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Research Note: Development and characterization of monoclonal antibodies specific for chicken interleukin-7 receptor α (CD127).

ABSTRACT: CD127, also named interleukin-7 receptor (IL-7R), is expressed on various cell types including naive and memory T cells, and plays a critical role in the differentiation and activation of T lymphocytes. The availability of poultry-specific immune reagents to identify and measure chicken CD127 response will enhance fundamental and applied research in poultry immunology. Mouse monoclonal antibodies (MAbs) against chicken CD127 (chCD127) were developed and characterized. More specifically, a 678 bp ectodomain of chCD127 gene was cloned in the pET28a (+) vector and expressed in BL21-AI E. coli competent cells. The recombinant chCD127 protein with a size of 30 KDa which was also recognized by a mouse anti-human CD127 MAb (Clone G-11) was used to immunize mice, and 6 new mouse MAbs which specifically detected chicken CD127 were developed and characterized. Availability of these new sets of chCD127-specific MAbs will facilitate the immunological studies on CD127 in poultry, especially in understanding effector and memory T immune cell responses in normal and diseased states.

SUBMITTER: Sun Z

PROVIDER: S-EPMC9396400 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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Research Note: Development and characterization of monoclonal antibodies specific for chicken interleukin-7 receptor α (CD127).

Sun Zhifeng Z Lu Mingmin M Kim Woohyun W Lillehoj Hyun H Panebra Alfredo A Zhao Hongyan H Li Charles C

Poultry science 20220709 10

CD127, also named interleukin-7 receptor (IL-7R), is expressed on various cell types including naive and memory T cells, and plays a critical role in the differentiation and activation of T lymphocytes. The availability of poultry-specific immune reagents to identify and measure chicken CD127 response will enhance fundamental and applied research in poultry immunology. Mouse monoclonal antibodies (MAbs) against chicken CD127 (chCD127) were developed and characterized. More specifically, a 678 bp ...[more]

PMID: 35973349

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Project description:BackgroundDengue virus (DV) infection causes a spectrum of clinical diseases ranging from dengue fever to a life-threatening dengue hemorrhagic fever. Four distinct serotypes (DV1-4), which have similar genome sequences and envelope protein (E protein) antigenic properties, were divided. Among these 4 serotypes, DV1 usually causes predominant infections and fast diagnosis and effective treatments are urgently required to prevent further hospitalization and casualties.MethodsTo develop antibodies specifically targeting and neutralizing DV1, we immunized mice with UV-inactivated DV1 viral particles and recombinant DV1 E protein from residue 1 to 395 (E395), and then generated 12 anti-E monoclonal antibodies (mAbs) as the candidates for a series of characterized assays such as ELISA, dot blot, immunofluorescence assay, western blot, and foci forming analyses.ResultsAmong the mAbs, 10 out of 12 showed cross-reactivity to four DV serotypes as well as Japanese encephalitis virus (JEV) in different cross-reactivity patterns. Two particular mAbs, DV1-E1 and DV1-E2, exhibited strong binding specificity and neutralizing activity against DV1 and showed no cross-reactivity to DV2, DV3, DV4 or JEV-infected cells as characterized by ELISA, dot blot, immunofluorescence assay, western blot, and foci forming analyses. Using peptide coated indirect ELISA, we localized the neutralizing determinants of the strongly inhibitory mAbs to a sequence-unique epitope on the later-ridge of domain III of the DV1 E protein, centered near residues T346 and D360 (346TQNGRLITANPIVTD360). Interestingly, the amino acid sequence of the epitope region is highly conserved among different genotypes of DV1 but diverse from DV2, DV3, DV4 serotypes and other flaviviruses.ConclusionsOur results showed two selected mAbs DV1-E1 and DV1-E2 can specifically target and significantly neutralize DV1. With further research these two mAbs might be applied in the development of DV1 specific serologic diagnosis and used as a feasible treatment option for DV1 infection. The identification of DV1 mAbs epitope with key residues can also provide vital information for vaccine design.

Dataset Information

Research Note: Development and characterization of monoclonal antibodies specific for chicken interleukin-7 receptor α (CD127).

Publications

Research Note: Development and characterization of monoclonal antibodies specific for chicken interleukin-7 receptor α (CD127).

Similar Datasets

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