Recommendations of the 7th European Meeting of Influenza and Its Prevention.
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Disease Outbreaks↗
Biomedical subjects
L R Haaheim
Publications and source records attributed to L R Haaheim.
An early humoral immune response in peripheral blood following parenteral inactivated influenza vaccination.
The enzyme-linked immunospot assay was used to examine the humoral immune response in 15 healthy volunteers immunized with either split or subunit inactivated trivalent influenza vaccine containing A/Beijing/353/89 (H3N2), A/Taiwan/1/86 (H1N1) and B/Yamagata/16/88. The rapidity of the individual B-cell and serum antibody response was examined in lymphocyte and serum samples collected at various time intervals after vaccination. A rapid serological response was detected with increases in antibody titre detected in the majority of volunteers by 7-8 days postvaccination. Influenza-specific plasma cells were detected as early as 4 days postvaccination, higher numbers of IgA and IgG antibody-secreting cells (ASC) were observed which peaked at 7-8 days postvaccination. The number of ASCs then declined, with low numbers of cells detected at 11 days postvaccination. Influenza-specific IgA ASCs were predominantly of the IgA1 subclass. This rapid immune response may have a bearing on future vaccination policies of unimmunized 'at risk groups' in times of high influenza activity.
Adult↗
Autoantibodies to myelin basic protein are not present in the serum and CSF of MS patients.
Myelin basic protein (MBP) is one of the main constituents of the CNS myelin sheaths, and an autoimmune response directed against MBP may be crucial in the demyelination process in patients with multiple sclerosis (MS). In this study sera and cerebrospinal fluid (CSF) from 25 MS patients, 25 patients with other neurological diseases and 16 healthy controls were examined for antibodies against MBP by using radio immunoblot, western blot, radio immunoassay and enzyme-linked immunosorbent assay. No evidence for the presence of antibodies to MBP was found in sera or CSFs in either the MS patients, or in the control groups tested.
Adult↗
Monoclonal antibodies--their diagnostic potential.
Considering the long and excellent performance of 'classical' immune sera in a range of diagnostic applications, the advent of 'tailor-made' highly specific monoclonal antibodies has given even higher hopes of diagnostic accuracy for the future. Understanding what monoclonal antibodies are and how they compare to polyclonal sources of antibodies is absolutely essential in order to appreciate their potential. This paper will briefly focus on the production, characterization and use of monoclonal antibodies and look critically at the future prospects. Specificity and cross-reactivity are key terms which are explained with examples in this paper. It can be anticipated that diagnostic kits in the future will be composite assays using highly specific monoclonals (or oligoclonal cocktails) in conjunction with high-avidity polyclonal sera, thus taking advantage of the best features of both systems.
Antibodies, Monoclonal↗
Fine molecular specificity of linear and assembled antibody binding sites in HIV-1 p24.
A set of seven murine monoclonal antibodies were generated against a chemically synthesized 11-kDa 104-mer peptide covering the C-terminal residues 270-373 of the p24 gag protein (HIV-1BRU strain). All monoclonal antibodies recognized HIV-1IIIB infected MOLT3 cells by fluorescence and gave positive Western blot signals with viral gag peptides (p55 and/or p24). Oligopeptide binding regions were located with competitive enzyme-linked immunosorbent assays. Detailed epitope scanning analyses (the Geysen technique) were performed by serological testing of the monoclonal antibodies against 99 overlapping hexapeptides which corresponded to the entire 104-mer region. The antibodies bound to p24 peptide sequences located within the 275-293 and 351-368 regions. One antibody (LH104-B) which reacted with residues 357-362 bound to p55 alone. In contrast, another antibody (LH104-I), which recognized the residues 358-363, i.e. with five out of six residues in common with antibody LH104-B for its epitope region, reacted exclusively with p24. At least two of the antibodies (LH104-C and -A) which bound to p24 alone, apparently recognized conformational epitopes. They gave positive reactions with the regions 288-293/351-356 and 284-289/351-356, respectively. This work shows that chemical synthesis of large peptides is a viable alternative approach to immunochemical studies of viral proteins.
Amino Acid Sequence↗
Monoclonal antibodies reacting with the exopolysaccharide xanthan from Xanthomonas campestris.
