Transcriptional Heterogeneity of IgM+ Cells in Rainbow Trout (Oncorhynchus mykiss) Tissues

Abós, Beatriz; Castro, Rosario; Pignatelli, Jaime; Luque, Alfonso; González, Lucia; Tafalla, Carolina

doi:10.1371/journal.pone.0082737

Cited by 58 publications

(51 citation statements)

References 62 publications

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“…After stimulation, leukocytes were collected from the wells by gentle scraping, resuspended in phosphate-buffered saline (PBS), and incubated for 30 min on ice with a specific anti-trout IgM monoclonal antibody (MAb) (1.14) coupled to phycoerythrin (PE) (25). Following two washing steps, cells were resuspended in PBS, and IgM-positive cells were isolated by fluorescence-activated cell sorting (FACS) in a BD FACSAria III instrument (BD Biosciences), first by using their forwardscatter (FSC)/side-scatter (SSC) profiles (to exclude the granulocyte gate) and then on the basis of the fluorescence emitted by the anti-trout IgM antibody (16). IgM ϩ and IgM Ϫ cells were then collected into different tubes for RNA isolation.…”

Section: Methodsmentioning

confidence: 99%

“…Reverse transcription was also performed by using the Power SYBR green Cells-to-Ct kit (Invitrogen) according to the manufacturer's instructions. To evaluate the levels of transcription of the different genes, real-time PCR was performed with a LightCycler 96 System instrument (Roche), using SYBR green PCR core reagents (Applied Biosystems) and specific primers described previously (16,(26)(27)(28) and summarized in Table 1. The efficiency of amplification was determined for each primer pair by using serial 10-fold dilutions of pooled cDNA, and only primer pairs with efficiency values of between 1.95 and 2 were used.…”

Section: Methodsmentioning

confidence: 99%

“…Accordingly, rainbow trout (Oncorhynchus mykiss) IgM-positive (IgM ϩ ) cells from different tissues, including kidney, spleen, blood, intestine, gills, and liver, have relatively high constitutive transcription levels of TLR1, TLR2, TLR3, TLR5, TLR7, TLR8a, TLR9, and TLR22 (16 (17,18).…”

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confidence: 99%

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Early Activation of Teleost B Cells in Response to Rhabdovirus Infection

Abós

Castro

Granja

et al. 2015

J Virol

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To date, the response of teleost B cells to specific pathogens has been only scarcely addressed. In this work, we have demonstrated that viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus, has the capacity to infect rainbow trout spleen IgMpositive (IgM ؉ ) cells, although the infection is not productive. Consequently, we have studied the effects of VHSV on IgM ؉ cell functionality, comparing these effects to those elicited by a Toll-like receptor 3 (TLR3) ligand, poly(I·C). We found that poly(I·C) and VHSV significantly upregulated TLR3 and type I interferon (IFN) transcription in spleen and blood IgM In mammals, Toll-like receptors (TLRs) recognize highly conserved structures of viral (TLR3, -7, -8, and -9) and bacterial (TLR1, -2, -4, -5, -6, -7, -8, and -9) origins. While TLR1, -2, -4, -5, and -6, together with TLR11 and TLR12 in mice and TLR10 in humans, are mostly expressed on the cell surface, a second group of TLRs, including TLR3, -7, -8, and -9, are localized within endosomal compartments and detect foreign nucleic acids (1). Recognition of pathogen-associated molecular patterns (PAMPs) through TLRs and other pattern recognition receptors (PRRs) leads to the activation and maturation of innate immune cells such as macrophages or dendritic cells (DCs). Additionally, once the presence of several TLR receptors on distinct populations of human and murine B cells was verified, further investigations concluded that B cells have evolved to directly sense microbes and that this TLR-mediated activation of B cells contributes to the establishment of an adequate humoral response (2). However, controversy remains as to what degree TLR signaling in B cells conditions the antibody response. On one hand, early studies showed that mice lacking B cell TLR signaling failed to mount an efficient antibody response (3). However, subsequent studies suggested a slightly different model in which these receptors play a role in the regulation of antibody class switching and in sustaining antibody secretion at late times after immunization in B cells (4), contributing to the amplification of the humoral response but not being completely responsible for it (5). In support of these observations, further studies demonstrated that the primary responses of some immunoglobulin (Ig) subclasses (i.e., IgG2a or IgG2c) were absolutely dependent on signaling through the adaptor protein MyD88, used by most TLRs, whereas other Ig classes were not (IgG1 and IgG3) or were much less (IgG2b and IgA) dependent on the MyD88 signaling cascade (6, 7). Interestingly, the conditional deletion of MyD88 in either DCs or B cells revealed that the antibody response to virus-like particles required TLR signaling in B cells, while the response to a soluble antigen was dependent on TLR signaling on DCs (8). This result reveals an ability of B cells to discriminate among antigens based on their physical form.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Early Activation of Teleost B Cells in Response to Rhabdovirus Infection

