Journal: Journal of Immunology Research

Article Title: Decreased GZMB , NRP1 , ITPR1 , and SERPINB9 Transcripts Lead to Reduced Regulatory T Cells Suppressive Capacity in Generalized Vitiligo Patients

doi: 10.1155/2022/3426717

Figure Lengend Snippet: Gating strategy for CD4 + CD25 + FOXP3 + Treg cells. Estimation of protein levels of CD4, and CD25. (a) Lymphocytes were gated on the basis of size and morphology. (b) CD3 + CD4 + T cells were gated on the basis of CD3 and CD4 expression.(c) Treg cells were gated on the basis of CD3, CD4, CD25, and FOXP3 expression. The purity of isolated CD3 + CD4 + CD25 + FOXP3 + Treg cells was found to be 94.22%. (d) Expression of CD3 in T cells. Representative graph shows the amount of CD3 in the T cells as mean fluorescence intensity (MFI). (e) Expression of CD4 in T cells. Representative graph shows amount of CD4 in the T cells as mean fluorescence intensity (MFI). (f) Expression of CD25 in T cells. Representative graph shows amount of CD25 in the T cells as mean fluorescence intensity (MFI). (g) Expression of FOXP3 in T cells. Representative graph shows amount of intracellular FOXP3 in the T cells as mean fluorescence intensity (MFI).

Article Snippet: CD4 + CD25 + Treg cells and CD4 + T cells were isolated from three-milliliter peripheral blood of 52 GV patients and 48 controls using MACSxpress® whole blood Treg isolation kit (Miltenyi Biotec, Auburn, CA) as mentioned previously [ ].

Techniques: Expressing, Isolation, Fluorescence

Journal: Journal of Immunology Research

Article Title: Decreased GZMB , NRP1 , ITPR1 , and SERPINB9 Transcripts Lead to Reduced Regulatory T Cells Suppressive Capacity in Generalized Vitiligo Patients

doi: 10.1155/2022/3426717

Figure Lengend Snippet: Correlation of in vitro Treg suppression assay with ITPR1 , GZMB , NRP1 , and SERPINB9 transcripts in Tregs of GV patients. The correlation of in vitro Treg suppression assay with ITPR1 , GZMB , NRP1 , and SERPINB9 was analyzed by Spearman's correlation analysis. (a) GZMB transcripts were positively correlated with in vitro Treg mediated suppression of CD4 + T cells in GV patients' Tregs ( r = 0.61; p = 0.0012). (b) NRP1 transcripts were positively correlated with in vitro Treg-mediated suppression of CD4 + T cells in GV patients' Tregs ( r = 0.55; p = 0.021). (c) SERPINB9 transcripts were positively correlated with in vitro Treg-mediated suppression of CD4 + T cells in GV patients' Tregs ( r = 0.56; p = 0.002). (d) ITPR1 transcripts were positively correlated with in vitro Treg-mediated suppression of CD4 + T cells in GV patients' Tregs ( r = 0.54; p = 0.001). (e) GZMB transcripts positively correlated with in vitro Treg-mediated suppression of CD8 + T cells in GV patients' Tregs ( r = 0.58; p = 0.004). (f) NRP1 transcripts positively correlated with in vitro Treg-mediated suppression of CD8 + T cells in GV patients' Tregs ( r = 0.52; p = 0.0022). (g) SERPINB9 transcripts positively correlated with in vitro Treg-mediated suppression of CD8 + T cells in GV patients' Tregs ( r = 0.48; p = 0.024). (h) ITPR1 transcripts positively correlated with in vitro Treg-mediated suppression of CD8 + T cells in GV patients' Tregs ( r = 0.49; p = 0.032).

Article Snippet: CD4 + CD25 + Treg cells and CD4 + T cells were isolated from three-milliliter peripheral blood of 52 GV patients and 48 controls using MACSxpress® whole blood Treg isolation kit (Miltenyi Biotec, Auburn, CA) as mentioned previously [ ].

Techniques: In Vitro, Suppression Assay

Journal: Journal of Immunology Research

Article Title: Decreased GZMB , NRP1 , ITPR1 , and SERPINB9 Transcripts Lead to Reduced Regulatory T Cells Suppressive Capacity in Generalized Vitiligo Patients

doi: 10.1155/2022/3426717

Figure Lengend Snippet: Role of GZMB , NRP1 , SERPINB9 , and ITPR1 transcripts in GV pathogenesis. (a) The decreased ITPR1 transcripts could lead to impaired calcium-NFAT signalling pathway, which might result in decreased GZMB and NRP1 transcripts. Further, the decreased SERPINB9 transcripts may result in increased granzyme B-mediated endogenous apoptosis of Tregs. Overall, the decreased GZMB , NRP1 , SERPINB9 , and ITPR1 transcripts result into decreased Treg suppressive capacity, which could lead to unchecked CD4 + and CD8 + T cells and thereby results into melanocytes' destruction contributing to GV pathogenesis, progression, and severity. (b) Upon calcium treatment, ITPR1 mRNA expression is increased which may lead to intracellular Treg calcium influx and calcium-NFAT signalling pathway, thereby results into increased GZMB and SERPINB9 transcripts, leading to increased Treg suppressive capacity. The increased Treg suppressive capacity controls the CD8 + and CD4 + T cells proliferation and IFN- γ production and thereby contributes to melanocytes survival.

Article Snippet: CD4 + CD25 + Treg cells and CD4 + T cells were isolated from three-milliliter peripheral blood of 52 GV patients and 48 controls using MACSxpress® whole blood Treg isolation kit (Miltenyi Biotec, Auburn, CA) as mentioned previously [ ].

Techniques: Expressing