We have prepared murine hybridomas secreting monoclonal antibodies against the exopopolysaccharide xanthan from Xanthomonas campestris pv. campestris 646 after fusing NSO myeloma cells and spleen cells from BALB/c mice immunized with xanthan. Four hybridomas, secreting antibodies designated A6 (IgM kappa), B3 (IgM kappa), D1 (IgM kappa), and D3 (IgG2A kappa), were selected for further studies. All antibodies reacted with a range of different xanthans. Competition studies using variants of the exopopolysaccharide as competitors suggested that specificity was mainly against the side-chain. One of the antibodies (B3) appeared to require the fully acylated side-chain with the pyruvylated terminal mannose as the immunodominant part. The three others were assumed to be directed against the nonsubstituted trisaccharide with the inner mannose-glucuronic acid being immunodominant. None of the antibodies reacted with cellulose (the xanthan backbone). Using immunoblotting techniques on nitrocellulose paper both a mixture of monoclonal antibodies, and also polyclonal ascitic fluid, could detect xanthan quantities of approximately 0.1 microgram.
Animals↗
Antibodies in sera from Norwegian multiple sclerosis patients and the general population reacting with HTLV-1. A pilot study.
By using enzyme-linked immunosorbent assay, Western blot and passive agglutination we have performed a pilot study involving the testing of 349 sera from the general population of 3 coastal regions in southern Norway, 230 sera from multiple sclerosis patients from western Norway, 109 age- and sex-matched controls and 34 sera from patients with other neurological diseases, for antibodies to HTLV-I. About 2% and 4% of sera from the general population had antibodies reacting with HTLV-I in ELISA and Adult T-cell Leukemia Antigen by agglutination, respectively, whereas about 8% and 5% of the multiple sclerosis sera had such antibodies. In the general population antibodies were more frequent among young individuals and there appeared to be some prevalent regional differences. Western blot confirmatory tests showed that in some instances antibodies reacted with the gag proteins, in particular p19. The results obtained in this study do not indicate an association between HTLV-I and multiple sclerosis, although at this stage we cannot rule out that another retrovirus is involved.
Agglutination Tests↗
Antibodies to staphylococcal peptidoglycan and its peptide epitopes, teichoic acid, and lipoteichoic acid in sera from blood donors and patients with staphylococcal infections.
Antibodies to the staphylococcal antigens peptidoglycan, beta-ribitol teichoic acid, and lipoteichoic acid, as well as to the peptidoglycan epitopes L-Lys-D-Ala-D-Ala, L-Lys-D-Ala, and pentaglycine, were found over a wide range of concentrations in sera from both blood donors and patients with verified or suspected staphylococcal infections. The patient group was heterogeneous with regard to both age and type of staphylococcal infections, being representative for sera sent to our laboratory. In single-antigen assays antibodies to pentaglycine had the highest predictive positive value (67%), although only 32% of the patients had elevated levels of such antibodies. Combinations of test antigens could yield positive predictive values as high as 100%, but then the fraction of positive sera was low. Indeed, the fraction of patient sera which was positive in multiple-antigen tests never exceeded 61%. The clinical usefulness of these seroassays for identifying Staphylococcus aureus as a causative agent was limited, owing to the considerable overlap in the range of antibody concentrations between patient and blood donor sera.
Adolescent↗
Staphylococcus aureus exopolysaccharide in vivo demonstrated by immunomagnetic separation and electron microscopy.
Staphylococcus aureus strains were separated from mastitis milk samples without cultivation by using monodisperse magnetic polymer particles coated with polyclonal antiserum against an encapsulated S. aureus strain. Exopolysaccharide was verified by transmission electron microscopy and the serum soft-agar culture technique. Capsular polysaccharide was found on virtually all clinical isolates. Surface protein A and S. aureus-specific cell wall components were masked when the strains were cultured on an exopolysaccharide-promoting medium. Masking of surface determinants was dependent on their concentration on the bacterial surface as well as on exopolysaccharide abundance. The polysaccharide layer on in vivo bacteria was reduced markedly after just one transfer from milk to blood agar plates but was reexpressed after culturing was done on a capsule-generating medium.
