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PubMed1994

Nucleotide sequence analysis of Marek's disease virus (MDV) serotype 2 homolog of MDV serotype 1 pp38, an antigen associated with transformed cells.

Marek's disease (MD) virus (MDV) serotype 1 (MDV1) pp38, an antigen associated with MD-transformed cells, has been identified as an MDV1 serotype-specific antigen by analyses using specific monoclonal antibodies (MAbs). In the present study, we determined the region in the MDV serotype 2 (MDV2) genome that has homology with the MDV1 pp38 gene. As a result of sequence analysis, it was determined that an open reading frame (ORF) existed in this region. Furthermore, Northern blot analysis and reverse transcriptase polymerase chain reaction confirmed the existence of an RNA transcript related to the ORF in MDV2-infected cells. Computer analyses revealed that MDV1 pp38 could be divided into two parts; one part was highly homologous and the other part was not homologous with the peptides deduced from the ORF of MDV2, indicating that the epitope recognized by a MDV1 pp38-specific MAb is located in the latter part.

Amino Acid Sequence↗
PubMed1994

Synthesis of recombinant baculoviruses expressing the outer capsid protein VP2 of five BTV serotypes and the induction of neutralizing antibodies to homologous and heterologous BTV serotypes.

DNA representing RNA segment L2 of 5 different bluetongue virus (BTV) serotypes (BTV-1, -2, -11, -13 and -17) corresponding to the gene that codes for the BTV neutralization antigen VP2, have been inserted individually into baculovirus transfer vectors in lieu of the 5' coding region of the polyhedrin gene of Autographa california nuclear polyhedrosis virus (AcNPV). After co-transfection of Spodoptera frugiperda cells with wild-type AcNPV DNA in the presence of the derived transfer vector DNAs, polyhedrin-negative recombinant baculoviruses were recovered. When S. frugiperda cells were infected with each of these viruses, protein was made similar in size and antigenic properties to the authentic BTV VP2 protein. To evaluate the biological activities of these proteins, antibodies were raised to them in guinea pigs. The neutralization capabilities of these antisera were tested against homologous and, for two of the higher titered sera, heterologous BTV serotypes. Each antiserum neutralized the infectivity of the homologous virus serotype. In heterologous virus challenges, the serum raised to the VP2 protein of BTV-13 also neutralized the infectivity of BTV-1, and to lesser extents BTV-3, -16, -8 and -9 (BTV-2, -10, -11, -15 were not tested). The serum raised to the VP2 of BTV-17 neutralized BTV-20 and -21, and to lesser extents BTV-4 and -8 (again, BTV-2, -10, -11, -15 were not tested).

Animals↗
PubMed1997

Lipo-oligosaccharides of Campylobacter jejuni serotype O:10. Structures of core oligosaccharide regions from a bacterial isolate from a patient with the Miller-Fisher syndrome and from the serotype reference strain.

Lipo-oligosaccharide (LOSa) was obtained by phenol-water extraction of bacterial cells of an isolate PG 836, identified as Campylobacter jejuni serotype O:10, from a patient who subsequently developed the Miller-Fisher syndrome (MFS). The product was separated into a water-insoluble gel of low Mr and a water-soluble component of high Mr. The structure of the core oligosaccharide region in LOSa is reported herein for comparison with LOSb from the C. jejuni O:10 reference strain, and is based on investigations carried out on: (1) O-deacylated LOSa; (2) the core oligosaccharide (OS 1a) liberated on acetic acid hydrolysis of the ketosidic linkages to lipid A, with accompanying loss of N-acetylneuraminic acid residues; (3) the product of the removal of phosphate residues from OS 1a to give OS 2a; and (4) the Smith degradation of OS 2a to yield a mixture of Os 3a and OS 4a. The results revealed that the core oligosaccharide region in LOSa from the MFS bacterial isolate had chains (1a), of which some were terminated by an N-acetylneuraminobiose [Neu5Ac(alpha 2-8)Neu5Ac] unit in a GD3 [Neu5Ac-Neu5Ac-Gal] epitope, and the inner regions of which were different from those of other C. jejuni serotypes. Similar experiments on LOSb from bacterial cells of the C. jejuni O:10 reference strain showed that the core oligosaccharide unit [1a, R = P (phosphoric monoester)] of LOSa from the MFS isolate was more uniformly complete than that of the O:10 reference strain [1b, R = AEP (2-aminoethylphosphate)] differing in the nature of the phosphate substituent at the inner heptose residue. The close structural relationship of LOSa from the MFS associated bacterium to LOSb from the O:10 reference strain runs parallel to that of the previously studied Guillain-Barré syndrome (GBS) associated bacterium typed as C. jejuni O:19 in comparison with the lipo-oligosaccharide from the reference strain. Preliminary studies on the high Mr components showed that those from the O:10 strains were indistinguishable from each other, but were structurally unrelated to those from the GBS associated C. jejuni serotype O:19 isolates and the O:19 reference strain [G.O. Aspinall, A.G. McDonald, and H. Pang, Biochemistry, 33 (1994) 250-255].

