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PubMed2000

Tyrosine phosphorylation of cytoplasmic proteins in proliferating, differentiating, apoptotic HL-60 cells and blood neutrophils.

Two-dimensional electrophoretic analysis was used to assess quantitative and qualitative changes in the expression and tyrosine phosphorylation of cytoplasmic proteins of proliferating, differentiating HL-60 cells and mature human blood neutrophils. The total tyrosine phosphorylation level of cytoplasmic proteins appeared approximately constant during the pre-commitment period, i.e., 6-24 h after induction of differentiation by 700 nM all-trans retinoic acid. At the time of granulocytic phenotype formation (48-120 h), the total level of tyrosine phosphorylation of cytoplasmic proteins increased significantly. Tyrosine phosphorylation of cytoplasmic proteins in matured blood neutrophils was significantly lower than that of cytoplasmic proteins of HL-60 cells differentiated for 96 h with retinoic acid. Immunoblotting with anti-Erk2 and anti-phosphotyrosine monoclonal IgG2bk antibodies showed that Erk2 was expressed and tyrosine-phosphorylated at different levels in HL-60 proliferating cells and in cells at all stages of differentiation. Our data showed that tyrosine phosphorylation of cytoplasmic proteins in differentiating HL-60 cells changes dramatically during the period of phenotype formation and is accompanied by increasing activity of Erk2. An increasing number of apoptotic cells appeared in the differentiating HL-60 cell population during the granulocyte maturation stage (48-120 h of differentiation). The appearance at this time of differentiation of a new set of tyrosine-phosphorylated cytoplasmic proteins (also distinctive for apoptotic HL-60 cells mediated by etoposide) together with an increasing number of apoptotic cells in the differentiating population strongly suggests that these proteins are associated with the apoptotic process.

Apoptosis↗
PubMed2003

Difference in cytoplasmic localization pattern of neutral mucin among lobular endocervical glandular hyperplasia, adenoma malignum, and common adenocarcinoma of the uterine cervix.

To investigate whether the cytoplasmic localization pattern of neutral mucin differs between lobular endocervical glandular hyperplasia (LEGH) and cervical adenocarcinoma (CxAd), including minimal-deviation adenocarcinoma (MDA), or adenoma malignum, alcian blue (pH 2.5)/periodic acid-Schiff (AB-PAS) staining was performed to formalin-fixed paraffin-embedded tissue sections of 13 lesions of LEGH and 53 tumors of CxAd, including 6 tumors of MDA. The cytoplasmic localization of neutral mucin was classified as a "whole cytoplasmic pattern," in which neutral mucin filled the cytoplasm entirely, or as an "apical pattern," in which neutral mucin was localized in the subsurface area only. Cytoplasmic neutral mucin patterns were detected in all 13 cases of LEGH and in 19 cases (36%) of CxAd, including five cases of MDA. The localization of neutral mucin was always the whole cytoplasmic pattern in 13 cases of LEGH, but was the apical pattern in these 19 cases of CxAd. The other 34 cases of CxAd, including 1 case of MDA, corresponded to the acid mucin pattern stained purple or blue or no staining pattern by AB-PAS. Among the 53 cases of CxAd, the apical neutral mucin pattern was an indicator of poorer patient prognosis by univariate and multivariate analyses. The examination of cytoplasmic localization of neutral mucin might be applicable, not only to differential diagnosis between LEGH and CxAd, including MDA, but also to estimate clinical aggressiveness of CxAd.

Adenocarcinoma↗
PubMed2002

Diagnostic role of anti-Saccharomyces cerevisiae mannan antibodies combined with antineutrophil cytoplasmic antibodies in patients with inflammatory bowel disease.

