Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Mar;123(3):426-37.
doi: 10.1111/j.1365-2567.2007.02707.x. Epub 2007 Oct 4.

Low-dose antigen-experienced CD4+ T cells display reduced clonal expansion but facilitate an effective memory pool in response to secondary exposure

Seong Ok Park  1 , Young Woo Han, Abi George Aleyas, Junu Abi George, Hyun A Yoon, John Hwa Lee, Ho Young Kang, Seong Ho Kang, Seong Kug Eo
Affiliations

Low-dose antigen-experienced CD4+ T cells display reduced clonal expansion but facilitate an effective memory pool in response to secondary exposure

Seong Ok Park et al. Immunology. 2008 Mar.

Abstract

The strength and duration of an antigenic signal at the time of initial stimulation were assumed to affect the development and response of effectors and memory cells to secondary stimulation with the same antigen. To test this assumption, we used T-cell receptor (TCR)-transgenic CD4+ T cells that were stimulated in vitro with various antigen doses. The primary effector CD4+ T cells generated in response to low-dose antigen in vitro exhibited reduced clonal expansion upon secondary antigenic exposure after adoptive transfer to hosts. However, the magnitude of their contraction was much smaller than both those generated by high-dose antigen stimulation and by naïve CD4+ T cells, resulting in higher numbers of antigen-specific CD4+ T cells remaining until the memory stage. Moreover, secondary effectors and memory cells developed by secondary antigen exposure were not functionally impaired. In hosts given the low-dose antigen-experienced CD4+ T cells, we also observed accelerated recall responses upon injection of antigen-bearing antigen-presenting cells. These results suggest that primary TCR stimulation is important for developing optimal effectors during initial antigen exposure to confer long-lasting memory CD4+ T cells in response to secondary exposure.

