Methods of Simultaneous Visualization of Cytoplasmic Enzyme Reactivity and Cell Surface Antigens (by Cytochemistry Combined with Immunocytochemistry) in Individual Hematopoietic and Lymphoid Cells

1988 ◽  
Vol 3 (4) ◽  
pp. 221-232 ◽  
Author(s):  
A. Gloghini ◽  
M. Cozzi ◽  
S. Sulfaro ◽  
R. Volpe ◽  
A. Carbone

Enzyme cytochemistry alone, and more recently, immunocytohistochemistry have been satisfactorily used by hematologists and hematopathologists for the study, diagnosis and classification of human hematological and lymphoproliferative disorders. To enhance the potential of these techniques, the possibility of combining immunocytohistochemical techniques with enzyme cytohistochemistry with simultaneous visualization of both reaction products has been examined by some investigators. This approach has been applied to normal, reactive and neoplastic material using mainly cell suspensions and frozen sections, with the aim of improving cell identification in specimens containing different cell types, of determining the cytochemical profiles of well-defined lymphocyte subpopulations and of establishing the cell surface phe-notypes of cells that are positive for certain enzymes. In this paper, published reports on this subject are reviewed and compared with the experience of our study group.

1982 ◽  
pp. 33-68 ◽  
Author(s):  
Robert Fox ◽  
Stephen Baird ◽  
Patrick Kung ◽  
Ron Levy ◽  
Ivor Royston

1993 ◽  
Vol 4 (5) ◽  
pp. 639-677 ◽  
Author(s):  
Irving Dardick ◽  
Aileen P. Burford-Mason

Because of their complexity and relative infrequency, salivary gland tumors commonly result in diagnostic problems. Histogenetic and morphogenetic concepts of tumorigenesis in these glands are reviewed and their relevance to routine diagnosis and classification of salivary gland tumors evaluated. Evidence is presented from animal and human studies that under steady-state and pathophysiological conditions, all cell types present in the normal gland, including acinar cells, are capable of rapidly entering the cell cycle and are, therefore, possible targets for neoplastic transformation.


Blood ◽  
1965 ◽  
Vol 25 (1) ◽  
pp. 63-72 ◽  
Author(s):  
HOWARD C. MEL ◽  
LINDA T. MITCHELL ◽  
BO THORELL

Abstract A single-cell suspension of normal rat bone marrow is prepared mechanically. This suspension is continuously fractionated in free solution, under sedimentation rate conditions, using 1 g. only. With a sample flow of 2.2 x 106 cells/minute and a 32-minute steady-state residence time in the stable-flow free boundary (STAFLO) flow-cell, the cells exit almost entirely into 7 of the 12 collection bottles. Maximum numbers of different cell types are observed, with good repeatability, in approximately descending order from top to bottom as follows: erythrocytes, "erythroblasts," "immatures," "myelocytes," and mature granulocytes. Major changes are effected relative to the starting marrow composition, and large relative enrichments are achieved for certain cell types. In addition to the rapid, mild, preparative aspect of this study, nominal sedimentation rates can be assigned for the different collection fractions, in the range of 3 x 105 to 4 x 106 svedbergs, thus making a start on this kind of simple physical classification of the cellular elements in this complex tissue.


2019 ◽  
Vol 30 (18) ◽  
pp. 2349-2357 ◽  
Author(s):  
Richik Nilay Mukherjee ◽  
Daniel L. Levy

Endoplasmic reticulum (ER) tubules and sheets conventionally correspond to smooth and rough ER, respectively. The ratio of ER tubules-to-sheets varies in different cell types and changes in response to cellular conditions, potentially impacting the functional output of the ER. To directly test whether ER morphology impacts vesicular trafficking, we increased the tubule-to-sheet ratio in three different ways, by overexpressing Rtn4a, Rtn4b, or REEP5. Only Rtn4a overexpression increased exocytosis, but not overall levels, of several cell surface and secreted proteins. Furthermore, Rtn4a depletion reduced cell surface trafficking without affecting ER morphology. Similar results were observed in three different mammalian cell lines, suggesting that Rtn4a generally enhances exocytosis independently of changes in ER morphology. Finally, we show that Rtn4a levels modulate cell adhesion, possibly by regulating trafficking of integrins to the cell surface. Taking the results together, we find that altering ER morphology does not necessarily affect protein trafficking, but that Rtn4a specifically enhances exocytosis.


1975 ◽  
Vol 64 (3) ◽  
pp. 719-724 ◽  
Author(s):  
GL Campbell ◽  
LT Goldstein ◽  
BB Knowles

Quantitation of the expression of cell surface antigens has hitherto been limited to analysis by either cytotoxicity tests or radioimmune assays (5, 15). We report here the use of a new methodology to analyze and quantitate the expression of mouse histocompabililty antigens (H-2 locus) in hybrid clones and parental cell types. The binding of fluorescein-tagged antibody is measured on a cell-to-cell basis in large viable cell populations using flow microfluorimetric techniques. These techniques have been used to measure hapten and immunoglobulin binding to lymphocyte populations (8, 9, 14). However, this is the first report in which these techniques have been used to examine the expression of the H-2 locus. The advantage of this approach is twofold: first, a large and statistically significant sample population may be analyzed one cell at a time, thus revealing the fine detail of heterogeneity in the expression of the cell surface markers within a population. Second, as has been demonstrated for analysis of specific components of the immune system, this method does permit fluorescence-activated sorting of cell types according to their different surface populations (8, 9, 14).


