Use of a New Dual-Antigen Enzyme-Linked Immunosorbent Assay To Detect and Characterize the Human Antibody Response to the Human Immunodeficiency Virus Type 2 Envelope gp125 and gp36 Glycoproteins

Marcelino, José Maria; Barroso, Helena; Gonçalves, Fátima; Silva, Sofia M. da; Novo, Carlos; Gómes, Perpétua; Camacho, Ricardo Jorge; Taveira, Nuno

doi:10.1128/jcm.44.2.607-611.2006

Cited by 19 publications

(21 citation statements)

References 47 publications

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“…To investigate the humoral immunogenicity of HIV-2ALI-derived envelope gp125t and rpC2-C3 polypeptide (37), BALB/c mice were inoculated with one or more of the following immunogens: rVV/ALIM2, gp125t or rpC2-C3 polypeptide (see Table S1 in the supplemental material). The binding antibody response to each immunogen was analyzed with the ELISA-HIV2 assay (37), which uses the rpC2-C3 polypeptide as antigen, and with a newly derived ELISA using gp125t as a capture antigen. All mice produced IgG antibodies reacting with gp125t or rpC2-C3 polypeptide (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A new recombinant vaccinia virus, rVV/ALIM2, was produced expressing high levels of a truncated version of gp125 from HIV-2ALI. Mice immunized with this virus elicited a binding IgG antibody response that is similar to natural HIV-2 infection (37,38) and elicited the production of low levels of NAbs. In contrast, animals vaccinated solely with monomeric gp125t or rpC2-C3, despite eliciting a binding IgG antibody response that was even stronger than that attained in natural infection (4.4 log 10 versus 3.3 log 10 ) (38), could not raise the production of NAbs.…”

Section: Discussionmentioning

confidence: 99%

“…These core Env regions contain highly antigenic and immunodominant HIV-2 epitopes, and most HIV-2-infected patients produce IgG and IgA antibodies reacting with them (37,38). We have recently shown that the V3 and C3 regions in the HIV-2 envelope are remarkably stable over the course of infection and that C2V3C3-specific IgG antibodies may contribute to reduce viral population size and limit the number of virus escape mutants (10).…”

Section: Discussionmentioning

confidence: 99%

“…Another two groups (IV and VI) of mice were immunized by the i.p. route with 10 g of the rpC2-C3 polypeptide (37) or soluble gp125t emulsified in complete Freund adjuvant (priming) or incomplete Freund adjuvant (boosts). Four mice were used in each group.…”

Section: Methodsmentioning

confidence: 99%

“…Enzyme-linked immunosorbent assay (ELISA) plates were coated with the rpC2-C3 polypeptide as described previously (37). To produce gp125t for ELISAs, 293 cells were infected with rVV/ALIM2 (4 PFU per cell).…”

Section: Methodsmentioning

confidence: 99%

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Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

Marcelino¹,

Borrego

Rocha

et al. 2010

J Virol

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Human immunodeficiency virus type 2 (HIV-2) infection affects about 1 to 2 million individuals, the majority living in West Africa, Europe, and India. As for HIV-1, new strategies for the prevention of HIV-2 infection are needed. Our aim was to produce new vaccine immunogens that elicit the production of broadly reactive HIV-2 neutralizing antibodies (NAbs). Native and truncated envelope proteins from the reference HIV-2ALI isolate were expressed in vaccinia virus or in bacteria. This source isolate was used due to its unique phenotype combining CD4 independence and CCR5 usage. NAbs were not elicited in BALB/c mice by single immunization with a truncated and fully glycosylated envelope gp125 (gp125t) or a recombinant polypeptide comprising the C2, V3, and C3 envelope regions (rpC2-C3). A strong and broad NAb response was, however, elicited in mice primed with gp125t expressed in vaccinia virus and boosted with rpC2-C3. Serum from these animals potently neutralized (median 50% neutralizing titer, 3,200) six of six highly divergent primary HIV-2 isolates. Coreceptor usage and the V3 sequence of NAb-sensitive isolates were similar to that of the vaccinating immunogen (HIV-2ALI). In contrast, NAbs were not reactive on three X4 isolates that displayed major changes in V3 loop sequence and structure. Collectively, our findings demonstrate that broadly reactive HIV-2 NAbs can be elicited by using a vaccinia virus vector-prime/rpC2-C3-boost immunization strategy and suggest a potential relationship between escape to neutralization and cell tropism.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

Marcelino¹,

Borrego

Rocha

et al. 2010

J Virol

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Genotypic and Phenotypic Diversity of Cyanobacteria in Biological Soil Crusts of the Succulent Karoo and Nama Karoo of Southern Africa

et al. 2013

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Biological soil crusts (BSCs) are communities of cryptogamic organisms, occurring in arid and semiarid regions all over the world. Based on both morphological identification and genetic analyses, we established a first cyanobacterial inventory using the biphasic approach for BSCs within two major biomes of southern Africa. The samples were collected at two different sites in the Succulent Karoo and one in the Nama Karoo. After cultivation and morphological identification, the 16S rRNA gene was sequenced from the cyanobacterial cultures. From the soil samples, the DNA was extracted, and the 16S rRNA gene sequenced. All the sequences of the clone libraries from soil and cultures were compared with those of the public databases. Forty-five different species were morphologically identified in the samples of the Succulent Karoo (observatories of Soebatsfontein and Goedehoop). Based on the genetic analyses, 60 operational taxonomic units (OTUs) were identified for the Succulent Karoo and 43 for the Nama Karoo (based on 95% sequence similarity). The cloned sequences corresponded well with the morphologically described taxa in cultures and sequences in the public databases. Besides known species of typical crust-forming cyanobacterial genera (Microcoleus, Phormidium, Tolypothrix and Scytonema), we found sequences of so far undescribed species of the genera Leptolyngbya, Pseudanabaena, Phormidium, Oscillatoria, Schizothrix and Microcoleus. Most OTUs were restricted to distinct sites. Grazed soils showed lower taxa numbers than undisturbed soils, implying the presence of early successional crust types and reduced soil surface protection. Our combined approach of morphological identification and genetic analyses allowed both a taxa inventory and the analysis of species occurring under specific habitat conditions.

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Development of an immunoassay for differentiating human immunodeficiency virus infections — from vaccine-induced immune response in Tiantan vaccine trials in China

Yuan¹,

Chen²,

Zhang³

et al. 2012

Clinical Biochemistry

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Use of a New Dual-Antigen Enzyme-Linked Immunosorbent Assay To Detect and Characterize the Human Antibody Response to the Human Immunodeficiency Virus Type 2 Envelope gp125 and gp36 Glycoproteins

Cited by 19 publications

References 47 publications

Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

Potent and Broadly Reactive HIV-2 Neutralizing Antibodies Elicited by a Vaccinia Virus Vector Prime-C2V3C3 Polypeptide Boost Immunization Strategy

Genotypic and Phenotypic Diversity of Cyanobacteria in Biological Soil Crusts of the Succulent Karoo and Nama Karoo of Southern Africa

Development of an immunoassay for differentiating human immunodeficiency virus infections — from vaccine-induced immune response in Tiantan vaccine trials in China

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