Cardiac myxoma: an immunoperoxidase study of histogenesis

Eleven surgically excised left atrial myxomas have been stained for factor VIII related antigen (VIII-RA) and actin by the peroxidase-antiperoxidase technique. Most tumour cells were negative for VIII-RA. However a population was identified forming cords and vascular channels which possessed a core or lining of tumour cells with an endothelial morphology which were positive. Tumour cells lining vessel-like invaginations of the surface also showed focal positive staining. The staining pattern for actin precisely mirrored that for VIII-RA. These findings support the established theory that cardiac myxomas arise from primitive endocardial or subendocardial mesenchymal cells with the potential to differentiate towards endothelial cells. They are incompatible with the theory that these tumours originate from cardiac endothelium.     

Epithelial markers in thyroid carcinoma: an immunoperoxidase study

Ten cases each of papillary, follicular, anaplastic and medullary carcinoma of the thyroid were stained for thyroglobulin, calcitonin, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA) and cytokeratin (CAM 5.2). Monoclonal or affinity purified polyclonal antibodies, and an indirect immunoperoxidase technique were used. All the papillary and follicular tumours, 5/10 anaplastic and 3/10 medullary carcinomas contained thyroglobulin. Only the 10 medullary carcinomas stained positively for calcitonin. Three out of 10 papillary, 1/10 follicular, 0/10 anaplastic and 10/10 medullary carcinomas were positive for CEA. Nine out of ten papillary, 7/10 follicular, 2/10 anaplastic and 3/10 medullary carcinomas were positive for EMA. Ten out of 10 papillary, 10/10 follicular, 5/10 anaplastic and 10/10 medullary carcinomas were positive for cytokeratin. The presence of calcitonin and CEA is of value in the diagnosis of medullary carcinoma, and enable its distinction from anaplastic thyroid carcinoma. Thyroglobulin is a useful marker in thyroid carcinomas.     

Cholesterol emboli to the kidney: an immunoperoxidase study

Cholesterol emboli (CE) are an increasingly recognised cause of renal impairment in the elderly population, especially following diagnostic vascular procedures and aortic surgery. They can present as part of a multisystem disease which can mimic many conditions, depending on the site of the emboli. As a pathological entity, it was described by Florey in 1945. Relatively few cases have been reported in the literature. At this stage there is no accepted treatment protocol for CE induced renal failure. Little is known about the precise nature of the cells involved in the proliferating tissue surrounding CE in the kidney. To date, all studies on CE have involved routine Haematoxylin and Eosin (H&E) stains. By studying the cellular interactions, hopefully this will contribute further to our understanding of CE in the kidney. Nine (n = 9) out of 1150 consecutive renal biopsies over a six year period were analysed. Standard three- and four-layered peroxidase-antiperoxidase techniques were employed. A panel of antibodies to specific cells were used. The particular cells analysed were the myofibroblast, smooth muscle, endothelium, macrophage, neutrophil, T cell and B cell. All vessels in the biopsy specimen containing CE were analysed. T cell, B cell, macrophage and neutrophil infiltrates were counted and expressed as cells/mm2 using a 0.022 mm2 graticule under 400 (10x40) magnification. The vessel and perivascular space were analysed. The myofibroblast, smooth muscle and endothelial cell proliferation were graded semiquantitatively. Vessels without evidence of CE were used as controls. The data were subjected to statistical analysis using the unpaired non-parametric Mann-Whitney Two Sample test. P values <0.05 were accepted as significant. Histological sections demonstrated the host response in the vessel to CE involve a significant response including the myofibroblast, endothelium, T cell and macrophage. The B cell response was absent and the smooth muscle cell response was not significantly different. The perivascular responses were not different for the cells studied. This study has characterised the host response to CE in the human kidney by demonstrating the presence of the myofibroblast, macrophage T cell and endothelial cell response. The myofibroblast is a cell which is increasingly being recognised in the host response of both granulation tissue and pathological tissue. The population at risk for CE is growing and the disease is increasingly iatrogenic in origin. Currently our only treatment is prevention.     

New marker for mesothelioma: an immunoperoxidase study.

