Large Numbers Of Positive Cells Research Articles

CLASSICAL SWINE FEVER VIRUS DETECTION IN FETAL SWINE TISSUES BY IMMUNOHISTOCHEMISTRY

The classical swine fever virus has the ability to cross the placental barrier, resulting in the infection of fetuses, which may consequently lead to persistent infection in piglets. The aim of this study was to report the lesions in fetuses naturally infected with CSFV during late gestation and clarify the nature of infected cells and the distribution of viral antigen in different tissues. A total of twenty-nine fetuses aged 82, 83 and 95 gestational days originating from three naturally CSFV infected sows were examined in this study. In all tested sows and their fetuses CSFV was detected using RT-PCR method. Immunohistochemistry method was used to detect viral antigen and monoclonal antibody WH303 was used on formalin fixed tissue samples of brain, spleen, heart, tonsils, kidney, ileocecal valve and umbilical cord. The most common lesions in the majority of fetuses were hyperemia, petechial haemorrhages in the skin, lymph nodes and kidneys. With the exception of myocardium, CSF viral antigen was detected in all the examined tissues. WH303 positive cells included endothelial cells, monocytes, macrophages and lymphocytes. The largest number of positive cells was found in kidneys in all of the examined fetuses. Reticular cells, macrophages, lymphocytes and endothelial cells in the spleen were also intensely and widely stained in most of the fetuses. These results showed that CSFV antigen can be detected in formalin-fixed, paraffin-embedded fetal tissue specimens originating from naturally CSFV infected sows by using monoclonal antibody WH303. Fetal kidneys proved to be a very useful organ for diagnosis of the CSF virus. Having that in mind, it is assumed that persistently infected piglets may shed a high amount of viral particles through urine. However, further research is needed to confirm this hypothesis.

Immunodetection of hepatic stellate cells in dogs with visceral leishmaniasis.

Hepatic stellate cells (HSC), or Ito cells, store vitamin A when at rest but undergo phenotypic changes in situations of liver injury, which may induce fibrosis, and they may participate in the immune response in the liver. The objective of the present study was to investigate the role of HSC in the livers of dogs with visceral leishmaniasis (VL). Twenty-eight livers from dogs infected with VL that were living in an area endemic for the disease were evaluated, among which 13 were asymptomatic (A) and 15 were symptomatic (S). A control group (C) was formed by five dogs from an area that was not endemic for VL. These organs were subjected to histopathological analysis (Masson's trichrome for fibrosis) and immunohistochemical analysis (Leishmania, smooth-muscle α-actin and TGF-β). In the livers from the symptomatic dogs, a moderate to severe granulomatous inflammatory reaction was observed in the capsule and in the portal, centrilobular and intralobular regions. In the asymptomatic dogs, there was slight to moderate presence of granulomas, and these were even absent in some dogs. The intensity of hepatic fibrosis was predominantly low in the infected dogs (A and S), and fibrosis was absent in the control group. The immunomarking of HSC in the infected groups (A and S) differed significantly (P = 0.0153) from that of the control group. The symptomatic dogs presented the largest number of positive cells. This group also presented a larger number of parasitized macrophages, but did not differ statistically from the asymptomatic group (P > 0.05). The cytokine TGF-β was only detected at low levels, and only in the infected animals, but this did not differ from the control group. Immunomarking for HSC was observed mainly in the nuclei of cells present in the hepatic granulomas of symptomatic dogs and in the sinusoids of the asymptomatic dogs. It was concluded that in the livers of dogs with VL, the HSC are activated and participate in the hepatic response to the parasite. The cytokine TGF-β may be involved in this activation, but in the chronic phase of the infection, this cytokine was detected at lower proportions. It is possible that HSC may also contribute towards chemotaxis of leukocytes for the hepatic compartment, along with other cell types such as Kupffer cells.

Immunolocalization of bone morphogenetic protein 2 during the early healing events after guided bone regeneration

The objective of this study was to evaluate the immunolocalization of bone morphogenetic protein 2 (BMP-2) after autogenous block grafting covered or not with an e-PTFE membrane. Forty-eight rats were divided into 2 groups, autogenous block graft (B) and autogenous block graft + e-PTFE membrane (MB), and were evaluated by immunohistochemistry at baseline and 3, 7, 14, 21, and 45 days. The largest number of positive cells in the recipient bed was observed after 3 days in both groups. At the graft border, the largest number of positive cells was seen after 7 days in group B and after 14 days in group MB. The highest proportion of staining in the graft was observed after 3 days in group B and after 21 days in group MB. High proportions of stain were related to intense revascularization and osteogenesis. Except for the interface, BMP-2 staining occurred later in group MB than in group B in all structures analyzed.

