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Biomedical subjects

D M Moore

Publications and source records attributed to D M Moore.

At least 73 records · Page 4Linked to original sources
PubMed1985

Radioimmunoassay for detection of VP1 specific neutralizing antibodies of foot and mouth disease virus.

A solid-phase radioimmunoassay was developed for the detection of antibodies against a specific region of the VP1 protein of the A24 and 01 serotypes of foot and mouth disease virus. The antibody titers from the radioimmunoassay showed a positive correlation with neutralizing antibody titers determined by a mouse protection assay. The specificity of the assay resides in the peptide used as antigen. The assay is rapid, reproducible and does not require the use of whole virions.

Animals↗
PubMed1985

Sequence variation in the gene for the immunogenic capsid protein VP1 of foot-and-mouth disease virus type A.

The nucleotide sequences have been determined and compared from cloned cDNA genes coding for the foot-and-mouth disease virus (FMDV) immunogenic capsid protein, VP1, from eight different A subtypes: A5 Westerwald/58, A12 119ab (large plaque variant), A22 550 USSR/65, A24 Cruzeiro Brazil/55, A27 Cundinamarca Colombia/76, A32 Venezuela/70, A Venceslau Brazil/76, and A Argentina/79. We have also found sequence variations among different cDNA clones of the A5 and A24 subtypes. There are regions of nucleotide sequence within the VP1 gene that vary considerably among the subtypes as well as other regions that remain relatively constant. One highly variable region (codons 130-171) encodes amino acids previously identified as being exposed on the virus surface and constituting an important immunogenic site of the virus. There potentially exist secondary structures within the viral RNA sequences that code for this immunogenic site that could decrease the fidelity of replication at this sequence. The rapid generation of FMDV variants encouraged by such structures in the RNA could work together with various selective pressures to explain the observed accumulation of immunologically distinct viruses of the FMDV A type.

Aphthovirus↗
PubMed1985

Nucleotide and amino acid sequence coding for polypeptides of foot-and-mouth disease virus type A12.

The coding region for the structural and nonstructural polypeptides of the type A12 foot-and-mouth disease virus genome has been identified by nucleotide sequencing of cloned DNA derived from the viral RNA. In addition, 704 nucleotides in the 5' untranslated region between the polycytidylic acid tract and the probable initiation codon of the first translated gene, P16-L, have been sequenced. This region has several potential initiation codons, one of which appears to be a low-frequency alternate initiation site. The coding region encompasses 6,912 nucleotides and ends in a single termination codon, UAA, located 96 nucleotides upstream from a 3'-terminal polyadenylic acid tract. Microsequencing of radiolabeled in vivo and in vitro translation products identified the genome position of the major foot-and-mouth disease virus proteins and the cleavage sites recognized by the putative viral protease and an additional protease(s), probably of cellular origin, to generate primary and functional foot-and-mouth disease virus polypeptides.

Amino Acid Sequence↗
PubMed1985

Dose-response evaluation of a genetically engineered foot-and-mouth disease virus polypeptide immunogen in cattle.

Four groups of 9 cattle each were vaccinated with 10, 50, 250, or 1,250 micrograms of foot-and-mouth disease (FMD) virus A12 VP1 fusion protein that was produced in Escherichia coli and emulsified in an oil adjuvant. The groups given the 10 and 50 micrograms of antigen were revaccinated at 15 weeks and were challenge exposed at 30 weeks; 5 of 9 and 7 of 9 cattle, respectively, were protected from FMD virus infection. The remaining 2 groups, vaccinated with 250 or 1,250 micrograms of antigen, were revaccinated at 32 weeks and were challenge exposed at 45 weeks; 8 of 9 and 9 of 9 cattle, respectively, were protected. The results indicated that the biosynthetic polypeptide FMD vaccine was effective using vaccination intervals frequently followed with conventional whole-virus vaccines.

Animals↗
PubMed1984

Purification and immunogenicity of fusion VP1 protein of foot and mouth disease virus.

