcinoma. Overall there was good correlation between cytologic and histologic specimens for the presence of PSA and PSAP, although metastases tended to show fewer positively stained cells than the primary tumor. There was no relationship between tumor grade and percentage of positively stained cells. Ninety-three percent of aspirated primary and secondary prostatic tumors stained positively for PSAP compared with 81% for PSA. In one of 3 patients, negative staining of neoplastic cells by both PSAP and PSA was helpful in confirming the existence of a second primary tumor.

Acid Phosphatase↗
PubMed1986

Requirement for cysteine in the color silver staining of proteins in polyacrylamide gels.

To determine whether cysteine residues have a contribution to the mechanism of color silver staining, we silver stained sodium dodecylsulfate polyacrylamide gel electrophoresis separations of proteins which have few or no cysteines. Proteins without cysteine stained negatively (yellow against a yellow background) with silver. Proteins with one or more cysteines stained orange, red, brown, or green/gray depending on the mole percentage of cysteine and whether they contained covalently attached lipids. The colors could not be correlated with the mole percentages of cysteine of these proteins indicating that some components other than cysteine affect the staining color of cysteine-containing proteins. Silver staining of amino acids, sugars, nucleotide bases, or lipopolysaccharide dot-blotted onto nitrocellulose paper implicated adenine, lipids, the basic amino acids, and glutamine, but not sugars or other amino acids in silver/protein complexes.

Bacterial Outer Membrane Proteins↗
PubMed1990

Prenatal development of human major salivary glands and immunohistochemical detection of keratins using monoclonal antibodies.

The major salivary glands were examined from 69 human fetuses ranging from 10 to 40 weeks of gestation. Prenatal growth curves of developing salivary glands could be established by histological scoring, and development was divided into the early developmental stage (EDS) from 10 to 18 weeks, early intermediate developmental stage (EIDS) from 19 to 24 weeks, late intermediate developmental stage (LIDS) from 15 to 32 weeks, late developmental stage (LDS) from 33 to 40 weeks. Characteristic morphogenesis and cytodifferentiation occurred in glandular duct cells during the period of EIDS and LIDS. In the LDS, acini and ducts of the salivary glands histologically developed into a mature state similar to adult glands. Immunohistochemical staining with monoclonal antibodies (MoAbs) PKK1, KL1, K8.12, K8.13, K4.62, RPN 1160, 1162, 1163, 1164, and 1165 was performed. During the fetal period, keratin expression as revealed by MoAbs PKK1, KL1, K8.12 was well established, and the staining pattern for each of these antibodies was comparable. Other antibodies showed rare or negative staining except K8.13 which had a diffuse, non-specific staining pattern. Accordingly, the proliferation and cytodifferentiation of fetal stage keratin staining in ductal cells as revealed by MoAbs PKK1, KL1, and K8.12 showed a heterogenic distribution in both luminal and basal cells. It is a characteristic finding that the cytodifferentiation of ductal luminal cells precedes ductal basal cells. Ductal basal cells stained with MoAb K8.12 and show heterogeneity of keratin distribution continuously until the full term of gestation. The keratin staining of oral epithelium was also examined to compare with distribution of salivary gland ductal cells and oral epithelial cells. In the present study, the developmental sequence of salivary gland cells and the immunohistochemical properties of keratin proteins in these cells were described in relation to the histogenesis of salivary gland tumours.

Antibodies, Monoclonal↗
PubMed2003

Extramedullary presentation of acute leukaemia: a case of myeloid/natural killer cell precursor leukaemia.