Campylobacter jejuni↗
PubMed1975

The serological relationship between Brucella spp., Yersinia enterocolitica serotype IX and Salmonella serotypes of Kauffmann-White group N.

The serological relationship between Brucella spp., Yersinia enterocolitica IX, and the group N salmonella serotypes S. godesberg, S. landau, S. morehead, S. neusdorf, S. soerenga and S. urbana was examined using agglutination, antiglobulin, complement fixation, immunodiffusion and fluorescent antibody methods. Antisera to the group N salmonella serotypes all reacted to significant titres in agglutination and complement fixation, but not antiglobulin or immunodiffusion tests with smooth brucella antigens. These antisera also reacted in agglutination, but not antiglobulin, tests with Y. enterocolitica IX. They did not react significantly in any tests with rough brucella antigens. Conversely, antisera to smooth Brucella spp. agglutinated group N salmonellas to low titre and Y. enterocolitica IX to titres similar to those given against the homologous strain. Antiserum to Y. enterocolitica IX on the other hand reacted with smooth brucella antigens to high titre in agglutination, complement fixation and antiglobulin tests, and with the group N salmonella antigens to substantial titres in agglutination tests. In direct fluorescent antibody tests, smooth Brucella strains and Y. enterocolitica IX reacted strongly with FITC-labelled antibody to Br. abortus whereas the group N salmonella strains reacted weakly. In tests with monospecific antisera to the A and M determinants of Br. abortus and Br. melitensis respectively, Y. enterocolitica IX reacted only with the antiserum to the A determinant whereas group N salmonellas reacted to low titre with both A and M antisera. The results of cross-absorption tests confirmed this relationship and suggested that the O30 antigens of group N salmonella serotypes contained antigenic determinants similar to, but not identical with, the antigenic structure shared by smooth Brucella spp. and Y. enterocolitica IX.

Absorption↗
PubMed1995

The distribution of serotype-specific plasmids among different subgroups of strains of Salmonella enterica serotype Enteritidis: characterization of molecular variants by restriction enzyme fragmentation patterns.

Four hundred and thirty-four isolates of Salmonella enterica serotype Enteritidis were studied. They were grouped into five subsets defined by either the collection criteria or the parameter which formed the basis for subsequent analysis. Seventy-seven per cent harboured the serotype-specific plasmid (SSP). In 55% of the isolates this was the sole plasmid. Molecular variation in the SSP was detected in 17 (5%) of the isolates on the basis of restriction enzyme fragmentation pattern (REFP) analysis using Pst I and Sma I. The SSP variants were further characterized using additional restriction enzymes chosen to optimize the information content and analysed using a coefficient of similarity. A variant SSP designated pOG690 showed greater resemblance to the SSP of Salmonella enterica serotype Typhimurium than Enteritidis; 89% and 68% respectively for Pst I and 79% and 55% respectively for Sma I. In respect of the Pst I data pOG690 shared at least 55 kb of DNA with the Typhimurium SSP and 37 kb with the SSP of Enteritidis. This variant was associated with poultry (duck, goose, chicken) and all isolates belonged to phage type 9b. Other variants were associated with phage types 4, 6, 6a, 9a, 11, 15 and 24. The epidemiological implications of these results are discussed.

