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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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The variable transcription of NK cell receptor genes in ex vivo NK cells. The transcription of 25 NK cell receptor and associated genes was determined by PCR in NK cells from 14 MHC class I–defined animals (4 A18, 4 A14, and 6 A31). Genes that showed consistent variation are not shown. The dots represent transcription in immediately ex vivo NK cells (black), NK cells stimulated for 7 d in recombinant bovine IL-2 (orange), recombinant bovine IL-12, and recombinant human IL-18 (gray) or recombinant human IL-15 (blue).
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fig02: The variable transcription of NK cell receptor genes in ex vivo NK cells. The transcription of 25 NK cell receptor and associated genes was determined by PCR in NK cells from 14 MHC class I–defined animals (4 A18, 4 A14, and 6 A31). Genes that showed consistent variation are not shown. The dots represent transcription in immediately ex vivo NK cells (black), NK cells stimulated for 7 d in recombinant bovine IL-2 (orange), recombinant bovine IL-12, and recombinant human IL-18 (gray) or recombinant human IL-15 (blue).

Mentions: Five genes showed variable mRNA expression patterns that were independent of MHC class I background (Fig. 2). NCR1 expression on CD3− cells currently defines cattle NK cells, but we observed that NCR1 transcription was not ubiquitous, and in cells from animal 982 we were not able to detect NCR1 transcription following isolation based on NCR1 expression. The activating receptor gene 2B4 was always transcribed by ex vivo NK cells but was partially or completely lost in cytokine-stimulated NK cells from eight animals. Conversely, TIM-3 was absent in the ex vivo populations from 10 of the 14 animals but was consistently transcribed by all animals after in vitro cytokine stimulation. CD8β was consistently transcribed by ex vivo and IL-2 and IL-15 in vitro stimulated NK cells, but in four animals, transcription was lost upon stimulation with IL-12/18. Only KIR3DXL6 appeared to have variable transcription with no discernible pattern (Fig. 2). Although KLRA appears differentially transcribed, two distinct allele lineages are found in cattle. We have previously genotyped this group of animals for the presence of each KLRA lineage, and these NK population differences are due to genotype rather than differential regulation.


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

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

The variable transcription of NK cell receptor genes in ex vivo NK cells. The transcription of 25 NK cell receptor and associated genes was determined by PCR in NK cells from 14 MHC class I–defined animals (4 A18, 4 A14, and 6 A31). Genes that showed consistent variation are not shown. The dots represent transcription in immediately ex vivo NK cells (black), NK cells stimulated for 7 d in recombinant bovine IL-2 (orange), recombinant bovine IL-12, and recombinant human IL-18 (gray) or recombinant human IL-15 (blue).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4543905&req=5

fig02: The variable transcription of NK cell receptor genes in ex vivo NK cells. The transcription of 25 NK cell receptor and associated genes was determined by PCR in NK cells from 14 MHC class I–defined animals (4 A18, 4 A14, and 6 A31). Genes that showed consistent variation are not shown. The dots represent transcription in immediately ex vivo NK cells (black), NK cells stimulated for 7 d in recombinant bovine IL-2 (orange), recombinant bovine IL-12, and recombinant human IL-18 (gray) or recombinant human IL-15 (blue).
Mentions: Five genes showed variable mRNA expression patterns that were independent of MHC class I background (Fig. 2). NCR1 expression on CD3− cells currently defines cattle NK cells, but we observed that NCR1 transcription was not ubiquitous, and in cells from animal 982 we were not able to detect NCR1 transcription following isolation based on NCR1 expression. The activating receptor gene 2B4 was always transcribed by ex vivo NK cells but was partially or completely lost in cytokine-stimulated NK cells from eight animals. Conversely, TIM-3 was absent in the ex vivo populations from 10 of the 14 animals but was consistently transcribed by all animals after in vitro cytokine stimulation. CD8β was consistently transcribed by ex vivo and IL-2 and IL-15 in vitro stimulated NK cells, but in four animals, transcription was lost upon stimulation with IL-12/18. Only KIR3DXL6 appeared to have variable transcription with no discernible pattern (Fig. 2). Although KLRA appears differentially transcribed, two distinct allele lineages are found in cattle. We have previously genotyped this group of animals for the presence of each KLRA lineage, and these NK population differences are due to genotype rather than differential regulation.

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