| Objective:Leprosy is a chronic infection where the skin and peripheral nervous system is invaded by Mycobacterium leprae.The infection and immune mechanism remain unknown.T helper 17(Th17)cells and regulatory T(Treg)cells are derived from naive CD4+T cells.The balance between Th 1 7 and Treg cells is crucial for immune homestatis and tolerance.Immunoreaction is the major factor in the pathogenesis of leprosy.It remains unclear that whether the balance between Th17 and Treg cells is disturbed in leprosy patients.In this study,the expression of transcription factors and cytokines was detected in Thl7 and Treg cells from skin lesions of leprosy patients for further investigating the potential role of transforming growth factor-beta(TGF-β), retinoid-related orphan receptor gamma t(RORyt),interleukin-10(IL-10),IL-17,and forkhead box P3(FOXP3)in the pathogenesis of leprosyMethods:Clinical specimen collection60 cases was collected with skin lesion paraffin section of leprosy patients who had visited Zhe Jiang Skin Disease Prevention and Treatment Hospital,The cases of each type are as follows:15 cases of tuberculous leprosy(TT),14 cases of borderline tuberculoid leprosy(BT),15 cases of borderline lepromatous leprosy(BL),and 16 cases of lepromatous leprosy(LL).15 cases was collected with skin tissue paraffin sections of healthy volunteers.ImmunohistochemistryAfter immunohistochemical(SP)was performed,the expression level of detected Th17/Treg cells transcription factor and related cytokines is that Th17 cells marked as transcription factor RORyt and Th17 cells secretion cytokines IL-17;Treg cells marked as transcription factor FOXP3 and Treg cells secretion cytokines IL-10,TGF-β.Result analysis and evaluationThe results were semi-quantitatively analysed and scored according to the intensity of immunochemical staining and the percentage of positive cells.The ratios of immuno-positive cells were divided into 5 degrees:0(negative),1(1～25%staining),2(26～50%staining),3(51～75%staining),and 4(>75%staining).The staining intensity of immuno-positive cells was divided into 4 degrees:0(negative),1(weak),2(moderate),and 3(strong).The final score was calculated as the sum of the two scores above.Statistical analysis of the relationship between the expression of Th17 and Treg cells Transcription Factor and Related Cytokines in different types of leprosy.Results:The expression in RORγt、IL-17、FOXP3、IL-10、TGF-β epidermis or dermal cells of leprosy patients,which was brown or tawny,and they were less expressed in skin tissues of healthy control group.Thl7 cell transcription factor RORyt’s positive expression score in BL,BT,TT tissues are 3.69±0.87,4.12±0.59,3.64±0.87,which was higher than normal tissue 2.42±0.54 and 2.65±0.86in LL tissue(p<0.05)Thl7 cell marker cytokine IL-17’s positive expression score in BL,BT,LL,TT are 4.29±0.65,3.98±0.80,4.46±0.67,4.02±1.15,and there was no significant difference with normal tissue 3.82±1.05(p>0.05).Treg cell transcription factor FOXP3’s positive expression score in BL,BT,LL,TT are 3.31±0.99,3.40±0.71,3.60±1.08,3.73±0.70,which was higher than normal tissue 2.69±0.77(p<0.05)Treg cell cytokine IL-10’s positive expression score in BL,LL,TT tissues are 4.31±0.61,4.13±0.51,4.24±0.90,which was higher than normal tissue 3.18±0.98 and 3.19±0.77in BT tissue(p<0.05)Compared with healthy control group.Treg cell cytokine TGF-β’s positive expression score in BL,BT,LL tissues are respectively 5.00±1.14,4.79±1.24,4.58±1.10,it was higher than normal tissue 3.71±0.49(p<0.05)Conclusions:1.The expression of Th17 cell transcription factor RORyt was increased in borderline lepromatous leprosy(BL),borderline tuberculoid leprosy(BT),and tuexistrculoid leprosy(TT)groups.Interestingly,RORyt expression was lower in lepromatous leprosy(LL)group than that in other types of leprosy.However,there was no significant difference in the expression of IL-17 between leprosy groups and healthy controls.These results suggest that Th17 cells may have a protective response to Mycobacterium leprosy infection,which is beneficial to pathogen clearance2.Treg cell transcription factor FOXP3 was significantly increased in all types of leprosy when compared with healthy controls.The expression of TGF-β was obviously increased in BL,BT,and LL groups.The expression of IL-10 was increased in BL,LL,TT groups.Besides,IL-10 expression was lower in BT group than that in other types of leprosy.These results suggest that Treg cells may be associated with clinical typing and immunological characteristics of leprosy.3.Changes in the number or function of Th17/Treg cells in leprosy lesions may be related to the immune pathogenesis of leprosy or the occurrence and development of the disease. |
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