Animals↗
PubMed1997

Nucleotide sequence analysis of two virulence-associated genes in Listeria monocytogenes serotype 1/2b and comparison with the same genes in other serotypes important in human disease.

The nucleotide sequences of the hly and plcA genes (encoding listeriolysin, a thiol-activated cytolysin and a non-specific phospholipase C, respectively) were determined for two strains of Listeria monocytogenes serotype 1/2b. The deduced amino acid sequences for listeriolysin were identical for the two strains and identical to that of listeriolysin from serotype 4b. The two serotype 1/2b strains differed in one amino acid (histidine vs asparagine) in the deduced amino acid sequence for phosopholipase C.

Cloning, Molecular↗
PubMedDate not supplied

VP2 gene based phylogenetic relationship of Indian isolates of Bluetongue virus serotype 1 and other serotypes from different parts of the world.

Bluetongue virus (BTV), a member of genus Orbivirus, family Reoviridae, is non-enveloped with double shelled structure and 10 segmented double stranded RNA genome. The RNA segment L2 encodes an outer capsid serotype specific viral protein VP2. BTV serotype 1 (BTV-1) specific novel primer pair, forward primer (1240-1271 bp) and reverse primer (1844-1813 bp), was designed using VP2 gene sequences available in GenBank to amplify 1240-1844 bp region because two hypervariable and three conserved regions have been reported within these 604 nucleotides. This primer pair successfully amplified cell culture adapted six Indian isolates of BTV-1. The 604 bp PCR product of VP2 gene of BTV-1 Avikanagar (A), Chennai (C) and Sirsa 3 (S3) Indian isolates were cloned in pPCR-Script Amp SK (+) vector and transformed into XL10-Gold Kan ultracompetent Epicurian coli cells. The positive clones selected by blue-white screening and colony touch PCR were sequenced. BTV-1A, C and S3 isolates revealed 99% nucleotide sequence identity within 1304-1844 bp region of VP2 gene. The partial VP2 gene sequences (1240-1844 bp region) revealed that BTV-1 Indian isolates were 89% identical with Australian (AUS) BTV-1 isolates while the identity with South African (SA) BTV-1 isolate was 75%. Phylogenetically, three BTV-1 Indian isolates formed one group which is closely related to BTV-1AUS isolates followed by BTV-1SA, BTV-2, 9, 23, 13, 17, 10 and 11 isolates from different parts of world. Based on partial VP2 gene sequences, it is concluded that Indian isolates of BTV-1 are closely related to BTV-1AUS isolates than BTV-1SA and other serotypes.

Animals↗
PubMed1985

Purification, characterization and immunological properties of the serotype-specific capsular polysaccharide of Pasteurella haemolytica (serotype T4) organisms.

The serotype-specific capsular polysaccharide from two strains of Pasteurella haemolytica serotype T4 organisms was purified and characterized by chemical analysis and NMR spectroscopy. The polymer, a teichoic acid, has the backbone structure ----(2-glycerol-l)----(phosphate)----(6-alpha-D-galactose-1)---- and is partially O-acetylated on the C2 and C3 galactose residues. Chemical removal of O-acetyl groups from the polysaccharide destroyed both its ability to precipitate with antiserum raised against killed whole serotype T4 organisms and its ability to adhere to sheep erythrocytes in passive haemagglutination experiments. Attempts to elicit antisera using the purified polymer were unsuccessful but a partially purified material was immunogenic.

Animals↗
PubMed1992

Structural identification of an epitope of antigenic factor 5 in mannans of Candida albicans NIH B-792 (serotype B) and J-1012 (serotype A) as beta-1,2-linked oligomannosyl residues.