PURPOSE: Perinuclear antineutrophil cytoplasmic autoantibody is known to be a marker for ulcerative colitis, and anti-Saccharomyces cerevisiae Mannan antibody is a serologic marker associated with Crohn's disease. The aim of this study was to assess the value of detecting perinuclear antineutrophil cytoplasmic autoantibody and/or anti-Saccharomyces cerevisiae Mannan antibody for the diagnosis of ulcerative colitis, Crohn's disease, Behçet's colitis, and tuberculous colitis. METHODS: Serum samples were obtained from 85 patients with Crohn's disease, 77 with ulcerative colitis, 36 with Behçet's colitis, 14 with tuberculous colitis, 20 healthy controls, and 21 first-degree relatives of patients with Crohn's disease. Determination of perinuclear antineutrophil cytoplasmic autoantibody and anti-Saccharomyces cerevisiae Mannan antibody was performed with the standardized indirect immunofluorescence technique and an enzyme-linked immunosorbent assay, respectively. RESULTS: A relatively high percentage of patients with Crohn's disease (49.4 percent), relatives of Crohn's disease patients (61.9 percent), and patients with Behçet's disease (41.7 percent) tested seropositive for anti-Saccharomyces cerevisiae Mannan antibody compared with normal controls (10 percent). In cases of ulcerative colitis, 44.2 percent tested seropositive for perinuclear antineutrophil cytoplasmic autoantibody, whereas the controls showed 0 percent positivity. The combination of a positive anti-Saccharomyces cerevisiae Mannan antibody test and a negative perinuclear antineutrophil cytoplasmic autoantibody yielded a sensitivity and specificity of 48.2 and 87 percent, respectively, for Crohn's disease. The combination of a positive perinuclear antineutrophil cytoplasmic autoantibody test and a negative anti-Saccharomyces cerevisiae Mannan antibody test yielded a sensitivity and specificity of 36.4 and 97.6 percent, respectively, for ulcerative colitis. CONCLUSION: Anti-Saccharomyces cerevisiae Mannan antibody may be associated with Crohn's disease and Behçet's disease and perinuclear antineutrophil cytoplasmic autoantibody with ulcerative colitis. A combination of both tests may aid the differential diagnosis of inflammatory bowel disease.

Adult↗
PubMed1984

Characterization of the ATPase activities of myosins isolated from the membrane and the cytoplasmic fractions of human platelets.

Myosin was purified from the membrane fraction and the cytoplasm of human platelets, and the K+(EDTA)- and Ca2+-dependent ATPase activities were studied under various experimental conditions. The ATPase activity of the myosin from the membrane fraction was slightly lower than that of its cytoplasmic counterpart, regardless of the different assay conditions (pH, ionic strength, and temperature). Both myosins showed the same pH optima and a similar ionic strength dependence for the two ATPase activities measured. In addition, they exhibited the same substrate specificity using ATP, CTP, and GTP as substrates. The activation energy of the Ca2+-dependent ATPase activity was essentially the same for the two myosins, while the activation energy of the K+(EDTA)-dependent ATPase activity of the membrane myosin was higher than that of the cytoplasmic myosin. The ATPase activity of the membrane myosin was found to be more sensitive to freezing and thawing than the cytoplasmic myosin. The alkylation of the thiol groups by N-ethylmaleimide or N-iodoacetyl-N-(5-sulfo-1-naphtyl)ethylenediamine, and the trinitrophenylation of the lysyl residues by 2,4,6-trinitrobenzenesulfonate caused a significant decrease in the K+(EDTA)-dependent ATPase activity of the two myosins. However, the membrane myosin was much less affected than the cytoplasmic myosin. Actin induced inhibition of the K+ (EDTA) ATPase of both myosins, and much smaller quantities of actin were needed to inhibit the cytoplasmic myosin ATPase compared to quantities needed to inhibit the myosin ATPase from the membrane fraction. This indicates that the membrane myosin has a lower affinity toward actin. The observed variations in the ATPase activity of the myosins isolated from the membrane and the cytoplasm fractions of human platelets may reflect differences in their respective physiological functions.

Actins↗
PubMed1989

Contribution of cytoplasmic storage triacylglycerol to VLDL-triacylglycerol in isolated rat hepatocytes.

The cytoplasmic triacylglycerol (TG) storage pool of isolated hepatocytes was labelled in order to evaluate its incorporation into very low density lipoproteins (VLDL). Rats were injected with [1-14C]oleate 2 min prior to surgery and cell incubations began 90-100 min thereafter. In keeping with the equilibration of the two TG pools (in smooth endoplasmic reticulum, SER, and cytoplasm) in 120 min (Stein, Y. and Shapiro, B. (1959) Am. J. Physiol. 196, 1238-1241) the bulk of radioactive TG at time zero was in the cytoplasm and TG specific activities were similar in cytoplasm and SER. Radioactive and total VLDL-TG secretions were greatly inhibited after 80 min by chloroquine which is assumed to block lysosomal hydrolysis of cytoplasmic TG. When the SER-TG pool was labelled by addition of [1-14C]oleate in vitro, chloroquine affected neither [1-14C]oleate uptake and esterification nor its incorporation into VLDL-TG from 15-20 min until 80 min. After 100 min, when [1-14C]oleate-TG was transferred back from cytoplasm to SER, chloroquine began to decrease radioactive VLDL-TG output and by 210 min caused the same inhibition as under the in vivo labelling condition. These results are consistent with an inhibition by chloroquine of the lysosomal hydrolysis of cytoplasmic TG resulting in a blockage of their back transfer to SER membranes whereas other steps of VLDL production were not affected, at least up to 100 min. This study also showed that stored TG is a quantitatively important VLDL precursor, sustaining VLDL production for several hours in the absence of exogenous fatty acids.