1994 ◽  
Vol 5 (7) ◽  
pp. 819-828 ◽  
Author(s):  
Y Wang ◽  
G M Fuller

Recent evidence has shown that members of the Jak kinase family are activated after IL-6 binds to its receptor complex, leading to a tyrosine phosphorylation of gp130, the IL-6 signal-transducing subunit. The different members of the IL-6 cytokine subfamily induce distinct patterns of Jak-Tyk phosphorylation in different cell types. Using monospecific antibodies to gp130, Jak2 kinase, and phosphotyrosine, we investigated the kinetics of IL-6 stimulation of members of this pathway in primary hepatocytes. Our findings show that Jak 2 is maximally activated within 2 min of exposure to IL-6, followed by gp130 phosphorylation that reaches its peak in another 2 min then declines to basal level by 60 min. In vitro phosphorylation experiments show that activated Jak 2 is able to phosphorylate both native gp130 and a fusion peptide containing its cytoplasmic domain, demonstrating gp130 is a direct substrate of Jak 2 kinase. Experiments designed to explore the cell surface expression of gp130 show that > or = 2 h are required to get a second round of phosphorylation after the addition of more cytokines. This finding suggests that activated gp130 is internalized from the cell surface after IL-6 stimulation. Additional experiments using protein synthesis inhibitors reveal that new protein synthesis is required to get a second cycle of gp130 phosphorylation indicating gp130 must be synthesized de novo and inserted into the membrane. These findings provide strong evidence that down regulation of the IL-6 signal in hepatocytes involves the internalization and cytosol degradation of gp130.


2007 ◽  
Vol 2007 ◽  
pp. 1-19 ◽  
Author(s):  
M. Jäger ◽  
C. Zilkens ◽  
K. Zanger ◽  
R. Krauspe

Cell-surface interactions play a crucial role for biomaterial application in orthopaedics. It is evident that not only the chemical composition of solid substances influence cellular adherence, migration, proliferation and differentiation but also the surface topography of a biomaterial. The progressive application of nanostructured surfaces in medicine has gained increasing interest to improve the cytocompatibility and osteointegration of orthopaedic implants. Therefore, the understanding of cell-surface interactions is of major interest for these substances. In this review, we elucidate the principle mechanisms of nano- and microscale cell-surface interactions in vitro for different cell types onto typical orthopaedic biomaterials such as titanium (Ti), cobalt-chrome-molybdenum (CoCrMo) alloys, stainless steel (SS), as well as synthetic polymers (UHMWPE, XLPE, PEEK, PLLA). In addition, effects of nano- and microscaled particles and their significance in orthopaedics were reviewed. The significance for the cytocompatibility of nanobiomaterials is discussed critically.


1979 ◽  
Vol 12 (2) ◽  
pp. 245-257 ◽  
Author(s):  
Robert L. Simmer ◽  
Joffre B. Baker ◽  
Dennis D. Cunningham

1981 ◽  
Vol 154 (3) ◽  
pp. 659-675 ◽  
Author(s):  
Y Obata ◽  
E Stockert ◽  
A B DeLeo ◽  
P V O'Donnell ◽  
H W Snyder ◽  
...  

A new cell surface antigen of the mouse related to xenotropic murine leukemia virus (MuLV) is described. The antigen, designated G(erld), is defined by cytotoxic tests with the B6-x-ray-induced ERLD and naturally occurring antibody. G(erld) is distinct from all previously defined cell surface antigens. Monoclonal antibody with the same specificity has been developed. Inbred mouse strains are classified as G(erld)+ or G(erld)- according to the presence of absence of the antigen on lymphoid cells. G(erld)+ strains differ with regard to quantitative expression of G(erld) on normal thymocytes. The emergence of G(erld)+ tumors in G(erld)- strains indicates the presence of genes coding for the antigen even in strains not normally expressing the antigen. G(erld) has the characteristic of a differentiation antigen in normal mice. In G(erld)+ strains, high levels of the antigen are found on thymocytes with lower levels being detected on cells of spleen, lymph nodes and bone marrow. No G(erld) was detected in brain or kidney or on erythrocytes. The segregation ratios for G(erld) expression on thymocytes in backcross and F2 mice of crosses between G(erld)+ (B6, 129, and B6-Gix+) and G(erld)- (BALB/c) strains suggest that G(erld) expression is controlled by a single locus in B6, by two unlinked loci in 129, and by three unlinked loci in B6-Gix+ mice. Induction of the antigen by MuLV infection of permissive cells in vitro indicates that G(erld) is closely related to xenotropic and dualtropic MuLV; all xenotropic and dualtropic MuLV tested induced the antigen, whereas the majority of ecotropic and the two amphotropic MuLV failed to do so. As dualtropic MuLV are thought to be recombinants between ecotropic and xenotropic MuLV sequences, G(erld) coding by dualtropic MuLV may signify the contribution of the xenotropic part in the recombinational event. Serological and biochemical characterization indicates that G(erld) is related to the gp 70 component of the MuLV envelope. The relation of G(erld) to the previously defined gp 70-related cell surface antigens (Gix, G(rada), and G(aksl2) is discussed, particularly with regard to their characteristics as differentiation antigens, the genetic origin of dualtropic MuLV, and the leukemogenicity of MuLV.


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