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PubMed1985

Double infection of the CNS with herpes simplex and mumps viruses. Serological and ultrastructural studies.

Clinical, serological and ultrastructural features are described in a case of acute encephalitis. The ultrastructural study of brain biopsy specimens revealed herpes-and paramyxovirus-like particles in neurons and glial cells that were characterized as herpes simplex and mumps viruses by serological study in serum and CSF. Of the few cases previously reported with double encephalitis, this is the only one in which ultrastructural and serological evidence of combined brain infection has been found.

Antibodies, Viral↗
PubMed1983

Serologic dissection of HLA-D specificities by the use of monoclonal antibodies. II. Distinction between HLA-Dw2 and HLA-Dw12.

Three cytotoxic monoclonal antibodies, HU-11, HU-32, and HU-33, specific for human Ia-like antigens were used to analyze the two HLA-DR2-associated HLA-D specificities, HLA-Dw2 and HLA-Dw12. In the HLA-Dw2, DR2, MB1 homozygous B-cell line EB-CMG, the binding of radiolabeled HU-32 and HU-33 was strongly inhibited by the addition of nonlabeled HU-11, whereas no inhibition occurred in the HLA-Dw12, DR2, MB1 homozygous B-cell line EB-KT. To confirm this differential inhibition pattern further, F(ab')2 fragments were prepared from HU-11, and their ability to inhibit complement-dependent lysis mediated by HU-32 and HU-33 was assessed against a total of five homozygous typing cell lines homozygous for HLA-Dw2, DR2, MB1 or HLA-Dw12, DR2, MB1, including EB-CMG and EB-KT. Here again, the same differential inhibition pattern as that observed in the radiobinding inhibition assays was obtained. Thus, the data suggest that the two kinds of HLA-DR2-positive homozygous typing cell lines with distinct HLA-D specificity can be distinguished from each other by using solely serologic methods. This is the first clear-cut serologic distinction made between homozygous typing cells defining HLA-Dw2 and those defining HLA-Dw12, since no serologic means that enables one to distinguish one from the other has been available.

Antibodies, Monoclonal↗
PubMed1984

Comparison of one-dimensional IEF patterns for serologically detectable HLA-A and B allotypes.

A new mouse monoclonal antibody (MoAb) 4E, which detects an epitope shared by HLA-B locus antigens, together with the MoAb W6/32, detecting a common HLA, B, C, determinant, and the MoAb4B, detecting HLA-A2 and A28, were used to isolate HLA-A and -B antigens in sequential immunoprecipitation. The HLA antigens obtained from metabolically labeled cell extracts of B-lymphoblastoid cell lines or from phytohemagglutinin (PHA)-activated peripheral blood lymphocytes were compared by one-dimensional isoelectric focusing (1D-IEF). The IEF banding patterns obtained with native HLA antigens segregated in a family with HLA. Neuraminidase treatment of isolated antigens reduced the number of bands to one or two, simplifying the analysis of characteristic patterns. Thus, we have cataloged IEF banding patterns for the majority of the serologically recognized HLA-A and -B allotypes obtained from 57 unrelated American Caucasians. While no HLA-A locus or HLA-B locus specific banding patterns were observed, the HLA-A antigens had, in general, slightly higher pI values than the HLA-B antigens. HLA-C antigens could not be detected in this assay system. The polymorphism detected by IEF banding patterns was as extensive as the serologically detected polymorphism identified by classical HLA serology. Moreover, variants for some HLA allotypes could be detected by this method. In addition to previously recognized A2 variants, new variants were identified for HLA-A1, A26, and Bw44. Each A1 and Bw44 variant was associated with particular haplotypes. The HLA-A2 antigens occurred on 43 HLA haplotypes in the unrelated Caucasian population. Only one of each A2 variants was identified in this population.

Alleles↗
PubMed1986

Polymorphism of the HLA DR1 haplotype in the Israeli population investigated at the serological, cellular, and genomic levels.