Abós

Castro

Granja

et al. 2015

J Virol

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“…After 30 days, four control and six infected fish were sacrificed and the head kidney was removed. Head kidney leukocytes were isolated as described before (19). ELISPOT plates containing Inmobilon-P membranes (Millipore) were activated with 70% ethanol for 30 s, coated with anti-trout IgM monoclonal antibody (MAb; clone 4C10) at 2 g/ml diluted in phosphate-buffered saline (PBS), and incubated overnight at 4°C.…”

Section: Table 1 Primers Used In This Studymentioning

confidence: 99%

Attenuated Infectious Hematopoietic Necrosis Virus with Rearranged Gene Order as Potential Vaccine

Rouxel

Tafalla

Mérour

et al. 2016

J Virol

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“…It might be possible that less evolved lymphoid organizations in mucosal tissues such as the gills or the digestive tract are also capable of initiating an immune response locally, because B and T cells have been reported in most fish tissues (9,10). In any case, there are many aspects of B cell biology still unknown in teleost fish.…”

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confidence: 99%

CCR7 Is Mainly Expressed in Teleost Gills, Where It Defines an IgD+IgM− B Lymphocyte Subset

Castro

Bromage

Abós

et al. 2014

The Journal of Immunology

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Chemokine receptor CCR7, the receptor for both CCL19 and CCL21 chemokines, regulates the recruitment and clustering of circulating leukocytes to secondary lymphoid tissues, such as lymph nodes and Peyer's patches. Even though teleost fish do not have either of these secondary lymphoid structures, we have recently reported a homolog to CCR7 in rainbow trout (Oncorhynchus mykiss). In the present work, we have studied the distribution of leukocytes bearing extracellular CCR7 in naive adult tissues by flow cytometry, observing that among the different leukocyte populations, the highest numbers of cells with membrane (mem)CCR7 were recorded in the gill (7.5 ± 2% CCR7+ cells). In comparison, head kidney, spleen, thymus, intestine, and peripheral blood possessed <5% CCR7+ cells. When CCR7 was studied at early developmental stages, we detected a progressive increase in gene expression and protein CCR7 levels in the gills throughout development. Surprisingly, the majority of the CCR7+ cells in the gills were not myeloid cells and did not express membrane CD8, IgM, nor IgT, but expressed IgD on the cell surface. In fact, most IgD+ cells in the gills expressed CCR7. Intriguingly, the IgD+CCR7+ population did not coexpress memIgM. Finally, when trout were bath challenged with viral hemorrhagic septicemia virus, the number of CCR7+ cells significantly decreased in the gills while significantly increased in head kidney. These results provide evidence of the presence of a novel memIgD+memIgM− B lymphocyte subset in trout that expresses memCCR7 and responds to viral infections. Similarities with IgD+IgM− subsets in mammals are discussed.

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Transcriptional Heterogeneity of IgM+ Cells in Rainbow Trout (Oncorhynchus mykiss) Tissues

Cited by 58 publications

References 62 publications

Early Activation of Teleost B Cells in Response to Rhabdovirus Infection

Early Activation of Teleost B Cells in Response to Rhabdovirus Infection

Attenuated Infectious Hematopoietic Necrosis Virus with Rearranged Gene Order as Potential Vaccine

CCR7 Is Mainly Expressed in Teleost Gills, Where It Defines an IgD+IgM− B Lymphocyte Subset

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