In previous articles, we reported the presence of phosphate-bound beta-1,2-linked oligomannosyl residues in the mannans of strains of Candida albicans serotypes A and B and Candida stellatoidea. To identify the antigenic factor corresponding to this type of oligomannosyl residue, a relationship between chemical structure and antigenic specificity in the mannans of C. albicans NIH B-792 (serotype B, B-strain) and C. albicans J-1012 (serotype A, J-strain) was investigated by using a combination of two-dimensional 1H nuclear magnetic resonance spectroscopy of H-1, H-2, and H-5 regions in the mannans and an enzyme-linked immunosorbent assay that employed concanavalin A-coated microtiter plates. It was shown in the present 1H nuclear magnetic resonance study that an examination of chemical shifts not only in the H-1 region but also in the H-5 region was useful for the quantitative determination of the phosphate-bound beta-1,2-linked oligomannosyl residues. In the enzyme-linked immunosorbent assay using concanavalin A-coated plates, it was revealed that, of factor sera 1, 4, and 5, only factor serum 5 showed a reactivity proportional to the densities of the beta-1,2-linked oligomannosyl residues of the mannan subfractions of different phosphate contents that had been prepared from the bulk B-strain mannan by DEAE-Sephadex chromatography. The above results indicate that the phosphate-bound beta-1,2-linked oligomannosyl residues, Manp beta 1----(2Manp beta 1----)n2Man (n = 0-5), correspond to antigenic factor 5.

Antigens, Fungal↗
PubMed1995

Comparative sequence analysis of the reovirus S4 genes from 13 serotype 1 and serotype 3 field isolates.

The reovirus sigma 3 protein is a major outer capsid protein that may function to regulate translation within infected cells. To facilitate the understanding of sigma 3 structure and functions and the evolution of mammalian reoviruses, we sequenced cDNA copies of the S4 genes from 10 serotype 3 and 3 serotype 1 reovirus field isolates and compared these sequences with sequences of prototypic strains of the three reovirus serotypes. We found that the sigma 3 proteins are highly conserved: the two longest conserved regions contain motifs proposed to function in binding zinc and double-stranded RNA. We used the 16 viral isolates to investigate the hypothesis that structural interactions between sigma 3 and the cell attachment protein, sigma 1, constrain their evolution and to identify a determinant within sigma 3 that is in close proximity to the sigma 1 hemagglutination site.

Amino Acid Sequence↗
PubMed2003

Substitution of the adenovirus serotype 5 knob with a serotype 3 knob enhances multiple steps in virus replication.

Adenovirus (Ad) serotype 5 (Ad5) continues to be the predominant vector used for cancer gene therapy. However, many tumor types are reported to be relatively refractory to Ad5 infection because of low surface expression of the native Ad5 receptor, CAR. The observation that many tumor cells are CAR deficient has necessitated the development of CAR-independent infection strategies, including the introduction of heterologous ligand sequences into the virus fiber gene and immunological or chemical modifications of the capsid proteins. Alternatively, native Ad5 tropism can be modified by substituting the knob region from other Ad serotypes such as Ad type 3 (Ad3) into the Ad5 knob region. To date, the effect(s) of tropism modification on the replication and oncolytic capacity of these chimeric Ad vectors has not been fully evaluated. To address this issue, Ad5 vectors and isogenically matched chimeric vectors with Ad3 tropism (Ad5/3) were compared in this study. Various parameters of virus infection were compared, including binding, nuclear translocation, E1A transcription, transgene expression, de novo virus production, and oncolysis. Overall, the chimeric Ad5/3 virus was progressively more efficient at each step of the replication cycle compared with its Ad5 counterpart. The higher replication efficiency of the chimeric Ad5/3 vector translated into improved therapeutic efficacy in a murine in vivo tumor rejection model. These findings suggest that in addition to the initial target cell interaction, multiple mechanisms contribute to the enhanced replication of the chimeric Ad5/3 vector. Furthermore, the data demonstrate that alternative Ad serotype receptors can be used to improve infection and subsequent oncolytic replication, which is particularly relevant in gene therapy applications for tumors that are inefficiently infected with Ad5.

Adenovirus E1A Proteins↗
PubMed1975

Broad antigenic relationships among rhinovirus serotypes revealed by cross-immunization of rabbits with different serotypes.

