Transneuronal spread of intraperitoneally administered herpes simplex virus type 1 from the abdomen via the vagus nerve to the brains of mice.

The MacIntyre strain of Herpes simplex virus type 1 was inoculated intraperitoneally into young male mice. Immunohistochemical detection of the virus antigen demonstrated the passage of virus from the gastro-intestinal myenteric plexus towards the brain and its distribution within the areas of brain related to the principal nuclei of the vagus nerve. Selective trapping of the virus in the autonomic nerves and subsequent localization of antigen behind the blood-brain barrier may prevent therapeutic effects of agents tested in this model.

Antiviral activity of a novel recombinant human interferon-alpha B/D hybrid.

The antiviral potential of a novel cross-species active, recombinant human interferon-alpha B/D hybrid (rHuIFN-alpha B/D), was evaluated for its efficiacy in cultured human monocytes and in several murine models of viral disease. When examined in 14-day-old human monocyte cultures, rHuIFN-alpha B/D was highly effective in preventing viral replication and cell destruction caused by herpes simplex virus type 1 (HSV-1/VR3). The effect observed with 100 units of this hybrid IFN was as good or higher than that observed with equivalent amounts of rHuIFN-alpha A or IFN-gamma. In addition, a single dose (5 X 10(7) U/kg) of rHuIFN-alpha B/D administered several hours after intranasal infection with HSV-1/VR3 suppressed pulmonary virus replication and prevented death due to interstitial pneumonia. Similarly, mice infected with a more aggressive strain of HSV-1 (McIntyre) were protected when this IFN preparation was administered at the time of virus infection and 1 day later. The anti-retroviral activity of rHuIFN-alpha B/D was examined in two murine leukemia retroviral models, Rauscher (RMLV) and Friend (FMLV), and a murine model of acquired immunodeficiency (LP-BM5). Treatment of RMLV or FMLV infected mice significantly prolonged mean survival times and the number of long-term FMLV survivors. These therapeutic effects were demonstrated when IFN was administered on the day of virus infection or as late as 3 days following infection. Transient reversal of the immunosuppressive effects induced by LP-BM5 infection was observed when rHuIFN-alpha B/D treatment was initiated at the time of virus infection. Moreover, when rHuIFN-alpha B/D was used together with azidothymidine (AZT), the effect of the combination was better than either drug alone.

Monoclonal antibodies to human renin: properties and applications.

A series of 11 different monoclonal antibodies generated against human kidney renin have been characterised. Their binding affinity, inhibition of renin activity, epitope distribution, crossreactivity with related enzymes and finally in vivo pharmacological effects were analysed. All antibodies were found to be specific for primate renin recognising 6 independent antigenic structures on the renin molecule. They expressed different effects on renin activity namely (1) no inhibition, (2) only partial, or (3) complete inhibition. Partially inhibiting antibodies demonstrated specific degrees of inhibition (30, 60 or 80%). One antibody, R-36-16, demonstrated an IC 50 of 1.3 X 10(-11) M/L and, when injected into marmosets, induced complete inhibition of plasma renin activity and reduction of blood pressure. Using a selected pair of antibodies a radioimmunoassay has been established providing a fast and highly reproducible determination of human and marmoset immunoreactive renin, detecting both active and inactive renin down to concentrations of 10 pg/ml (1.25 X 10(-17) moles of renin per 50 microliter sample).

Antagonism of interferon induction in spleen and adherent peritoneal cells of mice by the lipophilic antiviral muramyl peptide MTP-PE.

The immunomodulator muramyl tripeptide-phosphatidylethanolamine (MTP-PE) has been shown to enhance host resistance against a variety of experimental infections and to cure influenza virus infection in mice when given in a single dose, even at a late stage of the disease. Tests of its capacity to induce alpha/beta- and gamma-interferon (IFN-alpha/beta and -gamma) in vitro demonstrated that it is neither an inducer nor a primer of IFN synthesis. On the contrary, we found that it inhibits the induction of IFN-alpha/beta and -gamma by poly(rI:rC), Newcastle disease virus, lipopolysaccharide, or concanavalin A in adherent cells from the peritoneal cavity and spleen of mice. The antiviral activity of already induced or exogenously added murine IFN was, however, not impaired.