An antibody was raised against a protein isolated from the cytoplasm of mesothelioma cells. It was subsequently used in an immunoperoxidase procedure on formalin fixed, paraffin embedded tissue sections. A representative sample of benign and malignant tumours from all the systems of the human body was examined. All the tumours derived from coelomic surfaces (mesotheliomas of pleura, peritoneum, and ovary, and adenomatoid tumour of epididymis) reacted with the antibody. No other tumour tested in this study expressed the protein. These findings indicate that the antibody may be useful in the identification of mesothelioma cells in both histological and cytological diagnostic routine practice when morphological interpretation is in doubt.     

Hairy-cell leukaemia: an immunoperoxidase study of paraffin-embedded tissues.

Paraffin sections of a variety of tissues from 12 patients with typical hairy-cell leukaemia (HCL) were stained for immunoglobulin heavy and light chains by the peroxidase-antiperoxidase (PAP) technique. Plasma cells were frequent, particularly in a lymph node from a severely infected patient. The reactive nature of the plasma cells of HCL was suggested by the fact that there was no restriction of light-chain expression, although viable hairy cells were shown to express monoclonal surface immunoglobulin. This, together with the absence by both light and electron microscopy of forms intermediate between hairy cells and plasma cells and the lack of ribosome-lamella complexes in the plasma cells, suggested that hairy cells do not differentiate into plasma cells. Although hairy cells are known to contain immunoglobulin, this was not demonstrable in hairy cells in the paraffin-embedded tissue. The PAP technique was also useful for demonstrating abundant splenic macrophages in HCL.     

Anaplastic small cell neoplasms of the thyroid: an immunoperoxidase study

The cell origins of ten anaplastic small cell neoplasms of the thyroid gland were investigated using the immunoperoxidase technique. Sections of the neoplasms were examined for immunostaining for the tissue markers of B lymphocytes, thyroid follicular cells, and C cells by incubation with antisera to the lambda and kappa light chains, human thyroxine and human calcitonin, respectively. Six neoplasms were identified as malignant lymphomas, and two were identified as anaplastic small cell follicular carcinomas. The cell origins of the remaining two neoplasms could not be determined. The prognosis for patients with malignant lymphoma was favorable compared with the prognoses for patients in the other two groups. The prognosis for patients with anaplastic small cell follicular carcinomas was better than for those with small cell malignancies of undetermined cell origins. These findings suggest an important role for the immunoperoxidase technique in the precise classification of anaplastic small cell neoplasms of the thyroid.     

Isoantigen status in condyloma acuminata of the uterine cervix: an immunoperoxidase study

The immunoperoxidase technic was used to investigate the blood isoantigen status in condyloma acuminata, which is regarded as being caused by the human papillomavirus (HPV). Since HPV is associated with epithelial atypias and intra-epithelial neoplasia, and since epithelial malignant transformation is associated with isoantigen loss, the purpose of the study was to determine if koilocytotic atypias are associated with isoantigen loss. Complete isoantigen loss was seen in 33% of cases, partial loss in 47%, and retention in 20%. The significance of this study lies in being able to recognize those lesions that may be associated with malignant transformation (80%) as indicated by isoantigen loss. Isoantigen retention may identify those epithelial atypias that undergo spontaneous regression. Long range follow-up of such patients will help further elucidate the role of HPV in neoplastic transformation of condylomatous lesions. The immunoperoxidase technic can be used in retrospective studies of condylomata of the cervix.     

Dendritic reticulum cells in reactive lymph nodes and tonsils: an immunohistological study

There has recently been much interest in the patterns of follicular dendritic reticulum cells (DRC) in pathological lymph nodes, particularly in relation to the phenomenon of DRC break-up (thought to be pathognomonic of AIDS-related lymphadenopathies) and to progressive transformation of germinal centres (as a possible precursor of lymphocyte predominant Hodgkin's disease). In the present study we have immunostained twenty-nine reactive lymph nodes and five tonsils with monoclonal antibody R4/23 (DAKO-DRC) in order to evaluate the frequency of such changes in lymphoid tissue unaffected by AIDS or Hodgkin's disease. Most of the specimens contained typical secondary follicles with clearly defined germinal centres and mantle zones. There were two variants in lymph nodes showing follicular hyperplasia characterized by (i) progressive transformation of germinal centres and (ii) inclusions of nests of small lymphocytes within germinal centres. In each of these types of follicles the compact evenly-distributed meshwork of DRCs, as previously described, was seen. However there were considerable variations in DRC meshwork in each category (the pattern could not be predicted from the morphology) with examples in all three of the DRC break-up previously considered specific for the AIDS related lymphadenopathy. Since none of the lymph nodes and tonsils studied had any known relationship to either Hodgkin's disease or AIDS it is argued that none of the changes in the DRC meshwork observed are specific for these conditions.     