The rhinoviruses are a large group of at least 89 serotypes and extensive cross-relationships have been demonstrated among these serotypes using both rabbit and human antisera. The extent of these cross-relationships was further investigated by sequential intravenous immunization of rabbits with different virus serotypes, known to be both related and unrelated. In all cases in which a relationship had previously been demonstrated with monovalent antiserum, after heterotypic stimulation with the related virus, the antibody titer to the virus used as primary immunogen showed an anamnestic response, whereas the antibody response to the secondary immunogen was good, but resembled a primary response. In some instances when immunized rabbits were injected with an unrelated antigen, previously unsuspected relationships were revealed. A number of clusters of related rhinoviruses were demonstrated: 5, 17, 42; 3, 4, 6, 14; 9, 32, 67; and 13, 14, 41. Heterotypic immunization proved to be a powerful biologic probe for detecting relationships among the rhinoviruses, and together with newly available epidemiologic data it offers a possible approach for design of a vaccine for the common cold.

Animals↗
PubMed1990

Replicating Marek's disease virus (MDV) serotype 2 DNA with inserted MDV serotype 1 DNA sequences in a Marek's disease lymphoblastoid cell line MSB1-41C.

We characterized the properties of herpes-type viruses which grew well in a Marek's disease lymphoblastoid cell line, MSB1-41C, inducing cytopathic effect characterized by the formation of syncytial giant cells. Examination of the infectious virus by field inversion gel electrophoresis revealed the presence of DNA of about 180 kbp in both the culture fluid and cell fractions of the infected MSB-41C cells. The DNA was found to consist of Marek's disease virus (MDV) serotype 2 (MDV2) and MDV serotype 1 (MDV1) DNA by Southern blot hybridization. The MDV1 DNA consisted of sequences mainly from the long inverted repeats including multiple copies of 132 bp direct tandem repeats. Molecular cloning of BamHI digests of the MDV2 DNA revealed a fragment of MDV1 DNA and MDV2 DNA fused together, indicating that the recombinant MDV2 DNA had been generated by genetic recombination with the latent MDV1 DNA.

Animals↗
PubMed1995

Expression of Marek's disease virus (MDV) serotype 2 gene which has partial homology with MDV serotype 1 pp38 gene.

We constructed the recombinant baculovirus expressing the gene of non-pathogenic Marek's disease virus (MDV) serotype 2 (MDV2) which encodes a polypeptide with partial homology to MDV serotype 1 (MDV1) pp38, an antigen associated with transformed cells. The recombinant MDV2 protein was detected as a band of 32 kDa in immunoblot analysis with MDV2-infected chicken serum. Mouse serum against insect Spodoptera frugiperda cells infected with the recombinant baculovirus immunoprecipitated a 38 kDa molecule from the lysate of MDV2-infected chicken embryo fibroblasts (CEFs) but did not immunoprecipitate the MDV1 pp38 from the lysate of MDV1-infected CEFs. This result indicates that the recombinant MDV2 protein has no epitopes shared with the MDV1 pp38.

Animals↗
PubMed1995

The efficacy of a live Listeria monocytogenes combined serotype 1/2a and serotype 4b vaccine.

In each of two experiments, sheep and lambs were vaccinated by a subcutaneous injection of a test vaccine (consisting of a combined serotype 1/2a and serotype 4b live Listeria monocytogenes culture) and challenged 16 days later with a mixture of the homologous wild strains. After challenge, the mortality rate of vaccinated sheep was 28.1% and that of controls 71.9%; that of lambs was 25.0%, although these had been inoculated with the LD70 dose. Furthermore, in each of two field trials in Listeria-infected flocks, primiparous pregnant ewes were vaccinated. In the first field trial, 3 or 110 lambs died of listeriosis of those born of vaccinated (n = 564) or unvaccinated (n = 3345) ewes, respectively. In the second, the perinatal mortality rate of lambs born of vaccinated or unvaccinated ewes was 7.6 or 30.3%, respectively, and the mean birth weight of lambs born of vaccinated or control ewes was 2.2 or 1.8 kg, respectively; the mean milk production of vaccinated ewes was 106 and that of controls 83 l; no Listeria was isolated from milk samples of vaccinated ewes. It is concluded that the vaccine is efficacious for the protection of sheep from listeriosis.

