A monoclonal antibody recognising a surface marker on rainbow trout (Oncorhynchus mykiss) monocytes

Köllner, Bernd; Blohm, Ulrike; Kotterba, Günter; Fischer, Uwe

doi:10.1006/fsim.2000.0300

Cited by 34 publications

(16 citation statements)

References 40 publications

(42 reference statements)

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“…The majority of the GFP-positive cells were in the non-lymphocyte quadrant (Fig. 4, compare percentage in upper right quadrant to lower right quadrant), consistent with known properties of rainbow trout phagocytic cells (Kollner et al 2001, Moritomo et al 2003. Interestingly, there was an increase in the percentage of larger, granular cells in the spleen and peripheral blood of infected fish compared to sham injected controls, while there was a decrease in these types of cells in the anterior kidney.…”

supporting

confidence: 79%

“…4. Cells falling within the known lymphocyte, monocyte and neutrophil regions (Kollner et al 2001, Stafford et al 2001, Moritomo et al 2003 were included in the R1 gate. Spleen and anterior kid- 67: 267-272, 2005 ney tissues from both Y. ruckeri WT and YRNC10-gfp infections contained increased numbers of autofluorescent cells compared to sham-injected controls (data not shown).…”

Section: Detection Of Trout Cells Associated With Gfpexpressing Yersimentioning

confidence: 99%

Section: Detection Of Trout Cells Associated With Gfpexpressing Yersimentioning

confidence: 99%

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Construction of a virulent, green fluorescent protein-tagged Yersinia ruckeri and detection in trout tissues after intraperitoneal and immersion challenge

Welch

Wiens²

2005

Dis. Aquat. Org.

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A green fluorescent protein (GFP) expressing strain of Yersinia ruckeri was created by the transposition of a Tn10-GFP-kan cassette into the genome of Y. ruckeri Strain YRNC10. The derivative, YRNC10-gfp, was highly GFP fluorescent, retained the gfp-km marker in the absence of kanamycin selection, and exhibited in vitro growth kinetics similar to those of the wild type strain. YRNC10-gfp colonized and caused mortality in immersion and intraperitoneally challenged rainbow trout Oncorhynchus mykiss, although it was modestly attenuated compared to the wild type strain. The distribution and location of YRNC10-gfp in infected fish was visualized by epifluorescence microscopy. Abundant extracellular bacteria and a small number of intracellular bacteria were observed in the kidney, spleen and peripheral blood. To determine the percentage of trout cells containing intracellular bacteria, GFP fluorescence was measured by flow cytometry. A small population of GFP positive leukocytes was detected in peripheral blood (1.6%), spleen (1.1%) and anterior kidney (0.4%) tissues. In summary, this is the first report of the construction of a virulent, GFP-tagged Y. ruckeri, which may be a useful model for detecting and imaging the interactions between an aquatic pathogen and the natural salmonid host. KEY WORDS: Yersinia ruckeri · Green fluorescent protein · Flow cytometry · Epifluorescence microscopy Resale or republication not permitted without written consent of the publisherDis Aquat Org 67: [267][268][269][270][271][272] 2005 tagging allows in situ detection of bacteria-host interactions in real time. GFP fluorescence associated with tagged cells can easily be visualized and quantified by epifluorescence microscopy, flow cytometry, spectrofluorometry or fluorescence colony counting (Chalfie et al. 1994. More recently, artificial plasmids carrying the gene encoding the green fluorescent protein have been used to tag several fish pathogens (Ling et al. 2000, O'Toole et al. 2004, while these tagged strains have been useful for determining the hostpathogen interactions during the early stages of infection, the inherent instability of plasmid-mediated tags may limit their use in studies involving the long-term persistence of pathogens. Herein, we describe the use of a specialized transposon to construct a highly stable gfp-tagged derivative of Yersinia ruckeri and the use of this strain to visualize and quantify the interaction of this pathogen with trout immune cells. MATERIALS AND METHODSBacterial strains and culture conditions. Yersinia ruckeri Strain YRNC10 is a Serotype 1 strain that was isolated from a moribund rainbow trout collected at a fish farm in North Carolina. YRNC10 and its derivatives were cultured at 24°C in trypticase soy broth (TSB), or trypticase soy agar (TSA). Escherichia coli strains were grown at 37°C in Luria broth or Luria agar. When required, antibiotics (Sigma) were added at the following concentrations: ampicillin (100 µg ml -1 ); kanamycin (100 µg ml -1 for Y. ruckeri and 50 µg ml -1 for ...