Animals↗
Monoclonal antibodies evoked by the free oligopeptide (Gly)5 reacting specifically with peptidoglycan from staphylococci.
Murine monoclonal antibodies reactive with the Staphylococcus aureus peptidoglycan (PG) epitope (Gly)5 were obtained using the synthetic oligopeptide (Gly)5 in its free form as immunogen. The selected monoclonal antibodies were of the IgM kappa isotype and reacted specifically with PG from S. aureus and Staphylococcus epidermidis, but gave no reaction with PG from Streptococcus pyogenes, Bacillus subtilis and Micrococcus lysodeikticus. Affinity chromatography showed that the antibodies were reactive with the N-terminus of the (Gly)5 peptide. These monoclonal antibodies can be used for the detection of staphylococcal PG in solution.
Antibodies, Monoclonal↗
Intrathecal synthesis of antibodies to staphylococcal antigens in multiple sclerosis patients.
Human serum and cerebrospinal fluid (CSF) antibodies against three staphylococcal antigens were measured using the ELISA technique. Antibody levels in sera from patients with multiple sclerosis (MS) were lower than those found in sera from patients with other neurological diseases (OND) and from healthy individuals (N). The CSF antibody mean titres for patients with relapsing remittent MS were about twofold higher than those found for the other groups. Both serum and CSF antibodies were of the IgG class while IgM could only be detected in serum samples. Elevated IgG-indexes, indicating intrathecal immunoglobulin synthesis, were found only among individuals in the MS patient groups. Some of the patients also had elevated indexes of specific antibodies to the staphylococcal antigens used, i.e. lipoteichoic acid (LTA), peptidoglycan (PG) and beta-ribitol teichoic acid (beta-RTA). This indicates that some MS patients have synthesized staphylococcal antibodies intrathecally.
Adult↗
Antibodies to lipoteichoic acid from Staphylococcus aureus. Specificity of murine monoclonal and human antibodies.
Two monoclonal antibodies against staphylococcal lipoteichoic acid (LTA) were made by fusing P3X63Ag8 myeloma cells and splenocytes from mice immunized with purified LTA. Both were isotyped as being IgM kappa. Their specificities were determined by enzyme-linked immunosorbent assays indicating that both antibodies reacted with the glycerol-phosphate backbone, while one of them also had some affinity for the alanyl substituent. Antibodies in serum from 7 multiple sclerosis (MS) patients and serum and cerebrospinal fluid (CSF) from 7 non-MS patients apparently reacted with the sugar moiety of LTA. In contrast, CSF antibodies from 6 of the 7 MS patients and 1 of the 7 non-MS patients had affinity for the alanine residue. This non-MS patient also had serum antibodies against the alanine residue. None of the other sera tested appeared to contain such antibodies.
Animals↗
Monoclonal antibodies to the h1 agglutinogen from Staphylococcus aureus 17A. Serological testing with type strains.
Four monoclonal antibodies to the h1 agglutinogen were produced by conventional means, and slide agglutination of S. aureus type strains was performed with protein A affinity purified IgG1 antibodies. In accordance with Oeding's serotype system the type strains 17A and 670 were strongly and consistently agglutinated. In addition, however, several of the remaining twelve type strains investigated showed varying reaction patterns. Our results indicate that the h1 agglutinogen may be more widely distributed among S. aureus strains than previously assumed.
Agglutination Tests↗
A monoclonal antibody inhibiting human placental Fc gamma-receptor activity.
Fc gamma receptor (FcR) from human placenta was solubilized using EDTA and 2-mercaptoethanol and purified by affinity chromatography on human IgG-coated Sepharose 4B. BALB/C mice were immunized with FcR and monoclonal antibodies were obtained by growing hybridoma cells following fusion of spleen cells with P3 X 63Ag8 myeloma cells. Using an immunofluorescence technique, the IgG1 monoclonal antibody secreted by clone B1D6 stained the FcR-positive areas in sections of placental tissue. The endothelium of the foetal stem vessels stained more strongly than did the trophoblasts. The antibody also inhibited the haemadsorption to placental tissue of erythrocytes (E) sensitized with IgG antibodies (A), (EA), and inhibited the agglutination of EA by FcR. The data indicate that the monoclonal antibody reacts with the placental FcR at the binding site for IgG, or with an epitope close to the binding site. Apparently, the FcR in different anatomical areas in the placenta have a common antigenic determinant.
