Modulation of apoptosis by V protein mumps virus.

BACKGROUND: The Urabe AM9 vaccine strain of mumps virus contains two variants of V protein: VWT (of HN-A1081 viral population) and VGly (of HN-G1081). The V protein is a promoting factor of viral replication by blocking the IFN antiviral pathway. FINDINGS: We studied the relationship between V protein variants and IFN-α2b-induced apoptosis. V proteins decrease activation of the extrinsic IFN-α2b-induced apoptotic pathway monitored by the caspase 8 activity, being the effect greater with the VWT protein. Both V proteins decrease the activity of caspase 9 of the intrinsic apoptotic pathway. In a system without IFN, the VWT and VGly proteins expression promotes activation of caspases 3 and 7. However, when the cellular system was stimulated with IFN-α, this activity decreased partially. TUNEL assay shows that for treatment with IFN-α and ibuprofen of cervical adenocarcinoma cells there is nuclear DNA fragmentation but the V protein expression reduces this process. CONCLUSIONS: The reduction in the levels of caspases and DNA fragmentation, suggesting that V protein, particularly VWT protein of Urabe AM9 vaccine strain, modulates apoptosis. In addition, the VWT protein shows a protective role for cell proliferation in the presence of antiproliferative signals.

Endotoxin-neutralizing effects of histidine-rich peptides.

Inflammatory responses of human peripheral blood monocytes to the Gram-negative endotoxin lipopolysaccharide (LPS) are enhanced by structurally diverse substances, such as anionic polysaccharides or cationic polypeptides. Only a few substances are known to effectively blunt LPS-induced monocyte activation. We now show that synthetic poly-L-histidine (Hn) binds to LPS and abrogates the release of the proinflammatory cytokine interleukin-8 (IL-8) in LPS-stimulated human whole blood. LPS-induced stimulation of monocytes was strictly pH-dependent with only minor amounts of IL-8 secreted in acidic blood. Maximum levels of IL-8 secretion occurred at a strongly basic pH. Hn inhibition of the release of IL-8 from LPS-stimulated monocytes was observed under acidic, neutral and physiological conditions. With increasing alkalosis, the effectiveness of Hn was gradually lost, suggesting that protonated, but not deprotonated, Hn was effective in inhibiting LPS-induced monocyte responses. Histidine-rich protein 2 from the malaria parasite, Plasmodium falciparum, inhibited the ability of LPS to evoke an inflammatory response in CD14-transfected THP-1 cells. Further, a short synthetic peptide derived from human histidine- and proline-rich glycoprotein also exhibited LPS-inhibitory effects in CD14 transfectants. Taken together, these observations demonstrate the capacity of histidine-rich peptides, irrespective of their origin, to neutralize LPS-induced proinflammatory host responses.

Up-regulation of cytokine production in alveolar macrophages by histogranin, a novel endogenous pentadecapeptide.

Recently, histogranin (HN), a newly found pentadecapeptide, was shown to enhance tumor necrosis factor (TNF) production by alveolar macrophages (AM). In this study, we have investigated whether HN was present in tissues rich with immune cells and further explored the effect of HN and [Ser1]HN on the production of TNF and other key cytokines. Relatively high levels of immunoreactive (ir)-HN were found in rat lung (14.9 pmol/g) and spleen (12.3 pmol/g), indicating its localization in close proximity to macrophages/monocytes and lymphocytes. Furthermore, HN and [Ser1]HN (10(-8)-10(-7) M) stimulated basal and lipopolysaccharide (LPS)-induced interleukin 1 (IL-1) mRNA expression and IL-1 release from rat AM. [Ser1]HN also stimulated basal and LPS-induced interleukin-6 (IL-6) release. Although HN did not affect the kinetics of cytokine production, the maximal enhancing effect of HN was seen at 3 h for TNF, 6 h for IL-1 and 18 h for IL-6. These data indicate that HN can up-regulate a cytokine cascade involving TNF, IL-1 and IL-6 and suggest a role for this endogenous peptide in immune regulation.

gp330 associates with a 44-kDa protein in the rat kidney to form the Heymann nephritis antigenic complex.

Using antibodies isolated from glomeruli of nephritic rats we have previously identified a 330-kDa cell surface glycoprotein (gp330) as a major pathogenic antigen of Heymann nephritis (HN), an experimental model of human membranous glomerulonephritis. Recently, we have isolated a cDNA clone, C14, encoding a polypeptide that contains a pathogenic epitope of HN responsible for the initiation of the disease. Subsequently, another protein, alpha 2-macroglobulin receptor-associated protein (alpha 2-MRAP), which is a subunit of the receptor for human alpha 2-macroglobulin/low density lipoprotein receptor-related protein (LRP), was shown to possess a high degree of sequence homology to the C14 protein (C14p). In this report, we have investigated the relationship between gp330, C14p, and alpha 2-MRAP. Immunoprecipitation studies demonstrate that gp330 forms a heterodimeric association with a 44-kDa polypeptide that is stable to detergent extraction and long-term centrifugation. Further, immunoblotting analysis on the purified complex indicates that the 44-kDa associated protein shares immunological identity to C14p and alpha 2-MRAP. In addition, antibodies eluted from glomeruli of HN rats and antibodies to a C14 fusion protein immunoprecipitated gp330 and the 44-kDa protein, demonstrating that the epitopes responsible for the initial events of HN are accessible within the complex. Based on these data, three models are proposed to explain how pathogenic epitopes in the gp330-44-kDa, HN antigenic complex may be presented at the cell surface and initiate the onset of HN.

[Antigenic differences in the glycoproteins of the Newcastle disease virus]. / Antigenni razliki pri gliukoproteinite na virusa in Niukiasulskata bolest (NDV).

The two glucoproteins of the Newcastle disease virus - hemagglutinin-neuraminidase (HN) and F-protein - were isolated in a pure state from two strains of the virus: the La Sota apathogenic strain and the Roussev strain pathogenic for birds. It was found that the large HN glucoprotein of each of the strains was not identical antigenically with the F-protein. These seemed to be partial antigenic relatedness between the HN of the two strains. On the other hand, the F-proteins of the strains were identical.