scholarly journals IMMUNOFLUORESCENT STUDIES OF HUMAN CELL CULTURES AND CHICK EMBRYOS INOCULATED WITH THE AMEBOID CELL-"LIPOVIRUS" COMPLEX

1966 ◽  
Vol 124 (6) ◽  
pp. 1167-1180 ◽  
Author(s):  
Chien Liu ◽  
Patricia Rodina
Keyword(s):  
Phase Contrast ◽  
Early Stage ◽  
Immunofluorescent Staining ◽  
Chick Embryos ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Human Cell Cultures ◽  
Contrast Microscopy ◽  
Infected Cells ◽  
Ameboid Cell

Tissue culture cells of human origin (HeLa, Lich, and AV3) were inoculated with the AL complex. By immunofluorescent staining, AL complex antigen was detectable in the cytoplasm of infected cells as punctate fluorescent granules during the early stage and as homogeneous fluorescence during the late stage of infection. By combining fluorescence and phase-contrast microscopy, many infected cells with cytoplasmic AL complex antigen were shown to have a normal nuclear morphology indistinguishable from uninfected cells. Initiation of AL complex infection was interpreted as occurring by transfer of transmissible factor(s) from cell to cell by contact and mutiplication of such factor(s) therein. Chick embryos were susceptible to AL complex infection following allantoic or amniotic inoculations. Antigen and infectious AL complex were demonstrable in the liver, brain, intestines, lungs, and embryonic membranes. Further investigations on AL complex and its relation to human disease are suggested.

scholarly journals A CYTOCHEMICAL DESCRIPTION OF THE MULTIPLICATION OF MENGOVIRUS IN L-929 CELLS

1962 ◽  
Vol 12 (1) ◽  
pp. 1-15 ◽  
Author(s):  
Richard M. Franklin
Keyword(s):  
Phase Contrast ◽  
Basic Protein ◽  
Virus Production ◽  
Cytological Change ◽  
Virus Particles ◽  
Contrast Microscopy ◽  
Infected Cells ◽  
Cytological Changes ◽  
Perinuclear Area

A correlation of cytochemical changes with virus production has been studied in L cells infected with Mengovirus. After a latent period of about 2 hours, virus was produced rapidly, reaching maximum titers of up to 12,000 particles per cell in 6 to 8 hours. The earliest cytological change was in the nucleus and consisted of a slight condensation of chromatin. There is no evidence, however, for the multiplication of either the viral RNA or protein in the nucleus. RNA, of high molecular weight, accumulated in the perinuclear area of the cytoplasm and was later found in inclusions. The perinuclear RNA was digestible with RNase and may be located in or on ribosomes. The inclusion RNA was resistant to RNase but could be removed by pepsin or potassium permanganate; it is probably in completed virus particles. Viral antigen was first observed in a perinuclear location and later in the above-mentioned inclusions. Although the viral protein contains appreciable amounts of arginine and lysine, it is not a basic protein of the histone type. Phase-contrast microscopy of living cells clearly demonstrated the role of the inclusions in release of virus from infected cells. A comparison is made between these cytological changes in Mengo-infected cells and those which have been found by other workers in polio-infected cells. There are many very similar changes.

scholarly journals Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line.

1982 ◽  
Vol 93 (3) ◽  
pp. 576-582 ◽  
Author(s):  
J V Kilmartin ◽  
B Wright ◽  
C Milstein
Keyword(s):  
Monoclonal Antibodies ◽  
Myeloma Cell ◽  
Yeast Cells ◽  
Immunofluorescent Staining ◽  
Mitotic Spindles ◽  
Spleen Cells ◽  
Microtubule Network ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Parental Lines

Hybrid myeloma cell lines secreting monoclonal antibodies to tubulin have been prepared using rat myelomas and spleen cells from rats immunized with yeast tubulin. A comparison between the results obtained with the rat myeloma Y3-Ag 1.2.3., which secretes a light chain, and a new line, YB2/O, which does not, shows that they are both excellent parental lines and that the second produces hybrids with no myeloma chain components. The antitubulin antibodies in the serum of rats bearing two of the hybrid myeloma tumors gave titers of up to 1:10(6) from which large amounts of monoclonal antibodies could be easily purified. They recognized tubulin from yeast as well as from birds and mammals. The two antibodies gave clear immunofluorescent staining of yeast mitotic spindles as well as the interphase microtubule network of tissue culture cells. Some difference in the pattern of immunofluorescence staining of yeast cells and nuclei was observed between the two antibodies. The purified antibodies could be conjugated to colloidal gold particles and used for direct labeling of yeast microtubules for electron microscopy.

