Limits...
Gene set enrichment analysis identifies LIF as a negative regulator of human Th2 cell differentiation.

Ullah U, Tripathi P, Lahesmaa R, Rao KV - Sci Rep (2012)

Bottom Line: In this study we show that IL-4 is crucial during reinforcement window of human Th2 differentiation for optimal Th2 development.We have also shown here that during this stage, IL-4 helps in cellular decision-making process of differentiation versus proliferation.This approach can be generalized to analyze "omics" data to identify key regulatory modules.

View Article: PubMed Central - PubMed

Affiliation: Immunology Group International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India.

ABSTRACT
In this study we show that IL-4 is crucial during reinforcement window of human Th2 differentiation for optimal Th2 development. We have also shown here that during this stage, IL-4 helps in cellular decision-making process of differentiation versus proliferation. We have combined computational and experimental methods to analyze Th2 transcription network to name novel players of the process of Th2 differentiation. Here we report that LIF through STAT3 negatively regulates Th2 differentiation. This approach can be generalized to analyze "omics" data to identify key regulatory modules.

Show MeSH

Related in: MedlinePlus

Continuous IL-4 is required for optimal Th2 development.Panel a shows results of IL-4 ELISPOT experiments. Culture conditions are mentioned at the bottom. Error bars represent standard error of mean and * denotes significant (p value 0.017) difference between the two sample (Two tailed paired sample t test). In panel b blue line represents IL-4 level detected in the supernatant at given time points and red line is a measure of IL-4R expression level at corresponding time points. Panel c shows a result obtained in a CFSE labeling experiment in ERK silenced cells. Red, blue and green colors represent not activated, TCR + IL-4 stimulated and TCR + IL-4 stimulated at day 1 followed by additional doses of IL-4 at day 3,4 and 5. Less fluorescence indicates more cell divisions because as the cells divide fluorescent label is diluted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3376453&req=5

f1: Continuous IL-4 is required for optimal Th2 development.Panel a shows results of IL-4 ELISPOT experiments. Culture conditions are mentioned at the bottom. Error bars represent standard error of mean and * denotes significant (p value 0.017) difference between the two sample (Two tailed paired sample t test). In panel b blue line represents IL-4 level detected in the supernatant at given time points and red line is a measure of IL-4R expression level at corresponding time points. Panel c shows a result obtained in a CFSE labeling experiment in ERK silenced cells. Red, blue and green colors represent not activated, TCR + IL-4 stimulated and TCR + IL-4 stimulated at day 1 followed by additional doses of IL-4 at day 3,4 and 5. Less fluorescence indicates more cell divisions because as the cells divide fluorescent label is diluted.

Mentions: IL-4 drives both the initiation and the reinforcement stages of the Th2 polarization response120. In view of the kinetics of the process though, we speculated that endogenous IL-4 would be specifically involved during the reinforcement/commitment of Th2 lineage specification. To test this we generated parallel cultures of either Th activation, or Th2 differentiation and added to these a combination of neutralizing antibodies against IL-4 and IL-4R. Antibodies were first added at day three after initiation of the cultures, and then supplemented every 24 h up to day 5. This timing of addition was based on the fact that IL-4 transcription peaks at around day3 post stimulation7. Neutralization of exogenous IL-4 significantly reduced the extent of Th2 cell generation (P ≤0.05, Figure 1a). Further, this inhibition was also observed in TCR activated T cells (“Data not shown”), supporting that this was likely due to depletion of the endogenously produced cytokine (Figure 1a). The partial attenuation seen may well derive from the possibility that the added antibodies were only partly effective at neutralizing the autocrine/paracrine action of the endogenously produced IL-4.


Gene set enrichment analysis identifies LIF as a negative regulator of human Th2 cell differentiation.

Ullah U, Tripathi P, Lahesmaa R, Rao KV - Sci Rep (2012)

Continuous IL-4 is required for optimal Th2 development.Panel a shows results of IL-4 ELISPOT experiments. Culture conditions are mentioned at the bottom. Error bars represent standard error of mean and * denotes significant (p value 0.017) difference between the two sample (Two tailed paired sample t test). In panel b blue line represents IL-4 level detected in the supernatant at given time points and red line is a measure of IL-4R expression level at corresponding time points. Panel c shows a result obtained in a CFSE labeling experiment in ERK silenced cells. Red, blue and green colors represent not activated, TCR + IL-4 stimulated and TCR + IL-4 stimulated at day 1 followed by additional doses of IL-4 at day 3,4 and 5. Less fluorescence indicates more cell divisions because as the cells divide fluorescent label is diluted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376453&req=5

f1: Continuous IL-4 is required for optimal Th2 development.Panel a shows results of IL-4 ELISPOT experiments. Culture conditions are mentioned at the bottom. Error bars represent standard error of mean and * denotes significant (p value 0.017) difference between the two sample (Two tailed paired sample t test). In panel b blue line represents IL-4 level detected in the supernatant at given time points and red line is a measure of IL-4R expression level at corresponding time points. Panel c shows a result obtained in a CFSE labeling experiment in ERK silenced cells. Red, blue and green colors represent not activated, TCR + IL-4 stimulated and TCR + IL-4 stimulated at day 1 followed by additional doses of IL-4 at day 3,4 and 5. Less fluorescence indicates more cell divisions because as the cells divide fluorescent label is diluted.
Mentions: IL-4 drives both the initiation and the reinforcement stages of the Th2 polarization response120. In view of the kinetics of the process though, we speculated that endogenous IL-4 would be specifically involved during the reinforcement/commitment of Th2 lineage specification. To test this we generated parallel cultures of either Th activation, or Th2 differentiation and added to these a combination of neutralizing antibodies against IL-4 and IL-4R. Antibodies were first added at day three after initiation of the cultures, and then supplemented every 24 h up to day 5. This timing of addition was based on the fact that IL-4 transcription peaks at around day3 post stimulation7. Neutralization of exogenous IL-4 significantly reduced the extent of Th2 cell generation (P ≤0.05, Figure 1a). Further, this inhibition was also observed in TCR activated T cells (“Data not shown”), supporting that this was likely due to depletion of the endogenously produced cytokine (Figure 1a). The partial attenuation seen may well derive from the possibility that the added antibodies were only partly effective at neutralizing the autocrine/paracrine action of the endogenously produced IL-4.

Bottom Line: In this study we show that IL-4 is crucial during reinforcement window of human Th2 differentiation for optimal Th2 development.We have also shown here that during this stage, IL-4 helps in cellular decision-making process of differentiation versus proliferation.This approach can be generalized to analyze "omics" data to identify key regulatory modules.

View Article: PubMed Central - PubMed

Affiliation: Immunology Group International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi-110067, India.

ABSTRACT
In this study we show that IL-4 is crucial during reinforcement window of human Th2 differentiation for optimal Th2 development. We have also shown here that during this stage, IL-4 helps in cellular decision-making process of differentiation versus proliferation. We have combined computational and experimental methods to analyze Th2 transcription network to name novel players of the process of Th2 differentiation. Here we report that LIF through STAT3 negatively regulates Th2 differentiation. This approach can be generalized to analyze "omics" data to identify key regulatory modules.

Show MeSH
Related in: MedlinePlus