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Cattle NK Cell Heterogeneity and the Influence of MHC Class I.

Allan AJ, Sanderson ND, Gubbins S, Ellis SA, Hammond JA - J. Immunol. (2015)

Bottom Line: Cytokine stimulation differentially influenced the transcription of five receptors at the cell population level.Using dilution cultures, we found that a further seven receptors were differentially transcribed, including five predicted to recognize MHC class I.Moreover, there was a statistically significant reduction in killer cell lectin-like receptor mRNA expression between cultures with different CD2 phenotypes and from animals with different MHC class I haplotypes.

View Article: PubMed Central - PubMed

Affiliation: The Pirbright Institute, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.

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Related in: MedlinePlus

Ex vivo NK cells are predominantly CD2high, and CD8 expression increases upon stimulation with IL-2. PBMCs from 14 MHC class I–defined cattle were assessed for the expression of NCR1, CD2, and CD8 immediately ex vivo (unstimulated, filled bars) or after 7-d in vitro IL-2 stimulation (open bars). (A) The percentage of CD2high and CD8+ cells within the NCR1+ population. Statistical significance was determined using a linear mixed model, with expression as the response variable and CD2/CD8 status and stimulated/unstimulated as fixed effects, plus an interaction between them and the animal as a random effect. (B) Overall change in the percentage of CD2high and CD8+ within the NCR1+ population after cytokine stimulation. Statistical significance was determined using paired t tests. *p < 0.001.
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fig01: Ex vivo NK cells are predominantly CD2high, and CD8 expression increases upon stimulation with IL-2. PBMCs from 14 MHC class I–defined cattle were assessed for the expression of NCR1, CD2, and CD8 immediately ex vivo (unstimulated, filled bars) or after 7-d in vitro IL-2 stimulation (open bars). (A) The percentage of CD2high and CD8+ cells within the NCR1+ population. Statistical significance was determined using a linear mixed model, with expression as the response variable and CD2/CD8 status and stimulated/unstimulated as fixed effects, plus an interaction between them and the animal as a random effect. (B) Overall change in the percentage of CD2high and CD8+ within the NCR1+ population after cytokine stimulation. Statistical significance was determined using paired t tests. *p < 0.001.

Mentions: As this study exploited a specially bred herd of MHC homozygous Holstein-Friesian cattle, it was important to establish that NK cells isolated from these animals were not substantively different from those used in previous studies (25, 26). Cattle NK cells are defined as NCR1+, with variable CD2 and CD8α expression (25, 26). We determined the cell surface expression of CD2 and CD8α on ex vivo and in vitro stimulated NCR1+ PBMCs from 14 MHC homozygous cattle. These animals represented three different haplotypes, four A18 containing only gene 6, four A14 containing a gene 1, 2 and 4 and six A31 containing a gene 1 and 2 but with different alleles to the A14. In all the animals, ∼80% of NCR1+ cells were CD2high ex vivo and CD2 expression was reduced nonsignificantly (p = 0.27) after stimulation with IL-2 (Fig. 1). In contrast, CD8α expression increased in vitro under IL-2 when compared with ex vivo cells (p < 0.001) (Fig. 1), as has been shown with other cytokines (25). These NK phenotypes are consistent with previously published data showing that NK cells from these homozygous cattle have a broadly conventional phenotype and are responsive to cytokine stimulation.


Cattle NK Cell Heterogeneity and the Influence of MHC Class I.

Allan AJ, Sanderson ND, Gubbins S, Ellis SA, Hammond JA - J. Immunol. (2015)

Ex vivo NK cells are predominantly CD2high, and CD8 expression increases upon stimulation with IL-2. PBMCs from 14 MHC class I–defined cattle were assessed for the expression of NCR1, CD2, and CD8 immediately ex vivo (unstimulated, filled bars) or after 7-d in vitro IL-2 stimulation (open bars). (A) The percentage of CD2high and CD8+ cells within the NCR1+ population. Statistical significance was determined using a linear mixed model, with expression as the response variable and CD2/CD8 status and stimulated/unstimulated as fixed effects, plus an interaction between them and the animal as a random effect. (B) Overall change in the percentage of CD2high and CD8+ within the NCR1+ population after cytokine stimulation. Statistical significance was determined using paired t tests. *p < 0.001.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4543905&req=5

fig01: Ex vivo NK cells are predominantly CD2high, and CD8 expression increases upon stimulation with IL-2. PBMCs from 14 MHC class I–defined cattle were assessed for the expression of NCR1, CD2, and CD8 immediately ex vivo (unstimulated, filled bars) or after 7-d in vitro IL-2 stimulation (open bars). (A) The percentage of CD2high and CD8+ cells within the NCR1+ population. Statistical significance was determined using a linear mixed model, with expression as the response variable and CD2/CD8 status and stimulated/unstimulated as fixed effects, plus an interaction between them and the animal as a random effect. (B) Overall change in the percentage of CD2high and CD8+ within the NCR1+ population after cytokine stimulation. Statistical significance was determined using paired t tests. *p < 0.001.
Mentions: As this study exploited a specially bred herd of MHC homozygous Holstein-Friesian cattle, it was important to establish that NK cells isolated from these animals were not substantively different from those used in previous studies (25, 26). Cattle NK cells are defined as NCR1+, with variable CD2 and CD8α expression (25, 26). We determined the cell surface expression of CD2 and CD8α on ex vivo and in vitro stimulated NCR1+ PBMCs from 14 MHC homozygous cattle. These animals represented three different haplotypes, four A18 containing only gene 6, four A14 containing a gene 1, 2 and 4 and six A31 containing a gene 1 and 2 but with different alleles to the A14. In all the animals, ∼80% of NCR1+ cells were CD2high ex vivo and CD2 expression was reduced nonsignificantly (p = 0.27) after stimulation with IL-2 (Fig. 1). In contrast, CD8α expression increased in vitro under IL-2 when compared with ex vivo cells (p < 0.001) (Fig. 1), as has been shown with other cytokines (25). These NK phenotypes are consistent with previously published data showing that NK cells from these homozygous cattle have a broadly conventional phenotype and are responsive to cytokine stimulation.

Bottom Line: Cytokine stimulation differentially influenced the transcription of five receptors at the cell population level.Using dilution cultures, we found that a further seven receptors were differentially transcribed, including five predicted to recognize MHC class I.Moreover, there was a statistically significant reduction in killer cell lectin-like receptor mRNA expression between cultures with different CD2 phenotypes and from animals with different MHC class I haplotypes.

View Article: PubMed Central - PubMed

Affiliation: The Pirbright Institute, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.

Show MeSH
Related in: MedlinePlus