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supporting

confidence: 79%

Section: Detection Of Trout Cells Associated With Gfpexpressing Yersimentioning

confidence: 99%

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Construction of a virulent, green fluorescent protein-tagged Yersinia ruckeri and detection in trout tissues after intraperitoneal and immersion challenge

Welch

Wiens²

2005

Dis. Aquat. Org.

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“…Because there are no Abs against extracellular pan-T cell markers available for rainbow trout, we used T cell-enriched cultures as responder cells. These T cell-enriched cultures from isogeneic and allogeneic splenocytes were obtained by depletion of splenocytes labeled with anti-IgM, anti-MHC class II mAbs and mAbs specific for thrombocytes (37) and myeloid cells (38), using flow sorting. The resulting negative population, representing ∼10% of splenocytes, was labeled with CellTrace Far Red (Life Technologies).…”

Section: Mlrmentioning

confidence: 99%

Identification of Teleost Skin CD8α+ Dendritic-like Cells, Representing a Potential Common Ancestor for Mammalian Cross-Presenting Dendritic Cells

Granja¹,

Leal²,

Pignatelli³

et al. 2015

The Journal of Immunology

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Although fish constitute the most ancient animal group in which an acquired immune system is present, the presence of dendritic cells (DCs) in teleosts has been addressed only briefly, and the identification of a specific DC subset in teleosts remained elusive because of the lack of specific Abs. In mice, DCs expressing CD8α+ in lymphoid tissues have the capacity to cross-present extracellular Ags to T cells through MHC I, similarly to tissue-derived CD103+ DCs and the human CD141+ DC population. In the current study, we identified a large and highly complex subpopulation of leukocytes coexpressing MHC class II and CD8α. This CD8α+ MHC II+ DC-like subpopulation constituted ∼1.2% of the total leukocyte population in the skin, showing phenotypical and functional characteristics of semimature DCs that seem to locally regulate mucosal immunity and tolerance in a species lacking lymph nodes. Furthermore, we identified trout homologs for CD141 and CD103 and demonstrated that, in trout, this skin CD8+ DC-like subpopulation expresses both markers. To our knowledge, these results provide the first evidence of a specific DC-like subtype in nonimmune tissue in teleosts and support the hypothesis of a common origin for all mammalian cross-presenting DCs.

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“…Mab against di#erent leucocyte populations used in this study were: mab N2 anti-trout IgM, B-lymphocytes [28], mab 7+mab 10 anti-trout 36 thrombocytes [28], mab 6-1 anti-trout granulocytes [29] and mab 45 anti-trout monocytes [30]. A mab against an antigen on Newcastle disease virus (a-NDV, mab 59) served as irrelevant mab control.…”

Section: Monoclonal Antibodies (Mabs)mentioning

confidence: 99%

Temperature dependent activation of leucocyte populations of rainbow trout, Oncorhynchus mykiss, after intraperitoneal immunisation with Aeromonas salmonicida

Köllner

Kotterba

2002

Fish & Shellfish Immunology

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A monoclonal antibody recognising a surface marker on rainbow trout (Oncorhynchus mykiss) monocytes

Cited by 34 publications

References 40 publications

Construction of a virulent, green fluorescent protein-tagged Yersinia ruckeri and detection in trout tissues after intraperitoneal and immersion challenge

Construction of a virulent, green fluorescent protein-tagged Yersinia ruckeri and detection in trout tissues after intraperitoneal and immersion challenge

Identification of Teleost Skin CD8α+ Dendritic-like Cells, Representing a Potential Common Ancestor for Mammalian Cross-Presenting Dendritic Cells

Temperature dependent activation of leucocyte populations of rainbow trout, Oncorhynchus mykiss, after intraperitoneal immunisation with Aeromonas salmonicida

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