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Immunofluorescent Labeling in Nasal Mucosa Tissue Sections of Allergic Rhinitis Rats via Multicolor Immunoassay
Published on: September 22, 2023
[Immunoglobulin domain and mucin domain-1 in helper T lymphocytes in allergic rhinitis]
Wen-kui Wu1, Yun-fang An, Chang-qing Zhao
1Department of Otolaryngology, Second Hospital of Shanxi Medical University, Taiyuan 030001, China.
Objective:
To investigate the significance of T cell immunoglobulin domain and mucin domain-1 (TIM-1) expression in helper T lymphocyte in allergic rhinitis (AR) patients.
Methods:
Peripheral blood samples were collected from 20 AR patients, 9 males and 11 females, aged 31.4 (20-54), and 20 healthy persons, 12 males and 8 females, aged 38, 6 (18-60). Peripheral blood mononuclear cells (PBMNs) were isolated and magnetic cell sorting technique was used to purify the CD4+ T cells and CD4 + CD45RA + T cells. Immunofluorescence staining and flow cytometry were used to detect the percentage of CD4 + CD45RA + T cells positive in TIM-1, a helper T lymphocyte marker. Phytohemagglutinin (PHA) was added into the culture media to stimulate the TIM-1 positive helper T lymphocytes. Dendritic cells (DCs) thus induced and differentiated were conditioned by standardized allergen from dust mite (Dermatophagoides pteronyssinus) or lipopolysaccharide (LPS), and then co-cultured with CD4 T cells. The percentage of TIM-1 positive CD4 + CD45RA + T cells was counted. ELISA was used to detect the IL-4 and interferon (IFN)-gamma levels in the supernatants of the culture media of the dust mite and LPS groups.
Results:
The percentage of TIM-1 positive CD4+ T cells of the AR patients was (9.48 +/- 1.51)%, significantly higher than that of the healthy controls [(3.12 +/- 0.32)%, P < 0.05], and the percentage of TIM-1 positive CD4 + T cells after PHA stimulation of the AR patients was (47.05 +/- 4.35)%, significantly higher than that of the healthy controls [(29.92 +/- 3.36)%, P < 0.05]. The percentage of TIM-1 positive CD4 + CD45RA + T cells of the AR patients was (26.34 +/- 4.0)%, significantly higher than that of the healthy controls [(18.4 +/- 8.1)%, P < 0.05], and the percentage of TIM-1 positive CD4 + CD45RA + T cells after PHA stimulation of the AR patients was (65.3 +/- 7.4)%, significantly higher than that of the healthy controls [(36.3 +/- 6.8)%, P < 0.05]. The percentage of TIM-1 positive CD4 + CD45RA + T cells co-cultured with the DCs conditioned by dust mite of the AR patients was (10.81 +/- 1.48)%, significantly higher than that of the LPS group [(6.08 +/- 1.45)%] and the blank control group [(1.56 +/- 2.30)%, both P < 0.05]. The IL-4 level of the DCs conditioned by dust mite allergen was (74.61 +/- 13.82) pg/ml, significantly higher than those of the LPS group and blank control group [(28.57 +/- 3.36) and (1.40 +/- 0.99) ng/ml respectively, both P < 0.05]. The IFN-gamma level of the DCs conditioned by dust mite allergen was (25.31 +/- 7.23) pg/ml, significantly lower than that of the LPS group [(163.41 +/- 82.37) pg/ml, P < 0.05], however, significantly higher than that of the blank control group [(1.58 +/- 0.53) pg/ml, P < 0.05].
Conclusion:
The percentage of TIM-1 positive helper T lymphocytes of the AR patients are abnormally high. After stimulation of allergen The DCs increase the number of TIM-1 expressing naïve Th cells and promote them to differentiate into Th2 cells. Promoting naïve Th cells to differentiate into Th2 cells, TIM-1 expression induces Th2 cytokine-based immune responses.
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