scholarly journals A New Model of SARS-CoV-2 Infection Based on (Hydroxy)Chloroquine Activity

2020 ◽  
Author(s):  
Robert J. Sheaff
Keyword(s):  
Oxidative Phosphorylation ◽  
Host Cell ◽  
Disease Progression ◽  
Therapeutic Benefit ◽  
Beta Oxidation ◽  
New Model ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Obesity And Diabetes ◽  
Infected Cells

Chloroquine and hydroxychloroquine [(H)CQ] are well known anti-malarial drugs, while their use against COVID-19 is more controversial. (H)CQ activity was examined in tissue culture cells to determine if their anti-viral benefits or adverse effects might be due to altering host cell pathways. Metabolic analysis revealed (H)CQ inhibit oxidative phosphorylation in mitochondria, likely by sequestering protons needed to drive ATP synthase. This activity could cause cardiotoxicity because heart muscle relies on beta oxidation of fatty acids. However, it might also explain their therapeutic benefit against COVID-19. A new model of SARS-CoV-2 infection postulates virus enters host cell mitochondria and uses its protons for genome release. Oxidative phosphorylation is eventually compromised, so glycolysis is upregulated to maintain ATP levels. (H)CQ could prevent viral infection and/or slow its replication by sequestering these protons. In support of this model other potential COVID-19 therapeutics also targeted mitochondria, as did tobacco smoke, which may underlie smokers protection. The mitochondria of young people are naturally more adaptable and resilient, providing a rationale for their resistance to disease progression. Conversely, obesity and diabetes could exacerbate disease severity by providing extra glucose to infected cells dependent on glycolysis. The description of (H)CQ function presented here, together with its implications for understanding SARS-CO-V2 infection, makes testable predictions about disease progression and identifies new approaches for treating COVID-19.

Phase Contrast Microscopy with Modified Hansel's Staining: Mast Cell Identification and Activity-Related Changes in Cell Membrane Refractivity in Allergic Nasal Smears

2003 ◽  
Vol 17 (2) ◽  
pp. 101-106
Author(s):  
Masatoshi Masuko ◽  
Fumiyuki Goto
Keyword(s):  
Cell Membrane ◽  
Phase Contrast ◽  
Early Stage ◽  
Japanese Cedar ◽  
Cell Types ◽  
New Technique ◽  
Morphological Identification ◽  
Phase Contrast Microscopy ◽  
Contrast Microscopy ◽  
Japanese Cedar Pollinosis

Background A new technique called phase contrast microscopy with modified Hansel's staining (used with bright field microscopy) was developed to identify mast cells (MCs) and granulocytes (eosinophil/neutrophil/basophil [E/N/B]). Methods Nasal scratching smears from 618 patients with Japanese cedar pollinosis were examined using this new technique. Results This technique permitted accurate morphological identification. MCs can be discriminated from E/N/Bs. The surface of the cell membrane appeared as low refractile (lr), moderately refractile (mr), or high refractile (hr). This was caused by the light, which is not related to the phase difference, but rather originated from the difference in the refraction of the direct light. In specimens from the onset stage (i.e., 1–3 days after onset), lr-MC and mr/hr-E/N/B were dominant. In specimens from the early stage (i.e., 4–7 days after onset), lr-E/N/B significantly increased in number. Conclusion The phospholipid bilayers of the cell membrane exhibit a phase transition after the onset, and phase refractivity of the cell membrane is closely related to the activity of the cell. This indicates that in the onset stage, MCs are already activated, whereas most of the E/N/Bs are not. In contrast, the latter cell types become activated subsequently in the early stage.

scholarly journals Identification of Necrotic Tissue by Phase–Contrast Microscopy at an Early Stage of Acute Myocardial Infarction

