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Biomedical subjects

A H Warfel

Publications and source records attributed to A H Warfel.

14 recordsLinked to original sources
PubMed1995

Synergism between interferon-gamma and cytokines or lipopolysaccharide in the activation of the HIV-LTR in macrophages.

Macrophages (møs) obtained from transgenic mice carrying the HIV long terminal repeat-chloramphenicol acetyl transferase (LTR-CAT) sequence were used to study the influence of various biologic response modifiers (BRMs) on the activation of LTR-directed CAT expression. It was found that LPS or IL-6 alone induced moderate levels of CAT expression, whereas IFN-gamma or TNF-alpha had no significant effect. Co-exposure of møs to IFN-gamma and either LPS, IL-6, or TNF-alpha led to significant synergistic increases in CAT levels. Levels were also synergistically augmented when møs were exposed first to IFN-gamma (priming), washed, and then exposed to either LPS, IL-6, or TNF-alpha. Although IL-6 was synergistic with subsequently added IFN-gamma, the reverse sequence of addition was more effective. LPS and TNF-alpha were inactive when added before IFN-gamma. Initial priming signals were rapid as exposure for 3 h to IFN-gamma was sufficient to prepare the cells for subsequent activation by other BRMs. These results suggest that the duration of latency and progression of HIV infections may be greatly influenced by events such as intercurrent infections that cause IFN-gamma production, thereby priming møs to respond to other cytokines that have been reported to be constitutively elevated during the course of infection with HIV (e.g., IL-6 and/or TNF-alpha).

Animals↗
PubMed1992

Specific ligation of surface alpha-D-galactosyl epitopes markedly affects the quantity of four major proteins secreted by macrophages.

Activated macrophages (M phi s) have terminal alpha-D-galactosyl (alpha D-Gal) residues on their membranes that are not apparent on resting cells. Ligation of these epitopes with Griffonia simplicifolia I-B4 (GSI-B4), a lectin that has specificity for alpha-D-Gal residues, alters selected M phi functions. To explore the mechanism(s) that may be responsible for some of the functional changes, alterations in the secretory pattern of [35S]methionine-labeled proteins were assessed when cells were cultured with or without this ligand. The proteins were identified by Western blots and quantitated. Interestingly, alpha-D-Gal ligation proved to decrease the secretion of some proteins while increasing the secretion of others. Some of the most significant changes were observed in four proteins: fibronectin and transglutaminase were down-regulated by 55 and 66% respectively, while plasminogen activator inhibitor type 2 was increased by 259% and collagenase was increased 1000-fold. These observations show that the emergence of new oligosaccharide epitopes, such as alpha-D-Gal, concomitant with M phi activation may serve to mediate the transduction of signals that cause quantitative changes in the elaboration of diverse M phi products. The biologic significance of the four identified proteins has been well established. Fluctuations in their levels are likely to play a role at sites of chronic inflammation.

Animals↗
PubMed1991

Macrophage membrane glycoproteins that bind Griffonia simplicifolia I-B4: effect on cytotoxicity and protein secretion.

Thioglycollate elicited peritoneal (TG-Møs) but not resident peritoneal Møs (R-Møs) were found to bind the lectin Griffonia simplicifolia isotype I-B4 (GSI-B4). This was demonstrated by ultrastructural studies and FACS analyses. Membranes from TG-Møs were isolated, separated on SDS-PAGE, electrotransferred onto nitrocellulose, and exposed to peroxidase-labeled GSI-B4. These procedures revealed two major membrane glycoproteins of molecular weights 180,000 and 94,000 daltons that bound the lectin GSI-B4 which has a specificity for recognizing terminal alpha-galactosyl residues. The presence of these epitopes on the two membrane glycoproteins was further substantiated by the fact that treatment of the membranes with alpha-galactosidase destroyed their capacity to bind GSI-B4 and that alpha-D-galactopyranoside but not N-acetyl-D-glucosamine competitively inhibited GSI-B4 from binding to the glycoproteins. Treatment of TG-Møs with GSI-B4 reduced the capacity of interferon (IFN) and lipopolysaccharide (LPS), or IFN alone, to induce Mø mediated cytotoxicity towards tumor cells by as much as 40%. GSI-B4 also caused alterations in the pattern of biosynthetically 35S-methionine labeled secreted proteins as early as 2 hours after contact with TG-Møs. Out of 35 discernible proteins on fluorograms of SDS-PAGE separated proteins, 5 were down-regulated and 9 were enhanced. It is suggested that the two novel Mø membrane proteins may play a role in regulating the response of Mø subpopulations to their humoral and cellular environments.

Animals↗
PubMed1991

Cystatin C and cathepsin B production by alveolar macrophages from smokers and nonsmokers.

