1,087 research outputs found
ACTIVE SUPPRESSION OF IMMUNOGLOBULIN ALLOTYPE SYNTHESIS : II. TRANSFER OF SUPPRESSING FACTOR WITH SPLEEN CELLS
The mechanism of chronic allotype suppression in (SJL x BALB/c)F1 mice has been investigated by means of cell transfer studies. These mice are phenotypically negative for serum Ig-1b, the paternal allotype determinant on γG2a immunoglubulin, as a result of perinatal exposure to maternal anti-Ig-1b. When spleen or bone marrow (B) cells from suppressed mice were injected into irradiated BALB/c "indicator" hosts, detectable levels of Ig-1b were demonstrated in the sera of a majority of the recipients early after transfer. These results indicate that Ig-1b-producing cells or their precursors are present in the lymphoid tissues of suppressed mice, even though they are not expressed. Within 5–7 wk, it was no longer possible to detect Ig-1b in the sera of these hosts, although cells producing another paternal allotype (Ig-4b) were shown to persist. Control BALB/c mice, injected with spleen and B cells from normal mice, continued to produce high levels of immunoglobulin carrying this allotype. The disappearance of serum, Ig-1b occurred most frequently in the recipients of suppressed spleen cells. Similar results were obtained using a mixture of spleen cells from normal and suppressed mice. Ig-1b production in the recipient mice ceased within a few weeks, even though the majority of cells in the mixture were obtained from normal (nonsuppressed) donors. The data are interpreted as evidence that chronic allotype suppression in mice is actively maintained by cells which are resident in the lymphoid tissues, splenic cells being the most effective. These cells are capable of proliferating in a new host and exerting their suppressive influence on Ig-1b-producing cells and/or their precursors
The regulation of CD5 expression in murine T cells
BACKGROUND: CD5 is a pan-T cell surface marker that is also present on a subset of B cells, B-1a cells.Functional and developmental subsets of T cells express characteristic CD5 levels that vary over roughly a 30-fold range. Previous investigators have cloned a 1.7 Kb fragment containing the CD5 promoter and showed that it can confer similar lymphocyte-specific expression pattern as observed for endogenous CD5 expression. RESULTS: We further characterize the CD5 promoter and identify minimal and regulatory regions on the CD5 promoter. Using a luciferase reporter system, we show that a 43 bp region on the CD5 promoter regulates CD5 expression in resting mouse thymoma EL4 T cells and that an Ets binding site within the 43 bp region mediates the CD5 expression. In addition, we show that Ets-1, a member of the Ets family of transcription factors, recognizes the Ets binding site in the electrophoretic mobility shift assay (EMSA). This Ets binding site is directly responsible for the increase in reporter activity when co-transfected with increasing amounts of Ets-1 expression plasmid. We also identify two additional evolutionarily-conserved regions in the CD5 promoter (CD5X and CD5Y) and demonstrate the respective roles of the each region in the regulation of CD5 transcription. CONCLUSION: Our studies define a minimal and regulatory promoter for CD5 and show that the CD5 expression level in T cells is at least partially dependent on the level of Ets-1 protein. Based on the findings in this report, we propose a model of CD5 transcriptional regulation in T cells
ISOLATION OF ANTIGEN-BINDING CELLS FROM UNPRIMED MICE : Demonstration of Antibody-Forming Cell Precursor Activity and Correlation between Precursor and Secreted Antibody Avidities
Cells binding DNP groups conjugated to fluoresceinated mouse gamma globulin (FDNP-MGG) were isolated from spleens of unprimed mice using a fluorescence-activated cell sorter (FACS). The isolated cells were specifically enriched at least 100-fold for anti-DNP precursor activity in an adoptive transfer assay as compared to unfractionated spleen. The fraction depleted of binding cells, although depleted of anti-DNP precursor activity, responded as well as unfractionated spleen when assayed for anticarrier (keyhole limpet hemocyanin [KLH]) precursor activity. High avidity binding cells were stained using low concentrations of FDNP-MGG. Medium and low avidity binding cells were stained using high concentrations of FDNP-MGG in the presence of free hapten which selectively blocked staining of the high avidity binding cells. Cells were supplemented with an excess of carrier-primed (KLH), nylon-purified splenic T cells and transferred to irradiated recipients. DNP-KLH was given at transfer and 5 days later. The anti-DNP plaque-forming cell (DNP-PFC) response and the avidities of the DNP-PFC in the irradiated recipients were measured by hapten inhibition of direct PFC plaque formation 12 days after transfer. At this time, very few indirect PFC were found. There was a positive correlation between the avidity of the DNP-binding cells and the avidity of the anti-DNP antibody secreted by their progeny. High avidity DNP-binding cells gave rise to predominantly high avidity anti-DNP-PFC. Medium and low avidity binding cells gave rise to medium and low avidity DNP-PFC
TOLERANCE AND IMMUNITY TO MATERNALLY DERIVED INCOMPATIBLE IgG2a-GLOBULIN IN MICE
Progeny mice were confronted with maternal γ-globulin of a different allotype by either back-cross mating, intercross mating, or by foster nursing. In all cases, many mice subsequently produced alloantibodies directed against the incompatible maternal type of IgG2a-globulin. In one series of experiments, immunologic tolerance to the maternally derived γ-globulin was demonstrated to exist in the period before formation of spontaneous antibody. The state of tolerance was then lost, unless maintenance injections of foreign γ-globulin were given. These studies demonstrate in a natural situation that maternally derived foreign proteins can first induce a state of immunological tolerance which is followed, after disappearance of the antigen, by a state of immunity. As such, this parallels the experimental induction of tolerance to foreign proteins by neonatal injections
IN VITRO STUDIES OF MAMMALIAN SOMATIC CELL VARIATION : I. DETECTION OF H-2 PHENOTYPE IN CULTURED MOUSE CELL LINES
The isoantigenic phenotype of the H-2 locus has been detected by isohemagglutinin absorption in a line of mouse lymphoma cells growing continuously in culture for 6 years and in two established lines of fibroblastic mouse cells growing continuously in culture for 1 year. Quantitative absorption studies suggest that the concentration of H-2 isoantigens is higher in the cultured lymphoma cells than in the other two fibroblastic cell lines
Cell sorting in a Petri dish controlled by computer vision.
Fluorescence-activated cell sorting (FACS) applying flow
cytometry to separate cells on a molecular basis is a widespread
method. We demonstrate that both fluorescent and unlabeled live
cells in a Petri dish observed with a microscope can be
automatically recognized by computer vision and picked up by a
computer-controlled micropipette. This method can be routinely
applied as a FACS down to the single cell level with a very
high selectivity. Sorting resolution, i.e., the minimum distance
between two cells from which one could be selectively removed
was 50-70 micrometers. Survival rate with a low number of 3T3
mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was
66 +/- 12% and 88 +/- 16%, respectively. Purity of sorted
cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures
were 95 +/- 2% and 62 +/- 7%, respectively. Hydrodynamic
simulations confirmed the experimental sorting efficiency and a
cell damage risk similar to that of normal FACS
An improved cell-volume analyzer
Design and operation of cell-volume analyzer friction, glaze ice, and studded tire effects on highway
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