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Peripheral residence of naïve CD4 T cells induces MHC class II-dependent alterations in phenotype and function.

Rane S, Das R, Ranganathan V, Prabhu S, Das A, Mattoo H, Durdik JM, George A, Rath S, Bal V - BMC Biol. (2014)

Bottom Line: It is not clear if these interactions result in alterations in their activation, survival and effector programming.Naïve CD4lo and CD4hi subsets of thymic single-positive CD4 T cells did not show differences whereas peripheral naïve CD4lo and CD4hi subsets of T cell receptor (TCR)-transgenic T cells did.Our findings also suggest the feasibility of potential pharmacological interventions for improved CD4 T cell responses during vaccination of older people via either anti-oxidant or DUSP inhibitor small molecules.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Immunology, New Delhi, 110067, India. sanketrn@nii.ac.in.

ABSTRACT

Background: As individual naïve CD4 T lymphocytes circulate in the body after emerging from the thymus, they are likely to have individually varying microenvironmental interactions even in the absence of stimulation via specific target recognition. It is not clear if these interactions result in alterations in their activation, survival and effector programming. Naïve CD4 T cells show unimodal distribution for many phenotypic properties, suggesting that the variation is caused by intrinsic stochasticity, although underlying variation due to subsets created by different histories of microenvironmental interactions remains possible. To explore this possibility, we began examining the phenotype and functionality of naïve CD4 T cells differing in a basic unimodally distributed property, the CD4 levels, as well as the causal origin of these differences.

Results: We examined separated CD4hi and CD4lo subsets of mouse naïve CD4 cells. CD4lo cells were smaller with higher CD5 levels and lower levels of the dual-specific phosphatase (DUSP)6-suppressing micro-RNA miR181a, and responded poorly with more Th2-skewed outcomes. Human naïve CD4lo and CD4hi cells showed similar differences. Naïve CD4lo and CD4hi subsets of thymic single-positive CD4 T cells did not show differences whereas peripheral naïve CD4lo and CD4hi subsets of T cell receptor (TCR)-transgenic T cells did. Adoptive transfer-mediated parking of naïve CD4 cells in vivo lowered CD4 levels, increased CD5 and reactive oxygen species (ROS) levels and induced hyporesponsiveness in them, dependent, at least in part, on availability of major histocompatibility complex class II (MHCII) molecules. ROS scavenging or DUSP inhibition ameliorated hyporesponsiveness. Naïve CD4 cells from aged mice showed lower CD4 levels and cell sizes, higher CD5 levels, and hyporesponsiveness and Th2-skewing reversed by DUSP inhibition.

Conclusions: Our data show that, underlying a unimodally distributed property, the CD4 level, there are subsets of naïve CD4 cells that vary in the time spent in the periphery receiving MHCII-mediated signals and show resultant alteration of phenotype and functionality via ROS and DUSP activity. Our findings also suggest the feasibility of potential pharmacological interventions for improved CD4 T cell responses during vaccination of older people via either anti-oxidant or DUSP inhibitor small molecules.

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Consequences of prolonged tonic signaling and absence of tonic signaling in naïve T cells. A. Numbers of adoptively transferred NCD4 OT-II cells recovered from spleens three days after transfer into WT and MHCII- mice (one of three independent experiments). B. Comparison of CD4 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. C. Pooled data, of paired samples showing CD4 MFI and mean ± SE (one of three experiments). D. Comparison of CD5 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. E. Pooled data, of paired samples showing CD5 MFI and mean ± SE (one of three experiments). F. Proliferation assay of donor NCD4 OT-II cells from WT or MHCII- recipient mice four days post-transfer in response to OVA-II peptide in the presence of irradiated DCs from WT mice. Data were normalized for 1,000 donor NCD4 OT-II cells (Mean ± SE of triplicate cultures; data representative of two experiments; three recipient mice per group per experiment). G. Representative DCFDA staining (plotted on log on left and linear scale on right) for donor OT-II cells from WT or MHCII- spleen cells seven days post-transfer. H. Pooled data, of paired samples showing DCFDA MFI values (Mean ± SE, one of three experiments) in WT or MHCII- recipients seven days after transfer. DCs, dendritic cells; DCFDA, 2-7-dichlorofluorescin diacetate; MFI, mean fluorescence intensity; NCD4, naïve CD4 T cells; SE, standard error; WT, wild type.
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Fig6: Consequences of prolonged tonic signaling and absence of tonic signaling in naïve T cells. A. Numbers of adoptively transferred NCD4 OT-II cells recovered from spleens three days after transfer into WT and MHCII- mice (one of three independent experiments). B. Comparison of CD4 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. C. Pooled data, of paired samples showing CD4 MFI and mean ± SE (one of three experiments). D. Comparison of CD5 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. E. Pooled data, of paired samples showing CD5 MFI and mean ± SE (one of three experiments). F. Proliferation assay of donor NCD4 OT-II cells from WT or MHCII- recipient mice four days post-transfer in response to OVA-II peptide in the presence of irradiated DCs from WT mice. Data were normalized for 1,000 donor NCD4 OT-II cells (Mean ± SE of triplicate cultures; data representative of two experiments; three recipient mice per group per experiment). G. Representative DCFDA staining (plotted on log on left and linear scale on right) for donor OT-II cells from WT or MHCII- spleen cells seven days post-transfer. H. Pooled data, of paired samples showing DCFDA MFI values (Mean ± SE, one of three experiments) in WT or MHCII- recipients seven days after transfer. DCs, dendritic cells; DCFDA, 2-7-dichlorofluorescin diacetate; MFI, mean fluorescence intensity; NCD4, naïve CD4 T cells; SE, standard error; WT, wild type.