Pediatric Chronic Sclerosing Sialadenitis: Küttner Tumor

Chronic sclerosing sialadenitis is an uncommon cause of salivary gland enlargement mainly occurring in the fifth and seventh decade of life. In the Western population, chronic sclerosing sialadenitis has been characterized as an IgG4-related disease. Although rare, this lesion occurs in children. To increase awareness about this entity in the pediatric age group, we report the case of an 11-year-old boy with a hard, 4.0-cm circumscribed mass in the right submandibular gland. Histologically there was marked distortion of the gland architecture by a dense lymphocytic infiltrate and extensive fibrosis with septa that crossed and distorted the gland, leaving atrophic acini and dilated, irregular ducts. The lymphoid infiltrate formed multiple follicles with active germinal centers, numerous plasma cells, and areas with diffuse arrangement. Immunophenotyping showed abundant CD20- and CD3-positive lymphocytes; cytokeratin AE1/AE3 highlighted the distorted architectural pattern; IgG staining showed large numbers of positive cells infiltrating the interstitium and surrounding the atrophic acini and ducts. IgG4 staining revealed a large proportion of positive infiltrating elements. Küttner tumor belongs to the group of IgG4-related sclerosing diseases. The differential diagnosis includes pleomorphic adenoma and other salivary gland neoplasms. Its recognition in children is important clinically because this entity is amenable to steroid treatment, and additional work up and follow up is warranted to stave off other IgG4-related diseases/complications.

IgG4-related sclerosing disease: a disease that may turn up in myriad (and unexpected) anatomic sites

IgG4-related sclerosing disease is a syndrome characterised clinically by mass lesions which commonly involve exocrine glands, elevated serum IgG4 titre, frequent presence of circulating auto-antibodies, and favourable clinical response to steroid therapy, and pathologically by lymphoplasmacytic infiltration, sclerosis, phlebitis and increased IgG4 + plasma cells. The histologic features can be broadly categorised into three patterns: pseudolymphomatous (Pattern I), mixed (Pattern II) and sclerosing (Pattern III). These patterns may not represent distinct histologic manifestations, but probably snap-shots in the morphologic evolution of the disease. Immunohistochemistry plays an important role in confirming the diagnosis: absolute number of IgG4 + plasma cells should be over 50 per high power field in areas with the largest number of positive cells, and the IgG4/IgG percentage should be over 40%. While the pancreas is the prototypic site of involvement (autoimmune pancreatitis), increasing sites are recognised to be involved by the disease, sometimes quite unexpectedly. Examples include hepatobiliary tree, lacrimal gland, salivary gland, lymph node, prostate, lung, kidney, soft tissue, retroperitoneum, mesentery, aorta, mediastinum, breast, pituitary, meninges, upper aerodigestive tract and skin. Most patients have disease involving two or more sites, in various combinations, with or without pancreatic involvement, although the disease can rarely be confined to one organ-system. IgG4-related sclerosing disease is a syndrome characterised clinically by mass lesions which commonly involve exocrine glands, elevated serum IgG4 titre, frequent presence of circulating auto-antibodies, and favourable clinical response to steroid therapy, and pathologically by lymphoplasmacytic infiltration, sclerosis, phlebitis and increased IgG4 + plasma cells. The histologic features can be broadly categorised into three patterns: pseudolymphomatous (Pattern I), mixed (Pattern II) and sclerosing (Pattern III). These patterns may not represent distinct histologic manifestations, but probably snap-shots in the morphologic evolution of the disease. Immunohistochemistry plays an important role in confirming the diagnosis: absolute number of IgG4 + plasma cells should be over 50 per high power field in areas with the largest number of positive cells, and the IgG4/IgG percentage should be over 40%. While the pancreas is the prototypic site of involvement (autoimmune pancreatitis), increasing sites are recognised to be involved by the disease, sometimes quite unexpectedly. Examples include hepatobiliary tree, lacrimal gland, salivary gland, lymph node, prostate, lung, kidney, soft tissue, retroperitoneum, mesentery, aorta, mediastinum, breast, pituitary, meninges, upper aerodigestive tract and skin. Most patients have disease involving two or more sites, in various combinations, with or without pancreatic involvement, although the disease can rarely be confined to one organ-system.

