Search PubMed⌕ Search

Biomedical subjects

E Norrby

Publications and source records attributed to E Norrby.

At least 127 records · Page 7Linked to original sources

Non-lethal infection of aminergic reticular core neurons: age-dependent spread of ts mutant vesicular stomatitis virus from the nose.

In order to induce a non-lethal infection restricted to central aminergic neurons projecting to the olfactory bulbs a series of temperature sensitive (ts) and G-protein monoclonal antibody escape mutants of vesicular stomatitis virus (VSV) were instilled into the nasal cavity of mice. In three-week (wk)-old NMRI mice four monoclonal antibody escape mutants caused an extensive infection of the olfactory epithelium and, like a wild type strain, a lethal brain infection after spread along olfactory pathways. Three ts mutant strains showed an attenuated pathogenic potential. Strain G31 caused a lethal infection with a somewhat prolonged course while the strain G11 failed to invade the nervous system. Strain G41 showed minimal invasion of central nervous system in three-wk-old mice and caused a lethal infection in newborn and one-wk-old mice. In contrast, two-wk-old mice survived infection with this mutant, which spread along olfactory pathways and rather selectively affected aminergic reticular core neurons in the diagonal band, the locus ceruleus and the raphe nuclei in the brainstem. Thus, an age-dependent virus infection of the olfactory pathways can cause restricted lesions in the brain providing a model for studies of virus-induced changes in aminergic neurotransmission.

Aging↗

Molecular cloning and characterization of six genes, determination of gene order and intergenic sequences and leader sequence of mumps virus.

mRNA isolated from mumps virus-infected Vero cells was converted into cDNA and cloned into the PstI site of the plasmid pBR322. After screening with 32P-labelled cDNA synthesized from poly(A)+ RNA of uninfected or mumps virus-infected Vero cells, five different groups of virus-specific clones were obtained. The virus specificity of the clones was confirmed by Northern blot analysis, in which the cDNA inserts from the five different groups hybridized to mRNAs of about 2100, 1500, 1450, 2000 and 2200 nucleotides. By the use of oligonucleotides synthesized on the basis of sequences obtained from the five cDNA clones and mRNAs, the sequence of the intergenic and surrounding areas was determined. During genome sequencing, a separate gene was identified between the fusion protein (F) gene and the haemagglutinin-neuraminidase protein (HN) gene. Using oligonucleotides synthesized on the basis of the new gene sequence, cDNA clones with poly(A) were isolated from the cDNA library. The gene order was determined to be 3' NC-P-M-F-SH-HN-L 5' (where NC, P, M, SH, and L represent the genes for the nucleocapsid, phosphoprotein or polymerase-associated, matrix or membrane, small hydrophobic and large proteins respectively). There is one nucleotide between the P and M (A), M and F (A), and HN and L genes (G), two between the NC and P (AA) and SH and HN (3'-CG) genes, and seven between the F and SH genes (3' GAUUUUA) as intergenic sequence. The leader sequence at the 3' end of the genome has been determined by sequencing the dicistronic leader-NC mRNA using oligonucleotide primers. The sequence from the 3' terminus to the NC gene start of the mumps virus genome is similar in length (55 nucleotides) to that present in Sendai virus, Newcastle disease virus and parainfluenza virus type 3, and the first five nucleotides are conserved in all negative-stranded RNA virus genomes sequenced to date.

Base Sequence↗

Protective effects of monoclonal antibodies against parainfluenza virus type 3-induced brain infection in hamsters.

A collection of monoclonal antibodies (MAbs) against the haemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of parainfluenza virus type 3 (PIV3) was used for passive immunization of intracerebrally infected newborn hamsters. A significant degree of protection was obtained with MAbs against both the HN and F viral proteins, but no individual antibody conferred complete protection against disease. MAbs against different epitopes provided different degrees of protection. A MAb to the Sendai virus HN protein was found to confer protection against PIV3 brain infection.

Animals↗

Respiratory syncytial virus: heterogeneity of subgroup B strains.

