Pharmacokinetics, tissue distribution, and cell localization of [35S]methionine-labeled recombinant human and murine alpha interferons in mice.

The pharmacokinetics, tissue distribution, cell localization, and penetration into tumor xenografts of recombinant [35S]methionine-labeled human alpha interferon (HuIFN-alpha) and murine alpha interferon (MuIFN-alpha) were examined in mice. Both interferons (IFNs) were removed from the blood in a rapid biphasic manner; HuIFN-alpha was cleared faster than MuIFN-alpha. Tissues were analyzed for radioactivity and over 90% of the IFNs was accounted for.

Characterization of interferons produced by peripheral blood mononuclear cells from healthy subjects in response to Corynebacterium parvum or poly I: poly C.

The biological significance of acid labile interferon alpha is presently unknown. We examined the putative production of acid labile interferon in vitro from human peripheral blood mononuclear cells induced with Corynebacterium parvum or poly I: poly C. Both agents induced up to 1200 IU/ml interferon, and the interferon was 80 to 90% acid labile.

Comparison of different ELISAs for the detection of monoclonal antibodies to human interferon-alpha. Implications for antibody screening.

Three enzyme-linked immunosorbent assays (ELISA) were compared in the initial screening of some 400 hybridoma supernatants for antibodies to a recombinant human interferon-alpha subtype, 4a (IFN-alpha 4a). In these assays, (i) the antigen was coated directly to polystyrene microtitre plates (ELISA-PS), (ii) the antigen was coated directly to nitrocellulose (ELISA-NC), or (iii) the antigen and mouse antibody were reacted in solution and the resulting complex immobilized to a solid support precoated with polyclonal rabbit anti-IFN-alpha antibody (ELISA-SW). The ELISA-PS detected eight antibodies, the ELISA-NC 15 and the ELISA-SW 18.

Interferons in rheumatoid arthritis: alterations in production and response related to disease activity.

Various markers associated with the production of and response to interferons (IFNs) were studied in patients with either inactive rheumatoid arthritis (RA) or active RA, and in healthy subjects. The IFN markers assessed were serum and synovial fluid (SF) levels, the activity in peripheral blood leukocytes (PBL) of (2'-5') oligoadenylate synthetase (OAS), and the production in vitro by PBL of IFN-alpha/beta in response to Sendai virus or Poly(I):Poly(C) as inducers, and of IFN-gamma using PHA or Con A as inducers. IFN activity, tested by antiviral assays using two different cell lines, was not demonstrable in the serum of any patient with RA.

Isolation and characterisation of monoclonal antibodies against hydrophobic membrane subunit 9 of the yeast mitochondrial H+-ATPase.

Five stable lines of myeloma-spleen cell hybrids, producing antibodies against the proteolipid subunit 9 of the yeast mitochondrial H+-ATPase F0-sector, have been isolated by immunizing mice with a proteolipid preparation in the presence of sodium dodecyl sulphate. One of these monoclonal antibodies also reacted with subunit 8 of the enzyme complex indicating a shared epitope. The antibodies did not react with the holo-H+-ATPase, suggesting that their epitopes are shielded by other subunits of the enzyme complex.

Natural killer cells in viral arthritis.

Changes in natural killer (NK) cell activity were studied in patients with polyarthritis associated with rubella or Ross River virus infections. In 30 of 32 Ross River virus patients, peripheral NK cell activity was depressed at some stage of the disease but returned to normal levels as patients recovered from arthritic symptoms. Similar changes did not occur in rubella patients and no difference was found between changes in peripheral NK activity and serum interferon (IFN) levels in rubella patients with arthritis and those without.

Antiviral and antiproliferative activities of interferon-alpha 1: the role of cysteine residues.

Site-specific in vitro mutagenesis was used to direct serine for cysteine substitutions within the sequence of human interferon-alpha 1 (IFN-alpha 1). Antiviral specific activities and antiproliferative activities of IFN-alpha 1 analogs, expressed in M13 as fusion proteins, were assessed following purification by monoclonal antibody affinity chromatography. Based on analysis of IFN-alpha 2, IFN-alpha 1 contains two disulfide bridges between cysteine residues 29 and 139 and cysteine residues 1 and 99.

Cellular localization of alpha-interferon in hepatitis B virus-infected liver tissue.

Cells expressing alpha 2-interferon were identified by indirect immunofluorescence using both a polyclonal and a monoclonal anti-alpha-interferon antibody reagent. In hepatitis B or delta virus infection, focal clusters of alpha-interferon-positive infiltrating mononuclear cells and (to a lesser extent) fibroblasts were regularly seen in liver sections from patients who had chronic active hepatitis and cirrhosis and evidence of virus replication, but in a minority of patients with chronic persistent hepatitis B and not in nonvirally infected livers. This report provides evidence for local alpha-interferon production near the site of virus replication in hepatitis B infection, identifies mononuclear cells and fibroblasts (but not hepatocytes) as the main cell types producing interferon in this infection and suggests that locally produced alpha-interferon may be a natural regulator of virus replication in HBsAg-positive chronic active hepatitis.

Characterization of epitopes of the yeast mitochondrial H+-ATPase complex recognized by monoclonal antibodies.

Nine monoclonal antibodies which react with the beta subunit of the yeast mitochondrial H+-ATPase and three which react with a 25 kDa subunit of the enzyme complex (P25) have been characterized. Competitive binding studies indicated the presence of at least four antigenic regions on the beta subunit of the enzyme complex. One antigenic region of the beta subunit is recognized by two monoclonal antibodies RH 57.