Immunohistochemical patterns for α- and β-catenin, E- and N-cadherin expression in ovarian epithelial tumors

Objectives. This study aims were to analyze and compare the E- and N-cadherin, β- and α-catenin expression in benign and malignant epithelial neoplasms of the ovary, correlating with tumor staging, histological grade, and presence of metastases during evolution. Methods. Immunohistochemical reactions were performed on paraffin-embedded tissues and evaluated according to the number of positive, stained cellular structures and reaction intensity for each molecule. Information about histological type and grade, tumoral stage, and disease evolution was obtained from the patients' clinical records. Results. Most of the carcinomas showed more intense β-catenin reaction ( P = 0.02). More than 50% of the endometrioid carcinomas showed increased β-catenin expression, with a large number of positive cells and more intense staining, being the same also observed for most of the serous benign tumors ( P < 0.01). E-cadherin membrane expression was frequently observed in carcinomas without metastasis, whereas cases with metastases in evolution were negative or showing E-cadherin expression only in the cytoplasm ( P = 0.04). N-cadherin expression differed according to histological type and grade and α-catenin was also related to histological type, but these findings were not conclusive. Conclusions. Increased β-catenin expression was more frequent in ovarian carcinomas, especially, but not only, in the endometrioid ones. The maintenance of E-cadherin expression in cellular membrane may be an independent marker of good prognosis in ovarian cancer. New studies about N-cadherin and α-catenin and their importance during ovarian carcinogenesis will be required.

The alteration of odor-induced c-Fos immunoreactivity in the rat olfactory bulb after olfactory nerve transection.

We used the rats in which one olfactory nerve had been transected and observed the odor (Propionic acid) -induced c-Fos immunoreactivity in the bulb at different times (2, 4, 8 weeks) after nerve transection. The exposure to odor produced a strong cluster of c-Fos positive cells in the mediodorsal region of the intact bulb. On the other hand, the transected bulb showed much less reactivity 2 weeks after neurectomy; however, a large number of positive cells were observed in the whole of the bulb from 4 weeks after neurectomy. Furthermore, we measured the levels of mRNA for tyrosine hydroxylase (TH), which was the marker of odor-induced olfactory nerve activity in the bulb, by using real-time PCR. The level of TH mRNA decreased on the transected side at 2 weeks but recovered to the level of the contralateral side at 4 weeks after neurectomy. We firstly demonstrated that projection mapping of odor receptors was altered after olfactory nerve transection by using an immunohistochemical method.

Differential expression of active, phosphorylation-dependent MAP kinases, MAPK/ERK, SAPK/JNK and p38, and specific transcription factor substrates following quinolinic acid excitotoxicity in the rat

Excitotoxicity is considered a major cell death inductor in neurodegeneration. Yet mechanisms involved in cell death and cell survival following excitotoxic insults are poorly understood. Expression of active, phosphorylation-dependent mitogen-activated extracellular signal-regulated kinases (MAPK/ERKs), stress activated c-Jun N-terminal kinases (SAPK/JNKs) and p38 kinases, as well as their putative active specific transcriptional factor substrates CREB, Elk-1, ATF-2, c-Myc and c-Jun, have been examined following intracortical injection of the glutamate analogue quinolinic acid (QA). Increased JNK P and p38 P immunoreactivity has been found in the core at 1 h following QA injection, whereas increased MAPK P immunoreactivity occurs in neurons and glial cells localised around the lesion and in neurons in remote cortical regions. This is accompanied by strong phosphorylated Ser63 c-Jun (c-Jun P) immunoreactivity in the core at 3 h, and by strong phosphorylated CREB, Elk-1 and ATF-2 (CREB P, Elk-1 P and ATF-2 P) immunoreactivity mainly in neurons around the core at 24 h following QA injection. Examination with the method of in situ end-labelling of nuclear DNA fragmentation has revealed large numbers of positive cells with no apoptotic morphology in the core at 24 h, thus indicating that JNK P, p38 P and c-Jun P over-expression precedes cell death. In contrast, MAPK P, CREB P, Elk-1 P and ATF-2 P, but not phosphorylated c-Myc (c-Myc P), over-expression correlates with cell survival. Examination of cleaved, active caspase-3 has shown specific immunoreactivity restricted to a few hematogenous cells in the area of injection. Since cleaved caspase-3 is not expressed by dying cells in the present paradigm, JNK P, p38 P and c-Jun P expression is not associated with caspase-3 activation. The present results demonstrate selective activation of specific MAPK signals which are involved either in cell death or cell survival triggered by excitotoxic insult.

