Flow cytometric-based protocols for assessing anti-MT-2 IgG1 reactivity: High-dimensional data handling to define predictors for clinical follow-up of Human T-cell Leukemia virus type-1 infection

Coelho-dos-Reis, Jordana Grazziela Alves; Peruhype-Magalhães, Vanessa; Pascoal-Xavier, Marcelo Antônio; Gomes, Matheus de Souza; Amaral, Laurence Rodrigues do; Cardoso, Ludmila Melo; Jonathan-Gonçalves, Juan; Ribeiro, Ágata Lopes; Starling, Ana Lúcia Borges; Ribas, João Gabriel Ramos; Gonçalves, Denise Utsch; Carneiro-Proietti, Anna Bárbara de Freitas; Teixeira-Carvalho, Andréa; Martins‐Filho, Olindo Assis; Giph,

doi:10.1016/j.jim.2017.02.006

Cited by 6 publications

(9 citation statements)

References 32 publications

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“…The binding magnitude to naturally occurring sequence variants from apes and Cercopithecus was almost identical for all plasma samples. A direct relationship between viral load and the level of the antibody response is frequently observed among individuals chronically infected with HIV-1 or HTLV-1 (25)(26)(27)(28)(29)(30). Thus, the lower antibody response to Cercopithecus SFV than to ape SFV in humans may be indirect evidence for lower replication levels of Cercopithecus SFV during either the primary or the chronic phase of the infection.…”

Section: Discussionmentioning

confidence: 99%

An Immunodominant and Conserved B-Cell Epitope in the Envelope of Simian Foamy Virus Recognized by Humans Infected with Zoonotic Strains from Apes

Lambert

Batalie

Montange

et al. 2019

J Virol

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Cross-species transmission of simian foamy viruses (SFVs) from nonhuman primates (NHPs) to humans is currently ongoing. These zoonotic retroviruses establish lifelong persistent infection in their human hosts. SFV are apparently nonpathogenic in vivo, with ubiquitous in vitro tropism. Here, we aimed to identify envelope B-cell epitopes that are recognized following a zoonotic SFV infection. We screened a library of 169 peptides covering the external portion of the envelope from the prototype foamy virus (SFVpsc_huHSRV.13) for recognition by samples from 52 Central African hunters (16 uninfected and 36 infected with chimpanzee, gorilla, or Cercopithecus SFV). We demonstrate the specific recognition of peptide N 96 -V 110 located in the leader peptide, gp18 LP . Forty-three variant peptides with truncations, alanine substitutions, or amino acid changes found in other SFV species were tested. We mapped the epitope between positions 98 and 108 and defined six amino acids essential for recognition. Most plasma samples from SFV-infected humans cross-reacted with sequences from apes and Old World monkey SFV species. The magnitude of binding to peptide N 96 -V 110 was significantly higher for samples of individuals infected with a chimpanzee or gorilla SFV than those infected with a Cercopithecus SFV. In conclusion, we have been the first to define an immunodominant B-cell epitope recognized by humans following zoonotic SFV infection. IMPORTANCE Foamy viruses are the oldest known retroviruses and have been mostly described to be nonpathogenic in their natural animal hosts. SFVs can be transmitted to humans, in whom they establish persistent infection, like the simian lenti-and deltaviruses that led to the emergence of two major human pathogens, human immunodeficiency virus type 1 and human T-lymphotropic virus type 1. This is the first identification of an SFV-specific B-cell epitope recognized by human plasma samples. The immunodominant epitope lies in gp18 LP , probably at the base of the envelope trimers. The NHP species the most genetically related to humans transmitted SFV strains that induced the strongest antibody responses. Importantly, this epitope is well conserved across SFV species that infect African and Asian NHPs.

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Section: Discussionmentioning

confidence: 99%

An Immunodominant and Conserved B-Cell Epitope in the Envelope of Simian Foamy Virus Recognized by Humans Infected with Zoonotic Strains from Apes

Lambert

Batalie

Montange

et al. 2019

J Virol

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“…Two parallel protocols of “FC-Duplex IgG1 (HTLV-1/2)” have been tested, referred to as “FIX” and “FIX & PERM”. Previous studies from our group, originally describing the innovative approach to detect anti-HTLV-1 IgG1 antibodies by flow cytometry, have tested the performance of a simplex protocol, using only the MT-2 cell line, applied to the diagnosis of HTLV-1 infection ( 20 ). The simplex system has been tested using both “FIX” and “FIX & PERM” protocols, demonstrating excellent performance for the diagnosis of HTLV-1 infection.…”