In the present report, we used serological, cellular, and restriction fragment length polymorphism (RFLP) to investigate the DR1 haplotype in the Israeli population. We describe an Israeli homozygous typing cell (HTC), HLA-Dw"LVA", which defines a new lymphocyte-activating determinant associated with Bw65, DR1 and distinct from Dw1. The parents of this donor, non-Ashkenazi Algerian Jews, are first cousins and share HLA-Cw8,Bw65,BfS,DR1,DQw1,DPw4. No specificity could be assigned to HLA-Dw"LVA" using the 91 Ninth Workshop HTCs. Two families and forty unrelated DR1 individuals were studied with Dw"LVA" and a panel of DR1/Dw1 HTCs. HLA-Dw"LVA" showed segregation as a single determinant within families. This new specificity was present in 24 out of 40 (60%) unrelated DR1 individuals, indicating that in the Israeli population Dw"LVA" is the main lymphocyte-defined determinant associated with the serologically defined DR1 specificity, in contrast to non-Jewish Caucasoids where DR1 is significantly associated with Dw1. The vast majority of Dw"LVA"-positive carriers were also Bw65 carriers, indicating that Bw65,DR1, Dw"LVA" may represent a typical allele combination in the Israeli population. The RFLP analysis established the correlation of certain RFLPs with Dw1 and Dw"LVA". In addition, we describe a cluster of FRLPs that may correspond to a new Dw subtype associated with DR1, for which no serological and cellular reagents have been described so far.

DNA Restriction Enzymes↗
PubMed1983

Serologic dissection of HLA-D specificities by the use of monoclonal antibodies.

To study the gene products of the HLA complex, we produced two monoclonal antibodies, termed HU-18 and HU-23. They were active in complement-dependent cytotoxicity and detected B-cell alloantigens encoded by a locus (or loci) linked to HLA. When three types of HLA-DR4 homozygous B-cell lines with different HLA-D specificities were tested for reactivity with HU-18 and HU-23, they displayed distinct reaction patterns depending on the HLA-D specificities they possessed: EBV-Wa (HLA-DYT homozygous), negative for both HU-18 and HU-23; KT2 and KOB (HLA-DKT2 homozygous), positive only for HU-18; and ER (HLA-Dw4 homozygous), positive for both. These differential reaction patterns were further confirmed by testing against a panel of 17 HLA-DR4-positive peripheral blood lymphocytes with known HLA-D specificities. Thus, these monoclonal antibodies allow us to identify HLA-DYT, HLA-DKT2, and HLA-Dw4 solely by serologic methods. This is the first clearcut serologic identification of these three HLA-DR4-associated HLA-D specificities, which have been indistinguishable by conventional serology and identified only by cellular techniques. It is hoped that immunochemical investigations using HU-18 and HU-23 will advance our understanding of the HLA-D region on a molecular level.

Antibodies, Monoclonal↗
PubMed1993

Modulation of the serological response of specific pathogen-free (EHV-free) foals to EHV-1 by previous infection with EHV-4 or a TK-deletion mutant of EHV-1.

EHV-1 was inoculated into specific pathogen-free (SPF) foals in order to study uncomplicated primary responses. Infection resulted in a strong serological response recognizing EHV-1-specific antigens; this contrasts with a previous publication where a weak response was recorded in SPF animals. Antibodies to EHV-1 were readily detected by four techniques (virus neutralization, complement fixation, Western blots and immune precipitation), yet there was comparatively little cross-reaction to EHV-4 target antigen. Re-inoculation with the same virus strain stimulated antibodies to EHV-1 but no additional antigens were recognized and antibodies cross-reacting with EHV-4 antigens were not enhanced. Having characterized the uncomplicated primary response to EHV-1 in SPF foals, further animals were exposed to either EHV-4 or a thymidine kinase-deficient mutant of EHV-1 prior to challenge with w/t EHV-1 to investigate how these infections might modulate the immune responses to EHV-1 or 4. Primary inoculation with EHV-4 or with a thymidine kinase-deficient mutant of EHV-1 produced productive infections as evidenced by virus shedding and pyrexia. In both these cases, however, in contrast to that with w/t EHV-1, the serological response was very weak. Re-infection of foals primed with either EHV-4 or TK-deficient EHV-1 with w/t EHV-1 resulted in a strong response to EHV-1 antigens detected by all four methods. In addition, in the foals given a primary inoculation with EHV-4, superinfection with EHV-1 resulted in a strong cross-reactive response to EHV-4 target antigens. The relevance of these observations to the interpretation of previously reported serological responses to EHVs in SPF and naturally reared animals is discussed.

Animals↗
PubMed1988

Coat protein of potyviruses. 4. Comparison of biological properties, serological relationships, and coat protein amino acid sequences of four strains of potato virus Y.