Mentions: The data so far suggested the hypothesis that CD4 levels and the responsiveness of NCD4 cells were reduced with increasing duration of peripheral residence. During peripheral residence, MHCII-mediated tonic signals are received by NCD4 cells. To test if the reduction in CD4 levels and the loss of responsiveness were related to MHCII-mediated tonic signaling, we ‘parked’ purified NCD4 OT-II cells by adoptive transfer into either WT or MHCII- recipient mice. When recipients were euthanized on day 3 post-transfer, the numbers of OT-II cells recovered per spleen from WT and MHCII- recipients were comparable (Figure 6A) suggesting that there is unlikely to be a major difference in cell survival or homeostatic proliferation in the two groups at this time post-transfer. However, NCD4 OT-II cells recovered from WT recipients had lower CD4 levels (Figure 6B, log and linear plot and 6C) and higher CD5 levels (Figure 6D log and linear plots, and 6E) than cells recovered from MHCII- recipients. When these cells were stimulated with the cognate OVA-II peptide, OT-II cells parked in MHCII- recipients responded better than the corresponding cells parked in WT recipients (Figure 6F).Figure 6


Peripheral residence of naïve CD4 T cells induces MHC class II-dependent alterations in phenotype and function.

Rane S, Das R, Ranganathan V, Prabhu S, Das A, Mattoo H, Durdik JM, George A, Rath S, Bal V - BMC Biol. (2014)

Consequences of prolonged tonic signaling and absence of tonic signaling in naïve T cells. A. Numbers of adoptively transferred NCD4 OT-II cells recovered from spleens three days after transfer into WT and MHCII- mice (one of three independent experiments). B. Comparison of CD4 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. C. Pooled data, of paired samples showing CD4 MFI and mean ± SE (one of three experiments). D. Comparison of CD5 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. E. Pooled data, of paired samples showing CD5 MFI and mean ± SE (one of three experiments). F. Proliferation assay of donor NCD4 OT-II cells from WT or MHCII- recipient mice four days post-transfer in response to OVA-II peptide in the presence of irradiated DCs from WT mice. Data were normalized for 1,000 donor NCD4 OT-II cells (Mean ± SE of triplicate cultures; data representative of two experiments; three recipient mice per group per experiment). G. Representative DCFDA staining (plotted on log on left and linear scale on right) for donor OT-II cells from WT or MHCII- spleen cells seven days post-transfer. H. Pooled data, of paired samples showing DCFDA MFI values (Mean ± SE, one of three experiments) in WT or MHCII- recipients seven days after transfer. DCs, dendritic cells; DCFDA, 2-7-dichlorofluorescin diacetate; MFI, mean fluorescence intensity; NCD4, naïve CD4 T cells; SE, standard error; WT, wild type.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig6: Consequences of prolonged tonic signaling and absence of tonic signaling in naïve T cells. A. Numbers of adoptively transferred NCD4 OT-II cells recovered from spleens three days after transfer into WT and MHCII- mice (one of three independent experiments). B. Comparison of CD4 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. C. Pooled data, of paired samples showing CD4 MFI and mean ± SE (one of three experiments). D. Comparison of CD5 levels (plotted on log on left and linear scale on right) of donor NCD4 OT-II cells after three days in WT and MHCII- mice. E. Pooled data, of paired samples showing CD5 MFI and mean ± SE (one of three experiments). F. Proliferation assay of donor NCD4 OT-II cells from WT or MHCII- recipient mice four days post-transfer in response to OVA-II peptide in the presence of irradiated DCs from WT mice. Data were normalized for 1,000 donor NCD4 OT-II cells (Mean ± SE of triplicate cultures; data representative of two experiments; three recipient mice per group per experiment). G. Representative DCFDA staining (plotted on log on left and linear scale on right) for donor OT-II cells from WT or MHCII- spleen cells seven days post-transfer. H. Pooled data, of paired samples showing DCFDA MFI values (Mean ± SE, one of three experiments) in WT or MHCII- recipients seven days after transfer. DCs, dendritic cells; DCFDA, 2-7-dichlorofluorescin diacetate; MFI, mean fluorescence intensity; NCD4, naïve CD4 T cells; SE, standard error; WT, wild type.
Mentions: The data so far suggested the hypothesis that CD4 levels and the responsiveness of NCD4 cells were reduced with increasing duration of peripheral residence. During peripheral residence, MHCII-mediated tonic signals are received by NCD4 cells. To test if the reduction in CD4 levels and the loss of responsiveness were related to MHCII-mediated tonic signaling, we ‘parked’ purified NCD4 OT-II cells by adoptive transfer into either WT or MHCII- recipient mice. When recipients were euthanized on day 3 post-transfer, the numbers of OT-II cells recovered per spleen from WT and MHCII- recipients were comparable (Figure 6A) suggesting that there is unlikely to be a major difference in cell survival or homeostatic proliferation in the two groups at this time post-transfer. However, NCD4 OT-II cells recovered from WT recipients had lower CD4 levels (Figure 6B, log and linear plot and 6C) and higher CD5 levels (Figure 6D log and linear plots, and 6E) than cells recovered from MHCII- recipients. When these cells were stimulated with the cognate OVA-II peptide, OT-II cells parked in MHCII- recipients responded better than the corresponding cells parked in WT recipients (Figure 6F).Figure 6

