Immune Netw.  2017 Jun;17(3):163-170. 10.4110/in.2017.17.3.163.

Clonal Expansion of Allergen-specific CD4⁺ T Cell in the Lung in the Absence of Lymph Nodes

Affiliations
  • 1Laboratory of Immune Regulation, Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 08826, Korea. yeonseok@snu.ac.kr
  • 2BK21 Plus Program, College of Pharmacy, Seoul National University, Seoul 08826, Korea.

Abstract

The expansion of allergen-specific CD4⁺ T cells is a critical step in inducing airway inflammation during allergic asthma. Such clonal expansion of T cells is initiated through the interaction between allergen-bearing dendritic cells and allergen-specific naïve T cells in the draining lymph nodes. Whether such T cell clonal expansion also occurs in the lung, the primary organ encountering inhaled allergens, remains unclear. Compared with wild-type mice, we found similar frequencies of CD4⁺ T cells in the lung of lymph node-deficient Rorgt(gfp/gfp) mice after repeated exposure to inhaled allergens. In addition, we observed an evident population of CD4⁺ T cells that underwent clonal expansion in the lung of allergen-challenged mice treated with an S1P antagonist FTY720 in an in vivo proliferation study with CFSE-labeled OT-II T cells. Moreover, the expansion of allergen-specific CD4⁺ T cells was significantly enhanced in the lungs of Rorgt(gfp/gfp) mice in comparison to that of wild-type mice. These results together demonstrate that the clonal expansion of allergen-specific CD4⁺ T cells occurs in the absence of the lymph nodes, indicating that the lung can act as a primary site of the clonal expansion of CD4⁺ T cells in response to inhaled allergens.

Keyword

Allergens; Lung; RORγt; CD4⁺ T cell; Proliferation

MeSH Terms

Allergens
Animals
Asthma
Dendritic Cells
Fingolimod Hydrochloride
Inflammation
Lung*
Lymph Nodes*
Mice
T-Lymphocytes
Allergens
Fingolimod Hydrochloride

Figure

  • Figure 1 CD4+ T cell population in the lung of Rorgtgfp/gfp mice in steady-state. (A) Representative pictures of the superficial cervical lymph nodes, the mLN, the mesenteric lymph nodes, the inguinal lymph nodes and the Peyer's patches of wild-type mice and Rorgtgfp/gfp mice. (B) The number of CD45.2+ immune cells in the lungs of Rorgt+/gtp and Rorgtgfp/gfp mice and (C) the proportion of CD4+ cells among CD45.2+ cells in the lung of Rorgt+/gtp and Rorgtgfp/gfp mice before intranasal challenge. n=3 mice per group (B and C).

  • Figure 2 CD4+ T cells are major immune cells in the lung of RORγt-deficient mice after intranasal challenges with allergens. (A) Experimental scheme. Wild-type mice and Rorgtgfp/gfp mice were intranasally injected with PAO/Ova every other day for 4 times and analyzed one day after the last challenge. (B) The proportion of CD4+ cells among CD45.2+ cells in the lung of wild-type mice and Rorgtgfp/gfp mice after intranasal challenges with PAO/Ova. Data are means±SEM. n=4 mice per group. Data are representative of two independent experiments.

  • Figure 3 Kinetics of allergen-specific CD4+ T cell proliferation in the BALT in response to intranasal allergen. (A) Experimental scheme. (B-D) Proliferation of OT-II T cells after intranasal PAO/Ova in the indicated tissue. CD45.1+ donor T cells were gated and analyzed for CFSE dilution by flow cytometry. Data are means±SEM. *p<0.05 and **p<0.01 in comparison with the mLN. Data are representative of three independent experiments.

  • Figure 4 Expansion of allergen-specific CD4+ T cell in the lung in mice treated with FTY720. (A) Experimental scheme. CFSE-labeled CD45.1+OT-II T cells were transferred into wild-type mice (CD45.2+/+) and the recipients were intranasally challenged with PAO/Ova. Six hours after the intranasal challenge, the mice were intraperitoneally injected with 1 mg/kg FTY720 or vehicle (n=3). (BD) Forty-eight hours later, the frequency and the dilution of CFSE of donor T cells were analyzed in the indicated lymphoid tissues or blood. Data are means±SEM. **p<0.01 in comparison with vehicletreated group. Data are representative of two independent experiments.

  • Figure 5 Expansion of allergen-specific CD4+ T cell in the lung of Rorgtgfp/gfp mice. (A) Experimental scheme (n=3). (B) Forty-eight hours after the challenge, the proportion of divided cells among CFSE-labeled donor T cells was analyzed by flow cytometry. Numbers on the line indicate the percentage of divided cells. (C) Seventy-two hours after co-culture of the indicated T cells with BMDC in the presence of anti-CD3, CFSE-dilution was measured to determine the proliferation of the T cells. Data are means±SEM. **p<0.01 in comparison with wild-type mice. Data are representative of two independent experiments.


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