Four strains of potato virus Y, PVY-D, PVY-10, PVY-18, and PVY-43, obtained from different Australian sources were compared on the basis of their biological, serological and coat protein structural properties. Each of the strains could be distinguished on the basis of their reactions on selected test plant species. Two of the PVY strains, PVY-D and PVY-10, induced symptoms similar to those produced by the PVYO strain group. The reactions of PVY-18 and PVY-43, although comparable to PVYN in some hosts, did not completely match the description of the PVYN strain group. In contrast to the other three strains, PVY-18 could not be transmitted by Myzus persicae in repeated tests. No difference was observed in the serological properties of the four PVY strains in different assay systems, using polyclonal antisera. The amino acid sequences of the coat proteins of PVY-10, PVY-18, and PVY-43 were obtained and compared with the coat protein sequences of pepper mottle virus (PeMV) [Dougherty WG, Allison RF, Parks TD, Johnston RE, Feild MJ, Armstrong FB (1985) Virology 146: 282-292] and PVY-D [Shukla DD, Inglis AS, McKern NM, Gough KH (1986) Virology 152: 118-125]. The homology between the PVY strains ranged from 96.3 to 99.3% and with the PeMV sequence, 91.4 to 92.9%. Based on this high sequence homology, and the previous observation that coat protein sequences of potyvirus strains are always greater than 90% identical, PeMV could be considered a strain of PVY. However, PVY and PeMV are reported to be only distantly serologically related and on this basis PeMV is currently considered to be an independent member of the Potyvirus group.

Amino Acid Sequence↗
PubMed1989

Serological and genetical relationships of three herpesvirus strains from salmonid fish.

Serological and genetical relationships of three salmonid herpesviruses [strain H-83, OO-7812 strain of Oncorhynchus masou virus (OMV) and Herpesvirus salmonis] were investigated. Although the serological relationships of these viruses were not distinctly demonstrated by cross-neutralization test, cross enzyme-linked immunosorbent assay revealed that strain H-83 and OMV appeared to be serologically more closely related to each other than to H. salmonis. Strain H-83 was genetically similar to OMV but distinct from H. salmonis by restriction endonuclease cleavage analysis and DNA-DNA hybridization test. These results indicated that strain H-83 should be classified under the same group as OMV, and H. salmonis belong to another group of salmonid herpesviruses.

Animals↗
PubMed1987

Serological and biochemical characteristics of intermediate adenovirus strains of subgenus D.

Ten serologically intermediate strains of subgenus D related to six different serotypes were characterized by neutralization and hemagglutination-inhibition and by DNA restriction analysis with the endonucleases BamHI, BglII, BstEII, HindIII, and SmaI. Each strain was a unique genome type. While the serologically identical strains showed only slight variations in their DNA, the comparison of the intermediate strains with the serologically related prototypes resulted in only moderate DNA relationship (53 to 73% comigrating fragments, with one exception). Hence the possibility that the intermediate strains have originated by recombination of two prototype-like strains is unlikely. One strain lost its cross-reactivity with AV19 in hemagglutination-inhibition after terminal dilution.

Adenoviruses, Human↗
PubMed1984

Serological analyses of different mink Aleutian disease virus strains.

Four different isolates of Aleutian disease virus have been compared electrophoretically and serologically. These were the DK and Utah 1 isolates, known as highly virulent strains, the Pullman isolate, known as a low virulent strain and the avirulent ADV-G isolate, which is the only strain grown in cell culture. ADV-G was shown to migrate in agarose electrophoresis 22 per cent slower than the other strains. Several murine monoclonal antibodies were prepared against each of the isolates. Each one reacted with all 4 of the isolates, but a few showed higher affinity for some of the isolates. Competitive RIA analyses were also performed, and these studies indicated some serological differences between the 4 strains. It is concluded that ADV-G polypeptides are chemically different but immunologically cross-reacting with the polypeptides present on the field ADV strains. It is suggested that the small serological differences seen between the field strains might be caused by slightly different in vivo proteolytic degradation of the viral capsid proteins and thus might not be taken as an indication of strain variation.

Aleutian Mink Disease Virus↗
PubMed1978

A longitudinal study of the serological and virological status of 18 women infected with cytomegalovirus.

18 women, of known seropositivity, have been followed for between 18 and 66 months (mean 50.2 months) for viral excretion and serological changes. CMV was isolated from 58/146 (39.7 per cent) cultures from various sites, mostly cervix. A total of 129 sera were obtained and each was titrated in the late antigen, early antigen and anti-complement immunofluorescence assays, as well as the complement fixation and microneutralisation tests. From 3 women virus was consistently re-isolated, from 3 others virus was never re-isolated and the remaining 12 women excreted virus intermittently during the period of study. No significant changes in antibody titres could be detected by any of the 5 serological assays in any woman during periods of viral excretion. This suggests that local reactivation of latent CMV infection is not associated with a demonstrable systemic antibody response. Inter-assay correlations were sought for each of the 10 possible combinations of pairs of tests. Significant correlations were found for most pairs of tests, with the notable exception fo those involving the early antigen test. This confirms that antibodies detected in this assay are distinct from those detected by the other 4 serological assays.