Animals↗
PubMed2003

A mosaic adenovirus possessing serotype Ad5 and serotype Ad3 knobs exhibits expanded tropism.

The efficiency of cancer gene therapy with recombinant adenoviruses based on serotype 5 (Ad5) has been limited partly because of variable, and often low, expression by human primary cancer cells of the primary cellular-receptor which recognizes the knob domain of the fiber protein, the coxsackie and adenovirus receptor (CAR). As a means of circumventing CAR deficiency, Ad vectors have been retargeted by utilizing chimeric fibers possessing knob domains of alternate Ad serotypes. We have reported that ovarian cancer cells possess a primary receptor for Ad3 to which the Ad3 knob binds independently of the CAR-Ad5 knob interaction. Furthermore, an Ad5-based chimeric vector, designated Ad5/3, containing a chimeric fiber proteins possessing the Ad3 knob, demonstrates CAR-independent tropism by virtue of targeting the Ad3 receptor. Based on these findings, we hypothesized that a mosaic virus possessing both the Ad5 knob and the Ad3 knob on the same virion could utilize either primary receptor, resulting in expanded tropism. In this study, we generated a dual-knob mosaic virus by coinfection of 293 cells with Ad5-based and Ad5/3-based vectors. Characterization of the resultant virions confirmed the incorporation of both Ad5 and Ad3 knobs in the same particle. Furthermore, this mosaic virus was able to utilize either receptor, CAR and the Ad3 receptor, for virus attachment to cells. Enhanced Ad infectivity with the mosaic virus was shown in a panel of cell lines, with receptor profiles ranging from CAR-dominant to Ad3 receptor-dominant. Thus, this mosaic virus strategy may offer the potential to improve Ad-based gene therapy approaches by infectivity enhancement and tropism expansion.

Adenoviruses, Human↗
PubMed1996

Invasive disease due to Haemophilus influenzae serotype f: clinical and epidemiologic characteristics in the H. influenzae serotype b vaccine era. The Haemophilus influenzae Study Group.

With the decline in the rate of infections caused by Haemophilus influenzae serotype b, H. influenzae serotype f (Hif) is becoming a relatively important cause of invasive disease due to H. influenzae. We identified 91 cases of invasive Hif infections in a multistate area over a 6-year period. The incidence of invasive Hif disease was 0.5 case per 1,000,000 population in 1989 and 1.9 cases per 1,000,000 population in 1994. The proportion of all invasive H. influenzae disease caused by Hif rose from 1% in 1989 to 17% in 1994. Seventy-two percent of cases occurred in adults, and 26% of cases occurred in children younger than 5 years of age. Respiratory tract infections accounted for 82% of adult cases, and most adults had significant underlying diseases. In children, pneumonia and meningitis each accounted for 40% of cases, respectively. Overall mortality was 30% among adults, and 21% among children. Molecular typing demonstrated limited overall diversity in Hif isolates. Continued surveillance is warranted to evaluate the trend toward the increasing incidence of Hif disease that was noted in this study.

Adult↗
PubMed1986

Purification, characterization and immunological properties of the serotype-specific capsular polysaccharide of Pasteurella haemolytica serotype A7 organisms.

The serotype-specific capsular polysaccharide from two strains of Pasteurella haemolytica serotype A7 organisms was purified and characterized by chemical analysis and by 1H and 13C NMR spectroscopy using one- and two-dimensional methods. The polymer has the repeating unit----3)-beta-2-acetamido-2-deoxygalactopyranose-(1----3)-alpha- 2-acetamido- 2-deoxy-6-O-acetyl-glucopyranose-(1-phosphate----. It was immunogenic (capable of eliciting antibodies) for sheep. Chemical removal of O-acetyl groups destroyed both the ability of the polymer to adhere to sheep erythrocytes at neutral pH and the ability to form immune precipitates with specific antisera. Studies using the protein A-gold technique in the electron microscope showed the polysaccharide to be peripherally localized on the bacterial surface.

Amino Acids↗