Human epidermal growth factor receptor-2 and estrogen receptor expression, a demonstration project using the residual tissue repository of the Surveillance, Epidemiology, and End Results (SEER) program.

BACKGROUND: In 2001, the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) program established Residual Tissue Repositories (RTR) in the Hawaii, Iowa, and Los Angeles Tumor Registries to collect discarded tissue blocks from pathologic laboratories within their catchment areas. To validate the utility of the RTR for supplementing SEER's central database, we assessed human epidermal growth factor receptor-2 (HER2) and estrogen receptor expression (ER) in a demonstration project. MATERIALS: Using a prepared set of tissue microarrays (TMAs) residing in the Hawaii Tumor Registry (HTR), we performed standard immunohistochemistry. Breast cancers in the TMA were diagnosed in 1995, followed through 2006, and linked to SEER's main database. RESULTS: The TMA included 354 cases, representing 51% of 687 breast cancers in the HTR (1995). The HTR and TMA cases were similar with respect to patient demographics and tumor characteristics. Seventy-six percent (76%, 268 of 354) of TMA cases were HER2+ and/or ER+, i.e., 28 HER2+ER-, 12 HER2+ER+, and 228 HER2-ER+. There were 67 HER2-ER- cases and 19 were unclassified. Age distributions at diagnosis were bimodal with dominant early-onset modes for HER2+ER- tumors and dominant late-onset modes for HER2-ER+ breast cancers. Epidemiologic patterns for concordant HER2+ER+ (double-positive) and HER2-ER- (double-negative) were intermediate to discordant HER2+ER- and HER2-ER+. CONCLUSION: Results showed contrasting incidence patterns for HER2+ (HER2+ER-) and ER+ (HER2-ER+) breast cancers, diagnosed in 1995. Though sample sizes were small, this demonstration project validates the potential utility of the RTR for supplementing the SEER program.

The comparative importance of E. coli antigen in granulomatous colitis of Boxer dogs.

Granulomatous colitis of Boxer dogs is characterized by mucosal and submucosal infiltration by abundant large macrophages and lymphocytes and plasma cells. Involved intestine is thickened, corrugated and ulcerated. The macrophages that occur in colon, cecum and regional lymph nodes are PAS-positive, lipid-rich, contain cholesterol, and some of the time can be seen to hold bacteria. Paraffin tissue blocks of formalin-fixed colon and colic lymph nodes from 10 cases were cut at 5 microm and immunostained by a streptavidin-biotin immunoperoxidase technique, employing primary antibodies against Escherichia coli, E. coli 0157: 2, Campylobacter, C. jejuni-coli, Yersinia pseudotuberculosis, Salmonella, Shigella, Pseudomonas and Lawsonia intracellularis. The macrophages in the lamina propria and submucosa, as well as those in aggregates in regional lymph nodes, showed immunoreactivity with polyclonal E. coli antibody in all 10 cases. Tissues lacking granulomas were negative, as were those reacted with the other eight antibodies, with the exception that there was rare focal staining for Campylobacter, Lawsonia and Salmonella in a few dogs. We believe these results identify the causative agent of this granulomatous disease of Boxer dogs, a disease with great histologic and etiologic similarity to granulomatous leptomeningitis of Beagle dogs, and malacoplakia and xanthogranulomatous cholecystitis of man. Macrophages that are immunopositive for E. coli antigen occur in Crohn's disease as well, where their significance is less well understood.

Atypical fibroxanthoma. A study of 14 cases emphasizing the presence of Langerhans' histiocytes with implications for differential diagnosis by antibody panels.

Fourteen consecutive cases of atypical fibroxanthoma (AFX) seen during a 4-year period were studied histologically; of these, 12 were further examined for the presence of immunocytochemically detectable cytokeratin (CK), vimentin (VIM), S-100 protein, melanocyte-associated antigen (MAA), muscle-specific actin (MSA), alpha-1-antitrypsin (A1AT), alpha-1-antichymotrypsin (A1ACT), and ferritin (FER). In four cases, electron microscopy was also performed. Tumor cells were nonreactive with antibodies directed against CK and MAA, strongly reactive with anti-VIM, and variably reactive with A1AT, A1ACT, MSA, and FER. Our findings are consistent with the current notion that these tumors are "fibrohistiocytic". However, in 11 of 12 cases studied, a subpopulation of cells with features of Langerhans' histiocytes (LH) was also identified. These were dendritic cells within the substance of the tumor that were strongly reactive with S-100 antibody and uniformally nonreactive with MAA antibody; ultrastructurally, they were seen to contain typical Birbeck granules. LH characteristically comprised no more than 5% of the overall cell population of the tumor; however, in restricted portions of some lesions, they sometimes accounted for up to 80% of tumor cells. The occurrence of LH in AFX, although previously reported, has not generally been emphasized. Awareness of their presence as an expected and sometimes extensive component of AFX can be important when interpreting differential immunocytochemical panels applied to malignant spindle cell tumors of skin.