Bottom Line: It is not clear if these interactions result in alterations in their activation, survival and effector programming.Naïve CD4lo and CD4hi subsets of thymic single-positive CD4 T cells did not show differences whereas peripheral naïve CD4lo and CD4hi subsets of T cell receptor (TCR)-transgenic T cells did.Our findings also suggest the feasibility of potential pharmacological interventions for improved CD4 T cell responses during vaccination of older people via either anti-oxidant or DUSP inhibitor small molecules.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Immunology, New Delhi, 110067, India. sanketrn@nii.ac.in.

ABSTRACT

Background: As individual naïve CD4 T lymphocytes circulate in the body after emerging from the thymus, they are likely to have individually varying microenvironmental interactions even in the absence of stimulation via specific target recognition. It is not clear if these interactions result in alterations in their activation, survival and effector programming. Naïve CD4 T cells show unimodal distribution for many phenotypic properties, suggesting that the variation is caused by intrinsic stochasticity, although underlying variation due to subsets created by different histories of microenvironmental interactions remains possible. To explore this possibility, we began examining the phenotype and functionality of naïve CD4 T cells differing in a basic unimodally distributed property, the CD4 levels, as well as the causal origin of these differences.

Results: We examined separated CD4hi and CD4lo subsets of mouse naïve CD4 cells. CD4lo cells were smaller with higher CD5 levels and lower levels of the dual-specific phosphatase (DUSP)6-suppressing micro-RNA miR181a, and responded poorly with more Th2-skewed outcomes. Human naïve CD4lo and CD4hi cells showed similar differences. Naïve CD4lo and CD4hi subsets of thymic single-positive CD4 T cells did not show differences whereas peripheral naïve CD4lo and CD4hi subsets of T cell receptor (TCR)-transgenic T cells did. Adoptive transfer-mediated parking of naïve CD4 cells in vivo lowered CD4 levels, increased CD5 and reactive oxygen species (ROS) levels and induced hyporesponsiveness in them, dependent, at least in part, on availability of major histocompatibility complex class II (MHCII) molecules. ROS scavenging or DUSP inhibition ameliorated hyporesponsiveness. Naïve CD4 cells from aged mice showed lower CD4 levels and cell sizes, higher CD5 levels, and hyporesponsiveness and Th2-skewing reversed by DUSP inhibition.

Conclusions: Our data show that, underlying a unimodally distributed property, the CD4 level, there are subsets of naïve CD4 cells that vary in the time spent in the periphery receiving MHCII-mediated signals and show resultant alteration of phenotype and functionality via ROS and DUSP activity. Our findings also suggest the feasibility of potential pharmacological interventions for improved CD4 T cell responses during vaccination of older people via either anti-oxidant or DUSP inhibitor small molecules.

Show MeSH
Related in: MedlinePlus