Cross-reactivity of monoclonal antibodies against CD4-1 and CD8α of ginbuna crucian carp with lymphocytes of zebrafish and other cyprinid species

Miyazawa, Ryuichiro; Matsuura, Yuta; Shibasaki, Yasuhiro; Imamura, Shintaro; Nakanishi, Teruyuki

doi:10.1016/j.dci.2016.12.002

Cited by 22 publications

(13 citation statements)

References 23 publications

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“…On the other hand, CD4-2-negative cells expressed CD3ε, CD4-1, TCRα, TCRβ, CD8α, CD8β, IgL and IgM transcripts. These results implied that the strong expression pattern of the CD8, IgL and IgM transcripts observed in the CD4-2-negative cells is distinctive in the CD4-negative T cell subsets in teleosts [25,33,34]. Additionally, these results revealed that the expression of CD4-2 lymphocytes is associated with cell surface expression of CD3ε and TCR, and CD4-2 lymphocytes are able to co-express with CD4-1 lymphocytes, as described in many previous studies [11,17,37].…”

Section: Discussionsupporting

confidence: 85%

“…Although antibodies recognizing cell surface proteins have been successfully developed, these may exhibit different reactivities depending on the cell preparation used, e.g., fixed cell, live cell or cell lysates [13,25]. However, the anti-CD4-2 mAb (3C8) produced in this study showed a good level of responsiveness to the fixed cells used by immunofluorescence staining, the naïve form of CD4-2 expressed on the surface of the live cells used in flow cytometry, and the denatured cell lysate from the leukocytes used in Western blotting.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of CD4-Positive Lymphocytes in the Antiviral Response of Olive Flounder (Paralichthys oliveceus) to Nervous Necrosis Virus

Jung

Chun

Lee

et al. 2020

IJMS

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The presence of CD4 T lymphocytes has been described for several teleost species, while many of the main T cell subsets have not been characterized at a cellular level, because of a lack of suitable tools for their identification, e.g., monoclonal antibodies (mAbs) against cell markers. We previously described the tissue distribution and immune response related to CD3ε and CD4-1 T cells in olive flounder (Paralichthys oliveceus) in response to a viral infection. In the present study, we successfully produce an mAb against CD4-2 T lymphocytes from olive flounder and confirmed its specificity using immuno-blotting, immunofluorescence staining, flow cytometry analysis and reverse transcription polymerase chain reaction (RT-PCR). Using these mAbs, we were able to demonstrate that the CD3ε T cell populations contain both types of CD4+ cells, with the majority of the CD4 T cell subpopulations being CD4-1+/CD4-2+ cells, determined using two-color flow cytometry analysis. We also examined the functional activity of the CD4-1 and CD4-2 cells in vivo in response to a viral infection, with the numbers of both types of CD4 T cells increasing significantly during the virus infection. Collectively, these findings suggest that the CD4 T lymphocytes in olive flounder are equivalent to the helper T cells in mammals in terms of their properties and function, and it is the CD4-2 T lymphocytes rather than the CD4-1 T cells that play an important role in the Th1 immune response against viral infections in olive flounder.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Characterization of CD4-Positive Lymphocytes in the Antiviral Response of Olive Flounder (Paralichthys oliveceus) to Nervous Necrosis Virus

Jung

Chun

Lee

et al. 2020

IJMS

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“…In a relatively small study, we showed that T cells control Proteobacteria (Vibrio) abundance in the zebrafish gut, providing evidence that like in mice the adaptive immune system plays a role in shaping the microbiota composition (25). T cells are present in the thymus by 4 dpf as shown by using CD4-1:mCherry transgenic zebrafish (26) and CD8a+ antibody staining (27). It was shown that T cells egress from the thymus as early as 10 dpf.…”

Section: Zebrafish Intestine: Structure Function and Microbiotamentioning

confidence: 86%

Feed, Microbiota, and Gut Immunity: Using the Zebrafish Model to Understand Fish Health

et al. 2020

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“…The study of fish T cell responses has been hampered by the lack of specific antibodies and molecular tools. Nowadays, several T cell specific antibodies have been developed for a few fish species such as rainbow trout ( Oncorhynchus mykiss ), spotted green pufferfish ( Tetraodon nigroviridis ), ginbuna crucian carp ( Carassius auratus langsdorfii ) or Japanese pufferfish ( Takifugu rubripes ) [ 29 – 34 ] and some showed cross-reaction with close species [ 30 ]. In addition, CD4 + T cell lines have recently been established for common carp [ 35 ] and rainbow trout [ 32 ] and also CD4-transgenic zebrafish have been created [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

Hints on T cell responses in a fish-parasite model: Enteromyxum leei induces differential expression of T cell signature molecules depending on the organ and the infection status

et al. 2018

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BackgroudEnteromyxum leei is a myxozoan parasite that produces a slow-progressing intestinal disease. This parasite invades the paracellular space of the intestinal epithelium and progresses from the posterior to the anterior intestine. The aim of the present study was to gain insights into fish T cell responses in the gilthead sea bream-E. leei infection model using a PCR-array with 30 signature molecules for different leukocyte responses in head kidney, spleen, anterior and posterior intestine.ResultsThe PCR-array results suggest that E. leei induced migration of T cells from head kidney to intestines where TH1, CTL and TH17 profiles were activated and kept in balance by the upregulation of regulatory cytokines. These results were partially validated by the use of cross-reacting antibodies and BrdU immunostaining to monitor proliferation. Zap70 immunostaining supported the increased number of T cells in the anterior intestine detected by gene expression, but double staining with BrdU did not show active proliferation of this cell type at a local level, supporting the migration from lymphohaematopoietic tissues to the site of infection. Global analyses of the expression profiles revealed a clear separation between infected and exposed, but non-infected fish, more evident in the target organ. Exposed, non-infected animals showed an intermediate phenotype closer to the control fish.ConclusionsThese results evidence a clear modulation of the T cell response of gilthead sea bream upon E. leei infection. The effects occurred both at local and systemic levels, but the response was stronger and more specific at the site of infection, the intestine. Altogether, this research poses a promising basis to understand the response against this important parasite and establish effective preventive or palliative measures.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-3007-1) contains supplementary material, which is available to authorized users.

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Cross-reactivity of monoclonal antibodies against CD4-1 and CD8α of ginbuna crucian carp with lymphocytes of zebrafish and other cyprinid species

Cited by 22 publications

References 23 publications

Characterization of CD4-Positive Lymphocytes in the Antiviral Response of Olive Flounder (Paralichthys oliveceus) to Nervous Necrosis Virus

Characterization of CD4-Positive Lymphocytes in the Antiviral Response of Olive Flounder (Paralichthys oliveceus) to Nervous Necrosis Virus

Feed, Microbiota, and Gut Immunity: Using the Zebrafish Model to Understand Fish Health

Hints on T cell responses in a fish-parasite model: Enteromyxum leei induces differential expression of T cell signature molecules depending on the organ and the infection status

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