Keratin and carcinoembryonic antigen in exfoliated mesothelial and malignant cells: an immunoperoxidase study

Immunoperoxidase technics were used to identify keratin and carcinoembryonic antigen (CEA) in exfoliated cells of fine-needle aspirates and body cavity fluids. Staining was evaluated in cytocentrifuge preparations from 27 malignant and 30 benign cytologic specimens. Most reactive mesothelial cell preparations were strongly positive for keratin and negative or only weakly positive for CEA. Diffuse, peripheral, and perinuclear concentration of staining for keratin was noted in exfoliated reactive mesothelial cells. Positive staining for keratin, predominantly diffuse, was noted in exfoliated cells from 56% of the adenocarcinomas. Sixty-nine per cent of adenocarcinoma preparations were strongly positive for CEA. These findings suggest that keratin proteins are not restricted to squamous cells and that keratin staining does not permit distinction between adenocarcinoma and mesothelial cells in cytologic specimens. Staining for CEA and keratin was compared in cytocentrifuge preparations and histologic sections of 12 adenocarcinomas and 7 lymphomas. In some adenocarcinomas, staining was detected only in cytologic preparations. Possible explanations for these differences are discussed. Variable staining for keratin was observed among exfoliated reactive mesothelial cells, possibly identifying different mesothelial cell populations. All reactive and neoplastic lymphoid cells were negative for keratin and CEA in cytologic and histologic preparations. Immunoperoxidase technics can be applied to rehydrated Papanicolaou-fixed and Papanicolaou-stained cytologic preparations with excellent preservation of cytologic detail.     

Distribution of monoclonal antibody-defined monosialoganglioside in normal and cancerous human tissues: an immunoperoxidase study

The immunoreactivity of a monosialoganglioside antigen defined by monoclonal antibody 116NS19-9 (19-9) was studied in neoplastic and normal glandular and mucosal epithelia using an indirect immunoperoxidase method. In neoplastic mucosae, the antigen was detected in the majority of colorectal and endometrial carcinomas, predominantly in a focal staining pattern. A substantial proportion of gastric and pancreatic tumors and an occasional breast carcinoma also reacted with the monoclonal antibody. Expression of the monosialoganglioside in normal colonic mucosa appeared to be restricted to areas adjacent to tumor tissue. In gastric mucosa, the antigen was confined to some areas showing intestinal metaplasia. The antigen was also detected in the epithelium of normal mucosa of the gall bladder and endocervix, as well as in some ductal epithelia of the pancreas and salivary glands. Most other mucosae were negative for antigen expression.     

Granular cell myoblastoma: an immunoperoxidase study using a variety of antisera to human carcinoembroyonic antigen

Immunoperoxidase staining using five antisera to human carcinoembryonic antigen (CEA), including a mouse monoclonal antibody, was performed to investigate the expression of CEA reactivity in ten cases of granular cell myoblastoma. The granular cells were negative with four of the antisera although control sections of CEA producing colon carcinoma were positive. The single positive antiserum gave intense granular cytoplasmic staining of all tumour cells in the ten specimens studied. This reactivity was abolished after absorption of the antiserum with a perchloric acid extract of human lung to remove cross-reacting antibodies against non-specific cross-reacting antigen (NCA); a procedure which did not affect the staining of colon carcinoma specimens. The results indicate that the granular cells do not contain CEA but express a related antigen and that care in the choice of primary antiserum is important if the immunocytochemical detection of this antigen is to be used as a diagnostic aid.     

An immunoperoxidase study of cytomegalovirus mononucleosis

Indirect immunoperoxidase method was used to evaluate retrospectively a case of cytomegalovirus (CMV) mononucleosis. Specific rabbit antiserum against CMV was used as primary reagent in routinely prepared paraffin sections of lymph node and in fixed slide preparations of the bone marrow aspirate. Staining specific for CMV antigen was observed in the lymphoid cells of the lymph node in the predominantly T-cell area and in the large lymphoid cells of the bone marrow. Bone marrow lymphoid cells with morphologic characteristics similar to those of the cells positive for CMV antigen showed positive immunoperoxidase reaction with specific rabbit antihuman T-cell serum. The data strongly indicate that CMV antigen was localized predominantly in the cells of lymphoid origin during the acute stage of the CMV mononucleosis; some data also suggest that infected lymphoid cells may be of T-cell origin. An immunoperoxidase technique is advantageous in both diagnosing and evaluating CMV mononucleosis.     