Section: Discussionmentioning

confidence: 99%

“…The MT-2 and MoT single-cell suspensions were submitted to the “FIX” protocol, as previously described by Coelho-Dos-Reis et al. ( 20 ), modified as follows: Aliquots of single-cell suspensions (1.0 × 10 6 cells/ml) were mixed with equal volume of FACS Fix Solution (10 g/L of paraformaldehyde; 10.2 g/L of sodium cacodylate; 6.63 g/L of sodium chloride, pH 7.2) and incubated overnight at 4°C. The “FIX” cell suspensions were washed once with PBS (400 × g , 10 min, 4°C) and maintained at 4°C until use in the “FIX” protocol.…”

Section: Methodsmentioning

confidence: 99%

“…In parallel, aliquots of MT-2 and MoT single-cell suspensions were submitted to the “FIX & PERM” protocol, as previously described by Coelho-Dos-Reis et al. ( 20 ), modified as follows: aliquots of “FIX” cells (1.0 × 10 6 cells/ml in PBS) were centrifuged and resuspended in equal volume of PBS supplemented with 0.5% bovine serum albumin plus 0.1% sodium azide, 0.5% saponin (PBS-P) and incubated for 10 min at room temperature. Following the “FIX & PERM”, cells were washed once (400 × g , 10 min, 4°C) with PBS supplemented with 0.5% bovine serum albumin plus 0.1% sodium azide (PBS-W) and maintained at 4°C until use in the “FIX & PERM” protocol.…”

Section: Methodsmentioning

confidence: 99%

“…Previous studies from our group have demonstrated the applicability and accuracy of flow cytometry-based methods for detecting specific anti-HTLV-1 IgG1 antibodies ( 19 , 20 ). Aiming at optimizing a single-step serological procedure to differentiate the infection with HTLV-1 and HTLV-2 types, in the present study, we have developed an innovative flow cytometry assay, referred to as “FC-Duplex IgG1 (HTLV-1/2)”, for universal and differential diagnosis of HTLV infections.…”

Section: Introductionmentioning

confidence: 99%

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A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection

et al. 2022

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In the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as “FC-Duplex IgG1 (HTLV-1/2)”, for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in a single competitive immunofluorescence platform by flow cytometry using fluorescently labeled MT-2/MoT cell line mix coupled to a highly sensitive development system (Biotin/Streptavidin/Phycoerythrin). The stability of fluorescent labeling and the antigenicity of MT-2 and MoT cell lines were confirmed upon storage at −20°C for 2, 6, and 12 months. The anti-HTLV-1/2 IgG1 reactivity, expressed as percentage of positive fluorescent cells (PPFC), was evaluated for each target antigen along the titration curve of test serum samples (1:32 to 1:4,096). Upon selection of target cell line and serum dilutions with higher segregation score between groups, the performance of “FIX” and “FIX & PERM” protocols was evaluated. The “FIX” protocol presented excellent performance indices (Se = 92%/Sp = 94%/AUC = 0.96; Se = 96%/Sp = 100%/AUC = 0.99) for the universal (HTLV-1/2 vs. NI) and differential (HTLV-1 vs. HTLV-2) diagnosis of HTLV-1 infection, respectively. Optimization of the “FIX” protocol using the principle of synchronous and asynchronous pairwise analysis further improved the performance of “FC-Duplex IgG1 (HTLV-1/2)”, using the “FIX” protocol for differential diagnosis of HTLV-1 and HTLV-2 infections (Se = 100%/Sp = 100%/AUC = 1.00). In conclusion, the “FC-Duplex IgG1 (HTLV-1/2)” method represents an innovation in the biotechnology segment with the potential to compose a serological kit for differential diagnosis of HTLV-1/2 infection for reference laboratories and blood centers.

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Establishing tools for early diagnosis of congenital toxoplasmosis: Flow cytometric IgG avidity assay as a confirmatory test for neonatal screening

Zacche-Tonini

Fonseca

Jesus

et al. 2017

Journal of Immunological Methods

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Flow cytometric-based protocols for assessing anti-MT-2 IgG1 reactivity: High-dimensional data handling to define predictors for clinical follow-up of Human T-cell Leukemia virus type-1 infection

Cited by 6 publications

References 32 publications

An Immunodominant and Conserved B-Cell Epitope in the Envelope of Simian Foamy Virus Recognized by Humans Infected with Zoonotic Strains from Apes

An Immunodominant and Conserved B-Cell Epitope in the Envelope of Simian Foamy Virus Recognized by Humans Infected with Zoonotic Strains from Apes

A New Flow Cytometry-Based Single Platform for Universal and Differential Serodiagnosis of HTLV-1/2 Infection

Establishing tools for early diagnosis of congenital toxoplasmosis: Flow cytometric IgG avidity assay as a confirmatory test for neonatal screening

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