Animals↗
PubMed1975

Aminoacylation of Phaseolus vulgaris cytoplasmic, chloroplastic and mitochondrial tRNAsMet and of Escherichia coli tRNAsMet by homologous and heterologous enzymes.

Met-tRNA synthetase from Paseolus vulgaris cytoplasm could be separated from its chloroplastic or mitochondrial counterpart by DEAE-cellulose chromatography, but the Met-tRNA synthetase from the two latter organelles could not be distinguished using DEAE-cellulose, hydroxyapatite or CM-Sephadex chromatography. As revealed by reverse-phase chromatography, bean cytoplasm contains 2 tRNAsMet; only one is charged by chloroplast, mitochondrial or Escherichia coli Met-tRNA synthetase. Mitochondria contain, in addition to the 2 cytoplasmic tRNAsMet, 3 mitochondria-spedific tRNAsMet; 2 can be formylated by the mitochondrial or the E. coli transformylase; all 3 are charged by mitochondrial, chloroplastic or E, coli Met-tRNA synthetase; none is charged by the cytoplasmic enzyme. Chloroplasts contain, in addition to the 2 cytoplasmic tRNAsMet, 3 chloroplast-specific tRNAsMet, different from the mitochondrial tRNAsMet; one is formylatable by the chloroplastic or the E. coli transformylase; all 3 are charged by chloroplastic, mitochondrial or E. coli Met-tRNA synthetase; only one is charged by the cytoplasmic enzyme. Of the 3 E. coli tRNAsMet, only the formylatable species can be charged by bean cytoplasmic, chloroplastic or mitochondrial Met-tRNA synthetase.

Amino Acyl-tRNA Synthetases↗
PubMed1990

Analysis of cytoplasmic activity dependent on the Drosophila terminal pattern gene torso.

Cytoplasm from wildtype Drosophila embryos was transplanted into torso (tor) mutant embryos to determine the distribution of terminal rescuing activity at the cleavage stage. Although posterior and lateral wildtype cytoplasm contained rescuing activity that restored posterior terminal (telson) structures Klingler et al. (1988, Nature (London) 335, 275-277) this rescuing activity was not found in anterior cytoplasm. Similarly, transplantation of anterior and lateral wildtype cytoplasm into the anterior of tor embryos rescued anterior terminal (acron) structures, whereas posterior cytoplasm did not. This failure of reciprocal rescue is due to the presence of the products of the anterior and posterior classes of genes, because anterior cytoplasm from bicoid mutant embryos restored the telson in the posterior as well as the acron in the anterior of tor embryos, and because posterior cytoplasm from nanos embryos rescued the acron in the anterior as well as the telson in the posterior of tor embryos. Therefore terminal rescuing activity is evenly distributed throughout the cleavage stage embryo as anticipated from molecular studies.

Animals↗
PubMed1984

Localization of HMG chromosomal proteins in the nucleus and cytoplasm by microinjection of functional antibody fragments into living fibroblasts.

