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1.
J Immunol Methods ; 334(1-2): 104-13, 2008 May 20.
Article in English | MEDLINE | ID: mdl-18394641

ABSTRACT

Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-alpha) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-alpha represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-alpha2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7x10(7) M(-1) to 1.4x10(10) M(-1). An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-alpha biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-alpha family, including the recombinant cytokines hIFN-alpha2a and hIFN-alpha2b and a heterogeneous collection of IFN-alpha produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-alpha subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-alpha expression.


Subject(s)
Antibodies, Monoclonal/immunology , Immunoglobulin Fragments/immunology , Immunoglobulin Variable Region/immunology , Interferon Type I/immunology , Interferon-alpha/immunology , Animals , Antibody Affinity , Cloning, Molecular , Epitope Mapping , Female , Humans , Immunoglobulin Fragments/isolation & purification , Immunoglobulin Variable Region/isolation & purification , Interferon-alpha/antagonists & inhibitors , Mice , Mice, Inbred BALB C , Neutralization Tests , Polymerase Chain Reaction , Recombinant Proteins
2.
Biochimie ; 90(3): 437-49, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18039474

ABSTRACT

Human alpha interferons (hIFN-alpha) comprise a family of closely related proteins that block viral infection, inhibit cell proliferation and modulate cell differentiation. Recombinant hIFN-alpha2 has proved useful for the treatment of a variety of human viral diseases and cancers. However, the clinical use of this cytokine has been restricted due to its short circulating half-life, which makes frequent dosing over an extended period necessary. To circumvent this problem, a glycoengineering strategy was carried out using site-directed mutagenesis. Fourteen mutants were constructed by the insertion of one N-glycosylation consensus sequence into different positions of the cytokine. Mutations were focused on amino acid positions that were believed not to be critical for the protein's structure or function. Taking into account the retained specific in vitro bioactivity and the higher carbohydrate content, five N-glycosylation positions were selected to be introduced into the molecule. Successive increases in molecular weight were observed after each addition of a functional consensus sequence, resulting in analogs with 4 and 5 N-linked carbohydrates (4N- and 5N-IFN) with increased size and charge, factors that reduce renal clearance of proteins. Pharmacokinetic experiments showed a similar behavior of 4N- and 5N-IFN variants, with a 25-fold increase in the elimination half-life and a 20-fold decrease in the systemic clearance rate compared with the non-glycosylated rhIFN-alpha2 following subcutaneous administration to rats. Besides, both distribution and elimination half-lives of the 4N analog were longer in comparison with the non-glycosylated cytokine, determining a 10-fold increase in the area under the curve after intravenous inoculation. Thus, herein we describe for the first time heavily glycosylated IFN analogs with a remarkable improvement in pharmacokinetic properties, which allow us to project drugs that combine less frequency of administration with enhanced therapeutic efficacy.


Subject(s)
Interferon-alpha/genetics , Interferon-alpha/pharmacokinetics , Animals , Binding Sites , CHO Cells , Cells, Cultured , Cricetinae , Cricetulus , Dogs , Glycosylation , Humans , Interferon alpha-2 , Interferon-alpha/metabolism , Mutation , Protein Engineering , Rats , Recombinant Proteins
3.
Vet Microbiol ; 103(1-2): 1-12, 2004 Oct 05.
Article in English | MEDLINE | ID: mdl-15381260

ABSTRACT

The equine herpesvirus 1 (EHV-1) was isolated in Argentina from an aborted equine foetus in 1979. This virus (SPv) has special restriction patterns (RP) in comparison with other Argentine isolates. In addition, SPv could be distinguished on the basis of its pathogenicity in baby mice inoculated intracerebrally. We studied the growth properties of the SPv in cell culture and its effects in a mouse respiratory and abortion model. We observed that SPv did not modify its capacity to grow in cell culture with respect to reference HH1 strain. Nevertheless, we found significant differences between the titres of the two strains at 8-14 h post-infection (PI). In this work we demonstrated that SPv showed low virulence in female at different stages of gestation, consistently, with results found in the mouse respiratory model. We considered that this low virulence of SPv could be related to its RP because the RP of HH1 strain are similar to those of the HVS25A strain and both showed effect on pregnant mice. More specific studies about genomic alterations to the SPv are necessary for identifying, more clearly, if the intra-strain variations have relation with the low virulence in the mouse respiratory and abortion model.