The capacity of alveolar macrophages (AM) obtained from smokers and nonsmokers to secrete cathepsin B and its inhibitor cystatin C was examined because of the concept that an imbalance in the production of proteolytic enzymes and/or their inhibitors could be responsible for the lung damage seen in smokers. Quantitation of immunoprecipitates on Western blots showed that the amount of total cystatin C secreted into the culture medium by AM of smokers was significantly greater than the amount secreted by cells obtained from nonsmokers, whereas the difference between the amount of cathepsin B secreted by the AM of smokers and that from nonsmokers did not appear significant. The cystatin C found in the medium conditioned by AM of nonsmokers appeared to be more heterogeneous in molecular size, presenting either as a single band of about 14 Kd or as a high-molecular-weight triplet of about 69 Kd, 63 Kd, and 57.3 Kd. Furthermore, in some cases there were single or doublet bands at 14 Kd as well as the high-molecular-weight triplets. In contrast, smokers AM-conditioned medium uniformly possessed both the low-and the high-molecular-weight cystatin C. Cathepsin B was not detected in Western blots at its reported molecular weights but was identified at the exact area occupied by the higher molecular weight cystatin C, i.e., at bands corresponding to 69 Kd, 63 Kd, and 57.3 Kd. Therefore, it is clear that in culture media of AM, cystatin C and cathepsin B are present as proteinase-antiproteinase complexes. The observation also suggests that in smokers an excess of cystatin C may be elaborated, which, if further substantiated, would show for the first time a likely role for this proteinase inhibitor in vivo.

Adult↗
PubMed1987

Constitutive secretion of cystatin C (gamma-trace) by monocytes and macrophages and its downregulation after stimulation.

Cystatin C (gamma-trace) was found to be a constitutively secreted protein of isolated human monocytes and mouse peritoneal macrophages, as well as the histiocytic lymphoma cell lines U937, P388D.1, and J774. This proteinase inhibitor is not uniquely secreted by monocytes/macrophages, but was also identified in the conditioned media from several primary cells, including brain cells, and diverse established cell lines. In vitro treatment of resident mouse peritoneal macrophages with either LPS or IFN-gamma caused a downregulation in cystatin C secretion. Elaboration of this protein was also diminished by macrophages that had been stimulated by thioglycollate in vivo, and treatment of these cells with LPS led to further decline. It is suggested that, under some inflammatory conditions, downregulation of cystatin C may contribute to tissue pathology.

Animals↗
PubMed1986

Down-regulation of macrophage lysozyme by lipopolysaccharide and interferon.

Lipopolysaccharide (LPS) treatment of resident mouse peritoneal macrophages (M phi) was found to suppress intracellular as well as secreted lysozyme (LZM). Interferon (IFN) had a similar effect. LZM was identified by the capacity of cell lysates or medium to lyse Micrococcus lysodeikticus, and by the presence of a 14.5 Kd protein band which co-migrated with human LZM in SDS-PAGE and which reacted positively in Western blots with antiserum to human LZM. The size of the 14.5 Kd band decreased sequentially with increasing concentrations of LPS to which the cells were exposed. Although the LPS influence on LZM levels was dose-dependent, the intracellular LZM pool responded more readily than secreted LZM. Maximal intracellular LZM suppression of 80% was obtained with 10 micrograms LPS, whereas secreted LZM was reduced by only 66%. An IFN concentration of 100 U reduced secreted LZM by 24%, whereas 10,000 U of IFN decreased the amount of LZM secreted by 71%. Thioglycolate-elicited M phi had 75% less intracellular LZM than untreated resident M phi. Moreover, thioglycolate-elicited M phi were hyporesponsive to the suppressive effects of LPS added in vitro. Because both LPS and IFN have been shown to stimulate numerous M phi functions, the data are of interest because they support the concept, based on other studies, that agents which are capable of enhancing some M phi activities may concomitantly down-regulate other functions.

Animals↗
PubMed1980

Production and initial characterization of guinea pig interferon.

Guinea pig cell cultures produced extremely low levels of interferon when induced with Newcastle disease virus (NDV), or double-stranded RNA, poly rI.poly rC. Priming guinea pig cells with mouse interferon or guinea pig serum interferon did not significantly enhance interferon production. However, intracardial injection of 10(9) plaque forming units of NDV into guinea pigs lead to interferon production to levels over 6000 units per ml of serum 6 hr after the inducer was administered. The antiviral agent in the serum had the characteristics of interferon. Guinea pig interferon showed low levels of activity on mouse and bovine cells, and no detectable activity on human or rabbit cells.

Animals↗
PubMed1978

Macrophage fusion factor elicited from BGG-sensitized lymphocytes.

Lymphocytes obtained from rabbit lymph nodes sensitized to bovine gamma globulin produce in vitro the lymphokine macrophage fusion factor (MFF) which mediates the fusion of approximately 100% of normal alveolar and oil-induced peritoneal macrophages. Giant cells (GC) of Langhans and foreign body type form large syncytia containing as many as several hundred nuclei per cell. Nuclei of GC appear more spherical and larger than those of the normal mononucleated macrophages, and they possess several prominent nucleoli. Giant cells of peritoneal macrophage origin show enhanced intracytoplasmic vacuolization. Normal macrophages cultured as a monolayer in MFF-rich supernatants form cell clusters which progressively fuse during the 24-hour incubation period. A signoid dose-response curve was obtained for cell fusion with MFF-rich supernatants possessing high titers, ie, the latter supernatants undiluted partially inhibited macrophage fusion. MIF-like activity was detected in MFF-rich supernatants as well as a factor(s) which inhibited 3H-thymidine uptake by giant cells.

Animals↗
PubMed1970

Macrophage membranes viewed through a scanning electron microscope.

When rabbit peritoneal macrophages were cultured in serum and Tyrode medium, numerous membranous structures varying in size and transparency were observed to cover the surface in complex cascading patterns. Modifications in the methods of fixation and drying probably accounted for the new perspective of macrophage surfaces gained in this study.

Animals↗