Adult↗
PubMedDate not supplied

Serology, clinical manifestations and treatment of brucellosis in different age groups.

We prospectively studied 339 patients diagnosed of brucellosis over a six year period in order to evaluate the clinical and serological characteristics of brucellosis in the elderly. 319 patients (94.1%) were under 65 years of age (group A), and 20 patients (5.9%) were older than 65 (group B). No patient in group B developed splenomegaly as opposed to 69 (21.6%) in group A (p less than 0.05). The percentage of positive blood cultures (Brucella melitensis isolated in all cases) was 72.1% in group A and 60% in group B. The mean titer of IgM antibodies measured by indirect immunofluorescence test was significantly lower in the elderly patients, with no other differences in serologic response between the two groups. 103 patients (32.2%) in group A and seven patients (35%) in group B developed some complications; spondylitis was more common and severe among group B patients. There was no therapeutic failure or relapse among patients over 65. We conclude that clinical, bacteriological and serological characteristics and the outcome of brucellosis in the elderly are similar to those seen in younger patients.

Adolescent↗
PubMedDate not supplied

Prevalence of Chlamydia trachomatis in cases of genital non-gonococcal infection according to microbiological and serological investigations.

An investigation on the prevalence of Chlamydia trachomatis was carried out on 231 patients (115 men and 116 women, mean age 31.6 years) with genital non-gonococcal infection in order to study the role of this microorganism in infertility and/or sterility in the Italian population. One hundred and sixty-six apparently healthy subjects (108 men and 50 women, mean age 32.4) were also included as control for serological comparison only. Persons with gonococcal infection were excluded. ELISA method was chosen for the direct C. trachomatis examination of "scraping" samples. Serological investigations were carried out by means of the indirect immunofluorescence test for IgM and IgG determination as well as the immunoperoxidase assay for IgA antibodies. A direct C. trachomatis positive test was demonstrated in 27.8% of men vs. 11.2% in women (p less than 0.01) in the patient group. Anti-C. trachomatis IgG specific antibodies were present respectively at any serum dilution in 50.2% of patients vs. 47.6% of normal subjects respectively (p greater than 0.5). The low presence of C. trachomatis (19.5%) at the direct test contrasts with the higher percentage of anti-C. trachomatis positive patients (50.2%) in the same group suggesting that serological tests may be more useful than the direct test in demonstrating an active C. trachomatis infection provided that IgM, IgG and IgA specific antibodies be contemporaneously investigated.

Adolescent↗
PubMed1992

Intensification of GVHD prophylaxis interferes with the effects of pretransplant herpes virus serology on the occurrence of grades II-IV acute graft-versus-host disease.

The effects of pretransplant herpes virus serology on the occurrence of grades II-IV acute graft-versus-host disease (GVHD) were studied in 262 recipients and their HLA-identical family donors. In 131 recipients on standard GVHD prophylaxis (either methotrexate or cyclosporin A) significant effects were observed for donor HSV serology (seropositivity associated with increased risk for GVHD) and donor EBV serology (seronegativity associated with increased risk). However, these effects were nonsignificant in the other 131 recipients on intensified GVHD prophylaxis (i.e., methotrexate combined with cyclosporin A, in vivo anti-T-cell monoclonal antibodies, or various procedures to reduce the T-cell numbers in the transplants.

Acute Disease↗
PubMedDate not supplied

Response of soluble IL-2 receptor, interleukin-2 and interleukin-6 in patients with positive and negative Borrelia burgdorferi serology.

This study was designed to investigate serum soluble interleukin-2 receptor (S-IL-2R), interleukin-2 (IL-2) and interleukin-6 levels (IL-6) in patients with either a positive or negative Borrelia burgdorferi serology. Serum samples from 101 individuals, divided in to five groups according to clinical symptoms and outcome of serology were analysed. Samples of cerebrospinal fluid (CSF) from nine of the individuals were also studied. The highest average serum S-IL-2R levels (1,180 +/- 1,140 U/ml) were found in patients with erythema migrans, the hallmark of Lyme borreliosis, followed by patients with symptoms closely related to Borrelia infection (900 +/- 1,200 U/ml) and with a strong positive serology. In two patients with central nervous system (CNS) involvement, increased levels of S-IL-2R of 920 and 620 U/ml respectively (normal value < 50 U/ml) were detected in the CSF. No statistically significant relationship between IgG or IgM antibody activity and serum S-IL-2R levels was found. Detectable levels of IL-2 were only found in three patients. Increased levels of IL-6 were found in sera from 14 patients. The highest concentration, 90 pg/ml (normal value < 10 pg/ml), was measured in a patient presenting with vasculitis. In conclusion, B. burgdorferi infection causes a moderate increase of serum S-IL-2R levels, although there is no relationship between the severity of the infection, as estimated by the antibody concentration or to serum IL-2 or IL-6 levels. Secondary complications of the infection, such as vasculitis, may cause an increased level of serum IL-6.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1982

Serological grouping of streptococci: a collective evaluation in six laboratories of three rapid methods.