Immunocytochemical analysis of estrogen and progesterone receptors in benign stromal lesions of the breast. Evidence for hormonal etiology in pseudoangiomatous hyperplasia of mammary stroma.

Five cases of pseudoangiomatous hyperplasia of mammary stroma, together with seven examples of mammary hamartoma, were probed with monoclonal antibodies H222 and KD68 to investigate the possible role of estrogen and progesterone receptor expression in the pathogenesis of these benign stromal proliferations. All five cases of pseudoangiomatous hyperplasia showed patchy, intense labelling of the stromal cells with progesterone receptor antibodies, a pattern contrasting markedly with the absence of immunoreactivity in normal (nongestational) mammary stroma or the stromal component of common juvenile mammary hyperplasia. The stroma of the hamartoma group labeled inconsistently, with the notable exception of three myoid hamartomas. Stromal immunoreactivity was diffuse and intense in two of these, and patchy and distinct in the remaining case. These findings (a) support the contention that pseudoangiomatous hyperplasia represents a localized form of stromal overgrowth with a hormonal (primarily progestagenic) etiology and (b) further highlight the heterogeneity of so-called mammary hamartomas by demonstrating dramatically different progesterone receptor immunoreactivity patterns in myoid lesions as compared with other hamartoma variants.

Treponema pallidum and Helicobacter pylori recovered in a case of chronic active gastritis.

A 49-year-old man complaining of epigastric pain underwent endoscopy, during which thickened stomach folds below the fundus were observed. Microscopic examination of gastric tissue biopsy specimens revealed chronic active gastritis. Dieterle stain revealed overwhelming numbers of "corkscrew-like" spirochetes. These were proved to be consistent with Treponema pallidum. A comprehensive study of the tissue revealed the added presence of Helicobacter pylori. This appears to be the first case report describing the involvement of H. pylori and T. pallidum together in a case of chronic active gastritis.

Viral RNA in duodenal bile of cirrhotic patients with chronic hepatitis C.

BACKGROUND: Hepatitis C virus (HCV) has been detected in blood, saliva, urine, semen, breast milk, and tears. To our knowledge, bile has not yet been investigated. We observed histologic immunoreactivity in bile with an antibody to c100 protein in four of five HCV-positive cirrhotic livers, but also in two HCV-negative controls owing to a focally present cross-reacting antigen. METHODS: We collected duodenal bile from 13 cirrhotic patients during endoscopic evaluation of varices (10 HCV, three controls) and assayed for HCV by reverse transcriptase polymerase chain reaction. RESULTS: Viral RNA was detected in the bile of 8 of 10 seropositive patients and in 0 of 3 seronegative controls. CONCLUSION: Hepatitis C virus RNA and an antigen immunoreactive with anti-c100 protein are present in bile in a proportion of cirrhotic patients with chronic HCV. It remains to be determined whether the virus is intact or degenerate, and whether it is shed into bile from hepatocytes or is a contaminant from blood or other secretions.

Immunoperoxidase staining of tumors by an antibody to Xenopus pNiXa.

pNiXa, a serpin from oocytes and embryos of Xenopus laevis, was tested as a tumor marker in human and rodent tissues. A peptide corresponding to the histidine-rich domain of pNiXa was conjugated and administered to rabbits to produce a polyclonal antibody, which was purified by antigen-affinity and used for immunoperoxidase staining of formalin-fixed, paraffin-embedded tissue sections. Staining with pNiXa-antibody was positive in 23/187 human tumors (12 percent) and negative in 119 specimens of normal human tissues. Positive reactions were more frequent in liver (38 percent) and colon (34 percent) tumors than breast (18 percent), prostate (9 percent), mesothelioma (20 percent) or lung (0 percent) tumors. Staining was negative in human tumors from other sites. Rodent tumors and preneoplastic foci induced by chemical carcinogens were surveyed for staining with pNiXa-antibody. Staining was positive in 10/10 hepatic lesions (hepatocellular foci, adenomas, carcinomas) induced in hybrid D2B6F1 mice by diethylnitrosamine and phenobarbital, whereas murine mammary tumors and thyroid, pituitary, renal, and colon tumors of F-344/CNr rats were negative. Thus, immunostaining with pNiXa-antibody identifies a subset of human and murine tumors; further studies are needed to determine if reactivity of pNiXa-antibody has diagnostic or prognostic significance.