We have used microinjection and cell fractionation to localize the chromosomal high mobility group proteins (HMG) in human fibroblasts. Electrophoretic analysis of nuclear and cytoplasmic fractions from the fibroblasts indicates that the concentration of HMG-1,2 in the cytoplasm is 2.9 times larger than in the nucleus indicating that the majority of the cellular HMG-1,2 is present in the cytoplasm. In contrast, HMG-17 remains predominant in the nuclear fraction. We conclude that the cellular distribution of HMG-1,2 is significantly different from that of HMG-17. To avoid possible artifacts due to cell fractionation, fluoresceinated HMG-1 and HMG antibodies were microinjected into living fibroblasts. The cellular distribution of the injected proteins was monitored using fluorescent microscopy. Fluoresceinated HMG-1 microinjected into the cytoplasm moves very rapidly into the nucleus and concentrates in the nucleolus of living human fibroblasts. However, some control non-nuclear proteins also migrated into the nucleus raising the possibility that exogenous injected proteins do not always distribute in the same pattern as the endogenous proteins. The localization of microinjected F(ab)2 fragments derived from anti-HMG-1 was compared to that of microinjected F(ab)2 derived from anti-histones. Whereas the anti-histone F(ab)2 when injected into the cytoplasm migrated into the nucleus, the anti-HMG-1 F(ab)2 remained in the cytoplasm. Microinjection of anti-HMG-17 and anti-histone inhibited transcription in living cells, anti-HMG-1,2 did not. We conclude that HMG-1,2 proteins are present in both the nucleus and cytoplasm of living fibroblasts.

Antibodies↗
PubMed1979

Production of mRNA in Chinese hamster cells: relationship of the rate of synthesis to the cytoplasmic concentration of nine specific mRNA sequences.

We constructed cloned DNA sequences complementary to unselected mRNAs [poly(A)+ cytoplasmic RNA] from Chinese hamster ovary cells and used them in RNA:DNA hybridization experiments. Each cloned DNA hybridized a single mRNA from 1.3-3.5 kb in length. The relative rates of labeling (transcription rates) of nuclear RNA complementary to each individual DNA segment varied approximately 10 fold. The relative cytoplasmic concentration of the same specific RNA sequences in the mRNA after an equilibrium labeling of the cells varied approximately 100 fold. In addition, we estimated the sizes of the nuclear RNA precursor molecules to these cytoplasmic mRNAs. Four main conclusions arise from these studies. First, the primary RNA transcripts, which range in size from 2.4-13.5 kb, are 2-6 times larger than the mRNAs; second, each cloned DNA segment is complementary to only one species of mRNA; third, for the RNA complementary to at least three of the nine cloned DNA segments, the relative cytoplasmic content is considerably different from the relative rate of nuclear RNA synthesis, suggesting the post-transcriptional events are involved in the determination of the cytoplasmic concentrations of some mammalian mRNAs; and fourth, the fraction of total nonribosomal nuclear RNA complementary to the nine cloned DNA segments is in most cases 10 fold less than the fraction of cytoplasmic mRNA complementary to the same cloned DNA segments, suggesting the synthesis of many hnRNA molecules that are qualitatively different from those which eventually contribute mRNA to the cytoplasm.

Animals↗
PubMed1992

Prognostic significance of cytoplasmic p53 oncoprotein in colorectal adenocarcinoma.

Mutation of p53, a tumour-suppressor protein, leads to overexpression of the protein and loss of its tumour-suppressive properties. In some tumours (eg, breast) p53 expression is related to well-known prognostic factors, but findings in colorectal tumours are equivocal. We have used the polyclonal antibody CM1 to investigate nuclear and cytoplasmic p53 expression in colorectal tumours and to assess their relations with prognosis. Of 293 colorectal adenocarcinomas, 71 (24%) showed p53 expression in the nucleus alone, 30 (10%) showed p53 in the cytoplasm alone, and 43 (15%) showed both nuclear and cytoplasmic expression. Nuclear p53 expression showed no relation with survival or Dukes' stage of the tumour. However, the frequency of cytoplasmic expression increased with advancing Dukes' stage (chi 2 for trend 11.18, 1 df, p = 0.0008) and cytoplasmic expression was associated with poor survival (rate ratio 2.3 [95% CI 1.6-3.3], p < 0.0001). Among tumours of Dukes' stage A-C, cytoplasmic expression showed prognostic value independent of nuclear staining, grade of differentiation, and Dukes' stage (2.3 [1.4-3.7], p = 0.0021). We conclude that cytoplasmic expression of p53 may be a useful biological indicator of prognosis in colorectal adenocarcinoma.

Adenocarcinoma↗
PubMed1985

Cytoplasmic IgM in leukemic B cells by flow cytometry.