PubMed Disclaimer

Figures

Figure 1
Figure 1
(a) Activation phenotypes of antigen-experienced CD4+ T cells generated in vitro with different antigen doses. Naive OVA-TCR CD4+ T cells enriched by passage over a nylon-wool column and treated with depleting antibodies and complement were stimulated with APC in the presence of low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide with the Th1-polarized cytokines. Following the 12-hr antigen stimulation, the expression profiles of activation phenotypic markers (CD25, CD69, CD44, and CD62L) of CD4+ KJ1.26+ T cells were determined by FACS analysis. Dot lines represent activation phenotypes of naïve antigen-specific CD4+ T cells that did not pass nylon-wool column. (b) Proliferation potential of antigen-experienced CD4+ T cells generated in vitro with different antigen doses. Following the 12-hr antigen stimulation, primary CD4+ T effector cells were applied to a nylon-wool column to remove further antigenic stimulation and stained with the fluorescence dye CFSE. CFSE-labelled cells were then rested for 3 days. The proliferation of antigen-experienced CD4+ T cells was estimated, based on the assumption that the CFSE signal is reduced by half at each cell division. The histograms are based on a gate specific for the antigen-specific CD4+ T cells (KJ1.26+ CD4+).
Figure 2
Figure 2
The migration of antigen-experienced CD4+ T cells generated in vitro with the different antigen doses into lymphoid and non-lymphoid tissues. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentration of OVA323−339 peptide were rested for 3 days, and then adoptively transferred into hosts via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the following day, the number of donor OVA-TCR CD4+ T cells was determined in lymphoid and non-lymphoid tissues by staining with specific antibodies and FACS analysis. Values represent the absolute number of KJ1.26 + CD4+ T cells and are shown as the mean ± SD of three to five replicates per experiment. *Statistically significant between high-dose antigen-stimulated and the indicated groups (P < 0·05). **Statistically significant between high-dose antigen-stimulated and the indicated groups (P < 0·01). ***Statistically significant between high-dose antigen-stimulated and the indicated groups (P < 0·001).
Figure 3
Figure 3
The behaviour of antigen-experienced CD4+ T cells generated in vitro with different antigen doses in response to secondary antigenic exposure. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide were rested for 3 days and then adoptively transferred into normal BALB/c mice via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the following day, the recipients were immunized via the hind footpad with OVA323−339 peptide (20 μg/mouse) emulsified with CFA. On 5, 10, and 35 days p.i., the behaviour of antigen-experienced CD4+ T cells was examined by staining with specific antibodies and FACS analysis in popliteal LNs. (a) The percentage of KJ1.26 + CD4+ T cells from representative recipients in popliteal LNs. (b) Total number of KJ1.26+ CD4+ T cells in the popliteal LNs of three to five recipients. (c) Number of KJ1.26+ CD4+ T cells in the popliteal LNs of three to five recipients normalized to the peak of expanded response (5 days p.i). Data are shown as the mean ± SD of three to five recipients per experiment. *Statistically significant between low-dose and high-dose antigen-experienced groups (P < 0·05).
Figure 4
Figure 4
The production of polarized cytokine IFN-γ and IL-2 by antigen-experienced CD4+ T cells generated in vitro with different antigen doses. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide were rested for 3 days and then adoptively transferred into normal BALB/c mice via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the following day, the recipients were immunized via the hind footpad with OVA323−339 peptide (20 μg/mouse) emulsified with CFA. On 5, 10, and 35 days p.i., the cytokines secreted by antigen-specific CD4+ T cells in the total popliteal LN cells were determined by a standard ELISA after brief stimulation with OVA323−339 peptide (1 µg/ml). Data are shown as the mean ± SD of three to five recipients per experiment. P-values in graphs were statistically calculated by Student's t-test.
Figure 5
Figure 5
Phenotypes of antigen-specific CD4+ T memory cells developed by primary effector cells in response to secondary antigenic exposure. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide were rested for 3 days and then adoptively transferred into normal BALB/c mice via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the next day, the recipients were immunized via the hind footpad with OVA323−339 peptide (20 μg/mouse) emulsified with CFA. On 35 days p.i., the expression of phenotypic markers (CD44 and CD62L) of CD4+ KJ1.26 + T cells obtained from the popliteal LNs was determined by FACS analysis. The histograms based on a gate specific for the antigen-specific CD4+ T cells (KJ1.26+ CD4+) are representative of three to five recipients per experiment and values in histograms are the mean percent of experiments.
Figure 6
Figure 6
Recall responses of antigen-specific CD4+ T memory cells developed by primary effector cells generated in vitro with the different antigen doses in response to secondary antigen exposure. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide were rested for 3 days and then adoptively transferred into normal BALB/c mice via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the next day, the recipients were immunized via the hind footpad with OVA323−339 peptide (20 μg/mouse) emulsified with CFA. On 60 days p.i., the recipients were injected with OVA323−339 peptide-pulsed APC via tail vein (2·0 × 107 cells/mouse) and then the recall responses of antigen-specific CD4+ T cells were examined 5 days after injection of antigen/APCs. (a) The percentage of KJ1.26+ CD4+ T cells in the spleen of representative recipients re-challenged with antigen/APC. (b) Total number of KJ1.26+ CD4+ T cells expanded with antigen/APC in the spleen of three to five recipients. P-values in graph were statistically calculated by Student's t-test. (c) Enumeration of proliferating antigen-specific CD4+ T cells during recall response with antigen/APC. The recipients injected with antigen/APC were administered BrDU (0·8 mg/ml) in the drinking water. On day 5, BrDU incorporation of antigen-specific CD4+ KJ1.26+ T cells was determined by FACS analysis. The histograms based on a gate specific for the antigen-specific CD4+ T cells (KJ1.26+ CD4+) are representative of three to five recipients and values in histograms are the mean ± SD of each experiment. The dot-lines of histogram denote isotype control. (d) The expression profiles of early activation markers (CD25 and CD69) on the cell surface of antigen-specific CD4+ T cells. The histograms are representative of three to five recipients.
Figure 7
Figure 7
The production of polarized cytokines IFN-γ and IL-2 during recall response by antigen-specific CD4+ T memory cells developed by primary effector cells generated in vitro with different antigen doses. Antigen-experienced CD4+ T cells generated in vitro with low (0·05 μg/ml) and high (5·0 μg/ml) concentrations of OVA323−339 peptide were rested for 3 days and then adoptively transferred into normal BALB/c mice via i.v. injection (2·5 × 106 CD4+ KJ1.26+ cells/mouse). On the next day, the recipients were immunized via the hind footpad with OVA323−339 peptide (20 μg/mouse) emulsified with CFA. On At 60 days p.i., the cytokine levels were determined by a standard ELISA in the splenocytes of recipients 5 days after re-challenging with antigen/APC. Data are shown as the mean ± SD of three to five recipients per experiment. P-values in graphs were statistically calculated by Student's t-test.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Swain SL, Croft M, Dubey C, Haynes L, Rogers P, Zhang X, Bradley LM. From naive to memory T cells. Immunol Rev. 1996;150:143–67. - PubMed
    1. Sprent J, Surh CD. T cell memory. Annu Rev Immunol. 2002;20:551–79. - PubMed
    1. Davis MM, Boniface JJ, Reich Z, Lyons D, Hampl J, Arden B, Chien Y. Ligand recognition by alpha beta T cell receptors. Annu Rev Immunol. 1998;16:523–44. - PubMed
    1. van der Merwe PA, Davis SJ. Molecular interactions mediating T cell antigen recognition. Annu Rev Immunol. 2003;21:659–84. - PubMed
    1. Iezzi G, Karjalainen K, Lanzavecchia A. The duration of antigenic stimulation determines the fate of naive and effector T cells. Immunity. 1998;8:89–95. - PubMed

Publication types

LinkOut - more resources