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Frequency and phenotype of natural killer cells and natural killer cell subsets in bovine lymphoid compartments and blood

View Article: PubMed Central - PubMed

ABSTRACT

Natural killer (NK) cells are widely distributed in lymphoid and non‐lymphoid tissues, but little is known about the recirculation of NK cells between blood and tissues. This is relevant to understanding recirculation in the steady‐state and also for determining the roles for NK cells in vaccine‐induced immunity and responses to infection. Therefore, the percentage of NK cells and their phenotype across peripheral blood, afferent lymph and lymph nodes in steady‐state conditions was investigated in cattle using the pseudo‐afferent lymphatic cannulation model. CD2+ CD25lo NK cells were the predominant subset of NK cells within the blood. In contrast, CD2− CD25hi NK cells were the main subset present within the skin‐draining afferent lymphatic vessels and lymph nodes, indicating that CD2− NK cells are the principal NK cell subset trafficking to lymph nodes via the afferent lymphatic vessel. Furthermore, a low percentage of NK cells were present in efferent lymph, which were predominantly of the CD2− subset, indicating that NK cells can egress from lymph nodes and return to circulation in steady‐state conditions. These compartmentalization data indicate that NK cells represent a population of recirculating lymphocytes in steady‐state conditions and therefore may be important during immune responses to vaccination or infection.

No MeSH data available.


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CD2− natural killer (NK) cells are the predominant subset of NK cells present within skin‐draining afferent lymph (AL). Lymphocytes derived from peripheral blood (PB), AL and the lymph nodes (LNs) of seven calves were labelled with monoclonal antibodies to NKp46, CD3 and CD2 and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD3 by PB‐, AL‐ and LN‐derived lymphocytes (a). FACS plots from one representative animal illustrate the expression of NKp46 and CD2 by PB‐, AL‐ and LN‐derived lymphocytes (b). Gates were set based on Fluorescence Minus One controls. Pooled data from seven calves indicate the average percentage of NKp46+ CD3− NK cells ± SD present in PB (circles), AL (squares) and LNs (triangles) (c). Pooled data from seven calves illustrate the average percentage of CD2+ (lighter bars) and CD2− (darker bars) NK cells ± SD within the total gated NKp46+ NK cell population from PB, AL or the LNs (d). Data were normally distributed (P > 0·05). Significance between PB and AL was assessed using a paired t‐test and significance between PB and LN was assessed using a two‐sample t‐test. P < 0·001***. [Colour figure can be viewed at wileyonlinelibrary.com]
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imm12708-fig-0001: CD2− natural killer (NK) cells are the predominant subset of NK cells present within skin‐draining afferent lymph (AL). Lymphocytes derived from peripheral blood (PB), AL and the lymph nodes (LNs) of seven calves were labelled with monoclonal antibodies to NKp46, CD3 and CD2 and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD3 by PB‐, AL‐ and LN‐derived lymphocytes (a). FACS plots from one representative animal illustrate the expression of NKp46 and CD2 by PB‐, AL‐ and LN‐derived lymphocytes (b). Gates were set based on Fluorescence Minus One controls. Pooled data from seven calves indicate the average percentage of NKp46+ CD3− NK cells ± SD present in PB (circles), AL (squares) and LNs (triangles) (c). Pooled data from seven calves illustrate the average percentage of CD2+ (lighter bars) and CD2− (darker bars) NK cells ± SD within the total gated NKp46+ NK cell population from PB, AL or the LNs (d). Data were normally distributed (P > 0·05). Significance between PB and AL was assessed using a paired t‐test and significance between PB and LN was assessed using a two‐sample t‐test. P < 0·001***. [Colour figure can be viewed at wileyonlinelibrary.com]

Mentions: First, to determine if NK cells were present in skin‐draining afferent lymphatic vessels and to compare their frequency to those present in PB and LNs, the abundance of NKp46+ CD3− NK cells within PB, AL and LNs of seven calves was investigated. The gating strategy used to identify lymphocytes throughout this study is illustrated in the Supplementary material (Fig. S1). NKp46+ CD3− NK cells represented 5·1% (2·05–10·3%; SD = 3·1), 4·8% (1·39–7·68%; SD = 1·9) and 6% (3·03–11·7%; SD = 2·9) of lymphocytes in PB, AL and the LNs, respectively (representative dot plots are shown in Fig. 1a). No significant differences were evident between the percentages of NK cells present across the three compartments (Fig. 1c). CD2+ NK cells were the predominant subset in PB, but CD2− NK cells were the principal NK cell subset found in the skin‐draining afferent lymphatic vessels and LNs (representative dot plots are shown in Fig. 1b). Furthermore, CD2− NK cells were present in AL (P < 0·001) and LNs (P < 0·001) at a significantly higher proportion compared with PB (Fig. 1d).