The presence of intracellular cytoplasmic immunoglobulin M (IgM) in leukemic cells from patients with acute lymphocytic leukemia (ALL) and chronic lymphocytic leukemia (CLL) was investigated by flow cytometry. The objective of the study was to develop a reproducible flow cytometric method. A Burkitt's lymphoma-derived B-cell line, Daudi, and a pre-B ALL, Nalm-6, served as prototypes. Normal B cells and cells from patients with chronic myelogenous leukemia (CML) were used as negative controls. Cytoplasmic mu was expressed in 77.3 +/- 7.5% (n = 10) of Nalm-6 cells. CALLA+ ALL and CML cells lacked cytoplasmic mu. The surface-membrane immunoglobulin on the viable B cells was blocked with purified goat anti-human IgM. Subsequently, the B cells were fixed in cold absolute methanol and stained with a fluorescein-conjugated goat anti-human IgM to demonstrate cytoplasmic IgM. After the surface-membrane IgM was blocked, normal B cells had no cytoplasmic IgM (0.3-0.5% positive cells) detectable by flow cytometry. However, the peripheral blood lymphocytes from five patients with CLL and the Daudi cells contained cytoplasmic IgM, ranging from 7.8 to 76.7% and 45.7 to 89.3%, respectively. We conclude that cytoplasmic mu in Nalm-6, CLL, and Daudi cells can be easily and rapidly demonstrated by flow cytometry.

Animals↗
PubMed1991

Evaluation of four methods for scoring cytoplasmic grains in the in vitro unscheduled DNA synthesis (UDS) assay.

The in vitro unscheduled DNA synthesis (UDS) assay measures DNA repair (incorporation of [3H]thymidine) following in vitro treatment of rat primary hepatocytes. The autoradiographic method was used to detect UDS by counting developed silver grains in the photographic emulsion overlaying nuclei and cytoplasmic areas of the hepatocytes. In this communication we report results using 4 scoring methods: (1) the 2 most heavily labeled cytoplasmic areas adjacent to the nucleus (our standard method), (2) the cytoplasmic area left of the nucleus, (3) the cytoplasmic areas left and right of the nucleus, and (4) 2 cytoplasmic areas whose positions were selected at random. Rat primary hepatocyte cultures treated with a medium control, a solvent control (dimethyl sulfoxide) and 5 known genotoxic chemicals (2-acetylaminofluorene, dimethylnitrosamine, diethylnitrosamine, methyl methanesulfonate and ethyl methanesulfonate) were scored using these 4 methods. The average or maximum cytoplasmic grain count was subtracted from the nuclear grain count to yield net grains/nucleus (NG). In general, NG counts for Methods 2, 3 and 4 were similar, although shifted about 3-10 grains higher than Method 1 for controls and most treated groups. Methods 2, 3 and 4 showed more experiment-to-experiment variability in sensitivity for detecting statistically significant increases in treated groups than did our standard method. Thus, the alternative methods afforded no consistent improvements in sensitivity or reduction of variability for this assay. Subtraction of the average or the highest cytoplasmic count had virtually no effect on the sensitivity of the assay, but simply requires an appropriate adjustment of the criteria for a positive response.

Animals↗
PubMed1979

Regulation of rat lung adenylate cyclase by cytoplasmic factor(s) during development.

Basal adenylate cyclase activity in rat lung homogenate was low prenatally but increased several-fold after birth and remained elevated to maturity. The results also demonstrate the appearance of some factor(s) in the lung cytoplasm at a certain age which markedly activated adenylate cyclase. During late gestation and early neonatal life, when the cytoplasmic factor(s) was low or absent, basal adenylate cyclase activity was low and norepinephrine and NaF produced maximum activation of the enzyme. However, when the cytoplasmic factor(s) appeared in the adult lungs, basal adenylate cyclase activity was elevated and both norepinephrine and NaF produced little or no activation of the enzyme. These data suggest a role for the cytoplasmic factor(s) in regulating rat lung adenylate cyclase. The cytoplasmic factor(s) appeared to be a protein since it was inactivated by trypsin digestion and by heating to 75 degrees C. Activation of adenylate cyclase was not due to small ions or other low molecular weight components of the cytoplasm as dialysis of the supernatant did not alter its activation of adenylate cyclase. The cytoplasmic factor(s) did not appear to be either GTP or calcium-dependent regulator of cyclic AMP phosphodiesterase as these did not activate the rat lung adenylate cyclase.

Adenylyl Cyclases↗
PubMed1993

Cytoplasmic segregation and cytoskeletal organization in the electric catfish giant electromotoneuron with special reference to the axon hillock region.