Antibodies, Monoclonal↗
Strain specificity of serum antibody to the haemagglutinin of influenza A (H3N2) viruses in children following immunization or natural infection.
The specificity of serum anti-HA antibody from children immunized or infected with A/Victoria/75 (H3N2 or A/Texas/77 (H3N2) virus was examined using the single radial haemolysis test together with adsorption of antibody with three antigenic variants A/Hong Kong/68 (H3N2), A/Port Chalmers/73 (H3N2) and A/Victoria/75 (H3N2). The majority of young children reacted to vaccination or infection by producing strain-specific (SS) antibody to the homologous virus. A small proportion of children's sera contained cross-reacting (CR) antibodies capable of reacting with the haemagglutinins of all antigenic variants of the sub-type including A/HK/1/68. In contrast, most adults reacted immunologically to either vaccination or infection by producing CR antibody, reacting with all variants of the antigenic subtype including the prototype virus A/HK/1/68 (H3N2).
Adolescent↗
Antibodies to the strain-specific and cross-reactive determinants of the haemagglutinin of influenza H3N2 viruses. Antiviral activities of the antibodies in biological systems.
Antibodies to the strain-specific (SS) and cross-reactive (CR) antigenic determinants of the haemagglutinin (HA) of influenza H3N2 viruses were prepared, and their antiviral properties in experimentally infected mice and in eggs were investigated. The SS antibodies were considerably more efficient than the CR antibodies in direct virus neuralization tests. The difference in the activities of the two types of antibody preparations was less pronounced in virus growth inhibition studies in the allantois-on-shell system and in passive protection studies in mice give antibody before challenge with virus. Nevertheless, even in these tests SS antibody was approximately four-fold more effective than was CR antibody in protection against fatal infection. Passive administration of CR or SS antibody preparations to mice challenged with sublethal virus doses, reduced the peak pulmonary titres of virus on day three post-infection compared with the control animals. The degree of reduction in titre exerted by the SS antibodies was greater than for the CR antibodies. In contrast, the rate of clearance of pulmonary virus 9-12 days after infection in the mice treated with SS antibody was slower than for the CR treated animals. Also, in SS antibody treated animals viruses isolated from mouse lungs showed evidence of minor antigenic variation away from the parent virus.
Animals↗
Haemagglutination-inhibition antibodies in human sera to an antigenic mutant of influenza A/Texas/1/77 (H3N2) virus obtained in vitro.
An antigenic mutant of the wild influenza strain A/Texas/1/77 (H3N2) was obtained in vitro by growing virus of the allantosis-on-shell system in the presence of a mouse monoclonal antibody preparation to the Texas haemagglutinin (HA) antigen. A total of 204 human sera, from a population of randomly collected sera during the fall of 1979, were examined by the haemagglutination-inhibition (HI) tests against the Texas wild strain and the antigenic mutant. It was shown that sera from young children (0-5 years) reacted poorly, or not at all, with this mutant strain, whereas sera from adults (15 years +) reacted equally well with both viruses. It can be assumed that the antigenic site recognised by this particular monoclonal antibody preparation (185/1) may be important for the human immune response to the HA antigen of A/Texas/1/77 virus.
Adolescent↗
Antibodies to the strain-specific and cross-reactive determinants of the haemagglutinin of influenza H3N2 virus. 3. Selection of antigenic variations in vitro and in vivo.
Antigenic variants of influenza A/Hong Kong/1/68 (H3N2) were obtained in vitro by letting virus multiply in the allantosis-on-shell system in the presence of anti-haemagglutinin antibodies, prepared from immune goat serum to purified haemagglutinin antigen, and in vivo by giving mice antibody intraperitoneally one day before challenge with a sublethal dose of live virus. In both systems it was shown that the most narrowly reacting strain-specific antibodies selected antigenic variants at an apparently higher rate than a more cross-reactive preparation of antibodies.
Animals↗