A procedure has been developed to purify foot and mouth disease virus (FMDV) VP1 surface antigens from recombinant Escherichia coli. The VP1 antigens are expressed as fusion proteins derived from the E. coli Trp operon and VP1 surface protein of FMDV. The procedure is capable of recovering greater than 96% of the desired product at a purity of greater than 96%. The resulting antigens induce significant levels of virus-neutralizing antibody in guinea pigs and cattle as determined by a mouse protection assay [Skinner, H.H. (1952) Proc. Int. Vet. Congr., 15th 1, 195]. E. coli contaminants have a deleterious effect on ion-exchange chromatography as well as immunogenicity of the expressed fusion VP1 antigens. The method presented removes significant E. coli contaminants, yielding fusion VP1 proteins which are immunogenically potent. In particular, virus neutralization titers at 100-micrograms dosage of the fusion VP1 proteins of the O1 and A24 serotypes are similar to that induced by the natural VP1 proteins isolated from FMD virions.

Antigens, Viral↗
PubMed1984

Location of neutralizing epitopes defined by monoclonal antibodies generated against the outer capsid polypeptide, VP1, of foot-and-mouth disease virus A12.

The epitopes of six monoclonal antibodies generated against type A12 foot-and-mouth disease virus (FMDV) VP1 or its largest cyanogen bromide fragment (13 kd) were characterized. Five of these monoclonal antibodies neutralized viral infectivity. Solid-phase and competitive antigen binding assays using virion-derived antigens or a biosynthetic VP1 polypeptide identified two distinct neutralizing epitopes. One epitope was located between amino acid residues 145-168 of VP1 and the other between amino acids 169-179. The results indicate that antibodies reacting with two distinct areas of the VP1 polypeptide are capable of neutralizing FMD virus.

Animals↗
PubMed1984

Biochemical map of polypeptides specified by foot-and-mouth disease virus.

Pulse-chase labeling of foot-and-mouth disease virus-infected bovine kidney cells revealed stable and unstable viral-specific polypeptides. To identify precursor-product relationships among these polypeptides, antisera against a number of structural and nonstructural viral-specific polypeptides were used. Cell-free translations programmed with foot-and-mouth disease virion RNA or foot-and-mouth disease virus-infected bovine kidney cell lysates, which were shown to contain almost identical polypeptides, were immunoprecipitated with the various antisera. To further establish identity, some proteins were compared by partial protease digestion. Evidence for a membrane association of the polypeptides coded for by the middle genome region is also presented. A biochemical map of the foot-and-mouth disease virus genome was established from the above information.

Animals↗
PubMed1984

A simple technique for blood collection in the opossum (Didelphis virginiana).

The left and right brachiocephalic veins were used for rapid collection of 5-10 ml blood samples in adult opossums. These vessels were also used to collect 0.5-1 ml blood samples in 120-day-old opossums. The animals were anaesthetized with 30 mg/kg of ketamine hydrochloride given intramuscularly prior to blood collection.

Anesthesia↗
PubMed1983

Identification of amino acid and nucleotide sequence of the foot-and-mouth disease virus RNA polymerase.

Foot-and-mouth disease virus (FMDV) RNA polymerase was purified from the polyethylene glycol (PEG)-treated supernatant of infected cell media by a combination of ion-exchange chromatography, membrane molecular filtration, and affinity chromatography. The purified RNA polymerase which migrated as a single band of 56,000 molecular weight on a polyacrylamide gel was subjected to automated Edman degradation and the sequence of the first 30 amino acid residues established. On the basis of previous evidence, which indicated that the RNA polymerase was the most 3'-translated polypeptide, plasmids containing cDNA mapping at the 3' end of the genome were characterized by restriction enzyme analysis and nucleotide sequencing. These investigations definitively established the derived amino acid sequence by confirmation of 28 of the amino terminal residues determined by amino acid sequence analysis; the location of the FMDV RNA polymerase coding region at the extreme 3' end of the genome, 96 nucleotides from the poly(A) tail; and the N-terminal cleavage point of the RNA polymerase from its precursor P100 was found to be a glutamic acid-glycine bond.