Chronic antigen-specific immune-system activation may potentially be involved in the loosening of cemented acetabular components.

Previous studies have attempted to determine whether aseptic loosening and osteolysis are caused by a T cell-mediated type IV hypersensitivity reaction or a nonspecific foreign body reaction involving phagocytic macrophages. The purpose of this study was to examine the role of the B7-CD28 costimulatory pathway (which is indicative of an activated immune response) in loosening and osteolysis of total joint replacements (TJRs). We harvested periprosthetic tissues from 24 loose, cemented, all polyethylene, acetabular components in patients undergoing revision total hip replacement surgery for aseptic loosening. Prostheses were classified radiographically as to whether ballooning, scalloping osteolysis was present or not. Monoclonal antibodies were used to identify macrophages, antigen presenting cells (APCs) expressing B7-1 or B7-2, total T lymphocytes, and T cells expressing CD28 or CTLA-4. The large numbers of positive cells, including macrophages, T cells, and APCs in both groups are substantially higher than previously reported. Macrophages constituted the predominant cell type, the majority of which were APCs. B7-1 was expressed by 18.3% of all cells, and B7-2 was expressed by 61.0% of cells. Despite the fact that there were no statistically significant differences in expression of proteins in the B7-CD28 pathway between the osteolytic and nonosteolytic groups, the magnitude of positive staining suggests that the process of aseptic loosening (not osteolysis) may involve proteins of the B7-CD28 pathway, particularly B7-2. One possible antigenic stimulus is protein-coated particulate wear debris from prosthetic materials.

Differential Expression of Collapsin Response Mediator Proteins (CRMP/ULIP) in Subsets of Oligodendrocytes in the Postnatal Rodent Brain

The family of collapsin response mediator protein/Unc-33-like protein (CRMP/Ulip), composed of four homologous members, is specifically and highly expressed in the nervous system during embryonic neuronal development and dramatically down-regulated in the adult. Members of this family have been proposed to be part of the semaphorins signal transduction pathway involved in axonal outgrowth. Here, we show by in situ hybridization and immunohistochemistry that CRMP2/Ulip2, and to a lesser extent CRMP3/Ulip4, are expressed in immature and mature oligodendrocytes, but not in astrocytes. Transcripts encoding the other CRMP/Ulip members are also detectable by RT-PCR in highly purified mature oligodendrocytes. Interestingly, in the adult, the protein CRMP2/Ulip2 is mainly detectable in subsets of oligodendrocytes distributed according to an increasing rostrocaudal gradient, with the largest number of positive cells being present in the brain stem and spinal cord. In cultures of highly purified oligodendrocytes, however, CRMP2/Ulip2 was detectable in all the cells. Addition of Sema3A in the culture medium completely inhibited the emergence of oligodendrocyte processes suggesting that, as in neurons, a Sema3A signaling pathway mediated via CRMP2/Ulip2 may be involved in the regulation of oligodendroglial process outgrowth.

Optimization of sorting conditions for the selection of stable, high-producing mammalian cell lines.

The production of Green Fluorescent Protein in recombinant NIH3T3 mouse fibroblast cells was used as a model to determine the optimal conditions for the rapid isolation of high-producing cell lines with a fluorescence-activated cell sorter. "Bulk sorting", that is, sorting of a large number of positive cells, did not result in a stable, high-producing cell line due to overgrowth of high-producing cells by low- or nonproducing cells. The production kinetics and expression of GFP during batch culture was found to differ between NIH3T3 cells and HepG2 hepatoma cells, even though the same plasmid was used for transfection. The kinetics of product formation need therefore to be determined from case to case to select the optimal timepoint for analysis and sorting. Subcloning of sorted cells into microtiter plates only resulted in high-producing subclones when 1 or 2 cells were seeded per well. Higher seeding rates again resulted in overgrowth of low- or nonproducers. By subcloning, two high-producing cells lines could be isolated. They had a 10- and 15-fold higher fluorescent signal compared to the negative control. While one of these subclones started to decrease it's GFP expression after 2 months, the other clone stably expressed GFP for 4 months.