The cytoplasm of the highly polarized nerve cell is permanently segregated into domains with differing organellar composition. The mechanisms maintaining this segregation are largely unknown. In order to elucidate the potential role of cytoskeletal elements in this process we compared the cytoplasmic segregation within the giant electromotoneuron of the electric catfish (Malapterurus electricus) with the distribution of binding sites for antibodies against elements of the cytoskeleton. Most prominent cytoplasmic segregations include the formation of a subplasmalemmal cortical structure free of Nissl bodies and Golgi cisternae, the separation within the soma of domains containing rough endoplasmic reticulum and filament-rich domains, and the soma-axon transition. The cytoplasmic transition at the axon hillock forms a distinct borderline where Nissl bodies, Golgi cisternae and the bulk of lysosomes abruptly terminate and are excluded from the axoplasm. Synaptic vesicles and mitochondria are free to pass compartmental borders. Tropomyosin, spectrin, and alpha-actinin reveal a rather homogeneous immunofluorescence throughout the neuron. In contrast, neurofilament protein and tubulin display a distinctly increased immunofluorescence in the subplasmalemmal cortical layer, in dendrites as well as in the axon. The increase in immunofluorescence at the axon hillock exactly depicts the small transition zone from the somatic cytoplasm rich in Nissl bodies, Golgi cisternae and lysosomes to the differently structured axoplasm. The picture is similar for beta-tubulin, tyrosinylated and detyrosinylated alpha-tubulin. Detyrosinylated tubulin (glu-tubulin, which is contained in microtubules of increased stability) shows the most prominent enrichment in the axon. The distribution of myosin is comparable to that of neurofilament protein but there is less difference in immunofluorescence between the domains. Our results would be compatible with a role of microtubules together with (the closely associated) neurofilaments in the segregation of neuronal cytoplasmic domains. Active transport as well as stable binding to the somatic cytoskeleton might counteract a homogeneous cytoplasmic distribution of the various classes of organelles by diffusion.

Actinin↗
PubMed1996

Prognostic significance of p53 nuclear and cytoplasmic overexpression in right and left colorectal adenocarcinomas.

The prognostic significance of nuclear and cytoplasmic p53 protein, detected immunocytochemically using CM1 and PAb 1801 antibodies, was evaluated in right-sided and left-sided colorectal adenocarcinomas from 293 patients. CM1 nuclear and cytoplasmic p53 accumulation occurred in 38 and 25% of cases, respectively. PAb 1801 nuclear staining occurred in 18%, with no cytoplasmic staining. CM1 expression either in the nucleus or in the cytoplasm was positively related to PAb 1801 expression (P < 0.001 and P = 0.009, respectively). The incidence of CM1 nuclear and cytoplasmic expression was more frequent in right-sided tumours (P = 0.023 and P = 0.034, respectively), while PAb 1801 nuclear staining was more common in left-sided tumours (P = 0.011). In survival analyses, CM1 nuclear overexpression in the right-sided tumours (P = 0.016) and CM1 cytoplasmic overexpression in left-sided tumours (P = 0.04) were prognostic indicators, independent of Dukes' stage, DNA ploidy, PAb 1801 expression and each other. Further analysis showed that the prognostic value of CM1 nuclear expression was greater in right-sided tumours than in left-sided tumours (P = 0.018). The nuclear and cytoplasmic p53 protein detected with CM1 and PAb 1801 may play different roles in tumour progression and provide prognostic indicators for right- and left-sided colorectal tumours.

Adenocarcinoma↗
PubMed2005

Surface and cytoplasmic immunoglobulin expression in B-cell chronic lymphocytic leukemia (CLL).

Flow cytometric analysis of abnormal lymphocyte populations in chronic lymphocytic leukemia (CLL) has been widely reported to show weak expression of surface immunoglobulin (sIg). The international scoring system to help discriminate between CLL and other B-cell lymphoproliferative disorders lists this as the first of 5 criteria worth 1 point each. In the present study, 30 cases of CLL were studied for surface and cytoplasmic Ig expression. 23 of these 30 (76.7%) cases were positive for sIg. Of these 23, 14 cases (60.9%) showed moderate to bright sIg expression. All of these 23 cases were positive for CD5 and CD23; all were negative for CD10 and only 6 (26.1%) were positive for FMC7. 27 of 30 (90.0%) cases expressed cytoplasmic immunoglobulin (cIg) compared to 5% reported by others. This shows that cytoplasmic Ig occurs in a much greater percentage of cases than reported previously. 23 of 30 (76.7%) cases showed positivity for both surface and cytoplasmic Ig, with 15 showing kappa light chain restriction and 8 showing lambda light chain restriction. Six expressed cytoplasmic Ig only, with 4 showing kappa light chain restriction and 2 showing lambda light chain restriction. One case expressed neither cytoplasmic nor surface Ig. CD38 positivity portends a worse prognosis. Of the 29 cases tested for CD38, 13 (44.8%) were positive. Of these 13 cases, 12 were in the surface Ig/cytoplasmic Ig+ group and 1 in the cytoplasmic Ig+ group only. Also, of the 23 cases tested for CD22 expression, 16 (69.6%) were positive. These data question the use of both "weak" surface Ig expression and lack of CD22 expression as valid scoring criteria for CLL.