AIM: Myeloid/natural killer (NK) cell leukaemia is characterised by coexpression of myeloid with natural killer cell antigens, a high incidence of extramedullary disease and an aggressive clinical course. METHODS: We report a case of a 28-year-old woman with myeloid/NK cell precursor acute leukaemia. Clinical presentation was correlated with leukaemic blast morphology, immunophenotype, cytogenetic analysis, molecular studies for clonal rearrangements and histological review. RESULTS: The patient had noted skin lesions and a breast infiltrate 4 months prior to the diagnosis. Bone marrow biopsy at the time of presentation revealed characteristic morphological features with a dense infiltrate of bizarre, pleomorphic blast cells with marked nuclear invagination and reniform shapes. Immunophenotypic analysis of the blasts displayed coexpression of myeloid and natural killer cell antigens with a relatively immature phenotype: CD34-,HLADR+, CD33+, CD56+, CD16-, CD57-, MPO-. Cyto- genetic analysis revealed a complex karyotype: del(6)(q21);-12 and add(19)(p13). Histological review of the previous breast biopsy was consistent with granulocytic sarcoma of the breast with a phenotype corresponding to the circulating blasts (positive cytoplasmic staining for myeloid markers, CD68 and CD31, and the NK cell marker CD56, with negative staining for MPO). Skin biopsy confirmed leukaemia cutis. CONCLUSION: Although nodal extramedullary disease is common in the myeloid/NK cell leukaemias, this is the first description of myeloid/NK cell leukemia primarily involving breast and skin. We speculate that CD56 may predispose to extramedullary localisation of tumour.

Adult↗
PubMed1980

On gap junction structure.

We have studied the stain distribution within rat liver gap junctions for specimens prepared by thin sectioning and negative staining. Pools of stain molecules exist in two specific locations with respect to the distinctive morphological units (connexons) of the junction. One pool of stain surrounds the connexons and is restricted to the extracellular space in the gap between the adjacent plasma membranes. The other pool of stain is located along in the central axis of each connexon, measures 1-2 nm in diameter and 4-5 nm in length, and is restricted to the gap region. On rare occasions, barely discernible linear densities seem to extend from this latter pool of stain and traverse the entire width of the junction. The data indicate the existence of a hydrophilic cavity along the central axis of te connexon which, in most instances, is restricted to the gap region. However, the precise depth to which this cavity may further extend along the connexon axis is still uncertain.

Animals↗
PubMed1988

Granular cell leiomyosarcoma of the skin.

A case is presented of a multifocal malignant neoplasm involving the skin of the upper back in a 10-year-old boy following radiation therapy to the head and neck for a cerebellar medulloblastoma. Histologically, the neoplastic cells were remarkable for the presence of abundant periodic acid-Schiff (PAS)-positive diastase-resistant intracytoplasmic eosinophilic granules. Immunoperoxidase procedures revealed strong positive staining of the tumor cells with desmin, vimentin, and smooth muscle myosin antibodies, and negative staining for myoglobin, S-100 protein and keratin, thus supporting a smooth muscle line of differentiation for this neoplasm. Electronmicroscopy demonstrated numerous intracytoplasmic autophagic vacuoles that corresponded to the granules observed under the light microscope. Leiomyosarcoma should be entertained in the differential diagnosis of poorly differentiated cutaneous neoplasms histologically characterized by a proliferation of cells containing abundant granular eosinophilic cytoplasm.

Child↗
PubMed2003

Removal of mercuric chloride deposits from B5-fixed tissue will affect the performance of immunoperoxidase staining of selected antibodies.

B5 clearing is a step used before immunoperoxidase staining to remove the precipitated mercuric chloride deposits caused by B5 fixation of tissue. In the B5 clearing procedure, the slides are treated with Lugol's iodine and 5% sodium thiosulfate before antigen retrieval and the application of the primary antibody. The goal of this project was to study the effect of the B5 clearing protocol on immunoperoxidase staining on paraffin-embedded tissue, which has not been previously reported in a series of antibodies. We evaluated 75 antibodies using the 2-step clearing protocol and performed paired immunoperoxidase staining on the Ventana ES instrument, with and without the clearing protocol. We found that among 75 antibodies studied, 3 (CD5, CD30, and synaptophysin) showed total obliteration of reactivity, and 3 (ALK, Ulex, and GFAP) showed partial reduction of the staining compared with the controls. Pathologists must be aware of the possible false-negative staining effect caused by the routinely used B5 clearing protocol. Control tissues must receive the same clearing protocol (i.e., placed on case slides) to ensure detection of this effect.