Frequency and phenotype of natural killer cells and natural killer cell subsets in bovine lymphoid compartments and blood
CD2− natural killer (NK) cells are the predominant subset of NK cells present within skin‐draining afferent lymph (AL). Lymphocytes derived from peripheral blood (PB), AL and the lymph nodes (LNs) of seven calves were labelled with monoclonal antibodies to NKp46, CD3 and CD2 and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD3 by PB‐, AL‐ and LN‐derived lymphocytes (a). FACS plots from one representative animal illustrate the expression of NKp46 and CD2 by PB‐, AL‐ and LN‐derived lymphocytes (b). Gates were set based on Fluorescence Minus One controls. Pooled data from seven calves indicate the average percentage of NKp46+ CD3− NK cells ± SD present in PB (circles), AL (squares) and LNs (triangles) (c). Pooled data from seven calves illustrate the average percentage of CD2+ (lighter bars) and CD2− (darker bars) NK cells ± SD within the total gated NKp46+ NK cell population from PB, AL or the LNs (d). Data were normally distributed (P > 0·05). Significance between PB and AL was assessed using a paired t‐test and significance between PB and LN was assessed using a two‐sample t‐test. P < 0·001***. [Colour figure can be viewed at wileyonlinelibrary.com]
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imm12708-fig-0001: CD2− natural killer (NK) cells are the predominant subset of NK cells present within skin‐draining afferent lymph (AL). Lymphocytes derived from peripheral blood (PB), AL and the lymph nodes (LNs) of seven calves were labelled with monoclonal antibodies to NKp46, CD3 and CD2 and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD3 by PB‐, AL‐ and LN‐derived lymphocytes (a). FACS plots from one representative animal illustrate the expression of NKp46 and CD2 by PB‐, AL‐ and LN‐derived lymphocytes (b). Gates were set based on Fluorescence Minus One controls. Pooled data from seven calves indicate the average percentage of NKp46+ CD3− NK cells ± SD present in PB (circles), AL (squares) and LNs (triangles) (c). Pooled data from seven calves illustrate the average percentage of CD2+ (lighter bars) and CD2− (darker bars) NK cells ± SD within the total gated NKp46+ NK cell population from PB, AL or the LNs (d). Data were normally distributed (P > 0·05). Significance between PB and AL was assessed using a paired t‐test and significance between PB and LN was assessed using a two‐sample t‐test. P < 0·001***. [Colour figure can be viewed at wileyonlinelibrary.com]
Mentions: First, to determine if NK cells were present in skin‐draining afferent lymphatic vessels and to compare their frequency to those present in PB and LNs, the abundance of NKp46+ CD3− NK cells within PB, AL and LNs of seven calves was investigated. The gating strategy used to identify lymphocytes throughout this study is illustrated in the Supplementary material (Fig. S1). NKp46+ CD3− NK cells represented 5·1% (2·05–10·3%; SD = 3·1), 4·8% (1·39–7·68%; SD = 1·9) and 6% (3·03–11·7%; SD = 2·9) of lymphocytes in PB, AL and the LNs, respectively (representative dot plots are shown in Fig. 1a). No significant differences were evident between the percentages of NK cells present across the three compartments (Fig. 1c). CD2+ NK cells were the predominant subset in PB, but CD2− NK cells were the principal NK cell subset found in the skin‐draining afferent lymphatic vessels and LNs (representative dot plots are shown in Fig. 1b). Furthermore, CD2− NK cells were present in AL (P < 0·001) and LNs (P < 0·001) at a significantly higher proportion compared with PB (Fig. 1d).

View Article: PubMed Central - PubMed

ABSTRACT

Natural killer (NK) cells are widely distributed in lymphoid and non&#8208;lymphoid tissues, but little is known about the recirculation of NK cells between blood and tissues. This is relevant to understanding recirculation in the steady&#8208;state and also for determining the roles for NK cells in vaccine&#8208;induced immunity and responses to infection. Therefore, the percentage of NK cells and their phenotype across peripheral blood, afferent lymph and lymph nodes in steady&#8208;state conditions was investigated in cattle using the pseudo&#8208;afferent lymphatic cannulation model. CD2+ CD25lo NK cells were the predominant subset of NK cells within the blood. In contrast, CD2&minus; CD25hi NK cells were the main subset present within the skin&#8208;draining afferent lymphatic vessels and lymph nodes, indicating that CD2&minus; NK cells are the principal NK cell subset trafficking to lymph nodes via the afferent lymphatic vessel. Furthermore, a low percentage of NK cells were present in efferent lymph, which were predominantly of the CD2&minus; subset, indicating that NK cells can egress from lymph nodes and return to circulation in steady&#8208;state conditions. These compartmentalization data indicate that NK cells represent a population of recirculating lymphocytes in steady&#8208;state conditions and therefore may be important during immune responses to vaccination or infection.

No MeSH data available.


Related in: MedlinePlus