Abstract

Differentiation of porcine T helper cells is still poorly investigated, partly due to a lack of monoclonal antibodies (mAbs) specific for molecules involved in this process. Recently, we identified a mAb specific for porcine CD27 and showed that CD27 is expressed by all naïve CD8α- T helper cells but divides CD8α+ T helper cells into a CD27+ and a CD27- subset. In the present study, detailed phenotypical and functional analyses of these T-helper cell subpopulations were performed. Naïve CD8α-CD27+ T helper cells predominantly resided in various lymph nodes, whereas higher proportions of CD8α+CD27+ and CD8α+CD27- T helper cells were found in blood, spleen and liver. Both CD8α+CD27+ and CD8α+CD27- T helper cells were capable of producing IFN-γ upon in vitro polyclonal stimulation and antigen-specific restimulation. Experiments with sorted CD8α-CD27+, CD8α+CD27+ and CD8α+CD27- T-helper cell subsets following polyclonal stimulation revealed the lowest proliferative response but the highest ability for IFN-γ and TNF-α production in the CD8α+CD27- subset. Therefore, these cells resembled terminally differentiated effector memory cells as described in human. This was supported by analyses of CCR7 and CD62L expression. CD8α+CD27- T helper cells were mostly CCR7- and had considerably reduced CD62L mRNA levels. In contrast, expression of both homing-receptors was increased on CD8α+CD27+ T helper cells, which also had a proliferation rate similar to naïve CD8α-CD27+ T helper cells and showed intermediate levels of cytokine production. Therefore, similar to human, CD8α+CD27+ T helper cells displayed a phenotype and functional properties of central memory cells.

Highlights

  • A peculiarity of porcine T helper cells is the expression of CD8α on a substantial proportion of these cells in blood and secondary lymphatic organs [1,2]

  • CD8α-CD27+ T helper cells abounded in lymph nodes (> 57.5%), peripheral blood mononuclear cells (PBMCs) (55.3%) and spleen (45.8%)

  • Helper cells dominated (70.6%), and the frequency of this population descended from tonsil (35.0%) to PBMCs (27.2%)

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Summary

Introduction

A peculiarity of porcine T helper cells is the expression of CD8α on a substantial proportion of these cells in blood and secondary lymphatic organs [1,2]. In vitro stimulation by superantigens or mixed leukocyte reactions causes an up-regulation of CD8α expression on porcine T helper cells [1,3], and it was reported that CD8α+ T helper cells proliferate in response to stimulation with recall antigen [4,5,6]. Differentiation of T helper cells is commonly defined by i) the expression of receptors for lymph node homing, ii) the expression of co-stimulatory molecules and iii) the capability to produce certain cytokines. With regard to the lymph node homing receptors CD62L and CCR7, two functionally distinct memory subsets have been defined: CD62L+CCR7+ central memory and CD62L-CCR7- effector memory T helper cells. Regarding the expression of co-stimulatory molecules, T helper cells initially express CD27, a member of the tumor necrosis factor receptor (TNFR) family, which contributes to proliferation, survival and cytokine production. During T-cell differentiation, CD27 expression undergoes down-regulation and is lost on terminally differentiated effector cells [9,10]

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