Antibodies↗
PubMed2000

Hyalinizing Spitz nevus.

A seventeen-year-old Korean girl had a reddish-brown papular lesion on the nose. Histopathologically, it proved to be a "hyalinizing Spitz nevus" with the characteristic features of a discohesive growth pattern of nevus cells and hyalinized stroma. Immunohistochemical stains showed positive reactivity of nevus cells with S-100 protein and Vimentin and negative stainings with HMB45, CD68, CEA and low molecular weight cytokeratin. Hyalinizing Spitz nevus may represent a variant in the spectrum of Spitz nevus.

Adolescent↗
PubMed1993

Expression of nm23-H1 and nm23-H2 proteins in prostate carcinoma.

The nm23 gene products/nucleoside diphosphate (NDP) kinase expression in prostate carcinomas and benign hyperplasias was evaluated immunohistochemically. Monoclonal antibodies against nm23-H1 and nm23-H2 proteins were prepared using the corresponding proteins fused with glutathione S-transferase as immunogens. Of the 80 cases of nonmetastatic prostate carcinoma examined, 74% (59/80) and 60% (48/80) were immunoreactive for nm23-H1 or nm23-H2 protein, respectively. Negative staining for nm23-H1 occurred in 83% of metastatic lesions, while 34% were negative for nm23-H2. All primary tumors corresponding to the metastases examined showed positive immunostaining for nm23-H1, indicating an inverse relationship between expression of this protein and metastatic status. nm23-H2 protein was detected in 83% of primary tumors and its expression appeared to be significantly correlated to the degree of histological differentiation. In contrast, all cases of benign prostatic hyperplasia showed elevated levels of both nm23-H1 and nm23-H2 expression. These data suggest that the nm23/NDP kinase may play a role in suppressing the expression of malignant potential in prostate carcinomas.

Adenocarcinoma↗
PubMed1995

Production and characterisation of an antimelanoma monoclonal antibody KBA.62 using a new melanoma cell line reactive on paraffin wax embedded sections.

AIMS--To generate new monoclonal antibodies directed against melanoma associated antigens using a new melanoma cell line, KAL. METHODS--The melanoma cell line was established in culture from a lymph node metastasis of malignant melanoma. Normal Balb/c mice were immunised with KAL cells. Splenocytes were used for fusion experiments using standard techniques. Hybridoma supernatants were tested for antibody binding activity using an indirect immunoperoxidase method on frozen sections from KAL tumour cells xenografted onto nude mice and human tonsils. KBA.62 was selected because of its reactivity with melanocytic proliferations on both frozen and paraffin wax sections. RESULTS--On immunoblotting, KBA.62 reacted with three bands of 140, 135 and 128 kD and two weak bands of 88 and 73 kD. In normal human tissues basal melanocytes in the epidermis did not react with this antibody and only occasional labelling of endothelial cells was noted. Of the human tumours, KBA.62 reacted strongly and uniformly with the majority of benign (21/21) and malignant (75/86) melanocytic proliferations. Staining was localised predominantly to the cell membrane with little or no cytoplasmic reactivity. Negative staining was observed in the majority of human non-melanocytic neoplasms, the exceptions being some carcinomas (11/89), particularly the well differentiated squamous cell type. This, however, was not thought to present a diagnostic problem. CONCLUSIONS--KBA.62 appears to be potentially useful in ascertaining the immunomorphological diagnosis of malignant melanoma in routinely processed paraffin wax sections.

Animals↗
PubMed2003

Prognostic role of apoptotic, Bcl-2, c-erbB-2 and p53 tumor markers in salivary gland malignancies.