The use of monoclonal antibody R92F6 and polymerase chain reaction to confirm the presence of parvovirus B19 in bone marrow specimens of patients with acquired immunodeficiency syndrome.

BACKGROUND: Parvovirus B19 infection is a cause of chronic anemia and red cell aplasia in patients with acquired immunodeficiency syndrome (AIDS) and in other immunocompromised hosts. Anemia in AIDS patients has a multifactorial etiology, with parvovirus B19 infection being an infrequent but nevertheless treatable cause. Therapy with intravenous immune globulin can result in rapid improvement of parvovirus-induced anemia. This treatment is expensive, therefore accurate and rapid confirmation of parvovirus infection is important in providing appropriate and cost-effective therapy. METHODS: Bone marrow samples from 2 AIDS patients with severe anemia and reticulocytopenia were studied. Bone marrow morphology and serologic studies were evaluated for parvovirus B19 infection. An immunohistochemical method using a monoclonal antibody, R92F6, to B19 capsid proteins was utilized on decalcified, B5-fixed, paraffin-embedded bone marrow biopsies. Bone marrow aspirate cells were examined by electron microscopy for evidence of viral particles. In addition, polymerase chain reaction (PCR) studies using a nested PCR assay to the parvovirus B19 viral genome were performed in a case for which fresh cells were available. RESULTS: Bone marrow findings included marked erythroid hypoplasia with characteristic giant pronormoblasts and intranuclear inclusions. Serologic studies were negative in one case, while the second case showed positive parvovirus B19 immunoglobulin M antibody. Immunohistochemical studies for parvovirus B19 were positive in both cases. The presence of intranuclear virions was demonstrated by electron microscopy and was confirmed by PCR analysis. Both patients were treated with intravenous immune globulin, and subsequent improvement was noted. CONCLUSIONS: Both immunohistochemistry and PCR studies on bone marrow specimens from AIDS patients with anemia are rapid and sensitive methods for the confirmation of parvovirus B19 infection. They are valuable tools, particularly when serologic studies are negative. When PCR is not available, immunohistochemical methods can be useful. The rapid confirmation of parvovirus B19 infection will allow for early and cost-effective therapy.

Primary PNET of kidney: report of two cases and review of literature.

PNET of the kidney is a rare tumor with only a few published reports. In view of poorer prognosis and different therapeutic approach, renal PNET should therefore be differentiated from other primary renal neoplasma such as Wilms tumor, renal neuroblastoma and malignant rhabdoid tumor which on histology resemble renal PNET. Two cases of renal PNET have been described in this report. Cut surface of the tumor in both cases was greyish white lobulated, with multiple tiny cystic areas. Histologically, tumor consisted of loosely cohesive sheets of small to medium sized monomorphic cells with round nuclei and little cytoplasm. Tumor cells showed diffuse strong membrane positivity for MIC2 and focal weak to moderate positivity for NSE and vimentin. Renal PNET should therefore be included in differential diagnosis of rapidly enlarging renal lumps presenting with local infiltration and aggressive behaviour, particularly in children and young adults. Diffuse strong membrane positivity for MIC2 in PNET is helpful in differentiating it from other primary renal neoplasms.

Atypical seminoma--histologic features, immunohistochemical characterization, and correlation with survival.

Classical seminoma and embryonal carcinoma are two points in the spectrum of histologic differentiation in testicular germ cell tumors. The validity of an intermediate category, ie, atypical seminoma (AS) is questionable. Histopathologic and clinical data on 42 patients treated for primary testicular germ cell tumor from 1975 to 1985 were reviewed. Twenty-seven cases were identified as classical seminoma and nine were embryonal carcinoma. The remaining six cases were somewhat problematic to classify, combining the growth pattern of seminoma with cytologic features of embryonal carcinoma. Immunocytochemically, four of these tumors suggested some progression towards the embryonal carcinoma phenotype on the basis of cytokeratin expression. Survival for classical seminoma, AS, and embryonal carcinoma were 90%, 80%, and 63% respectively (mean follow-up, 8.6 years). Although the survival differences were not statistically significant, when considered with morphologic and selected immunocytochemical data, they tend to support the concept of an AS as an intermediate lesion between classical seminoma and embryonal carcinoma.

Case report: an autopsy study of a rare T-cell lymphoma clinically misdiagnosed as fulminant hepatic disease.