Enhancement of host resistance against virus infections by MTP-PE, a synthetic lipophilic muramyl peptide--I. Increased survival in mice and guinea pigs after single drug administration prior to infection, and the effect of MTP-PE on interferon levels in sera and lungs.

Muramyl tripeptide-phosphatidylethanolamine (MTP-PE, CGP 19835) displays prophylactic antiviral activity in mice infected with influenza viruses A and B, parainfluenza 1 virus or herpes simplex type 1 viruses (HSV/1) and in guinea pigs infected with herpes simplex type 2 viruses (HSV/2). MTP-PE is effective when given in a single intranasal dose as early as 1-4 weeks before infection. In the case of HSV/2 infections, prophylactic effectiveness can be demonstrated after a single topical application into the vagina seven days before infection. Antiviral effects are observed in response to doses as little as 0.001 mg/kg bodyweight. The activity of the substance seems to be inversely related to the size of the viral inoculum, but poor dose-effect relation is demonstrable in a dose-range extending over four to five orders of magnitude. Furthermore, the compound is devoid of antiviral effects in vitro. MTP-PE does not induce interferon (IFN) in serum and lung, nor does it influence kinetics or quantity of serum and lung IFN content in the course of viral infections. However, when given intranasally 7 days before an oral dose of tilorone, increased levels of IFN in lung suspensions are observed.

Immunodiagnosis of schistosomiasis outside endemic areas: use of histamine release from basophils by schistosomal antigens and radioallergosorbent test.

Specific allergic histamine release from leucocytes, radioallergosorbent tests (RAST) and indirect fluorescent antibody tests (IFAT) were applied for immunodiagnosis of schistosomiasis in patients outside endemic area. Of 10 parasitologically verified cases--all of them exhibiting a low and irregular egg output--8 were detected by histamine release, whereas 4 patient with filariasis, 3 with trichuriasis and 31 parasitologically normal controls were negative in this respect. By a combination of the histamine release test, RAST and IFAT all 10 cases were diagnosed. RAST and IFAT applied to 29 patients with active or treated schistosomiasis were positive in 14 (48%) and 20 (69%) cases, respectively.

Formation of antibodies to synthetic human calcitonin during treatment of Paget's disease.

Low-titre antibodies to synthetic human calcitonin (hCT) were detected in a 69-year-old woman suffering from Paget's disease who was treated for 16 months with hCT and the diphosphonate EHDP. Levels of antibody were highest between 10 and 18 months after commencement of therapy, slowly decreased after completion of treatment and were later no longer measurable. There was no immunological response to a single re-injection of hCT 14 months after discontinuation of therapy. Formation of antibodies to hCT in man is a very rare event, this being the first recorded case of an immune response to synthetic human calcitonin, whereas synthetic salmon calcitonin induces an immune response in a high percentage (up to 78%) of the patients treated with this hormone.

Nor-MDP, saponin, corynebacteria, and pertussis organisms as immunological adjuvants in experimental malaria vaccination of macaques.

Vaccination of primates against malaria using antigen derived from erythrocytic parasite stages has been most successful where Freund's complete adjuvant has been employed. Since this adjuvant is clinically unacceptable its replacement is a matter of urgency.In the present work a muramyldipeptide derivative (nor-MDP) given in mineral oil has proved to be partially effective as an adjuvant for merozoite vaccination of Macaca mulatta against Plasmodium knowlesi, and saponin has proved to be effective in similar vaccination of M. fascicularis.

Radioallergosorbent and indirect fluorescent antibody tests in immunodiagnosis of schistosomiasis.

Radioallergosorbent tests for specific IgE antibodies to Schistosoma haematobium and S. mansoni were positive in 82 and 72%, respectively, of sera from 136 African schistosomiasis patients. 99% of sera from controls (5 free of parasitic disease, 75 infested with hookworm, Ascaris, Trichuris, and/or Onchocerca) were negative. When titres in indirect fluorescent antibody tests were set at the same level of specificity only 38% of patients' sera were positive. A cut-off at a higher concentration increased sensitivity to 65% but lowered specificity (10% controls positive).

Synthetic human calcitonin: analysis of antibodies obtained from various animal species and determination of immunoreactive hormone in human sera.