Subject(s)
Fetal Death/veterinary , Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/pathogenicity , Horse Diseases/virology , Animals , Antibodies, Viral/blood , Argentina , Body Weight , DNA, Viral/chemistry , DNA, Viral/genetics , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Fetal Death/pathology , Fetal Death/virology , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Herpesvirus 1, Equid/genetics , Herpesvirus 1, Equid/growth & development , Herpesvirus 1, Equid/isolation & purification , Horse Diseases/pathology , Horses , Immunohistochemistry/veterinary , Lung/pathology , Lung/virology , Mice , Mice, Inbred BALB C , Polymerase Chain Reaction/veterinary , Pregnancy , Specific Pathogen-Free Organisms , Virulence , Virus Replication
4.
Biotechnol Lett ; 25(18): 1545-8, 2003 Sep.
Article in English | MEDLINE | ID: mdl-14571980

ABSTRACT

The peptide, Ala-Pro-Ala-Arg (APAR), was selected from the screening of a tetrapeptide combinatorial synthetic library as the ligand for affinity purification of an anti-Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) monoclonal antibody (Mab) developed in mouse ascitis. The affinity chromatographic matrix obtained by attachment of APAR to agarose, having a peptide density of 0.5 micromol ml(-1), showed a maximum capacity of 9.1 mg Mab ml(-1) and a dynamic capacity of 3.9 mg Mab ml(-1). A 95% yield of electrophoretically pure anti-GM-CSF was obtained in a single step.


Subject(s)
Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/isolation & purification , Chromatography, Affinity/methods , Combinatorial Chemistry Techniques , Granulocyte-Macrophage Colony-Stimulating Factor/chemistry , Peptide Library , Peptides/chemistry , Peptides/isolation & purification , Antibodies, Monoclonal/immunology , Granulocyte-Macrophage Colony-Stimulating Factor/immunology , Ligands , Peptides/immunology
5.
Rev Sci Tech ; 22(3): 1029-33, 2003 Dec.
Article in English | MEDLINE | ID: mdl-15005559

ABSTRACT

This paper describes the first isolation of equine arteritis virus (EAV) in Argentina. The virus was isolated from the semen of an imported seropositive stallion held in isolation at a breeding farm in Tandil in the Buenos Aires Province. In addition, viral nucleic acid was detected in seminal plasma using the reverse-transcription polymerase chain reaction. The isolated virus was propagated in cell cultures and confirmed as EAV by indirect immunofluorescence and virus neutralisation, using a serum specific for the reference Bucyrus strain of EAV. As far as the authors are aware, this is the first time that EAV has been isolated in South America. The equine industry is very important for Argentina and international movement of horses is very intensive. This finding may have effects on the international trade of horses and semen from Argentina.


Subject(s)
Arterivirus Infections/veterinary , Equartevirus/isolation & purification , Horse Diseases/virology , Semen/virology , Animals , Antigens, Viral/analysis , Argentina , Arterivirus Infections/virology , Cell Line , Cytopathogenic Effect, Viral , DNA, Complementary/analysis , Equartevirus/genetics , Equartevirus/immunology , Fluorescent Antibody Technique/veterinary , Horses , Male , Neutralization Tests/veterinary , RNA, Viral/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction/veterinary
6.
Article in English | MEDLINE | ID: mdl-11471844

