Immunosorbent Assay Based on Recombinant Hemagglutinin Protein Produced in a High-Efficiency Mammalian Expression System for Surveillance of Measles Immunity

Bouche, Fabienne B.; Ammerlaan, Wim; Berthet, Françoise; Houard, Sophie; Schneider, François

doi:10.1128/jcm.36.3.721-726.1998

Cited by 25 publications

(9 citation statements)

References 50 publications

(71 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…No false-negative serum was detected, and increases between paired sera were as significant as they were with MV-ELISA. In light of these results and the overall high sensitivity and specificity of H-ELISA for IgG (3,4), we conclude that this assay is as reliable for the diagnosis of measles in paired sera as an ELISA based on whole MV. Despite the longer persistence of maximal levels of IgM in MV-ELISA and the early waning of H-specific IgM, the initial time courses of H-and MV-specific IgGs are similar.…”

Section: Discussionmentioning

confidence: 76%

“…The nucleoprotein has been described as a suitable antigen to detect specific IgG and IgM (16,38,39). More recently we have shown that recombinant hemagglutinin (H) protein produced in a high-yield mammalian expression system can be used for the surveillance of measles immunity in late convalescents (3) and in vaccinees (4). The high sensitivity and specificity of this assay was due to high levels of detectable H-specific IgG antibodies in both of these cohorts.…”

mentioning

confidence: 99%

“…(iii) IgG detection. Specific IgG was measured by H-ELISA following the procedure described previously (3). In this assay, the H antigen and the negative control antigen were directly coated onto microtiter plates.…”

mentioning

confidence: 99%

“…In this assay, the H antigen and the negative control antigen were directly coated onto microtiter plates. The threshold for positivity (210 mOD) was defined with sera that were negative for HI and NT, as described previously (3).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

et al. 1998

Self Cite

View full text Add to dashboard Cite

Recombinant hemagglutinin (H) of the measles virus (MV) expressed in a mammalian high-expression system based on the Semliki Forest virus replicon was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG in patients with acute-phase measles. One hundred twelve serum specimens from 70 patients with measles were analyzed. Case definition was based on a commercial IgM ELISA that utilizes MV-infected cells (MV-ELISA) (Enzygnost; Behring Diagnostics); the clinical criteria of the Centers for Disease Control and Prevention (Atlanta, Ga.); and/or the increase in hemagglutinin test titers, neutralization test titers, and levels of MV-specific IgG whenever paired sera were available. The initial time courses of the IgM signal after the onset of rash are similar in the H- and MV-ELISAs. On days 0 to 19, both ELISAs detected IgM in 67 of 68 (98.5%) sera. Average maximal levels of IgM seem to persist, however, about 10 days longer in the MV-ELISA (up to day 25) than in the H-ELISA (day 15). From days 20 to 29 and 30 to 59, the H-ELISA detected only 64.3 (9 of 14) and 19.2% (5 of 26), respectively, of sera that were IgM positive by MV-ELISA. At least up to day 30, the performance of the H-ELISA seemed to be similar to that reported for commercial ELISAs based on whole MV. Our results demonstrate that MV H-specific IgM can be used to diagnose most measles cases from a single serum specimen collected within 19 days after the onset of rash and that the recombinant protein used in this study is suitable for this purpose.

show abstract

Section: Discussionmentioning

confidence: 76%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

et al. 1998

Self Cite

View full text Add to dashboard Cite

show abstract

“…The coding sequence corresponding to the measles virus hemagglutinin (MV-H) protein (Bouche et al, 1998a) was subcloned into the expression cassette of the pRTL2 vector at the NcoI-BamHI sites (Restrepo et al, 1990). In this vector, the MV-H sequence was under the control of the constitutively expressed cauliflower mosaic virus (CaMV) 35S promoter fused to the tobacco etch virus (TEV) 5 -untranslated region, a translational enhancer, and the CaMV 35S terminator (Odell et al, 1985;Pietrzak et al, 1986).…”

Section: Construction Of a Plant Expression Vectormentioning

confidence: 99%

Untitled

Marquet-Blouin

Bouche

Steinmetz

et al. 2003

Plant Molecular Biology

View full text Add to dashboard Cite

Although edible vaccines seem to be feasible, antigens of human pathogens have mostly been expressed in plants that are not attractive for human consumption (such as potatoes) unless they are cooked. Boiling may reduce the immunogenicity of many antigens. More recently, the technology to transform fruit and vegetable plants have become perfected. We transformed carrot plants with Agrobacterium tumefaciens to generate plants (which can be eaten raw) transgenic for an immunodominant antigen of the measles virus, a major pathogen in man. The hemagglutinin (H) glycoprotein is the principle target of neutralizing and protective antibodies against measles. Copy numbers of the H transgene were verified by Southern blot and specific transcription was confirmed by RT-PCR. The H protein was detected by western blot in the membrane fraction of transformed carrot plants. The recombinant protein seemed to have a 8% lower molecular weight than the viral protein. Although this suggests a different glycosylation pattern, proper folding of the transgenic protein was confirmed by conformational-dependent monoclonal antibodies. Immunization of mice with leaf or root extracts induced high titres of IgG1 and IgG2a antibodies that cross-reacted strongly with the measles virus and neutralized the virus in vitro. These results demonstrate that transgenic carrot plants can be used as an efficient expression system to produce highly immunogenic viral antigens. Our study may pave the way towards an edible vaccine against measles which could be complementary to the current live-attenuated vaccine.

show abstract