| Background and objectives:Vitiligo is an acquired pigmentary anomaly of the skin manifested by depigmented white patches surrounded by a normal or a hyperpigmented border. More and more evidences indicated that immune theory is close to the morbidity of vitiligo. Some scholars consider that the breakdown of immunological tolerance may play an important role in vitiligo pathogenesis.The relationship between the level of CD4+CD25+regulatory T cells (CD4+CD25+T cells) expressed in human beings and autoimmune diseases has become a hot spot in the research nowadays. TGF-βis a molecle functioning in immunological regulation and immune suppression. TGF-βcontributes to the maintenance of immunological self-tolerance. Abnormal expression of CD4+CD25+regulatory T cells and related cytokines may be involved in the pathogenesis of vitiligo. Therefore, study on CD4+CD25+T cells and related cytokines may be helpful with illuminating the mechanism of immunological regulation on the molecular level.In the previous study, we examined the proportion of CD4+CD25+T cells in the peripheral lymphocytes from patients with progressing vitiligo and found that the number of CD4+CD25+T cells and the expression of Foxp3 were lower than the normal controls, suggesting that the decrease of CD4+CD25+T cells or abnormality of CD4+CD25+T cells may be correlated with the development of vitiligo. The aim of the present study is to investigate the the level of TGF-β1 from serum and CD4+CD25+T cells in patients with vitiligo and analyze its clinical significance, in order to study the role of CD4+CD25+T cells and TGF-β1 in vitiligo. The relevance with the age,sex and course of disease as well as lesion area were conducted by correlation analysis.Methods:Forty six patients with vitiligo from out-patient clinic and 25 age-matched and sex-matched healthy control subjects were included in the study. Twenty six of the patients were in active period,20 in stable phase. Lymphocytes were separated from patients with vitiligo and normal group, and serum samples were collected from vitiligo patients and healthy controls. CD4+CD25+regulatory T cells were separated from the human PBMC in two steps by magnetic cell sorting (MACS) system. CD4+T cells were negatively sorted by biotin-antibody cocktail and antibiotic microbeads, and then CD4+CD25+T cells were positively sorted by CD25 microbeads. The purity and the survival rate of the sorted cells were measured by flow cytometry, and then stimulated with anti-CD3mAb and anti-CD28mAb.The CD4+CD25+T cells on day 5 after culture was collected. The levels of TGF-P both in the serum and supernatant in the medium of cultured CD4+CD25+T cells were detected by ELISA. In order to study the role of CD4+CD25+T cells and TGF-βin vitiligo, the relations with the age, sex and course of disease as well as lesion area were conducted by correlation analysis using spss software.Results:Analysis of.Serum TGF-β1 levels:1.Serum TGF-β1 levels(1019.49±276.34pg/ml) were significantly decreased in the progressing vitiligo group compared with the control group (7068.32±755.81pg/ml)(P<0.05);Serum TGF-β1 levels(1019.49±276.34pg/ ml) were significantly decreased in the progressing vitiligo group compared with the stable vitiligo group(6839.74±235.25pg/ml)(P<0.05).No difference was detected between the stable vitiligo group(6839.74±235.25 pg/ml) and control group (7068.32±755.81pg/ml) in mean levels of serum TGF-β1 (P >0.05).2. The level of serum TGF-β1 was not correlated with the age, sex and course of vitiligo(P>0.05), while positively correlated with area of skin lesions(r=0.337, P<0.05).Analysis of. TGF-β1 levels secreted by CD4+CD25+T: 1. The levels of (57.78±2.87pg/ml) TGF-β1 secreted by CD4+CD25+T cells were decreased in the progressing vitiligo group compared with the control group (71.97±4.42pg/ml)(P<0.05).The levels of(57.78±2.87pg/ml) TGF-β1 secreted by CD4+CD25+T cells were decreased in the progressing vitiligo group compared the stable vitiligo group (71.33±5.50pg/ml) (P<0.05).The levels of (71.33±5.50) TGF-β1 secreted by CD4+CD25+T cells was no difference in the stable vitiligo group compared with the control group (71.967±4.42pg/ml) (P>0.05).2. The level of TGF-β1 secreted by CD4+CD25+T cells was not correlated with the age, sex and course of vitiligo(P>0.05),and positively correlated with area of skin lesions(r=0.34, P<0.05).Conclusions:1.TGF-β1 levels both from serum and CD4+CD25+T cells were decreased in the progressing vitiligo group compared with the control group.TGF-β1 levels both from serum and CD4+CD25+T cells were decreased in the progressing vitiligo group compared with stable vitiligo group. There was no difference between the stable vitiligo group and control group in.TGF-β1 levels both from serum and CD4+CD25+T cells The level of serum TGF-β1 was not correlated with the age, sex and course of vitiligo, while positively correlated with area of skin lesions.2. The level of TGF-β1 secreted by CD4+CD25+T cells was not correlated with the age, sex and course of vitiligo, while positively correlated with area of skin lesions.
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