ABSTRACT

Equine herpesvirus 1 (EHV-1) is the causative agent of abortion, perinatal foal mortality, neurological and acute respiratory diseases in horses. Conventional laboratory diagnosis involving viral isolation from aborted foetuses is laborious and lengthy and requires processing of samples within 24 h of collection, which is problematic for samples that come from long distances. The aim of this study was to develop a polymerase chain reaction (PCR) assay useful in Argentina to detect DNA sequences of EHV-1 in different tissues from aborted equine foetuses with variable quality of preservation and without the use of conventional DNA fenolic extraction. Several DNA extraction protocols and primers were evaluated. The amplification method was standardized and its specificity was analysed using 38 foetal samples of variable quality of preservation. Of the 38 different foetal tissues, nine livers, six spleens and two lungs in good preservation and eight livers, one spleen and four lungs in a poor state of preservation were positive for PCR. EHV-1 was recovered only from the nine livers, five spleens and two lungs in good preservation. No virus was isolated from the samples that were poorly preserved. Viral isolation was confirmed by cytopathic effect and indirect immunofluorescence. The specificity of the PCR results was confirmed by the restriction endonuclease digestion of PCR products and hybridization.


Subject(s)
Abortion, Veterinary/virology , Fetus/virology , Herpesviridae Infections/veterinary , Herpesvirus 1, Equid/isolation & purification , Horse Diseases/virology , Abortion, Veterinary/embryology , Animals , DNA, Viral/analysis , Herpesviridae Infections/embryology , Herpesviridae Infections/virology , Herpesvirus 1, Equid/genetics , Horse Diseases/embryology , Horses , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/veterinary , Sensitivity and Specificity
7.
Rev Argent Microbiol ; 32(3): 109-15, 2000.
Article in English | MEDLINE | ID: mdl-11008701

ABSTRACT

In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of suid herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.


Subject(s)
DNA, Viral/isolation & purification , Herpesvirus 1, Suid/isolation & purification , Polymerase Chain Reaction , Pseudorabies/diagnosis , Swine Diseases/diagnosis , Swine/virology , Animals , Argentina/epidemiology , Blotting, Southern , Female , Pseudorabies/epidemiology , Pseudorabies/pathology , Pseudorabies/virology , Swine Diseases/epidemiology , Swine Diseases/pathology , Swine Diseases/virology , Time Factors
8.
Rev. argent. microbiol ; 32(3): 109-115, jul.-sept. 2000.
Article in English | BINACIS | ID: bin-6725

ABSTRACT

In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of suid herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.(AU)


Subject(s)
Animals , Female , RESEARCH SUPPORT, NON-U.S. GOVT , DNA, Viral/isolation & purification , Herpesvirus 1, Suid/isolation & purification , Polymerase Chain Reaction , Pseudorabies/diagnosis , Swine/virology , Swine Diseases/diagnosis , Argentina/epidemiology , Blotting, Southern , Pseudorabies/epidemiology , Pseudorabies/pathology , Pseudorabies/virology , Swine Diseases/epidemiology , Swine Diseases/pathology , Swine Diseases/virology , Time Factors
9.
Rev. argent. microbiol ; 32(3): 109-115, jul.-sept. 2000.
Article in English | LILACS | ID: lil-332528

ABSTRACT

In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of suid herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.


Subject(s)
Animals , Female , DNA, Viral , Swine Diseases/diagnosis , Herpesvirus 1, Suid , Polymerase Chain Reaction , Pseudorabies , Swine/virology , Argentina , Blotting, Southern , Swine Diseases/epidemiology , Swine Diseases/pathology , Swine Diseases/virology , Pseudorabies , Time Factors
10.
J Vet Diagn Invest ; 12(3): 266-8, 2000 May.
Article in English | MEDLINE | ID: mdl-10826843

ABSTRACT

A blocking enzyme-linked immunosorbent assay (ELISA) using a urease conjugate (U-B-ELISA) was evaluated for screening sera for antibodies to pseudorabies virus under field conditions. A total of 764 serum samples were analyzed by U-B-ELISA. Of these, 264 were evaluated by both virus neutralization and U-B-ELISA, and the results were compared. U-B-ELISA showed 98.5% and 98.9% sensitivity and specificity, respectively. This test combines the sensitivity and specificity of the blocking ELISA format while allowing visual assessment of results.