Amino Acid Sequence↗
PubMed1983

Identification of an exposed region of the immunogenic capsid polypeptide VP1 on foot-and-mouth disease virus.

Iodination of intact foot-and-mouth disease virus results in the selective labeling of VP1, substantiating its exposed location on the virion. A comparison of tryptic peptides revealed that a single tyrosine-containing peptide was labeled with iodine on intact or protease-cleaved virus. The labeled peptide from intact and protease-cleaved virus was characterized by molecular weight sizing and sequence analysis. Carboxypeptidase digestion of intact VP1, limited trypsin-cleaved VP1, and VP1 purified from bacterially contaminated tissue cultures yielded carboxyterminal residues of leucine, valine-arginine, and serine-alanine, respectively. The correlation of these findings with previous data on the amino acid sequence derived from nucleotide sequencing of serotypes A12 and O1 of foot-and-mouth disease virus VP1 places the probable exposed antigenic region of VP1 in a serotype-variable region including residues 136 through 144.

Amino Acid Sequence↗
PubMed1983

Contagious ecthyma in lambs and laboratory personnel.

Contagious ecthyma, diagnosed in three lambs, was transmitted to two researchers having direct contact with oral secretions from these lambs. Intracytoplasmic viral particles were demonstrated by electron microscopy in gingival biopsies from one lamb. Lamb to lamb transmission was most likely caused by use of a contaminated gavage feeding tube. Concern for the effects of this disease and Q fever on patients having contact with contaminated medical researchers prompted the formulation of safety guidelines to prevent potentially disastrous zoonotic disease.

Animals↗
PubMed1983

Venipuncture sites in armadillos (Dasypus novemcinctus).

Several venipuncture sites were evaluated in nine-banded armadillos. Included were the jugular, subclavian, cephalic, saphenous, and ventral tail veins, as well as cardiac puncture. Animals were anesthetized with 25 mg/kg ketamine hydrochloride prior to sampling. All sites yielded adequate blood volumes for hematology and serum chemistry studies.

Animals↗
PubMed1982

Foot-and-mouth disease virus: immunogenicity and structure of fragments derived from capsid protein VP and of virus containing cleaved VP.

Peptide fragments were obtained from the immunogenic capsid protein VP3, ca. 24 kilodaltons (kd), of foot-and-mouth disease virus type A12 119ab by three procedures: (1) spontaneous proteolysis of in virion VP3 in tissue cultures to produce a 15 kd peptide, designated S fragment; (2) trypsin treatment of purified virus to produce a 16 kg peptide, designated T fragment; and (3) cyanogen bromide cleavage of purified VP3 to produce a 13 kd fragment. Following isolation and purification by gel electrophoresis, VP3 and each of the three fragments were immunogenic for livestock. Lyophilization appeared to impair the immunogenicity of VP3. In addition, viruses containing VP3 fragments produced either by the spontaneous- or trypsin-induced proteolysis were as immunogenic as virus with its VP3 intact. Amino acid sequencing of N-terminal regions revealed that the S fragment was homologous with the N-terminus of VP3, whereas the 13 kd fragment possessed a unique N-terminus. Thus, putative common immunogenic amino acid sequences would appear to reside within an overlap region of the 15 kd S and 13 kd fragments. Sequencing of cDNA prepared to viral genome RNA provided three kinds of information: it (1) placed the above overlap region in the second and third quarters of VP3; (2) demonstrated that the codons for the C-terminus of VP1 and N-terminus of VP3 are contiguous; and (3) supported earlier evidence that these same codons program a chain reversal where VP1 and VP3 are joined in the precursor polyprotein.

Amino Acid Sequence↗
PubMed1982

Neutralization of foot-and-mouth disease virus. II. Further parameters related to the sensitization of the 140S virion by antibody.

