Canine tissue-associated CD4+CD8α+ double-positive T cells are an activated T cell subpopulation with heterogeneous functional potential

Rabiger, Friederike V.; Bismarck, Doris; Protschka, Martina; Köhler, Gabriele; Moore, Peter F.; Büttner, Mathias; Buttlar, Heiner von; Alber, Gottfried; Eschke, Maria

doi:10.1371/journal.pone.0213597

Cited by 11 publications

(10 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We also characterized T cell subtypes: 68.65% were identified as CD3 + CD4 + Th cells, 8.5% as CD3 + CD8a + cytotoxic T cells, and 3.3% as CD3 + CD4 + CD8a + double-positive T cells. These double-positive cells may correspond to those previously reported to be present in the secondary lymphoid organs of healthy dogs ( 26 ). We found that 12.8% of Th cells had a Treg phenotype and 6.4% were positive for Bcl-6, suggesting the presence of regulatory and follicular T cells ( Fig.…”

Section: Resultssupporting

confidence: 89%

Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

Miguelena Chamorro,

Hameed,

Dechelette

et al. 2023

ImmunoHorizons

View full text Add to dashboard Cite

The oral route is effective and convenient for vaccine administration to stimulate a protective immune response. GALT plays a crucial role in mucosal immune responses, with Peyer’s patches (PPs) serving as the primary site of induction. A comprehensive understanding of the structures and functions of these structures is crucial for enhancing vaccination strategies and comprehending disease mechanisms; nonetheless, our current knowledge of these structures in dogs remains incomplete. We performed immunofluorescence and flow cytometry studies on canine PPs to identify cell populations and structures. We also performed single-cell RNA sequencing (scRNA-seq) to investigate the immune cell subpopulations present in PPs at steady state in dogs. We generated and validated an Ab specifically targeting canine M cells, which will be a valuable tool for elucidating Ag trafficking into the GALT of dogs. Our findings will pave the way for future studies of canine mucosal immune responses to oral vaccination and enteropathies. Moreover, they add to the growing body of knowledge in canine immunology, further expanding our understanding of the complex immune system of dogs.

show abstract

Section: Resultssupporting

confidence: 89%

Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

Miguelena Chamorro,

Hameed,

Dechelette

et al. 2023

ImmunoHorizons

View full text Add to dashboard Cite

show abstract

“…Furthermore, dp T cells have also been associated with increased production of IFN-γ in pigs ( 50 ), similar to previous results found in dogs with CanL ( 66 ). Moreover, the presence of CD4 + CD8 + CD25 + FoxP3 + T cell subset in the peripheral blood of sick dogs reveals a possible regulatory activity, as proposed by other authors ( 62 ), while the lymph node and bone marrow presented decreased percentages of CD25, reflecting a possible cytotoxic role ( 63 ) resulting from the infection with L. infantum . In turn, in the present study, the administration of either treatment led to a change in both profiles, with dp T cells in the blood losing the regulatory phenotype, possibly in order to fight the infection, while the lymph node and bone marrow apparently switching to a regulatory profile to nullify a possible excessive cytotoxic damage.…”

Section: Discussionsupporting

confidence: 64%

“…This subpopulation has been identified as being increased in chronic diseases, such as cancer, autoimmune diseases, and viral infections (57)(58)(59)(60)(61). Several studies have also reported the presence of CD25 and FoxP3 in dp T cells of dogs, revealing a possible regulatory activity among this subpopulation (62,63).…”

Section: Introductionmentioning

confidence: 97%

Immunophenotyping of Peripheral Blood, Lymph Node, and Bone Marrow T Lymphocytes During Canine Leishmaniosis and the Impact of Antileishmanial Chemotherapy

et al. 2020

View full text Add to dashboard Cite

Dogs are a major reservoir of Leishmania infantum , etiological agent of canine leishmaniosis (CanL) a zoonotic visceral disease of worldwide concern. Therapeutic protocols based on antileishmanial drugs are commonly used to treat sick dogs and improve their clinical condition. To better understand the impact of Leishmania infection and antileishmanial drugs on the dog's immune response, this study investigates the profile of CD4 + and CD8 + T cell subsets in peripheral blood, lymph node, and bone marrow of sick dogs and after two different CanL treatments. Two CanL groups of six dogs each were treated with either miltefosine or meglumine antimoniate combined with allopurinol. Another group of 10 clinically healthy dogs was used as control. Upon diagnosis and during the following 3 months of treatment, peripheral blood, popliteal lymph node, and bone marrow mononuclear cells were collected, labeled for surface markers CD45, CD3, CD4, CD8, CD25, and intracellular nuclear factor FoxP3, and T lymphocyte subpopulations were immunophenotyped by flow cytometry. CanL dogs presented an overall increased frequency of CD8 + and CD4 + CD8 + double-positive T cells in all tissues and a decreased frequency of CD4 + T cells in the blood. Furthermore, there was a higher frequency of CD8 + T cells expressing CD25 + FoxP3 + in the blood and bone marrow. During treatment, these subsets recovered to levels similar to those of healthy dogs. Nevertheless, antileishmanial therapy caused an increase of CD4 + CD25 + FoxP3 + T cells in all tissues, associated with the decrease of CD8 + CD25 − FoxP3 − T cell percentages. These findings may support previous studies that indicate that L. infantum manipulates the dog's immune system to avoid the development of a protective response, ensuring the parasite's survival and the conditions that allow the completion of Leishmania life cycle. Both treatments used appear to have an effect on the dog's immune response, proving to be effective in promoting the normalization of T cell subsets.