We describe a case of a 73-year-old male with a rare T-cell lymphoma that presented deceptively as progressive hepatic failure with fever, weight loss, pancytopenia, mental confusion, splenomegaly, and no lymphadenopathy. An alcoholic history supported the diagnosis of cirrhosis, but a liver biopsy was not performed. A bone marrow biopsy was considered unremarkable. Death occurred after a course of four months. Postmortem examination showed hepatic, splenic, lymph node, and marrow infiltration by characteristically sparse, isolated, bizarre, medium-to-large sized neoplastic cells with extensive hepatic centrilobular necrosis, steatosis, and predominant splenic involvement. Immunohistochemical markers indicated a T-cell lymphoma consistent with either an alpha/beta peripheral T-cell lymphoma or a gamma/delta lymphoma. Definitive immunotyping was not available. However, the pathologic features are most consistent with a gamma/delta T-cell lymphoma. This case is an example of a rare, rapidly progressive lymphoma, which is a recognized clinical entity, easily missed, and treatable. Its diagnostic consideration must be explicitly communicated to pathologists, because the isolated or sparse tumor cells in a lymph node, liver, or bone marrow biopsy may easily be mistaken for variants of megakaryocytes or histiocytes.

Detection of occult metastatic lobular carcinoma in axillary lymph nodes using anticytokeratin monoclonal antibodies.

To examine the importance of immunocytochemically detectable occult axillary lymph node metastases in patients with lobular carcinoma of breast, tumor registry data from 54 cases indexed as lobular carcinoma during the period 1973-82 were reviewed. Recurrences and/or deaths due to cancer were essentially confined to the group of patients with a component of invasive lobular carcinoma (ILC), therefore this subset was selected for further study. Seven of 20 cases had lymph node metastases diagnosed histologically at the time of mastectomy. Follow-up of these patients showed four dead of disease (DOD) at one, three, three, and seven years; one alive with disease (AWD) at one year; and two with no evidence of disease (NED) at four and five years. Eleven of 20 were node negative. Follow-up of this group showed nine NED and two DOD at two and four years. Two of 20 had unknown node status. Formalin-fixed, paraffin embedded lymph node blocks were available in 12 of 20 cases with a component of ILC. Of these, 4/12 cases had histologically positive nodes while 8/12 were originally diagnosed as negative. A cytokeratin monoclonal antibody cocktail (MAK-6, CAM 5.2 and AE1/AE3) was applied to all 12 cases. Cytokeratin immunoreactivity (CK-IR) was found in all four cases that were histologically positive. Five of eight histologically negative nodes lacked CK-IR, however the other three cases showed CK-IR in micrometastases. Review of newly prepared hematoxylin-eosin sections from the paraffin blocks failed to demonstrate metastases.(ABSTRACT TRUNCATED AT 250 WORDS)

Gene amplification as means for determining therapeutic strategies in human cancers.

Pharmacogenomic analysis aspires to identify individuals with specific genetic characteristics in order to predict a positive response or reduce a negative response to a therapeutic modality. While the search continues for the many single nucleotide polymorphisms which will be used in such genetic analyses, other genetic alterations in specific cell types have proven useful in determining the potential for response to therapy. One such genetic alteration is amplification of entire gene sequences which results in overexpression of a gene product or protein. Amplification of the HER2 (neu, erbB-2) oncogene is found in up to 35% of human breast cancers and is associated with a poor prognosis. In addition, this genetic alteration may predict response to various therapeutic modalities. Assays are available to detect the HER2 protein receptor or copies of the HER2 gene sequence to determine eligibility for Herceptin treatment or adriamycin treatment in node positive patients, respectively. This model represents a somatic event used in the functional determination of a therapeutic strategy.

Hodgkin's disease, lymphocyte-predominant type: immunoreactivity with B-cell antibodies.

Utilizing the monoclonal antibodies L26 (a new antibody possessing immunoreactivity with B-lymphocytes in paraffin-embedded tissue), LN1, LN2, and Leu-M1, 44 cases of Hodgkin's disease (HD) were examined for the presence of immunoreactivity in Reed-Sternberg (R-S) cells by the avidin-biotin-peroxidase complex (ABC) technique. In 16 cases of lymphocyte-predominant Hodgkin's disease (LPHD), the L&H variants of R-S cells exhibited a different pattern of staining compared to R-S cells in other histologic types (total, 28 cases: 11, mixed cellularity; 8, nodular sclerosing; 6, lymphocyte depleted; 3, unclassified). L&H variants in LPHD were immunoreactive for L26 and LN1 in 15 and 14 cases, respectively, whereas R-S cells in the remaining types were negative or rarely positive (3, L26; 2, LN1). Leu-M1 was strongly positive in 27 of 28 cases of non-LPHD versus only 4 of 16 in LPHD. LN2 was reactive in virtually all cases (43 of 44). These findings suggest the possibility that the R-S cells of LPHD are derived from a different lineage than R-S cells in other histologic types of HD or that the latter have somehow lost the ability to express the antigens defined by L26 and LN1. Finally, based on immunologic and morphologic findings in this study, the similarities seen between the nodular and diffuse subtypes of LPHD are felt to favor a close relationship between the two subtypes.