Subject(s)
Antibodies, Viral/blood , Enzyme-Linked Immunosorbent Assay/veterinary , Herpesvirus 1, Suid/isolation & purification , Pseudorabies/diagnosis , Swine Diseases/diagnosis , Animals , Argentina , Enzyme-Linked Immunosorbent Assay/methods , Herpesvirus 1, Suid/immunology , Horseradish Peroxidase/chemistry , Neutralization Tests/veterinary , Pseudorabies/blood , Pseudorabies/virology , Sensitivity and Specificity , Spectrophotometry/veterinary , Swine , Swine Diseases/virology , Urease/chemistry
11.
Rev Argent Microbiol ; 32(1): 39-43, 2000.
Article in Spanish | MEDLINE | ID: mdl-10785942

ABSTRACT

An indirect enzyme linked immunosorbent assay was developed. Infected and non infected allantoic fluids precipitated with polyetilenglycol 6000 were used as antigen and control antigen, respectively. Serum samples were diluted 1/20 and a commercial horse radish peroxidase-labelled rabbit anti-equine IgG was used as second antibody. The reaction was developed using azino-diethylbenzotyazol-sulfonate (ABTS). Cut-off was determined by ratio sample (Rs). The hemagglutination inhibition test was used as a reference test for the 391 samples analyzed. Of these, 301 sera were positive by hemagglutination inhibition test and indirect ELISA, 75 were negative by both techniques, and 15 were positive by indirect ELISA and negative by hemagglutination inhibition test. Using hemagglutination inhibition test as standard, the indirect ELISA showed a relative specificity and sensitivity of 83.3 and 100%, respectively. This indirect ELISA is useful as screening test.


Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Influenza A virus/isolation & purification , Influenza, Human/diagnosis , Orthomyxoviridae Infections/diagnosis , Animals , Hemagglutination Inhibition Tests , Humans , Rabbits , Sensitivity and Specificity
12.
Rev. argent. microbiol ; 32(1): 39-43, ene.-mar. 2000.
Article in Spanish | BINACIS | ID: bin-6714

ABSTRACT

An indirect enzyme linked immunosorbent assay was developed. Infected and non infected allantoic fluids precipitated with polyetilenglycol 6000 were used as antigen and control antigen, respectively. Serum samples were diluted 1/20 and a commercial horse radish peroxidase-labelled rabbit anti-equine IgG was used as second antibody. The reaction was developed using azino-diethylbenzotyazol-sulfonate (ABTS). Cut-off was determined by ratio sample (Rs). The hemagglutination inhibition test was used as a reference test for the 391 samples analyzed. Of these, 301 sera were positive by hemagglutination inhibition test and indirect ELISA, 75 were negative by both techniques, and 15 were positive by indirect ELISA and negative by hemagglutination inhibition test. Using hemagglutination inhibition test as standard, the indirect ELISA showed a relative specificity and sensitivity of 83.3 and 100, respectively. This indirect ELISA is useful as screening test.(AU)


Subject(s)
Humans , Animals , Rabbits , Enzyme-Linked Immunosorbent Assay/methods , Influenza, Human/diagnosis , Influenza A virus/isolation & purification , Orthomyxoviridae Infections/diagnosis , Hemagglutination Inhibition Tests , Sensitivity and Specificity
13.
Rev. argent. microbiol ; 32(1): 39-43, ene.-mar. 2000.
Article in Spanish | LILACS | ID: lil-332539

ABSTRACT

An indirect enzyme linked immunosorbent assay was developed. Infected and non infected allantoic fluids precipitated with polyetilenglycol 6000 were used as antigen and control antigen, respectively. Serum samples were diluted 1/20 and a commercial horse radish peroxidase-labelled rabbit anti-equine IgG was used as second antibody. The reaction was developed using azino-diethylbenzotyazol-sulfonate (ABTS). Cut-off was determined by ratio sample (Rs). The hemagglutination inhibition test was used as a reference test for the 391 samples analyzed. Of these, 301 sera were positive by hemagglutination inhibition test and indirect ELISA, 75 were negative by both techniques, and 15 were positive by indirect ELISA and negative by hemagglutination inhibition test. Using hemagglutination inhibition test as standard, the indirect ELISA showed a relative specificity and sensitivity of 83.3 and 100, respectively. This indirect ELISA is useful as screening test.