2000 ◽  
Vol 80 (6) ◽  
pp. 981-982 ◽  
Author(s):  
Till Neumann ◽  
Ina Konietzka ◽  
Anita van de Sand ◽  
Stephanie Aker ◽  
Rainer Schulz ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Phase Contrast ◽  
Early Stage ◽  
Phase Contrast Microscopy ◽  
Necrotic Tissue ◽  
Contrast Microscopy

scholarly journals Overexpression of the Rabies Virus Glycoprotein Results in Enhancement of Apoptosis and Antiviral Immune Response

2002 ◽  
Vol 76 (7) ◽  
pp. 3374-3381 ◽  
Author(s):  
Milosz Faber ◽  
Rojjanaporn Pulmanausahakul ◽  
Suchita S. Hodawadekar ◽  
Sergei Spitsin ◽  
James P. McGettigan ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Caspase 3 ◽  
Marked Decrease ◽  
Fluorescence Staining ◽  
Antiviral Immune Response ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Glycoprotein G ◽  
Antibody Titers ◽  
Infected Cells

ABSTRACT A recombinant rabies virus (RV) carrying two identical glycoprotein (G) genes (SPBNGA-GA) was constructed and used to determine the effect of RV G overexpression on cell viability and immunity. Immunoprecipitation analysis and flow cytometry showed that tissue culture cells infected with SPBNGA-GA produced, on average, twice as much RV G as cells infected with RV carrying only a single RV G gene (SPBNGA). The overexpression of RV G in SPBNGA-GA-infected NA cells was paralleled by a significant increase in caspase 3 activity followed by a marked decrease in mitochondrial respiration, neither of which was observed in SPBNGA-infected cells. Furthermore, fluorescence staining and confocal microscopy revealed an increased extent of apoptosis and markedly reduced neurofilament and F actin in SPBNGA-GA-infected primary neuron cultures compared with neuronal cells infected with SPBNGA, supporting the concept that RV G or motifs of the RV G gene trigger the apoptosis cascade. Mice immunized with SPBNGA-GA showed substantially higher antibody titers against the RV G and against the nucleoprotein than SPBNGA-immunized mice, suggesting that the speed or extent of apoptosis directly determines the magnitude of the antibody response.

scholarly journals IMMUNOFLUORESCENT STUDIES OF ADENOVIRUS 12 TUMORS AND OF CELLS TRANSFORMED OR INFECTED BY ADENOVIRUSES

1964 ◽  
Vol 120 (4) ◽  
pp. 577-588 ◽  
Author(s):  
John H. Pope ◽  
Wallace P. Rowe
Keyword(s):  
Tumor Cells ◽  
Cell Cultures ◽  
Human Cell ◽  
Adenovirus Type ◽  
Mouse Tumor ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Induced Tumors ◽  
Human Cell Cultures ◽  
Embryo Tissue

Complement-fixing (CF) antibody-positive sera from hamsters bearing adenovirus type 12 (Ad. 12)-induced tumors revealed specific immunofluorescent stainable antigens in essentially all Ad. 12 hamster tumor cells. The antigens were primarily in the form of cytoplasmic flecks; less frequent staining was seen as nuclear flecks or homogeneous staining of nucleus and cytoplasm of a small proportion of cells. Tumor cells did not stain with rabbit antisera to crude Ad. 12 virus or A and C antigens. The hamster serum also stained cytoplasmic flecks in an Ad. 12-induced BALB/c mouse tumor and Ad. 12-"transformed" hamster embryo tissue culture cells. The hamster serum also stained fleck-shaped antigens in hamster and human cell cultures inoculated with homologous and heterologous adenovirus types, although the hamster cells did not react with the rabbit Ad. 12 antiserum. Attempts to identify the fluorescent-stainable fleck-shaped antigens indicated that they are not previously recognized viral antigens and that the cytoplasmic antigens formed in hamster cell cultures inoculated with Ad. 12 are different from those in tumors and in acutely infected human cell cultures.

scholarly journals Studies on Antibody-Producing Cells by Hemolytic Plaque Method and Phase Contrast Microscopy

