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Type of publication: Diploma Thesis
Type of document:

Year: 2012

Authors: Talker, Stephanie

Title: Phenotypic maturation of T- and NK-cell subsets in blood of pigs pursued from birth to three months of age.

Other title: Verlaufsstudie zum Differenzierungsstatus ausgewählter T- und NK- Zellsubpopulationen im Blut von juvenilen Schweinen

Source: Diplomarbeit, Vet. Med. Univ. Wien, pp. 83.


Advisor(s):

Saalmüller Armin

Reviewer(s):
Joachim Anja

Vetmed Research Units:
Institute of Immunology


Graduation date: 16.01.12


Abstract:
The immune system of the newborn piglet is inexperienced and naïve due to its protected residence in the uterus prior to birth. After birth, challenge with environmental antigens results in the differentiation of naïve immune cells into effector cells and later, memory cells. The latter are essential for long-term protection and of particular interest in vaccine development. In the current study, 6-color flow cytometry was performed to investigate phenotypic changes related to differentiation following antigen contact for T-and NK cell subsets in blood of twelve pigs. Blood samples were taken from the day of birth to three months of age. After isolation of peripheral blood mononuclear cells, the following lymphocyte subsets were analyzed by a combination of lineage markers (LM) and differentiation markers (DM): NK cells (LM: CD2, CD3, CD8[alpha], DM: NKp46, perforin, MHC-II), [gamma-delta]-T cells (LM: TCR-[gamma-delta], DM: CD2, CD8[alpha], CD27, SWC5, MHC-II), T-helper cells (LM: CD3, CD4, DM: CD8[alpha], CD27, CD45RC, MHC-II), regulatory T cells (LM: Foxp3, CD4, CD25, DM: CD8[alpha], CD45RC, MHC-II) and cytolytic T cells (LM: CD3, CD8[alpha], CD8[beta], DM: CD27, perforin, MHC-II). Interestingly, already at the time of birth, all NK cells stained positive for the cytotoxicity mediating molecule perforin. For [gamma-delta] T cells it was notable that only within CD2+SWC5-[gamma-delta] T cells further subpopulations expressing CD8[alpha] and MHC-II but lacking CD27 were identified over time. T-helper cells isolated at the time of birth were CD8[alpha]-CD27+MHC-IIand mostly positive for CD45RC. With increasing age of the piglets, T-helper cells partially became CD8[alpha]+MHC-II+ and subpopulations expressing no CD45RC or CD27 could be identified. Cytolytic T cells (CTLs) were negative for perforin and MHC-II and positive for CD27 at the time of birth. At later time points, MHC-II was expressed on a fraction of CTLs. This was followed by the appearance of cells showing an intracellular accumulation of perforin and partially these perforin+ CTLs lost expression of CD27. By tracking the phenotypes of various immune cell subsets in the first three months of life, at least for [gamma-delta]-T cells, T-helper cells and CTLs, particular differentiation patterns became obvious over time. These data lay the foundation for future studies, where – in controlled infection experiments – such differentiation patterns can be investigated in more detail for antigen-specific T cells during the time course of an immune response.


Publication(s) resulting from University thesis:

Talker, SC; Käser, T; Reutner, K; Sedlak, C; Mair, KH; Koinig, H; Graage, R; Viehmann, M; Klingler, E; Ladinig, A; Ritzmann, M; Saalmüller, A; Gerner, W (2013): Phenotypic maturation of porcine NK- and T-cell subsets. Dev Comp Immunol. 2013; 40(1):51-68

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