Tumor Necrosis Factor Antibody Research Articles

Induction of an endogenous tumor necrosis factor in mice by murine recombinant interferon-gamma combined with a lipid A subunit analog (GLA-60) of low toxicity.

We have investigated the endogenous production of a serum cytotoxic factor when recombinant interferon-gamma (rIFN-gamma) is combined with synthetic lipid A subunit analogs of low toxicity (GLA compounds). The cytotoxic activity of the serum was measured by the crystal violet staining method with L929 cells as a target. Intravenous administration of rIFN-gamma followed by intravenous administration of lipopolysaccharide induced the endogenous production of a cytotoxic factor in the serum. The priming effect of rIFN-gamma appeared immediately and persisted for approximately 20 h after the injection. Administration of lipopolysaccharide as a trigger enhanced the production of the cytotoxic factor in the serum maximally 2 h after the injection. The cytotoxic activity in the serum was completely inhibited by anti-(mouse tumor necrosis factor) (TNF) antibody. A synthetic lipid A subunit analog (GLA-60), which is much less toxic in its endotoxin activities than lipopolysaccharide or synthetic lipid A (compound 506), induced the endogenous production of serum TNF in rIFN-gamma-primed mice. GLA-60 entrapped within liposomes induced the production of serum TNF in rIFN-gamma-primed mice more effectively than GLA-60 solubilized in phosphate-buffered saline. Intravenous or intranasal administrations of rIFN-gamma followed by intranasal administration of GLA-60 produced TNF in the lung washing fluid but not in the serum, indicating that TNF production can be induced locally rather than systemically by the alteration of the administration route of the primer and trigger. These results indicate that GLA-60, a lipid A subunit analog of low toxicity, is a beneficial triggering agent in the production of endogenous TNF, as well as having other immunopharmacological properties, and may provide a basis for cancer (metastases) treatment as a result of its ability to induce endogenous TNF.

Role of tumour necrosis factor in the enhanced sensitivity of mice to endotoxin after exposure to lead.

Heavy metals administered to animals, at doses which appear relatively innoxious by themselves, enhance susceptibility to endotoxin. The mechanisms which underly this phenomenon are not yet fully understood. In this study we investigated the role of the cytokine Tumour Necrosis Factor (TNF), an important mediator of the effects of endotoxin, in this phenomenon. First it was studied whether lead enhances sensitivity of mice to endotoxin and to TNF. Lead appeared to enhance sensitivity to both endotoxin and TNF resulting in mortality of mice at low endotoxin and TNF doses. Next we studied the influence of lead on serum TNF levels after stimulation by endotoxin. Lead treated mice showed lower TNF blood levels two hours after injection of endotoxin and lead. Six and eight hours after injection TNF levels of lead treated mice were higher compared to those of mice injected with endotoxin only. In the last part of our investigation, we studied the influence of a monoclonal hamster anti TNF antibody on the effect of combined lead-endotoxin exposure. Administration of the antibody prevents lethality completely. Our data indicate that TNF plays a central role in the phenomenon of the enhanced susceptibility of animals to endotoxin after exposure to lead. The enhanced susceptibility to endotoxin is caused by an enhanced susceptibility to TNF and possibly by a prolonged exposure to a higher level of TNF.

Inhibition of Chlamydia trachomatis replication in HEp-2 cells by human monocyte-derived macrophages.

Monocytes (M) and macrophages are important components of the immune response to foreign agents. Using an in vitro system, we studied the influence of human M and M-derived macrophages (MdM) on the replication of Chlamydia trachomatis (L2/434) in HEp-2 cells. M or MdM were added to infected cells at a ratio of 4:1, and the resultant chlamydial yield was evaluated in one-step growth experiments. Chlamydial DNA production was evaluated by dot hybridization. Both M and MdM reduced chlamydial yield and DNA production, but the reductions caused by MdM were more pronounced. Electron microscopy showed that while control HEp-2 cells at 48 h postinfection contained large inclusions in which most particles were elementary bodies, the infected HEp-2 cells exposed to MdM contained small vacuoles with abnormal reticulate bodies and very few typical elementary bodies. Separation of the MdM from the HEp-2 cells by a membrane reduced the inhibitory effect of the MdM relative to that of MdM in direct contact with the infected cells. Addition of tumor necrosis factor antibodies to C. trachomatis-infected HEp-2 cells exposed to MdM (either in direct contact or separated by a membrane from the infected cells) reduced the inhibition of chlamydial DNA production. These data suggest the possibility that MdM may modulate C. trachomatis replication in vivo.

Cytolytic heterogeneity of polymorphonuclear leukocytes in killing of murine tumor cells.

Murine inflammatory polymorphonuclear leukocytes (PMN) and inflammatory ascitic fluid were found to contain cytotoxin. This cytotoxin was not oxygen metabolites, but a protein with a molecular weight of 70 kilodaltons as determined by high‐performance liquid chromatography. However, the cytolytic activity was completely neutralized by anti‐murine tumor necrosis factor (TNF) antibody, suggesting that PMN cytotoxin is a TNF‐like molecule or TNF itself. Cytolytic heterogeneity of PMN was also demonstrated: immature PMN obtained from ascitic fluid 6 hr after caseinate injection showed stronger cytotoxicity than mature PMN obtained 3 hr after the injection. However, mature PMN (3 hr) contained more cytotoxin than immature PMN (6 hr). This discrepancy may be due to differences in the abilities of these PMN to synthesize and release cytotoxin. These results indicate that not only macrophages, but also inflammatory PMN can produce a TNF‐like cytotoxin and that cytolytic heterogeneity of PMN exists in tumor killing.

Inhibition of CMV replication in human fibroblasts by human monocyte-derived macrophages: implication for CMV persistent infection

The effect of monocytes (M) and monocyte-derived macrophages (MdM) on cytomegalovirus (CMV) replication in human fibroblasts (HF) was studied by one step growth experiments, plaque formation and dot hybridization with a labeled CMV DNA probe. HF were infected with CMV at multiplicities of infection (mol) of 0.001 to 1. After virus adsorption, M or MdM were added at an effector target ratio of 2:1. MdM reduced both infectious viral yield and the quantity of viral DNA and inhibited viral plaque formation. M, however, affected these parameters to a lesser extent. Electron microscopic studies showed that MdM treated CMF-infected HF, 4 days pi, contained only a few capsids in their nuclei and many vacuoles in their cytoplasm as compared to CMV infected HF (control). A reduction of CMV DNA inhibition was observed upon incubation of the infected HF cells with MdM separated from the infected cells by a membrane. Addition of tumor necrosis factor (TNF) antibody to CMV-infected HF incubated with MdM either in direct contact or separated by a membrane from the infected cells reduced the inhibition of CMV-DNA production. The results of this study suggest that MdM may modulate CMV replication in vivo and may also have a role in CMV persistence or chronic infection.