show abstract

“…With these features they clearly differ from their less numerous TCRγδ + CD4 − CD8α − dn counterparts, and even more from conventional TCRαβ + CD8α + sp T cells. On the other hand, we found phenotypic similarities to TCRαβ + CD4 + sp T cells, even though their activation state is remarkably different from the latter and rather comparable to canine TCRαβ + CD4 + CD8α + double-positive T cells (26, 27, 52).…”

Section: Discussionmentioning

confidence: 55%

“…Leukocytes from mLN, tLN, and spleen were isolated as previously described (26). In brief, tissue pieces were minced, passed through a 100 μm nylon cell strainer (BD Biosciences, Heidelberg, Germany) and resuspended in PBS followed by lysis of erythrocytes and cell counting as mentioned earlier.…”

Section: Methodsmentioning

confidence: 99%

Distinct Features of Canine Non-conventional CD4−CD8α− Double-Negative TCRαβ+ vs. TCRγδ+ T Cells

et al. 2019

Self Cite

View full text Add to dashboard Cite

The role of conventional TCRαβ+CD4+ or TCRαβ+CD8α+ single-positive (sp) T lymphocytes in adaptive immunity is well-recognized. However, non-conventional T cells expressing TCRαβ or TCRγδ but lacking CD4 and CD8α expression [i.e., CD4−CD8α− double-negative (dn) T cells] are thought to play a role at the interface between the innate and adaptive immune system. Dn T cells are frequent in swine, cattle or sheep and predominantly express TCRγδ. In contrast, TCRγδ+ T cells are rare in dogs. In this study, we identified a high proportion of canine dn T cells in the TCRαβ+ T cell population of PBMC, lymphatic and non-lymphatic organs. In PBMC, the frequency of this T cell subpopulation made up one third of the frequency of TCRαβ+CD4+ sp, and almost half of the frequency of TCRαβ+CD8α+ sp T cells (i.e., ~15% of all TCRαβ+ T cells). Among TCRαβ+CD4−CD8α− dn T cells of PBMC and tissues, FoxP3+ cells were identified indicating regulatory potential of this T cell subset. 80% of peripheral blood FoxP3+TCRαβ+CD4−CD8α− dn T cells co-expressed CD25, and, interestingly, also the FoxP3-negative TCRαβ+CD4−CD8α− dn T cells comprised ~34% CD25+ cells. Some of the FoxP3-positive TCRαβ+CD4−CD8α− dn T cells co-expressed GATA-3 suggesting stable function of regulatory T cells. The frequency of GATA-3 expression by FoxP3−TCRαβ+CD4−CD8α− dn T cells was even higher as compared with TCRαβ+CD4+ sp T cells (20.6% vs. 11.9%). Albeit lacking FoxP3 and CD25 expression, TCRγδ+CD4−CD8α− dn T cells also expressed substantial proportions of GATA-3. In addition, TCRαβ+CD4−CD8α− dn T cells produced IFN-γ and IL-17A upon stimulation. T-bet and granzyme B were only weakly expressed by both dn T cell subsets. In conclusion, this study identifies two dn T cell subsets in the dog: (i) a large (~7.5% in Peyer's patches, ~15% in lung) population of TCRαβ+CD4−CD8α− dn T cells with subpopulations thereof showing an activated phenotype, high expression of FoxP3 or GATA-3 as well as production of IFN-γ or IL-17A and (ii) a small TCRγδ+CD4−CD8α− dn T cell subset also expressing GATA-3 without production of IFN-γ or IL-17A. It will be exciting to unravel the function of each subset during immune homeostasis and diseases of dogs.

show abstract

Canine tissue-associated CD4+CD8α+ double-positive T cells are an activated T cell subpopulation with heterogeneous functional potential

Cited by 11 publications

References 58 publications

Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

Characterization of Canine Peyer’s Patches by Multidimensional Analysis: Insights from Immunofluorescence, Flow Cytometry, and Single-Cell RNA Sequencing

Immunophenotyping of Peripheral Blood, Lymph Node, and Bone Marrow T Lymphocytes During Canine Leishmaniosis and the Impact of Antileishmanial Chemotherapy

Distinct Features of Canine Non-conventional CD4−CD8α− Double-Negative TCRαβ+ vs. TCRγδ+ T Cells

Contact Info

Product

Resources

About