OBJECTIVE: The clinical characteristics and survival probability rate of 36 patients with salivary gland malignancies and 10 patients with benign salivary tumors were summarized in relation to the immunohistological analysis of the tumor, apoptotic-related markers and apoptosis rate. The expression of the markers examined - Bcl-2, c-erbB-2, p53 - was detected in paraffin sections of the tumors by the streptavidin-biotin peroxidase method following heat-induced antigen retrieval, and the apoptosis rate was determined by the TUNEL method. RESULTS: The overall 5-year survival probability was 61% for patients with malignant tumors and 100% for those with benign tumors. The survival probability of patients over 60 at diagnosis was significantly lower than that of younger patients. Patients whose malignant tumors were larger than 2 cm at diagnosis had worse survival than those with smaller tumors. The survival probability of patients whose malignant tumors were located in the submandibular glands was significantly lower than that of patients whose malignancies were located in the parotid and minor salivary glands. The survival probability of patients who demonstrated positive staining for c-erbB- 2 or TUNEL was lower than for those with negative staining. Gender, the existence of concomitant non-salivary malignancies and ethnic origin had no significant impact on survival. CONCLUSIONS: Our results demonstrated significant positive staining in the salivary tumorigenic tissue but not in the surrounding non-tumorigenic tissue examined for TUNEL, c-erbB-2, Bcl-2 and p53, pointing to a biological role for all four markers in the tumorigenic process which is yet to be elucidated. Significant reduction in survival was related to the specific location of the tumor in the submandibular gland, its size and older age of patient. Survival was also found to be significantly reduced when positive staining was demonstrated in the tumor tissue for TUNEL or c-erbB-2, more so for concomitant positive staining of both markers. Clinically, the most important result of the current study is that the survival rate of the patients examined with salivary tumors larger than 2 cm, with positive staining for both TUNEL and c-erbB-2, was 0 (p = 0.0001)!

Adolescent↗
PubMed2005

Mechanism of counterattack of colorectal cancer cell by Fas/Fas ligand system.

AIM: To determine the role of Fas/Fas ligand (FasL) in the immune escape of colon cancer cells. METHODS: Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients. In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assay and CD45 staining were performed to detect the apoptosis of tumor-infiltrating lymphocytes (TILs). Co-culture assays of colon cancer cells (SW480) and Jurkat cells (Fas-sensitive cells) were performed to observe the counterattack of colon cancer cells to lymphocytes. RESULTS: Of 53 cases of colon carcinomas, 23 cases (43.4%) expressed Fas which was significantly lower as compared to the normal colonic mucosa (73.3%, P<0.01), and 45 cases (84.9%) of colon carcinomas expressed FasL, whereas only two cases (3.75%) in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TILs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region (54.84+/-2.79% vs 25.73+/-1.98%, P<0.01). The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells. CONCLUSION: Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy.

Apoptosis↗
PubMed1992

[Establishment of EGF, EGF-R and FN-R positive human adenocarcinoma cell line (GAC-1)].

A human adenocarcinoma cell line designated as GAC-1, was established from ascites of the 56-year old male patient with rapidly progressive gastric cancer. The doubling time was about 18.5 hours in vitro, and cell cycle analysis using flow cytometry showed marked increase of S phase (46.1%). Immunohistochemical demonstration of GAC-1 cells revealed positive staining of TGF-alpha, EGF, EGF-R, FN-R, laminin and negative staining of fibronectin. Histogram of them indicated aneuploidy with modal number 57 and they formed tumors in nude mice.

Adenocarcinoma↗
PubMed1977

[Histochemical detection of HBsAg in liver biopsies using the orcein staining method].

Liver biopsies from 30 HBsAg seronegative and 33 HBsAg positive subjects affected by liver diseases were stained by orcein method for the histochemical detection of HBsAg. All biopsies from HBsAg negative cases were orcein negative. Staining was positive in 55% of seropositive patients. The frequency of the different patterns of the staining is analyzed and its value as a diagnostic and prognostic tool in chronic liver disease is discussed.

Antibody Formation↗
PubMed1997

[Bellini duct carcinoma of the kidney: a case report].