Amylase as an additional marker of salivary gland neoplasms. An immunoperoxidase study

The presence of amylase in normal and neoplastic salivary gland tissue was investigated by immunoperoxidase techniques. Apart from normal and inflamed parotid glands, different kinds of tumours were studied with regard to amylase: acinic cell tumours, adenocarcinomas, adenoidcystic carcinomas, salivary duct carcinomas, mucoepidermoid tumours and squamous cell carcinomas. Amylase could be seen in acinic cell tumours, but not in other neoplasms. The results were discussed with respect to the diagnostic implications.     

Russell bodies: a light and electron microscopic immunoperoxidase study

Russell bodies have been previously regarded as aggregates of immunoglobulin. Light and electron microscopic immunoperoxidase studies show no detectable immunoglobulin determinants in the Russell body cores. Denaturation of antigens during tissue preparation appears to be an unlikely explanation, since immunoglobulins in the cytoplasm of plasma cells are clearly demonstrated. The presence of immunoglobulins on the surface of small intracellular Russell bodies may represent the immunoglobulin determinants in the surrounding rough endoplasmic reticulum. It seems likely that Russell bodies contain non-immunoglobulin molecules, by-products of immunoglobulin synthesis, or some altered form of immunoglobulins that no longer can be recognized by the anti-immunoglobulin antibody. The non-uniform dye staining pattern of Russell bodies further suggests that Russell bodies may be heterogenous in nature.     

Keratin protein immunoreactivity of sarcomatoid and mixed types of diffuse malignant mesothelioma: an immunoperoxidase study of 30 cases

To define the role of keratin protein immunohistochemistry in the pathologic diagnosis of the sarcomatoid type of diffuse malignant mesothelioma (DMM), we examined 30 DMM (16 pure sarcomatoid type and 14 mixed sarcomatoid-epithelial type) by an indirect immunoperoxidase technique using three commercially available antibodies to keratin proteins. The sarcomatoid (spindle-cell) areas of all 30 cases of sarcomatoid DMM were immunoreactive for keratin proteins. In 14 of 16 cases of sarcomatoid DMM, 50% or more of the tumor cells were reactive with one or more antibodies; however, polyclonal bovine muzzle and monoclonal AE1/AE3 antibodies were distinctly superior to polyclonal human callus keratin antibody in the detection of spindle tumor cells. In contrast with the staining patterns observed for DMM, 39 spindle-cell malignancies and tumor-like processes of 10 histogenetic types were unreactive with the three antibodies. Those spindle-cell tumors and reactive mesothelial proliferations that may enter into the differential diagnosis of sarcomatoid DMM are discussed. We conclude that keratin protein immunohistochemistry is a sensitive and highly useful method for the pathologic diagnosis of the sarcomatoid type of DMM and its distinction from other spindle-cell neoplasms.     

Immunohistological study of human lungs by immunoperoxidase technique.

An unlabelled antibody peroxidase-antiperoxidase method for the detection of IgG, IgM, complement (C3 and Clq), fibrinogen and albumin was applied to routinely processed paraffin sections of lung from 27 cases. The results in 11 cases were compared with those obtained by immunofluorescence using frozen sections. Tissue was obtained from surgical specimens of cases with interstitial pneumonia comprising 10 of the usual type (UIP) and three of the desquamative type (DIP). Tissue was also obtained from the specimens of cases with sarcoidosis (two cases) and granulomatous inflammation of unknown cause (one case). There were 11 control cases, nine with primary carcinoma of the lung and two with metastatic tumours of the lung. Immunoglobulins of various types and complement were seen in diseased lung tissue. Although most of these deposits were probably due to a non-immunological mechanism there was evidence of the possible implication of immune complexes in three cases of UIP and in the interstitial pneumonia present in the two cases of sarcoidosis. The immunoperoxidase technique is a more sensitive method than immunofluorescence and has the additional advantage of the easy identification of the precise sites of the various deposits.