The reaction of foot-and-mouth disease virus (FMDV) with 12S subunit/140S virion cross-reactive (sensitizing) antibody was studied in order to elucidate the requirements for neutralization versus sensitization. The presence of sensitizing antibody in immune serum caused an atypical in vitro neutralization response curve and a non-neutralized fraction. Cell-associated (cytophilic) antibody was not present in the system. Dissociation of the immune complex was not a factor and sensitized virus adsorbed to host cells via the regular virus receptor site(s). This finding led to the conclusion that sensitizing antibody is specific for non-critical sites. Dosing of the neutralization reaction mixtures with fractionated antibody of alternative antigenic specificities had an antagonistic effect on the neutralization response, suggesting steric hindrance. Cell receptor sites were a factor in sensitization since different host systems had different susceptibilities for sensitized antigen. The results suggest that in vitro neutralization of FMDV requires the attachment of multiple antibody molecules as proposed by the multi-hit theory of neutralization. The in vitro measurement of serum neutralizing activity as an indication of the in vivo immune response is discussed.

Antibodies, Viral↗
PubMed1981

Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine.

A DNA sequence coding for the immunogenic capsid protein VP3 of foot-and-mouth disease virus A12, prepared from the virion RNA, was ligated to a plasmid designed to express a chimeric protein from the Escherichia coli tryptophan promoter-operator system. When Escherichia coli transformed with this plasmid was grown in tryptophan-depleted media, approximately 17 percent of the total cellular protein was found to be an insoluble and stable chimeric protein. The purified chimeric protein competed equally on a molar basis with VP3 for specific antibodies to foot-and-mouth disease virus. When inoculated into six cattle and two swine, this protein elicited high levels of neutralizing antibody and protection against challenge with foot-and-mouth disease virus.

Amino Acid Sequence↗
PubMed1981

Neutralization of foot-and-mouth disease virus. I. Sensitization of the 140S virion by antibody also reactive with the 12S protein subunit.

The in vitro interaction of foot-and-mouth disease virus (FMDV) with an immune serum resulted in a fraction of virus which failed to be neutralized. This inability of antibody to neutralize the entire population of a virus preparation was studied with emphasis on the antigenic specificity of the antibody-virus reaction. Antibody to FMDV recognized multiple antigenic determinants. Immunoabsorbent fractionation of the serum into 12S subunit cross-reactive and 140S virion-specific antibody revealed that these multiple antigenic determinants are factors in determining the neutralizing ability of the antibody. Antibody specific to the infective 140S virion neutralized virus effectively, whereas antibody reactive with both the 140S virion and 12S non-infective component did so ineffectively. Neutralization was independent of viral aggregation, strain, or type heterogeneity, dissociation of the immune complex, heterogeneity of antibody class, or incubation time. The non-neutralized fraction of virus was not due to insufficient antibody in the system, was demonstrated to be complexed with antibody ('sensitized') and could be neutralized with anti-globulin serum. The findings demonstrate the heterogeneity of antibody specificity of FMDV in serum preparations and relate the importance of antibody specificity to the neutralization of virus in vitro.

Animals↗
PubMed1981

Effects of canavanine treatment on herpesvirus morphogenesis in cultured cells.

L-Canavanine, a naturally occurring analog of arginine, effectively inhibited the morphogenesis of herpes simplex virus (HSV) and human cytomegalovirus (HCMV) when added at the time of infection, but allowed the expression of several cytopathic changes. Exposure to canavanine at progressively later times ultimately led to qualitatively normal virion maturation. Under no conditions were morphologically aberrant viral particles observed. However, HSV-infected cells treated at 3 h postinfection or later did contain distinctive cytoplasmic inclusions resembling HCMV dense bodies. Recovery experiments showed that HCMV-infected cells exposed to canavanine for 1-5 days could support normal viral morphogenesis when washed free of this agent.

Canavanine↗