A rare case of Bellini duct carcinoma of the kidney is reported. A 44-year-old woman with macroscopic hematuria was referred to our hospital. the clinical diagnosis was a right renal tumor with direct invasion to the liver. Radical nephrectomy and segmental hepatectomy were performed. Histopathological examination revealed papillary growth of atypical cells different from the usual histological pattern of renal cell carcinoma. The histological diagnosis of Bellini duct carcinoma was confirmed by the positive immunohistochemical staining with a collecting duct marker (UEA-1), and distal tubule marker (EMA) and negative staining with a proximal tubule marker (Leu-M1).

Adult↗
PubMed1997

Lectin histochemical characteristics of the epithelial surface of ileal Peyer's patches in 3-week-old pigs.

Distinct glycoconjugate expression between dome enterocytes and villus enterocytes in ileum from twelve 3-week-old conventional pigs was examined by the use of twenty one biotinylated lectins with avidin-biotin-peroxidase complex method. Three patterns of staining were seen. First, lectins bind to dome enterocyte and villus enterocyte to approximately equal staining, i.e. DSL, WGA, s-WGA and ConA. Second, lectins display a markedly higher affinity for villus enterocyte than dome enterocyte, i.e. DBA, SBA, RCA I, SJA, VVA, BSL II, LEL, PNA, Jacalin, and ECL. Third, lectins exhibit negative staining to dome enterocyte only, i.e. PSA, LCA, UEA-I, and STL. Four lectins; PSA, LCA, UEA-I, STL may be useful negative marker to differentiate glycoconjugate expression between dome enterocyte and villus enterocyte. The staining patterns are slightly different among these negative markers. Three lectins, PSA, LCA, and UEA-1, are negative marker to differentiate dome enterocytes and villus enterocytes. But STL is also negative to dome epithelial surface and moderately positive to villus brush border. It is suggested that the present lectin-binding studies provide the marker of dome enterocyte as compared with villus enterocytes.

Animals↗
PubMed1975

[Toluidine blue staining in laryngeal diseases (author's transl)].

The authors present the results of 83 consecutive in vivo stainings with toluidine blue in various laryngeal diseases. Thirty one (86%) of the 36 malignant tumors showed positive staining. (95% confidence limits: 70-95.) It is concluded that malignant tumors will nearly always fix the dye, and that a negative staining test seems to be evidence against cancer. (Predictive value of negative test: 0.89, 95% confidence limits: 76-96.

Carcinoma in Situ↗
PubMed1994

The frequency of major histocompatibility complex antigen abnormalities in urological tumours and their correction by gene transfection or cytokine stimulation.

There is increasing evidence that loss of major histocompatibility complex (MHC) antigens from tumors may be a factor in escape from immune surveillance. In an attempt to quantify this phenomenon in bladder tumors, frozen sections were stained immunochemically and cell lines were tested in a radiobinding assay before and after treatment with interferon gamma (IFN gamma) and after attempts to correct the defect by normal human leukocyte antigen (HLA) gene transfection. Study of 68 tumor sections with W6/32 antibody against monomorphic class I demonstrated that 42% had reduced or absent staining compared with the intensity of stromal staining. Ten percent of cases had complete absence with W6/32, all of which were also negative for beta 2-microglobulin (beta 2-m) expression. Use of polymorphic antibodies for A2, A3, Bw4, and Bw6 increased this frequency of defects to 73%. Study of 21 tumor cell lines with W6/32 demonstrated negative staining in five (23%) that could not be induced by IFN gamma and reduced staining in three (14%) that could be increased by IFN gamma, the remainder showing normal levels unaffected by IFN gamma. An additional six (28%) failed to express class II in response to IFN gamma, leading to an overall incidence of abnormality of 65%. In no case did cotransfection of one cell line with a defect in one case transfection of beta 2-m gene into a class I negative line of fully assembled MHC class I antigens. It is concluded that the majority of tumor cells demonstrate some form of MHC class I and II defects.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigens, Neoplasm↗