Nuclear DNA fragmentation in Creutzfeldt-Jakob disease: does a mere positive in situ nuclear end-labeling indicate apoptosis?

The method of in situ end-labeling of nuclear DNA fragmentation was used in the study of ten patients (two biopsies, eight autopsies) with sporadic Creutzfeldt-Jakob disease (CJD). All the patients had the typical morphological lesions including neuron loss, spongiform change and astrocytosis. Four of them also showed prion protein (PrP) deposits in the cerebral cortex, and two of them kuru-like plaques in the cerebellum. A few cells with DNA breaks were found in the two biopsy cases; one of them, suffering from a panencephalopathic form of the disease, showed positive nuclei not only in the cerebral cortex but also in the subcortical white matter. Variable numbers of positive nuclei were observed in the gray and white matter in the eight autopsy cases, in which, although the distribution of positive cells roughly correlated with the distribution of neuron loss, no clear relationship was found as regards the distribution and degree of cell labeling and the degree of neuron loss. Furthermore, large numbers of positive cells were concentrated in a particular area, whereas a few cells were seen in a neighboring equally affected area. Positive glial cells in the plexiform layer of the CA1 area of the hippocampus, and in the frontal white matter were frequently encountered. Staining of the cytoplasm in a minority of cells was interpreted as the result of nuclear DNA leakage. None of the stained cells had the typical morphology of apoptosis; most particularly, peripheral chromatin condensation and formation of apoptotic bodies were not seen in any case. PrP deposits did not result in an increase of nuclear DNA breaks either within the area or in adjacent regions. Although positive cells were also observed in autopsy cases of controls which were processed in the same way, positive labeling as a whole was higher in CJD than in age-matched controls. These results show that brain nuclear DNA is vulnerable in CJD, and suggest that increased DNA vulnerability has a role in cell death and neuron loss. Since nuclear shrinkage and positive nuclear staining with the method of in situ end-labeling of nuclear DNA fragmentation are not exclusive to apoptosis, further information is needed to categorize cell death in CJD as apoptosis. Necrosis or other forms of cell death, as well as increased DNA vulnerability to agonal changes of the individuals, and to postmortem delay in the fixation of the tissues, may account for additional positive staining in cases examined at autopsy.

Central giant cell granulomas of the jaws: phenotype and proliferation-associated markers.

Central giant cell granulomas (CGCGs) are jaw tumors of unknown origin that often exhibit an aggressive, though unpredictable, clinical course. The purpose of this study was to determine the immunoprofile of the mononuclear cells that seem to be responsible for the biologic behavior of these tumors. Numbers of cells in cell cycle were also determined and compared in clinically aggressive and non-aggressive CGCGs. Sixteen aggressive and 12 non-aggressive CGCGs were immunohistochemically stained with antibodies to CD34, CD68, factor XIIIa, alpha-smooth muscle actin, prolyl 4-hydroxylase, Ki-67, and p53 protein. Cell populations and numbers of cells in cell cycle were determined through microscopic quantitative assessment. CD34-positive cells were limited to support vessels. CD68-positive mononuclear cells constituted a small population of cells in all tumors. With two exceptions, factor XIIIa-positive cells were rarely seen. Alpha-smooth muscle actin staining was present in approximately half the tumors, and occasionally large numbers of positive cells were seen. Most mononuclear cells were positive for fibroblast-associated antigen. No phenotypic differences were detected between aggressive and non-aggressive tumors. P53 protein did not appear to be overexpressed in CGCGs. Ki-67 staining showed that only mononuclear cells were in cell cycle, and that there were no differences between aggressive and non-aggressive tumors. We conclude that CGCGs are primarily fibroblastic (and myofibroblastic) tumors in which macrophages appear to play a secondary role. Tumor cells show no differentiation toward endothelial cells or macrophage-related dendrocytes (factor XIIIa). Cellular phenotypes and numbers of cells in cell cycle are similar in both aggressive and non-aggressive tumors.