Blood ◽  
1969 ◽  
Vol 33 (2) ◽  
pp. 149-158
Author(s):  
IRIMAJIRI KIYOHIRO ◽  
ATSUSHI HORIOCHI ◽  
YUJI OKAMOTO ◽  
ICHITA AMAKI
Keyword(s):  
Lymph Node ◽  
Plasma Cell ◽  
Thoracic Duct ◽  
Phase Contrast ◽  
Early Stage ◽  
Phase Contrast Microscopy ◽  
Whole Process ◽  
Contrast Microscopy ◽  
Mature Lymphocyte ◽  
High Level

Abstract The numbers of the PFC in the thoracic duct and lymph node of the immunized rabbit were counted by the hemolytic plaque method and the morphologic features of the PFC in the lymph node were simultaneously observed by phase contrast microscopy. The number of the PFC in the lymph node maintained a relatively high level on the fourth to seventh days, and then rose to attain the maximum on the ninth to thirteenth days, and then decreased. The number of the PFC in the thoracic duct reached the maximum on the fourth day after the immunization and then dropped rapidly. The mechanism was discussed. From morphologic observations, the PFC were classified into lymphogonia, lymphoblast, basophilic mature lymphocyte and plasma cell. The lymphogonia and lymphoblast, which belonged to immature lymphocyte series, were frequent in the early stage and the plasma cell was increased in the later stage of the immunization. The basophilic mature lymphocyte constituted more than 55 per cent through the whole process and were most frequent of the plaque forming cells.

scholarly journals Junctional Adhesion Molecule 1 Is a Functional Receptor for Feline Calicivirus

2006 ◽  
Vol 80 (9) ◽  
pp. 4482-4490 ◽  
Author(s):  
Akiko Makino ◽  
Masayuki Shimojima ◽  
Takayuki Miyazawa ◽  
Kentaro Kato ◽  
Yukinobu Tohya ◽  
...  
Keyword(s):  
Adhesion Molecule ◽  
Vero Cells ◽  
Prototype Strain ◽  
Feline Calicivirus ◽  
Progeny Production ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
The Family ◽  
Infected Cells ◽  
Functional Receptor

ABSTRACT The life cycle of calicivirus is not fully understood because most of the viruses cannot be propagated in tissue culture cells. We studied the mechanism of calicivirus entry into cells using feline calicivirus (FCV), a cultivable calicivirus. From the cDNA library of Crandell-Rees feline kidney (CRFK) cells, feline junctional adhesion molecule 1 (JAM-1), an immunoglobulin-like protein present in tight junctions, was identified as a cellular-binding molecule of the FCV F4 strain, a prototype strain in Japan. Feline JAM-1 expression in nonpermissive hamster lung cells led to binding and infection by F4 and all other strains tested. An anti-feline JAM-1 antibody reduced the binding of FCV to permissive CRFK cells and strongly suppressed the cytopathic effect (CPE) and FCV progeny production in infected cells. Some strains of FCV, such as F4 and F25, have the ability to replicate in Vero cells. We found that regardless of replication ability, FCV bound to Vero and 293T cells via simian and human JAM-1, respectively. In Vero cells, an anti-human JAM-1 antibody inhibited binding, CPE, and progeny production by F4 and F25. In addition, feline JAM-1 expression permitted FCV infection in 293T cells. Taken together, our results demonstrate that feline JAM-1 is a functional receptor for FCV, simian JAM-1 also functions as a receptor for some strains of FCV, and the interaction between FCV and JAM-1 molecules may be a determinant of viral tropism. This is the first report concerning a functional receptor for the viruses in the family Caliciviridae.

scholarly journals A STUDY OF TISSUE CULTURE CELLS BY ELECTRON MICROSCOPY

1945 ◽  
Vol 81 (3) ◽  
pp. 233-246 ◽  
Author(s):  
Keith R. Porter ◽  
Albert Claude ◽  
Ernest F. Fullam
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Electron Micrographs ◽  
Cell Structure ◽  
Culture Technique ◽  
Chick Embryos ◽  
Tissue Culture Technique ◽  
Culture Cells ◽  
Tissue Culture Cells

By means of a tissue culture technique, cells from chick embryos were procured in a state which proved to be suitable for electron microscopy. The electron micrographs disclosed details of cell structure not revealed by other methods of examination.

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