Cell Membrane↗
PubMed2002

The precrystalline cytoplasmic granules of alveolar soft part sarcoma contain monocarboxylate transporter 1 and CD147.

Alveolar soft part sarcoma (ASPS) is an unusual tumor of young adults with the characteristic presence on ultrastructural analysis of rhomboid or rectangular cytoplasmic crystals. These membrane-bound crystals are known to form within specific PAS-diastase-resistant electron-dense cytoplasmic granules. The composition of these crystals and the dense granules from which they are derived has remained elusive. After the detection of strong discrete granular cytoplasmic immunoreactivity in ASPS for monocarboxylate transporter 1 (MCT1) in the course of a broad immunohistochemical characterization of an MCT1 antibody, we studied the expression of MCT1 and its interacting partner, CD147, in a panel of 10 ASPS cases using appropriate antibodies. MCT1 is one of a family of widely expressed proton-linked transporters for monocarboxylates such as lactate and pyruvate. In all normal and neoplastic tissues studied to date, MCT1 immunoreactivity is limited to the cell surface. We find that the periodic acid-Schiff-diastase-resistant cytoplasmic granules of ASPS are strongly immunoreactive for MCT1 and CD147. Specifically, intense cytoplasmic granular positivity for MCT1 and CD147 was found in 7 of 10 and 8 of 10 ASPSs, respectively. Ultrastructural immunohistochemistry with immunogold labeling confirmed that the MCT1 immunoreactivity localized to the cytoplasmic electron-dense granules in ASPS. Western blot analysis of several ASPS cases confirmed that the protein reactive with the MCT1 antibody and that reactive with the CD147 antibody both migrated at the size expected for MCT1 and CD147, respectively. Thus, ASPS cells seem to accumulate MCT1-CD147 complexes in the specific cytoplasmic granules known to undergo crystallization. The possible basis for the overproduction or impaired surface localization of these proteins in ASPS remains unclear.

Adult↗
PubMed2003

Relevance of nuclear and cytoplasmic von hippel lindau protein expression for renal carcinoma progression.

Alterations of the von Hippel-Lindau tumor-suppressor gene (VHL) on 3p25-p26 are frequent in clear-cell renal-cell carcinoma (RCC). The VHL protein (pVHL) is implicated in cell-cycle control and gene regulation, and requires transcription-dependent nuclear-cytoplasmic trafficking for its function. There are two biologically active VHL protein isoforms: pVHL(30) and pVHL(19). To study prevalence, subcellular expression and biological significance of pVHL in renal tumors, tissue microarrays with renal-cell carcinomas were immunohistochemically examined for pVHL expression. Antibodies against both protein isoforms (anti-pVHL(30)/pVHL(19)) and against pVHL(30) (anti-pVHL(30); Ig33) were used. The anti-pVHL(30)/pVHL(19) antibody showed nuclear and cytoplasmic pVHL expression, whereas the anti-pVHL(30) antibody (Ig33) detected cytoplasmic pVHL expression, suggesting that the distribution of VHL protein isoforms varies in the nuclear and cytoplasmic compartments of renal tumors. There were 175 of 398 primary clear-cell RCCs (44%) with both nuclear and cytoplasmic pVHL expression. Seventy-seven clear-cell RCCs (19%) showed only nuclear, 22 (6%) showed only cytoplasmic, and 124 tumors (31%) showed no pVHL expression. Notably, combined nuclear and cytoplasmic pVHL expression was associated with low histological grade (P < 0.0001), early tumor stage (P < 0.01), and better prognosis (P < 0.01). These results imply that alteration of subcellular pVHL trafficking is of potential relevance for the biological behavior of clear-cell RCC.

Alleles↗