In order to investigate further possible structural differences among the two subgroups of respiratory syncytial virus (RSV), we analysed the antigenic characteristics and size of structural proteins of 20 subgroup A and 43 subgroup B strains by their reactions with monoclonal antibodies (MAbs) directed against the proteins of RSV using immunofluorescence, ELISA and radioimmunoprecipitation assays. The latter test also enabled determination of the size of different structural components. The 37 MAbs employed were generated by immunization with both subgroup A and B strains. They represented specificities for distinct epitopes on five different structural proteins. The subgroup A strains proved to be relatively uniform. The fusion (F) protein, nucleoprotein (NP) and matrix (M) proteins of all strains tested had the same Mr and all except one strain had a phosphoprotein (P protein) of the same Mr. The F and P proteins were lower in Mr in B strains compared to A strains, which confirmed previous findings. The Mr of the large surface glycoprotein (G protein) of subgroup A strains varied slightly, probably on the basis of differing glycosylation. By contrast, the subgroup B strains exhibited substantial variation in the Mr of the G and also the P proteins and in reactivity with MAbs directed against the G and F proteins. Three size classes of the P protein were identified in B strains: 33K to 34K, 32K to 33K, and 31K to 32K. Twenty-seven subgroup B strains failed to react with four anti-G MAbs representing a single epitope, G2; the remaining 16 strains reacted with these MAbs. We designated these two sets of variants of B strains B1, which lacked the epitope, and B2, which had the epitope. The B1 strains also varied in the size of the G and P proteins. In contrast, all B2 strains had large G proteins and all except two strains had relatively large P proteins (33K to 34K). All subgroup B1 and B2 strains exhibited the same sizes of NP, F and M proteins. We conclude that the subgroup B strains of RSV include two variants, B1 and B2, and that the major difference between them resides in the G and P proteins.

Antibodies, Monoclonal↗

Follow-up of the antibody response to measles vaccine in a rural area of Guinea-Bissau.

One hundred and fourty-four children who either were already immune or had been successfully immunized against measles were reexamined after 16 months. All still had circulating Elisa antibodies at a clearly detectable level. Titres were higher in the group of children stated to have had measles prior to the immunization. None of the children had measles after immunization. Boostering by the wild virus may have occurred, whereas no evidence of a booster effect from the vaccine was found. About one third of the children were underweight. Plasmodium falciparum parasitaemia rate, and also its seasonality, varied with the location of the child's homestead. Even children exposed to mesoendemic P. falciparum malaria and moderate malnutrition can be successfully immunized with a conventional live attenuated measles vaccine from 8 months of age, which probably results in a lasting protection.

Antibodies, Viral↗

Site-directed ELISA with synthetic peptides representing the HIV transmembrane glycoprotein.

Two partially overlapping 19 and 22 amino acids long peptides representing a highly immunogenic site of the transmembranous glycoprotein (gp41) of human immunodeficiency virus (HIV) were used as antigen in ELISA tests. The results of antibody determination with this assay were compared with those of three or more conventional ELISAs and Western blot (WB) tests and radioimmunoprecipitation assay. Twenty-six sera from patients with AIDS or LAS and from asymptomatic carriers of HIV infection all showed a pronounced reaction in the peptide ELISA as well as positive results with other tests. In contrast, 27 sera from laboratory workers and blood donors were negative by all tests. A group of 39 blood donor sera, which had shown false positive or ambiguous results in the ELISAs and sometimes in WB tests employed for confirmation, also were negative in all cases with the peptide ELISA. Consecutive samples collected from individuals with primary HIV infection were also analyzed. In 6 out of 9 cases, the peptide ELISA revealed an antibody response within one month after onset of clinical symptoms and sensitivity for antibody detection equaled that of other ELISA tests. Eight sera from five West African persons infected with HIV-related viruses did not react in the peptide ELISA, reflecting differences in properties of the envelope components. The peptide ELISA used in this study appears to represent a simple technique employing chemically synthesized antigen for accurate and sensitive estimation of antibodies to the HIV group of nontransforming human retroviruses.

AIDS-Related Complex↗

The intrathecal immune response in the early stage of multiple sclerosis.

Sequential pairs of cerebrospinal fluid (CSF) and serum samples from 10 patients followed for 2.5-12 years after onset of unilateral optic neuritis (ON) were studied. Eight patients developed definite multiple sclerosis (MS) during the observation period. All patients had normal CSF protein patterns on agar or agarose gel electrophoresis at onset. Six patients developed oligoclonal immunoglobulin (Ig) bands in the CSF during the observation period. Imprint immunofixation of electrofocused specimens disclosed intrathecal synthesis of oligoclonal IgG antibodies to 1 or more of 6 viruses (measles, herpes simplex type 1, varicella-zoster, cytomegalo, mumps, rota) during the observation period in 8 patients. Changes in patterns of intrathecally synthesized viral antibodies, characterized by the appearance of "new" antibody populations and the waxing or waning of others were observed in 6 patients. The results suggest that the early stage of MS in some patients is associated with transient as well as permanent recruitment of B cell clones producing viral antibodies of different specificities.

Antibodies, Viral↗

F1 polypeptides of two canine distemper virus strains: variation in the conserved N-terminal hydrophobic region.