Production of a monoclonal antibody to human interferon-alpha (IFN-alpha) and its use to identify IFN-alpha-producing cells in virus infection in vivo.

A monoclonal antibody to human interferon-alpha (IFN-alpha) was produced using affinity-purified IFN-alpha, that reacted with recombinant human IFN-alpha 2, but not with IFN-alpha 1, IFN-alpha M1 or IFN-beta. Indirect immunofluorescence using this monoclonal (designated 6C3) and anti-IFN-alpha polyclonal antibodies identified cells expressing IFN-alpha. After Sendai virus induction of normal human buffy-coat cells the proportion of monocytes and lymphocytes expressing IFN-alpha rose progressively from 0% to 50% and 34% respectively, preceding peak IFN-alpha titres in the culture supernatants.

Single amino acid substitutions at conserved residues of human interferon-alpha can effect antiviral specific activity.

Site-specific in vitro mutagenesis was used to direct various amino acid substitutions at conserved positions within the sequence of human interferon-alpha 1 (IFN-alpha 1). The antiviral specific activity of IFN-alpha 1, expressed in M13 as a fusion protein [IFN-alpha 1 (phi WT)], could be altered by single amino acid substitutions. The substitution of glycine for tyrosine at position 123 results in a loss of more than 99% of the antiviral specific activity on human cells, but causes no significant change in the antiviral specific activity on primary bovine cells.

Monoclonal antibodies against subunits of yeast mitochondrial H+-ATPase.

Fourteen stable lines of myeloma-spleen cell hybrids producing antibodies against the mitochondrial H+-ATPase have been isolated. One reacted with the alpha-subunit of the enzyme complex (Mr 56000), nine with the beta-subunit (Mr 54000), and four with a 25 kDa subunit which has not been previously characterized. These antibodies are inhibitory or stimulatory or have no effect upon the enzyme activity.

Improved conditions for the production of monoclonal antibodies to carcinogen-modified DNA, for use in enzyme-linked immunosorbent assays (ELISA).

The methodology for the production of monoclonal antibodies to chemical carcinogen-modified DNA has been improved to provide high yields of hybridomas, using guanine-imidazole ring-opened aflatoxin B1-modified DNA as an example (iro-AFB1 DNA). The percentage of immunised mice which responded to iro-AFB1 DNA-protein immunisation and the number of specific hybridomas produced was dependent on the level of modification of DNA. One in three BALB/c mice had detectable (but low) antibody titre when 0.

[Diaphragmatic eventration in adults. Apropos of 20 cases].

Twenty adults with a mean age of 49 were operated on between 1972 and 1980 for eventration of the diaphragm. The etiology was probably traumatic in 11 cases; it was congenital in 2 and degenerative in 7. The functional signs were usually respiratory (55%) or digestive (10%).

The release of 4,4'-diaminobiphenyls from azodyes in the rat.

Six azodyes derived from benzidine, o-tolidine or o-dianisidine were separately administered orally by gavage to rats. Urine was collected over a 24 h period. Following dichloromethane extraction, urines were analysed by h.

Production of monoclonal antibodies to guanine imidazole ring-opened aflatoxin B1DNA, the persistent DNA adduct in vivo.

Monoclonal antibodiess were produced following immunisation of mice with guanine imidazole ring-opened aflatoxin B1 DNA (iro AFB1 DNA), coupled electrostatically to methylated keyhole limpet haemocyanin. Three monoclonal hybridoma lines producing antibodies specific for iro AFB1 DNA were grown as ascites tumours and suitable dilutions of the ascitic fluid (1:8000-1:50,000) used in a competitive enzyme linked immunosorbent assay (ELISA) to measure reactivity of the antibodies to a variety of aflatoxin and nucleic acid-related compounds. These antibodies recognise AFB1 bound to DNA at levels 10(4)-10(5) times lower concentration than unmodified calf thymus DNA or 8,9-dihydro-8,9-dihydroxy-aflatoxin B1: and show 2-5 times the affinity to iro AFB1 DNA compared to AFB1 DNA.

The binding of 7,12-dimethylbenz[a]anthracene to mammary gland macromolecules in the hamster:effects of Enovid feeding or transplacental exposure to diethylstilboestrol.

The covalent binding of 7,12-[3H]dimethylbenz[a]anthracene ([3H--DMBA) to mammary gland macromolecules was studied in hamsters fed a contraceptive mixture, Enovid, those exposed transplacentally to diethylstilboestrol (DES), and controls. Compared with rats, hamsters are relatively resistant to DMBA mammary carcinogenesis, but susceptibility is increased by either of the above treatments with Enovid or DES. The amount of DMBA bound to DNA and protein ws 4-5 times greater than to RNA, but only DNA binding was persistent.

The apparent inhibition of urothelial DNA synthesis in neonatal rats by dietary saccharin.

[3H]Thymidine ([3H]TdR) incorporation into urothelial DNA of male neonatal rats was measured autoradiographically at birth and during the first 3 weeks of life. The rats were derived from control parents and those fed saccharin (1, 3, 5 and 7.5%) in the diet from before pregnancy.

[Neurogenic endothoracic tumours (author's transl)].

123 nervous thoracic tumours have been operated without immediate postoperative death. From 1964, 86 are the subject of a new study. 80 % were situated in the posterior gutter and were involving the sympathetic chain and intercostal-nerves.