Biomedical subjects

S M Laster

Publications and source records attributed to S M Laster.

23 records · Page 2Linked to original sources

PubMed1988

Tumor necrosis factor can induce both apoptic and necrotic forms of cell lysis.

TNF is a protein toxin which is secreted by activated macrophages and monocytes. Although the cytotoxic activity of TNF has been well documented, the mechanism of TNF-induced lysis is not well understood. The goal of this investigation was to determine whether TNF caused one of the classic forms of cell death, i.e., apoptosis, which is characterized by nuclear disintegration and cytoplasmic "boiling," or necrosis, which is characterized by the formation of a "balloon-like" plasma membrane and a lack of nuclear disintegration. Therefore, to distinguish apoptosis from necrosis, we have used time-lapse video microscopy to observe the death of several TNF-sensitive target cell lines while measuring the release of Na2(51)CrO4 and [3H]TdR from cytoplasmic and nuclear compartments, respectively. As targets we selected two spontaneously sensitive cell lines, F17 and L-M, and one resistant cell line, C3HA, which was sensitized by treatment with cycloheximide or by infection with the adeno-SV40 hybrid virus Ad2+ND2. We find that the type of cell death observed depends on the cell being tested. For example, in F17 cells we found that TNF treatment induced a classical form of apoptosis. In contrast, TNF induced a necrotic form of cell death in L-M cells, similar to the lysis induced by antibody and C. Finally, we found that sensitized C3HA cells displayed a novel cytolytic phenotype which resembled apoptosis but did not include DNA fragmentation. These results emphasize the complex nature of the TNF-induced cytotoxic response.

Adenoviruses, Human↗

PubMed1988

Target-induced changes in macrophage migration may explain differences in lytic sensitivity among simian virus 40-transformed fibroblasts.

Time-lapse video microscopy has been used to study macrophage movement in the presence of SV40-transformed fibroblasts. In all, five different SV40-transformed cell lines (an uncloned line and four clones derived from it) were tested for their affects on the movement of LPS-activated macrophages (AM). Conditions for video microscope recording were designed to simulate, as best as possible, the conditions used in a 51Cr-release cytotoxicity assay. Our analysis shows that these targets had a range of effects on macrophage migration, from stimulation to complete inhibition of movement. Two of the targets we tested (SV-COL and SV-COL-E8) both highly sensitive to lysis, stimulated macrophage movement, inducing an "agitated" response. Two other targets (SV-COL-F11 and SV-COL-H5), both of intermediate sensitivity had, for the most part, a suppressive effect on macrophage movement. Finally, we found that one target, clone SV-COL-F5, which is completely resistant to lysis by AM, was able to completely inhibit macrophage movement. This inhibitory effect was accompanied by a "pathologic" change in the structure of the macrophage cytoplasm suggesting that this target escapes lysis in vitro by secreting a factor that incapacitates the effector cell. Overall, these observations suggest that target cell products that can affect the behavior of the AM may be important in determining the lytic sensitivity of transformed target cells.

Animals↗

PubMed1987

Platelet-derived growth factor stimulates phagocytosis and blocks agonist-induced activation of the neutrophil oxidative burst: a possible cellular mechanism to protect against oxygen radical damage.

The effect of platelet-derived growth factor (PDGF) on agonist-induced activation of the superoxide-generating oxidative burst in human neutrophils was tested. PDGF had no effect on the resting level of superoxide generation but inhibited both the rate and the extent of fMet-Leu-Phe-stimulated superoxide production in a dose-dependent manner. The concentration required to inhibit the response by 50% was 95 +/- 26 pM (n = 10). PDGF also blocked activation by other receptor-mediated agonists such as the complement protein C5a and opsonized zymosan, but not by phorbol myristate acetate or arachidonate, both of which may act at postreceptor sites. The growth factor, however, had no effect on the binding of fMet-Leu-Phe to its receptor. PDGF in concentrations that blocked the oxidative burst stimulated phagocytosis of opsonized latex particles. Thus, PDGF functions as a heterologous "down-regulator" of receptor-mediated activation of the neutrophil oxidative burst and an activator of phagocytosis. A model for a feedback regulatory loop between platelets and neutrophils is proposed.

Blood Platelets↗

PubMed1985

Dynamic gene interactions in the evolution of rabbit VH genes: a four codon duplication and block homologies provide evidence for intergenic exchange.

Two rabbit VHa-negative genes, RVH831 and RVH832, were isolated from a single genomic fragment selected by hybridization with the mouse VHIII gene S107V1. RVH831 is a pseudogene with a frameshift mutation in FR3 and a 19 bp deletion within the VH-D splice site. In contrast, RVH832 has an open reading frame and an intact VH-D splice site and thus may be functional. However, RVH832 displays a unique 4 codon duplication/insertion in FR1 that may be the result of an unequal exchange event between two ancestral VH genes. Sequence comparisons between these and other rabbit VH genes reveal patterns of shared blocks of nucleotide substitutions, suggestive of gene conversion. A high overall homology (greater than or equal to 73%) between the compared VH nucleotide sequences suggests that rabbit VH genes may not be organized in clearly divergent families or subgroups.

Animals↗

PubMed1983

Response of the rabbit VH subgroups a and y following antigenic stimulation.

In order to test the relative immune competence of a minor VH subgroup (y), the concentrations of rabbit Ig bearing the VH subgroups a and y were determined in experimentally immunized rabbits. Three groups of three rabbits each were immunized with either Micrococcus luteus, bovine serum albumin or an alloantigenic determinant (b4) of the rabbit kappa light chain. The ratios of VHy to VHa Ig in the pre-immune sera ranged from 1:8 to 1:283. In five of the nine rabbits, the VHy to VHa ratios in affinity purified antibody (Ab) closely reflected the ratios of VHy to VHa in pre-immune serum. Of the remaining four rabbits, three produced disproportionately low amounts of VHy Ab, when compared to pre-immune values, while one rabbit produced virtually no VHy Ab. Regardless of the levels of VHy and VHa Ab detected, the kinetics of Ab production by each subgroup during the course of the immune response were similar to each other. When total serum levels of the subgroups were analysed, all rabbits displayed increases (6-450%) in total serum levels of both VHa Ig and total IgG following immunization, while in eight out of nine rabbits the level of VHy Ig remained unchanged. Our observations suggest that the immune repertoire of the minor VHy subgroup is sufficiently diverse to generate VH regions which react with a variety of antigens, and in five out of nine rabbits, VHy Ab-producing clones compete well with VHa Ab-producing clones. In addition, it appears that clones of the major and minor VH subgroups, once triggered, are under similar regulatory controls with respect to specific Ab, but differ with respect to non-specific Ig production.

Animals↗