The fusion protein of canine distemper virus was isolated by immunoadsorption from two virus strains, the rapidly growing Onderstepoort strain (forming large plaques) and the Convac vaccine strain (forming microplaques). The F1 subunits of the two fusion proteins were purified by preparative polyacrylamide gel electrophoresis. Direct amino acid sequence analysis revealed that 36-residue N-terminal regions of the proteins from the two strains are identical except at position 9, where Ala in the Convac strain is substituted by Val in the Onderstepoort strain. The two sequences show high homology with the previously determined N-terminal sequence of the F1 polypeptide of measles virus, and moderate homology with corresponding sequences of five paramyxoviruses, emphasizing the occurrence of an extensive conservation of these structures.

Amino Acid Sequence↗

Induction of class I and class II transplantation antigens in rat brain during fatal and non-fatal measles virus infection.

Measles virus induced a marked increase in the expression of MHC-coded class I and class II antigens as detected by immunostaining during both fatal and non-fatal brain infections in rats. The distribution of these molecules in the brain was much more widespread than the occurrence of viral antigen suggesting a soluble factor for their induction. In 14-day-old rats with a non-fatal infection there was a marked infiltration of T lymphocytes of 'cytotoxic/suppressor' phenotype in the brain parenchyma, whereas T 'helper' cell phenotypes mainly were located perivascularly. In brains from newborn rats with a fatal infection no or only few lymphocytes were detected.

Animals↗

Measurement of polypeptide- and antigenic site-specific antibodies to measles virus using a competitive enzyme immunoassay.

A competitive enzyme immunoassay (EIA) for measuring polypeptide- and antigenic site-specific antibodies against measles virus (MV) was developed. Three monoclonal antibodies (MAbs) against the fusion (F) and one MAb against the hemagglutinin (H) protein of MV were labelled with biotin and used in biotin-streptavidin EIA system. The inhibition of the binding of the labelled MAbs by the bound serum samples was measured in EIA and the titers were defined by reading at the level of 25% inhibition. The titers increased from acute to early convalescent measles sera in all the tests and antibody levels to the F protein increased earlier than antibodies to the H protein. The ratio of antibodies against 2 different antigenic sites on the F protein was significantly higher in subacute sclerosing panencephalitis patients than in normal blood donors. Similarly, SJL mice immunized with MV antigen had higher ratios of the corresponding antibodies than Balb/c mice. The results indicate that antibody response to different polypeptides and antigenic sites on the same polypeptide can be studied with the developed method and that the responses against different antigenic sites are independent.

Animals↗

Site-directed serology with synthetic peptides representing the large glycoprotein G of respiratory syncytial virus.

A set of 23 nested 15-amino-acid-long peptides with overlaps of 5 amino acids, representing the complete extramembranous part of the large glycoprotein G of respiratory syncytial (RS) virus, was analyzed in ELISA against different sera containing virus-specific antibodies. Seven of the peptides reacted with rabbit hyperimmune sera against purified virions. In contrast, only one of these seven peptides reacted with murine monoclonal antibodies specific for G. In connection with RS virus infections in humans, increase of antibody titers against three peptides was found in about one-third of the cases. These three peptides were included among those identified by both murine and rabbit antibodies. The present findings may open possibilities for site-directed clinical serology in the case of RS virus infections.

Antibodies, Monoclonal↗

Radioimmunoprecipitation and Western blotting with sera of human immunodeficiency virus infected patients: a comparative study.

The sensitivity and specificity of radioimmunoprecipitation assay (RIPA) and Western blot (WB) test were compared by use of a collection of 183 sera, representing different categories of individuals, noninfected or infected with the human immunodeficiency virus (HIV). The sera were subdivided on the basis of their reactivity in at least two anti-HIV enzyme-linked immunosorbent assays (ELISA); 53 sera were negative and 61 sera were positive in both tests, whereas 69 sera showed ambiguous reactions. The reaction patterns in RIPA could be divided into 6 different groups. The same grouping could to some extent be applied to the results of WB test. RIPA provided the most efficient means for identification of the large viral envelope glycoproteins, gp120/160, whereas gp41 was detected more effectively by WB. Internal virus components reacted to a varying extent with specific antibodies in the two tests. The reaction with pol products was more pronounced with the WB tests in which extracellular material was used as antigen. In a few WB tests, however, the reaction with internal components did not reflect a prior HIV infection. No such ambiguity was observed with RIPA, reflecting the advantage of a test that uses a minimally denaturated antigen and provides appropriate conditions for identification of the large viral glycoproteins. The practical choice of confirmatory tests to be used in diagnostic laboratories requires evaluation of both the sensitivity and specificity of the tests but also their economy and convenience.

Acquired Immunodeficiency Syndrome↗

A new human retrovirus isolate of West African origin (SBL-6669) and its relationship to HTLV-IV, LAV-II, and HTLV-IIIB.