A clinicopathologic and immunohistochemical study of 16 patients with small intestinal leiomyosarcoma. Limited utility of immunophenotyping.

Clinicopathologic data including immunophenotypic expression of 16 new cases of small intestinal leiomyosarcoma are presented. Patient age ranged from 27 to 87 years (mean, 61 years) with a 2.2:1 male-female ratio. Eighty-eight percent of tumors occurred distally, i.e., in the jejunoileum. Considerable histologic heterogeneity was noted including epithelioid, myxoid, and sclerotic variants. Stratification by histologic grade suggests important clinical subsets. Only in grade I tumors is surgical extirpation alone potentially curative. Grade II tumors are associated with meaningful survival usually measured in years, however, nearly all patients can be expected to die from their tumors. Grade III sarcomas are fully malignant and rapidly progressive: except for one case with limited follow-up, all patients with Grade III sarcomas died of disease with a mean survival of only nine months (median, 2 months). Immunostaining with antibodies directed against vimentin, muscle-specific actin, desmin, and S-100 protein were performed. Only antivimentin was uniformly reactive. Sixty-three percent of cases tested showed actin immunoreactivity, and no tumors revealed immunohistochemically detectable desmin or S-100 protein. Although many small bowel stromal sarcomas appear "poorly differentiated" by immunohistochemical methods presently available, that they are indeed leiomyosarcomas seems most likely from their intramural location and light histologic study. The limited utility of immunophenotyping and the importance of histologic grading are stressed.

Utilization of monoclonal antibody L26 in the identification and confirmation of B-cell lymphomas. A sensitive and specific marker applicable to formalin-and B5-fixed, paraffin-embedded tissues.

Immunophenotypic analysis of paraffin-embedded tissues of lymphoproliferative disorders has been facilitated by recent developments of monoclonal antibodies that react with epitopes that survive histologic processing. Leukocyte common antigen (LCA) antibody has made a significant contribution to the immunocytochemical separation of non-Hodgkin's lymphomas from nonlymphoid neoplasms. However, a small percentage of lymphomas, particularly some large cell or immunoblastic B-cell tumors, will not label with LCA antibody. Other antibodies, directed against B lymphocytes, experience problems of specificity and a lack of sensitivity when applied to formalin-fixed specimens. The authors recently investigated a monoclonal antibody (L26) that demonstrates excellent specificity and sensitivity for B lymphocytes, and tumors derived from them, in formalin- and B5-fixed, paraffin-embedded tissue. The avidin-biotin peroxidase complex (ABC) technique was utilized for immunostaining 95 cases of malignant lymphoproliferative disorders and a variety of normal and neoplastic nonlymphoid tissues. When applied to sections of benign lymphoid tissue, the L26 antibody labeled germinal center cells, mantle zone and scattered interfollicular lymphocytes, but not histiocytes or plasma cells. L26 marked 100% (44/44) of the large cell and immunoblastic B-cell lymphomas, along with 1 case of pre-B cell lymphoblastic lymphoma. This included 8 cases that were LCA-negative. None of the T-cell lymphomas or plasma cell tumors studied demonstrated L26 immunostaining. No normal, benign, or neoplastic nonlymphoid tissues examined stained with this antibody. L26 successfully labels B lymphocytes and B-cell lymphomas in routinely processed tissues, often with greater sensitivity and intensity than LCA. This antibody should prove invaluable in the investigation of atypical lymphoid proliferations and the identification of B-cell derived lymphomas, when fresh or frozen tissue is unavailable for analysis.

Estrogen and progesterone receptors in primary eccrine carcinoma.

Primary eccrine carcinoma and metastatic mammary duct carcinoma are histologically similar and histogenetically related neoplasms that may on occasion be mistaken for one another. Mammary duct carcinomas, in primary or metastatic sites, will often express estrogen and progesterone receptors (ER and PR), and these may be detected by either biochemical or immunocytochemical methods. The expression of ER and PR in primary eccrine carcinoma, however, has not been studied. Three eccrine carcinomas were probed with antibodies to ER and PR. Immunoreactivity for hormone receptors was identified in all three cases. The recognition of this immunophenotype may be important in avoiding misdiagnosing receptor-positive eccrine carcinoma as metastatic breast cancer.