Subject(s)
Humans , Animals , Rabbits , Enzyme-Linked Immunosorbent Assay , Orthomyxoviridae Infections/diagnosis , Influenza, Human , Influenza A virus/isolation & purification , Hemagglutination Inhibition Tests , Sensitivity and Specificity
14.
Rev. argent. microbiol ; 32(1): 39-43, 2000 Jan-Mar.
Article in Spanish | BINACIS | ID: bin-39904

ABSTRACT

An indirect enzyme linked immunosorbent assay was developed. Infected and non infected allantoic fluids precipitated with polyetilenglycol 6000 were used as antigen and control antigen, respectively. Serum samples were diluted 1/20 and a commercial horse radish peroxidase-labelled rabbit anti-equine IgG was used as second antibody. The reaction was developed using azino-diethylbenzotyazol-sulfonate (ABTS). Cut-off was determined by ratio sample (Rs). The hemagglutination inhibition test was used as a reference test for the 391 samples analyzed. Of these, 301 sera were positive by hemagglutination inhibition test and indirect ELISA, 75 were negative by both techniques, and 15 were positive by indirect ELISA and negative by hemagglutination inhibition test. Using hemagglutination inhibition test as standard, the indirect ELISA showed a relative specificity and sensitivity of 83.3 and 100


, respectively. This indirect ELISA is useful as screening test.

15.
Rev. argent. microbiol ; 32(3): 109-15, 2000 Jul-Sep.
Article in English | BINACIS | ID: bin-39796

ABSTRACT

In Argentina pseudorabies is an endemic disease. Routine diagnosis is made by virus isolation. It is a very long procedure to carry out and gives variable results depending on the quality of sample, hence the need for effective techniques, which are rapid and not dependent on the isolation of infectious virus. The polymerase chain reaction (PCR) technique has provided a sensitive, specific and rapid mean to detect DNA sequences. This study describes a PCR method for detection of pseudorabies virus sequences in swine tissues. In order to determine the presence of suid herpesvirus-1 DNA and antigens, 36 tissue samples collected from 19 dead pigs, with signs of pseudorabies infection, were examined by PCR, virus isolation and indirect immunofluorescence, respectively. Fifteen out of 19 pigs were positive at least for one tissue by PCR (15/19) while only three pseudorabies virus strains were isolated (3/19). All the amplified products were identified by digestion with Sa/l and hybridization. The method described herein circumvents tedious viral isolation and DNA purification and would be a valuable tool for rapid diagnosis, since it would take less than 5 h to reach an accurate result even in poorly preserved tissue samples.

16.
Zentralbl Veterinarmed B ; 46(7): 453-6, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10528541

ABSTRACT

The genomes of 10 bovine herpesvirus 1 and 5 strains isolated in Argentina from 1989 to 1994, recovered from animals showing different clinical signs, and two reference strains (Los Angeles and A663) were compared by restriction endonuclease analysis. Four restriction enzymes, HindIII, BamHI, EcoRI and PstI, were used and analysis of the restriction patterns used to assign the isolate to either the BHV-1.1, BHV-1.2 or BHV-5 genotype. There was a correlationship between restriction pattern and clinical signs in six out of ten Argentinian isolates.