A new human retrovirus of West African origin (SBL-6669) has been isolated from a patient with immunological and clinical signs of immunodeficiency. Using radioimmunoprecipitation assays (RIPA) and Western blot (WB) tests with human sera, the new virus isolate has been compared with HTLV-IV, LAV-II, and the HTLV-IIIB prototype strain of the human immunodeficiency virus (HIV). The West African isolates appeared to be members of the same virus group since their glycoproteins were antigenically indistinguishable. West African sera showed no detectable cross reaction with HTLV-IIIB glycoproteins. The external glycoprotein in the different virus strains only showed minor variations in size. The size of the transmembranous protein was not unambiguously defined. In the West African virus isolates a 30-35 kD protein was seen similar to the protein previously described possibly to represent this component. However, in SBL-6669 a distinct 41 kD protein was also identified. There were interstrain variations in the size of several viral proteins among the West African virus isolates. Only minor differences were seen between SBL-6669 and LAV-II. The variations were most pronounced in two core proteins corresponding to the 19 kD and 24 kD proteins of HTLV-IIIB. In addition, West African human retroviruses appear to differ in pathogenicity. LAV-II and SBL-6669 are associated with immunodeficiency, whereas HTLV-IV was isolated from healthy individuals. Since further spread of these viruses to other parts of the world is imminent, it is necessary to consider their antigenic and immunogenic properties in serodiagnosis of HIV infections and in planning for immunoprophylactic interventions.

Antigens, Viral↗

Visualization of defective measles virus particles in cerebrospinal fluid in subacute sclerosing panencephalitis.

Measles virus particles were visualized in the CSF of two patients with verified subacute sclerosing panencephalitis (SSPE) by using scanning electron microscopy. Immunologic identification of the accumulated particles was performed with monoclonal antibodies, directly conjugated to carboxylated microspheres, specific for different measles virus antigens. The beads were amassed on the filter surface after a 1-hr incubation in the CSF. Spherical particles with a diameter ranging between 150 and 500 nm were detected. Such particles bound specifically to latex beads covered by monoclonal antibodies to measles virus hemagglutinin but not to beads conjugated with monoclonal antibodies specific for nucleoprotein. Adding the two monoclonal antibodies to measles virus hemagglutinin to the CSF agglutinated the virus particles in a dose-dependent way. Further, no particles in the CSF bound to microspheres conjugated with monoclonal antibodies to non-related antigens of Sendai virus, cytomegalovirus, or human immunodeficiency virus. Similarly sized particles were also identified by transmission electron microscopy after concentrating the CSF.

Antigens, Viral↗

Preparation and characterization of monoclonal antibodies directed against five structural components of human respiratory syncytial virus subgroup B.

Mouse hybridomas producing antibodies against the structural proteins of strain WV4843, a subgroup B strain of respiratory syncytial (RS) virus, were produced by fusion of Sp2/0 myeloma cells with spleen cells from BALB/c mice immunized with purified preparations of the virus. After immunoprecipitation test with [35S]methionine-labelled extracellular virions, 35 clones found to produce antibodies against the fusion (F) protein, six against the member (M) protein, 21 against the nucleocapsid (NP) and eight against the phospho- (P) protein were further characterized. Immunoprecipitation with [3H]glucosamine-labelled intracellular virus polypeptides detected nine hybridoma cell lines producing antibodies against the large glyco- (G) protein of the virus. By competitive binding ELISA tests with monoclonal antibodies against each of the structural components, a minimum of two, 24, four, 15 and three epitopes were detected on the G, F, M, NP and P proteins, respectively. Eleven monoclonal antibodies directed against nine epitopes of the F protein could neutralize the infectivity of the virus. In contrast, none of the nine monoclonal antibodies against G could neutralize the infectivity of the virus. In order to find out more about the antigenic relationship between human and bovine RS virus strains all monoclonal antibodies were reacted with subgroup A RS virus and also with three different strains of bovine RS virus and one strain of caprine RS virus in immunofluorescence, ELISA and immunoprecipitation tests. In addition, 31 previously developed monoclonal antibodies against subgroup A virus were reacted with the bovine and caprine strains. The numbers of monoclonal antibodies of subgroup B specific for the B type of the two human subgroups were 9/9, 3/35, 0/6, 0/21, 0/8, for the G, F, M, NP and P proteins, respectively. No antigenic variations were found between the three bovine strains and the caprine strain. They did not react with the nine monoclonal antibodies against the G protein of subgroup B, nor did they react with nine monoclonal antibodies against subgroup A. Most but not all of the monoclonal antibodies against the other structural proteins of the two human RS virus subgroups reacted with the four strains. All 11 monoclonal antibodies against the F protein of subgroup B that could neutralize the infectivity of subgroup B also reacted with the bovine strains and neutralized their infectivity. It is concluded that although the bovine strains share many epitopes with the two human subgroups, they are antigenically distinct from the human viruses.

Antibodies, Monoclonal↗