Antibodies to synthetic human calcitonin (hCT) were developed in rabbits, goats and mice. The free peptide (32 amino-acid residues, Mwt. 3418) was administered together with adjuvant, and the effect of various immunization procedures, as well as of different dose-levels, was evaluated comparatively. Synthetic hCT was found to be a good immunogen for the three animal species examined. The relative importance of various structural parts of the hCT molecule with regard to immunological specificity was determined by reference to the inhibition of the specific binding of 125I-hCT to antibodies by peptide fragments of hCT. All the antisera studied were directed to structural and/or conformational properties of the 11-28 or 11-32 amino acid sequence of hCT. Six different antisera from rabbits and goats were selected for radioimmunological assay of hCT on the basis of their inhibitory dose50-values and immunological specificity. To improve the sensitivity of the radioimmunoassay (RIA), we studied the preparation of radioiodinated hCT and assessed various parameters determining the sensitivity of the assay. Despite all the efforts, CT in human plasma from healthy subjects could not be determined with certainty. The difficulties encountered in the determination of normal levels of circulating CT are discussed in terms of the sensitivity of RIA and non-specific interference of serum factors with RIA.

Human parathyroid hormone. Immunological characterization of antibodies against a glandular extract and the synthetic amino-terminal fragments 1-12 and 1-34 and their use in the determination of immunoreactive hormone in human sera.

Antibodies to a urea-trichloroacetic acid extract [hPTH-(TCA)] of human parathyroid tumors and to the synthetic NH(2)-terminal fragments of human parathyroid hormone hPTH-(1-12) and -(1-34) were developed in goats to characterize immunochemically various PTH preparations and to estimate immunoreactive PTH (iPTH) in human sera. They were quantitated on the basis of their capacity to bind [(131)I]-hPTH-(1-12), [(131)I]hPTH-(1-34) or [(131)I]bovine PTH (bPTH-(1-84)). The quality of the antibodies was assessed by reference to inhibition of their interaction with labeled peptides by synthetic hPTH comprising 34 NH(2)-terminal amino acid residues or fragments thereof [hPTH-(1-12), -(13-34), -(18-34), -(25-34), -(18-24)] or by the Sephadex G-100-purified full-length peptide hPTH-(1-84) [hPTH-(1-84)G-100]. The synthetic peptides used in this work correspond in their structure to the NH(2)-terminal amino acid sequence 1-34, as elucidated by Brewer and collaborators (1972. Proc. Natl. Acad. Sci. U. S. A.69: 3583-3588). Inhibition studies were also carried out with bPTH-(1-34) and bPTH-(1-84). Anti-hPTH-(TCA) exhibited specificities directed to determinants in the COOH-terminal and NH(2)-terminal part of hPTH-(1-84) and exhibited cross-reactivity with bPTH-(1-84). Anti-hPTH-(1-34), on the other hand, showed immunological specificities mainly directed to antigenic determinants located in the COOH-terminal half of hPTH-(1-34). In addition, some reactivity with the NH(2)-terminal hPTH-(1-12) and with the extractive full-length peptides of human and bovine origin was observed. Antibodies to hPTH-(1-12) cross-reacted with hPTH-(1-34) and -(1-84)G-100.IPTH WAS RADIOIMMUNOLOGICALLY DETERMINED IN HUMAN SERA BY THE FOLLOWING SYSTEMS: (a) [(131)I]bPTH-(1-84), anti-hPTH-(TCA) and hPTH-(1-84)G-100 as standard; (b) [(131)I]hPTH-(1-34), anti-hPTH-(1-34) and hPTH-(1-34) as standard. With system (a), COOH-terminal fragments of hPTH-(1-84) having a molecular weight of approximately 7,000 were detected, and there was an almost total discrimination of serum iPTH levels in normal and in hyperparathyroid subjects. With system (b), on the other hand, several molecular species of iPTH were detected, including a component larger than hPTH-(1-84) and others similar to hPTH-(1-84) and to a fragment co-eluting with the NH(2)-terminal fragment hPTH-(1-34). When serum iPTH was assayed in system (b), there was a large overlap of iPTH levels in control subjects and in patients with primary hyperparathyroidism.