Subject(s)
Cattle Diseases/virology , Herpesviridae Infections/veterinary , Herpesviridae/genetics , Herpesvirus 1, Bovine/genetics , Restriction Mapping/veterinary , Animals , Argentina , Cattle , DNA Restriction Enzymes/chemistry , DNA, Viral/chemistry , Herpesviridae/classification , Herpesviridae Infections/virology , Herpesvirus 1, Bovine/classification
17.
Braz J Med Biol Res ; 31(6): 771-4, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9698821

ABSTRACT

The genomes of 10 equine herpesvirus 1 (EHV-1) strains isolated in Argentina from 1979 to 1991, and a Japanese HH1 reference strain were compared by restriction endonuclease analysis. Two restriction enzymes, BamHI and BglII, were used and analysis of the electropherotypes did not show significant differences among isolates obtained from horses with different clinical signs. This suggests that the EHV-1 isolates studied, which circulated in Argentina for more than 10 years, belong to a single genotype.


Subject(s)
Bacterial Proteins , Genetic Variation , Genome , Herpesvirus 1, Equid/genetics , Argentina , Deoxyribonuclease BamHI , Deoxyribonucleases, Type II Site-Specific , Electrophoresis , Herpesvirus 1, Equid/isolation & purification
18.
Braz. j. med. biol. res ; 31(6): 771-4, jun. 1998. ilus
Article in English | LILACS | ID: lil-210964

ABSTRACT

The genomes of 10 equine herpesvirus 1 (EHV-1) strains isolated in Argentina from 1979 to 1991, and a Japanese HH1 reference strain were compared by restriction endonuclease analysis. Two restriction enzymes, Bam HI and Bgl II, were used and analysis of the electropherotypes did not show significant differences among isolates obtained from horses with different clinical signs. This suggests that the EHV-1 isolates studied, which circulated in Argentina for more than 10 years, belong to a single genotype


Subject(s)
Genetic Variation , Genome , Herpesvirus 1, Equid/genetics , Argentina , Deoxyribonuclease BamHI , Deoxyribonucleases, Type II Site-Specific , Electrophoresis , Herpesvirus 1, Equid/isolation & purification
19.
Viral Immunol ; 10(3): 159-64, 1997.
Article in English | MEDLINE | ID: mdl-9344338

ABSTRACT

In the present study, five mouse monoclonal antibodies (MAbs) to the pseudorabies virus (PRV) Yamagata-81 strain were produced. The MAbs were used in cross-neutralization tests and cross-indirect enzyme-linked immunosorbent assay (ELISA) against three PRV viral strains isolated in Argentina and another four obtained from the United States, Japan, France, and Sweden. Four of five MAbs needed the presence of complement to produce or enhance neutralization activity. No differences were observed by ELISA. The MAbs showed different neutralizing activity against PRV strains, suggesting phenotypic heterogeneity among them.


Subject(s)
Antibodies, Viral/immunology , Herpesvirus 1, Suid/classification , Animals , Antibodies, Monoclonal/immunology , Antigens, Viral/immunology , Cattle , Cell Line , Enzyme-Linked Immunosorbent Assay , Herpesvirus 1, Suid/immunology , Herpesvirus 1, Suid/isolation & purification , Mice , Neutralization Tests , Viral Envelope Proteins/immunology
20.
J Gen Virol ; 77 ( Pt 9): 2031-5, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8811000

ABSTRACT

We report the nucleotide sequence and genetic diversity of part of the envelope (env) gene of four strains of feline immunodeficiency virus (FIV) isolated from Argentine domestic cats. The DNA encoding the V3 to V5 regions of the env gene of the FIV isolates were amplified by PCR, cloned and sequenced. Phylogenetic analysis revealed that the Argentine isolates did not cluster into a single group; one isolate clustered with subtype B FIV isolated in the USA and Japan, whereas the others formed a new cluster of FIV which might represent a prototype sequence for subtype E.


Subject(s)
Genes, env , Genetic Variation , Immunodeficiency Virus, Feline/genetics , Amino Acid Sequence , Animals , Argentina , Base Sequence , Cats , Immunodeficiency Virus, Feline/classification , Immunodeficiency Virus, Feline/isolation & purification , Molecular Sequence Data , Phylogeny , Sequence Homology, Amino Acid
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