Generation of Small Fusion Genes Carrying Phleomycin Resistance andDrosophilaAlcohol Dehydrogenase Reporter Properties: Their Application in Retroviral Vectors

We have used theDrosophilaADH cDNA to engineer new fusion genes carrying both reporter activity and bleomycin/phleomycin resistance (Sh ble). Cassettes of ADH::Sh ble, Sh ble::ADH, or ADH::Sh ble::ADH with or without polyadenylation signals were constructed. Placed under the control of the strong CMV promoter, these constructs induced intense ADH substrate staining and phleomycin resistance, whatever the position of the ADH gene, in avian or mammalian cell lines. SNV-based nonreplicative retroviral vectors were constructed and introduced into the appropriate packaging cell line. Titers up to 106ADH forming units/ml of viral supernatant were obtained except for the ADH::Sh ble::ADH construct, which reached 105ADH forming units. These retroviral vectors were inoculated to the E3 chick embryo via the coelom. Three days later, cells from different organs were put in culture for 24 h and stained to detect ADH activity. A large number of positive cells were found in cultures from all organs. The new fusion genes described here are, to our knowledge, the smallest (1.1 kb) published to date that carry both reporter and drug resistance properties. These genes represent the basis of a new retroviral vector model with three distinct properties in two genetic units; their advantage is to reduce the size and increase the efficiency of the vector.

Mapping the origin of the avian enteric nervous system with a retroviral marker

The enteric nervous system is largely formed from the vagal neural crest which arises from the neuroaxis between somites 1-7. In order to evaluate the contribution of different regions of the vagal crest to the enteric nervous system, we marked crest cells by injecting somites 1-10 with a replication-defective spleen necrosis virus vector which contains the marker gene lacZ. After incubation in X-gal, lacZ-positive blue cells were found in the wall of the gut in three locations. Most were found at the peripheral edge of the developing circular muscle and within the developing submucosa, sites characteristic of developing ganglia. LacZ-positive cells in these ganglionic sites were always surrounded by HNK-1 immunostained cells, confirming their neural crest origin. LacZ-positive cells were also seen in a third location, the circular muscle layer of the esophagus and crop, and were separated from the HNK-1 positive ganglionic elements. These cells in the circular muscle are probably muscle cells derived from labeled mesodermal cells of the somite. Injection of somites 3, 4, 5, and 6 resulted in the largest percentage of preparations with lacZ-positive crest-derived cells and in the largest number of positive cells in the gut. After injection of these somites, lacZ-positive crest-derived cells were found in all regions of the gut from the proventriculus to the rectum. Very few positive crest-derived cells were found in the esophagus.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo.

To optimize the use of modified adenoviruses as vectors for gene delivery to the myocardium, we have characterized infection of cultured fetal and adult rat cardiac myocytes in vitro and of adult cardiac myocytes in vivo by using a replication-defective adenovirus carrying the chloramphenicol acetyltransferase (CAT) reporter gene driven by the cytomegalovirus promoter (AdCMVCATgD). In vitro, virtually all fetal or adult cardiocytes express the CAT gene when infected with 1 plaque-forming unit of virus per cell. CAT enzymatic activity can be detected in these cells as early as 4 hr after infection, reaching near-maximal levels at 48 hr. In fetal cells, CAT expression was maintained without a loss in activity for at least 1 week. Using in vitro studies as a guide, we introduced the AdCMVCATgD virus directly into adult rat myocardium and compared the expression results obtained from virus injection with those obtained by direct injection of pAdCMVCATgD plasmid DNA. The amount of CAT activity resulting from adenovirus infection of the myocardium was orders of magnitude higher than that seen from DNA injection and was proportional to the amount of input virus. Immunostaining for CAT protein in cardiac tissue sections following adenovirus injection demonstrated large numbers of positive cells, reaching nearly 100% of the myocytes in many regions of the heart. Expression of genes introduced by adenovirus peaked at 5 days but was still detectable 55 days following infection. Adenoviruses are therefore a very useful tool for high-efficiency gene transfer into the cardiovascular system.

