Cytofluorometric analysis of macrophages activated <i>in vivo</i> or <i>in vitro</i>

Stadler, Beda M.; Weck, A.L. de

doi:10.1002/eji.1830080405

Cited by 13 publications

(3 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Generally, macrophage activation is not a property of this cell type, but rather one that is acquired only following exposure to stimuli encountered in the microenvironment [Ohmori and Hamilton, 1994]. Their activation is based in cellular and ultrastructural changes in morphology and increased functional activities [Stadler and Weck, 1978]. These characteristics are results from the transcriptional profile of the cells and therefore the pattern of gene expression in a cell can provide detailed information about its state [DeRisi et al, 1997].…”

mentioning

confidence: 98%

Gene expression profiling of macrophages following mice treatment with an immunomodulator medication

Oliveira

Góes

et al. 2008

J of Cellular Biochemistry

View full text Add to dashboard Cite

Canova (CA) is a complex homeopathic medication used in diseases where the immune system is depressed. Previous studies demonstrated that it is neither toxic nor mutagenic and activates macrophages. We now evaluate CA effects on cytokine production and gene expression from mice macrophages. The global view of changes in expression of genes with known functions can provide a vivid picture of the way in which cell adapts to a changing environment or a challenge. We found a decrease in IL-2 and IL-4 production and a differential expression in 147 genes from CA group. These genes are mainly involved in transcription/translation, cell structure and dynamics, immune response, cytoprotection, enzymatic process, and receptors/ligands. With gene expression analysis we state that this medication provokes a reaction that involves alterations in gene expression profile mainly in the ones involved with macrophages activation, corroborating the laboratorial research and the clinical data.

show abstract

mentioning

confidence: 98%

Gene expression profiling of macrophages following mice treatment with an immunomodulator medication

Oliveira

Góes

et al. 2008

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…Cell flow analysis also confirmed this notion, showing that several populations of macrophages could be distinguished as a function of their content in RNA (17,18) and in various enzymes (5) or according to their affinity for certain antimacrophage monoclonal antibodies (15).…”

mentioning

confidence: 79%

Characterization and sorting of mouse cytotoxic macrophages by their light scattering properties

et al. 1983

View full text Add to dashboard Cite

The light scattering properties of mouse activated macrophages were analyzed by flow cytometry. Peritoneal adherent cells from B. abortus treated animals were found to segregate into two subpopulations as a function of their foward angle and 90" angle light scatter. The cell subpopulations were separated by automatic sorting. The strongly scattering ones contained an elevated proportion of large volume and acid phosphatase rich cells. Their nonspecific cytotoxic activity against tumor cells was more important than that of weakly light scattering cells. Thus, flow cytometry might be helpful to characterize and isolate cytotoxic macrophage populations.Key terms: Flow cytometry, light scattering, cell sorting, activated macrophages, cytotoxicityThe morphological and functional heterogeneity of macrophages has been evidenced in a number of experimental systems. Cell flow analysis also confirmed this notion, showing that several populations of macrophages could be distinguished as a function of their content in RNA (17,18) and in various enzymes (5) or according to their affinity for certain antimacrophage monoclonal antibodies (15).Using cell sorting techniques, flow cytometry also offers the possibility to study biological properties of isolated cell subpopulations. In the present work, mouse activated macrophages were analyzed and sorted as a function of their light scattering properties. Strongly and weakly light scattering cells were separated and their cytotoxic activity, cell volume, and acid phosphatase content were measured. MATERIALS AND METHODS Preparation of Peritoneal Exudate CellsCells were obtained by washing the peritoneal cavity of (C57BL6 x DBAB) F1 hybrid mice injected IP 3 days before with 500 pg of heat-inactivated B. abortus organisms as described previously (10,ll). The cells were left to adhere on microexudate-coated culture flasks as described by Mantovani et al (9). After 20 min of incubation the nonadherent cell fraction was separated. Adherent cells were detached by a 10-min exposure to ethylene-diamine-tetracetate (EDTA). Their viability was 92% as demonstrated by a Trypan blue exclusion test. Cells were suspended at 40 x 106/ml in RPMI 1640 medium (Difco) before cell flow measurements and sorting. Flow CytometryThe cytofluorograf system 50 H (Ortho Diagnostic Instruments; Westwood, MA) was uscd. The apparatus was standardized daily for scatter using a suspension of glutaraldehyde-fixed calf thymocytes. The suspensions of fresh peritoneal cells, maintained at 4"C, were previously filtered through 50-pm nylon filter. An argon laser beam of wavelength 488 nm and output energy 500 mW was used to illuminate the cells. Light scattering at 90" and at low forward angle (2"-20") was measured. Right angle scatter (RAS) and forward angle scatter (FAS) signals were displayed as a cytogram on a multichannel distribution analyzer (Model 2103, Digital Data Processing) that could also display frequency distribution histograms of cell number against one of the parameters FAS or RAS. Ten or twenty thousan...

show abstract

“…Sera were collected from healthy donors and animals and stored at -20 C. The fetal calf sera were obtained from various companies. The particular batch of FCS (Microbiological Associates, 101 89239) used throughout this suidy was chosen by the following criteria: (a) ability lo support nuirinc lymphtxrytc cultures stimulated liy mitogens or various antigens al lowest possible concentration; (b) ability to support primary immunization to sheep erythrocytes in vitro and (c) ability to support macrophage activation in vitro, assessed by RNA synthesis [17], Stock solutions of concanavalin A (Con A; Pharmacia Fine Chemicals AB, Sweden), lipopotysacxharidc(LPS)and phytohacmaggkitinjn (PHA-P; Difco Laboratories, Mieh.) were stored at -20 C until use.…”

Section: Methodsmentioning

confidence: 99%

The Influence of Serum on Lymphocyte Cultures

1981

View full text Add to dashboard Cite

Serum exerts several effects in lymphocyte cultures, one of them being manifested very early. The presence of fetal calf serum (FCS) results in a markedly higher thymidine uptake within a few minutes, as compared with serum‐free cultures. On the other hand, the uptake of uridine and other purine bases seem to be little influenced by serum. Experiments comparing the uptake of thymidine into the cytoplasm or into trichloroacetic acid‐precipitable material suggested that the intracellular thiymidine pool increases in size when FCS is added. Using a Lineweaver Burt plot for thymidine and uridine uptake over a 4‐h period, no changes in uridine uptake were observed in the presence of FCS; on the contrary, serum induced an increased Vmax for thymidine, whereas Km remained constant. Cytofluorographic quantitation of G0 and G1 cells indicated that cells disappear more rapidly from the G1 phase in the presence of FCS. The addition of hydroxyurea to the cultures prevented this disappearance. The results taken together strongly suggest that serum contains a factor promoting the shift of G1 cells into the S phase. When a 4‐h culture period was used, all sera from differed species tested at low concentrations appeared to contain this activity.

show abstract

Cytofluorometric analysis of macrophages activated in vivo or in vitro

Cited by 13 publications

References 14 publications

Gene expression profiling of macrophages following mice treatment with an immunomodulator medication

Gene expression profiling of macrophages following mice treatment with an immunomodulator medication

Characterization and sorting of mouse cytotoxic macrophages by their light scattering properties

The Influence of Serum on Lymphocyte Cultures

Contact Info

Product

Resources

About