Three serological methods for streptococci grouping: latex agglutination after pronase extraction (LAP), latex agglutination after nitrous acid extraction (LAN) and counterimmunoelectrophoresis after nitrous acid extraction (CIEN) were evaluated in six laboratories with 310 strains of streptococci. To reduce the possible influence of a particular laboratory, each serological procedure was performed in two different laboratories with identical protocols and strains. Between the three procedures, a total agreement of 81.3% with six identical results was obtained. Since only one discordant result was obtained in 12.2% (CIEN 4.8%-LAN 5.8%-LAP 1.6%) and two discordant results with two different serological methods in 2.6%, it could be concluded that agreement between the three procedures was 96%. For B (99 strains tested) and D(100 strains tested) groups, total agreements of respectively 95% and 92% were achieved. In conclusion, LAP, LAN, and CIEN gave equivalent results for streptococci grouping, but LAN was the most simple needing no heating, centrifugation, or electrophoresis equipment and lacking cross-reactivity of group C antibody.

Counterimmunoelectrophoresis↗
PubMed2005

The phylogenetic structure of the cluster of tobamovirus species serologically related to ribgrass mosaic virus (RMV) and the sequence of streptocarpus flower break virus (SFBV).

Ribgrass mosaic virus (RMV), turnip vein-clearing virus (TVCV) and Youcai mosaic virus (YoMV; formerly designated as oilseed rape mosaic virus; ORMV) belong to the genus Tobamovirus and are arranged in one out of three subgroups because of their common host range, serological cross-reactivity and amino acid composition of their coat proteins. The recently defined species Wasabi mottle virus (WMoV) is closely related to the same subgroup. The distinction of the four species is difficult and the lack of sequence information of a wide range of isolates has led to an unclear nomenclature. To clarify this situation we sequenced the coat protein genes from 18 isolates which were serologically related to members of the species of this cluster. The size of the coat protein was conserved with the exception of one isolate which revealed an N-terminal extension due to the mutation of three stop-codons. Phylogenetic analysis of these CP ORFs resulted in a tree with three clusters each containing at least one of the approved species RMV, TVCV and 1ptYoMV/WMoV in which our isolates were distributed. The tree was congruent and did support the present taxonomic status of species within this subgroup. For practical purpose we developed a subgroup 3 specific primer pair and a species differentiating restriction fragment length polymorphism (RFLP). Sequencing of the genome of Streptocarpus flower break virus (SFBV) which is serologically distantly related to the subgroup 3 viruses revealed a distinct genome organization. Therefore we propose that this virus should be regarded as a member of a species not belonging to any of the subgroups so far established.

Base Sequence↗
PubMed2000

Molecular characterization and coat protein serology of watermelon leaf mottle virus (Potyvirus).

A cDNA library was generated from purified RNA of watermelon leaf mottle virus (WLMV) (Genus Potyvirus). Two overlapping clones totaling 2,316 nucleotides at the 3' terminus of the virus were identified by immunoscreening with coat protein antiserum. The sequence analyses of the clones indicated an open reading frame (ORF) of 2,050 nucleotides which encoded part of the replicase and the coat protein, a 243-nucleotide non-coding region (3'UTR), and 23 adenine residues of the poly (A) tail. The taxonomic status of WLMV was determined by comparisons of the sequence of the cloned coat protein gene and 3'UTR with potyvirus sequences obtained from GenBank. The nucleotide sequence identities of WLMV compared with 17 other potyviruses ranged from 55.6 to 63.5% for the coat protein, and from 37.2 to 48.3% for the 3'UTR. Phylogenetic analyses of the coat protein region and the 3'UTR indicated that WLMV did not cluster with other potyviruses in a clade with high bootstrap support. The coat protein gene was expressed in Escherichia coli and a polyclonal antiserum was prepared to the expressed coat protein. In immunodiffusion tests, WLMV was found to be serologically distinct from papaya ringspot virus type W, watermelon mosaic virus 2, zucchini yellow mosaic virus, and Moroccan watermelon mosaic virus. In Western blots and ELISA, serological cross-reactivity with other cucurbit potyviruses was observed. Serological and sequence comparisons indicated that watermelon leaf mottle virus is a distinct member of the Potyvirus genus.

3' Untranslated Regions↗