Expression of the Alpha Subunit of Human Chorionic Gonadotropin in Normal and Neoplastic Neuroendocrine Cells

The alpha subunit of human chorionic gonadotropin (HCG) was localized Immunohistochemically in paraffin sections of normal human tissues and neuroendocrine tumors. A small subset of dispersed neuroendocrine cells was positive in normal adult tissues, including gastric antrum, urachal remnant, anal glands and prostate. Positive cells were consistently present in perinatal lung but rare in adult lung. In contrast, the beta subunit was absent from these cells. Seventy-two of 151 extrapituitary neuroendocrine tumors (48%) were alpha subunit-positive. Thirty-three of 37 bronchial carcinoids (92%) were immunoreactive, with a high percentage of the tumors (54%) containing moderate to large numbers of positive cells. The alpha subunit was further demonstrated in 9 of 45 small cell lung carcinomas (20%), 19 of 35 extrapulmonary carcinoids (54%), 3 of 11 islet cell tumors (27%) and 8 of 13 medullary thyroid carcinomas (62%). Two of three malignant islet cell tumors were positive. Positive cells were usually few in number, except for two small cell lung carcinomas, two rectal carcinoids, one thymic carcinoid and one malignant islet cell tumor. Pheochromocytomas (n = 10) were negative. Eleven of 19 pulmonary tumorlets (58%) were alpha subunit-immunoreactive. A few beta subunit-positive cells were detected in only 6 lung lesions. The physiological significance of the imbalance of expression of HCG subunits by certain neuroendocrine cells and their tumors remains unknown.

A comparative study of the estrogen receptor ratio in myometrium and uterine leiomyomas

Estrogen receptor (ER) content in myoma and myometrium was determined by radio-receptor assay (RRA), and ER staining was performed by immunocytochemical staining, i.e. the peroxidase-antiperoxidase (PAP) method, in uterine samples from eight patients with uterine leiomyoma. ER content in myoma tended to be hither than that in myometrium, but the difference was not significant. The distribution pattern of ER staining was thought to be an important criterion. When the data were compared on the basis of the number of cells in the site where the largest number of positive cells aggregated, the number for myoma was significantly higher than that for myometrium (P < 0.05). These results suggest that myoma contains more ER positive cells to obtain estrogen effects than myometrium.

Die immunhistochemische Wachstumsfraktion (Ki-67) von Mammakarzinomen: Beziehungen zur Tumorausdehnung, Tumormorphologie und Rezeptortestung

245 breast cancer tissues were analysed immunohistochemically, using monoclonal antibody Ki-67, which spezifically reacts with a nuclear antigen of proliferating cells and represents the so-called growth fraction. According to the number of positive cells, three Ki-score categories (I-III) were established. The results were compared with prognostic variables (Histological grading, DNA-cytophotometry), staging parameters, immunocytochemical (ER-ICA, mPRI), and biochemical (DCC) hormone receptor assay. We found a good correlation between Ki-67 and grading (Bloom and Richardson): G 1-tumors had small growth fractions, in contrast to G-III carcinoma, which showed a high percentage of Ki-Score III. Receptor negative cases had a large growth fraction and receptorpositive ones showed an uniform distribution. The comparison between Ki-67 and tumor spread or DNA-Cytophotometry showed no correlation. The majority of 31 recurrent tumors not only had a large number of positive cells, but also a definite dependence was found between Ki-Score and length of the disease-free period. Ki-67 seems to be an important immunohistochemical marker of breast cancer, which contributes to the individual assessment of the disease and can possibly give predictions on tumor development and response to radiation and systemic therapy.

The Possibility of Distinguishing a Subset of Antigen‐Presenting Cells from Macrophages by Means of Anti S‐100 Protein Sera

Macrophages may be distinguished from interdigitating cells and from Langerhans cells in paraffin sections, the latter cells being positive when an antiserum against brain S-100 protein is used. This antiserum was utilized to conduct a retrospective analysis of 10 cases of lichen planus, including both early and late lesions. In addition, staining of macrophages was carried out by means of anti-lysozyme, anti-alpha-1-antitrypsin and anti-alpha-1-antichymotrypsin. The anti-S-100 protein staining by immunoperoxidase methods showed large numbers of positive cells. Few macrophages were noted in the early lesions, but the ratios were reversed in the older lesions, in which macrophages predominated over dermal S-100-positive cells. Both Langerhans cells-interdigitating cells and macrophages could play important roles in various cutaneous disorders. The involvement of Langerhans cells-interdigitating cells or, on the other hand, of macrophages could distinguish among different pathological processes. Even in different evolutionary stages of the same lesion, as lichen planus, a different Langerhans cells-interdigitating cells/macrophages ratio could be important in explaining the pathogenetic development of the disease.