2021Cytometry Part AOpen access

Kinetics of activation marker expression after in vitro polyclonal stimulation of chicken peripheral T cells

Mohammad Naghizadeh, Nasim Hatamzade Esfahani, Frederik T. Larsen, Rikke Brødsgaard Kjærup, Eva Wattrang, Tina Sørensen Dalgaard

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Abstract

Abstract A comprehensive analysis of T cell activation markers in chicken is lacking. Kinetics of T cell activation markers (CD25, CD28, CD5, MHC‐II, CD44, and CD45) in response toin vitrostimulation of peripheral blood mononuclear cells with concanavalin A (Con A) were evaluated between two chicken lines selected for high and low levels of mannose‐binding lectin in serum (L10H and L10L, respectively) by flow cytometry. L10H chickens showed a stronger response to Con A based on the frequency of T cell blasts in both the CD4+and CD8+compartment. The majority of the proliferating CD4+and CD8+T cells expressed CD25. Proliferating T cells were seen both in the CD4+ MHC‐II+/−and CD8+ MHC‐II+/−population. For both CD4+and CD8+T cells, frequencies of CD25+and MHC‐II+T cells were increased 24 h after stimulation. CD28+frequencies were only increased on CD8+T cells 48 h after stimulation. An increase in the relative surface expression based on mean fluorescence intensity (MFI) upon activation was observed for most markers except CD5. For CD4+T cells, CD28 expression increased 24 h after stimulation whereas MHC‐II expression increased after 48 h. For CD8+T cells, a tendency toward an increase in CD25 expression was observed. CD28 expression started to increase 24 h after stimulation and only a transient peak in MHC‐II expression on CD8+T cells was observed after 24 h. CD44 and CD45 expressed on CD4+and CD8+T cells increased 24–72 h after stimulation. In summary, the frequency of CD25+and MHC‐II+T cells were shown to be early markers (24 h) forin vitroactivation of both CD4+and CD8+T cells. Frequency of CD28+T cells was a later marker (48 h) and only for CD8+T cells. Surface expression of all markers (MFI) increased permanently or transiently upon activation except for CD5.

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Abstract A comprehensive analysis of T cell activation markers in chicken is lacking. Kinetics of T cell activation markers (CD25, CD28, CD5, MHC‐II, CD44, and CD45) in response toin vitrostimulation of peripheral blood mononuclear cells with concanavalin A (Con A) were evaluated between two chicken lines selected for high and low levels of mannose‐binding lectin in serum (L10H and L10L, respectively) by flow cytometry. L10H chickens showed a stronger response to Con A based on the frequency of T cell blasts in both the CD4+and CD8+compartment. The majority of the proliferating CD4+and CD8+T cells expressed CD25. Proliferating T cells were seen both in the CD4+ MHC‐II+/−and CD8+ MHC‐II+/−population. For both CD4+and CD8+T cells, frequencies of CD25+and MHC‐II+T cells were increased 24 h after stimulation. CD28+frequencies were only increased on CD8+T cells 48 h after stimulation. An increase in the relative surface expression based on mean fluorescence intensity (MFI) upon activation was observed for most markers except CD5. For CD4+T cells, CD28 expression increased 24 h after stimulation whereas MHC‐II expression increased after 48 h. For CD8+T cells, a tendency toward an increase in CD25 expression was observed. CD28 expression started to increase 24 h after stimulation and only a transient peak in MHC‐II expression on CD8+T cells was observed after 24 h. CD44 and CD45 expressed on CD4+and CD8+T cells increased 24–72 h after stimulation. In summary, the frequency of CD25+and MHC‐II+T cells were shown to be early markers (24 h) forin vitroactivation of both CD4+and CD8+T cells. Frequency of CD28+T cells was a later marker (48 h) and only for CD8+T cells. Surface expression of all markers (MFI) increased permanently or transiently upon activation except for CD5.

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Available abstract

Abstract A comprehensive analysis of T cell activation markers in chicken is lacking. Kinetics of T cell activation markers (CD25, CD28, CD5, MHC‐II, CD44, and CD45) in response toin vitrostimulation of peripheral blood mononuclear cells with concanavalin A (Con A) were evaluated between two chicken lines selected for high and low levels of mannose‐binding lectin in serum (L10H and L10L, respectively) by flow cytometry. L10H chickens showed a stronger response to Con A based on the frequency of T cell blasts in both the CD4+and CD8+compartment. The majority of the proliferating CD4+and CD8+T cells expressed CD25. Proliferating T cells were seen both in the CD4+ MHC‐II+/−and CD8+ MHC‐II+/−population. For both CD4+and CD8+T cells, frequencies of CD25+and MHC‐II+T cells were increased 24 h after stimulation. CD28+frequencies were only increased on CD8+T cells 48 h after stimulation. An increase in the relative surface expression based on mean fluorescence intensity (MFI) upon activation was observed for most markers except CD5. For CD4+T cells, CD28 expression increased 24 h after stimulation whereas MHC‐II expression increased after 48 h. For CD8+T cells, a tendency toward an increase in CD25 expression was observed. CD28 expression started to increase 24 h after stimulation and only a transient peak in MHC‐II expression on CD8+T cells was observed after 24 h. CD44 and CD45 expressed on CD4+and CD8+T cells increased 24–72 h after stimulation. In summary, the frequency of CD25+and MHC‐II+T cells were shown to be early markers (24 h) forin vitroactivation of both CD4+and CD8+T cells. Frequency of CD28+T cells was a later marker (48 h) and only for CD8+T cells. Surface expression of all markers (MFI) increased permanently or transiently upon activation except for CD5.

Key concepts: Polyclonal antibodies, In vitro, Stimulation, Molecular biology, Kinetics, Peripheral, Chemistry, Cell biology

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