Decreased circulating Th1 cells and elevated Th2 cells in patients with UCC in different stages
We first analyzed the expression of Th1 and Th2 cells based on the cytokine patterns after in vitro activation by PMA/ionomycin in short-term cultures. Among the 38 UCC patients, 22 belonged to stage I, and the percentage of Th1 cells was 10.06 ± 1.24%. The other 16 patients belonged to stage II, and the percentage of Th1 cells was 7.77 ± 0.8%. Compared with healthy controls, a lower proportion of Th1 cells was seen in patients with UCC; the higher was the stage they belonged to, the lower was their Th1 cell percentage (P < 0.001) (Figure 1 A). The percentage of Th2 cells in UCC patients was 8.28 ± 1.44% at stage I but 11.82 ± 1.07% at stage II. The percentage of Th2 cells increased with higher stages, and the difference between the two groups was significant (P < 0.001) (Figure 1 A). Remarkably, Th1/Th2 imbalance was observed in UCC patients.
Elevated circulating Th17 cells and Treg cells in patients with UCC at different stages
Compared with healthy controls, Th17 cells were increased in UCC patients, particularly at stage II, as showed in Figure 1 A. The percentage of Th17 cells in patients was 11.11 ± 1.46% at stage I and 14.21 ± 1.37% at stage II (P < 0.001). The percentage of Treg cells in UCC patients was 5.02 ± 1.21% at stage I and 5.88 ± 0.84% at stage II (P = 0.019). The percentage of Treg cells increased marginally in patients at stage II compared with stage I. The differences of Th17 and Treg cell percentages between the two groups were all significant (Figure 1 A). An evident imbalance of Th17/Treg was also observed in UCC patients.
The percentages of Th1, Th2, Th17, and Treg cells were associated with the tumor size, lymph node metastases, and vasoinvasion
Tumor size was expressed as the diameter measured by pathologists. In Figure 2, we used horizontal coordinates to represent the tumor diameter and vertical coordinates to represent the cell percentage. The figure showed a negative correlation between Th1 cell percentage in the serum and the tumor diameter. The positive correlation existed between the tumor diameter and percentage of Th2 cells, Th17 cells, and Treg cells.
Among the 38 UCC patients, 30 patients had no lymph node metastases and eight patients had lymph node metastases. Thus, we accordingly separated these patients into positive and negative groups. As shown in Figure 1B, the percentage of Th1 cells was 8.07 ± 1.80% in the positive group and 9.51 ± 1.59% in the negative group (P = 0.013). The percentage of Th2 cells was 10.56 ± 1.81% in the positive group and 9.33 ± 2.01% in the negative group (P = 0.025). The percentage of Th17 cells was 13.50 ± 1.93% in the positive group and 11.93 ± 2.10% in the negative group (P = 0.010). The percentage of Treg cells was 5.10 ± 1.44% in the positive group and 5.48 ± 1.83% in the negative group. Only the difference of Treg cells between the two groups was not significant (P = 0.151).
Thirty-one UCC patients had no vasoinvasion, and seven cases were detected as having vasoinvasion. Thus, we accordingly separated the patients into positive and negative groups. As shown in Figure 1C, the percentage of Th1 cells was 7.79 ± 1.74% in the positive group and 9.53 ± 1.62% in the negative group (P = 0.009). The percentage of Th2 cells was 10.50 ± 1.72% in the positive group and 9.25 ± 2.06% in the negative group (P = 0.034). The percentage of Th17 cells was 13.90 ± 2.04% in the positive group and 11.21 ± 2.14% in the negative group (P = 0.005). The percentage of Treg cells was 5.03 ± 2.11% in the positive group and 5.52 ± 1.78% in the negative group. Only the difference of Treg cells between the two groups was not significant (P = 0.252).
The balance of Th1/Th2 and Th17/Treg cells recovered after radical hysterectomy in UCC patients
To understand the relationship of the balance of Th1/Th2 and Th17/Treg cells with our therapy, we estimated Th1, Th2, Th17, and Treg cell levels with or without therapy. Patients with UCC had a lower proportion of Th1 cells (10.27 ± 1.51%) compared with healthy controls (23.12 ± 2.81%)(P<0.001). However, after the operation, the proportion of Th1 cells recovered quickly one month later (20.69 ± 3.19%) (P = 0.092)and at six months (21.56 ± 2.39%)(P = 0.055) (Figure 1D, Figure 3). The percentage of Th2 cells in UCC patients also recovered at six months after the operation (2.67 ± 0.56%)(P = 0.309). However, at one month after the operation, the Th2 cell percentage was still significantly higher in UCC patients (6.34 ± 1.76%) than in the control group (2.11 ± 0.99%) (P < 0.001) (Figure 1D, Figure 3).
Compared with controls (1.23 ± 0.41%), patients with UCC had an evidently higher proportion of Th17 cells (11.25 ± 1.77%) (P<0.001). After the operation, Th17 cells recovered at six months (2.27 ± 0.81%) (P=0.056). One month postoperatively, Th17 cells had decreased to (7.07 ± 1.19%)(P<0.001) (Figure 1D). The percentage of Treg cells in UCC patients also recovered at six months after the operation (2.37 ± 0.89%)(P=0.153). However, at one month postoperatively, the Treg cell percentage was still evidently higher in UCC patients (3.34 ± 2.07%)(P=0.025) than in the control group (Figure 1D, Figure 3).
We found the balances began to recover at one month postoperatively and almost reach normal levels at six months postoperatively.
The balance of Th1/Th2 and Th17/Treg cells recovered after cervical conization in CIN patients
Among the 61 CIN patients, only 21 patients had CIN III and were treated with cold knife conization. To understand the relationship of cervical conization with the balance of Th1/Th2 and Th17/Treg cells, we estimated Th1, Th2, Th17, and Treg cell levels before and after the operation. CIN III Patients had a lower proportion of Th1 cells (13.94 ± 2.11%) compared with controls (23.12 ± 2.81%)(P<0.001). However, after conization, the proportion of Th1 cells recovered quickly at one month later (22.01 ± 2.5%)(P=0.073) (Figure 1E). Moreover, the percentage of Th2 cells also recovered quickly at one month later in CIN III patients (2.76±1.90%) (P=0.058) (Figure 1E).
Compared with controls (1.23 ± 0.41%), CIN III patients had a significantly higher proportion of Th17 cells (9.49 ± 0.93%) (P<0.001). After the operation, Th17 cells recovered at six months (1.49 ± 0.52%)(P=0.391). At one month, the proportion of Th17 had already deceased to 1.61 ± 0.69%. The percentage of Treg cells in CIN III patients also reached normal levels at six months after the operation (2.18 ± 0.71%) (P=0.205) (Figure 1E). We found the balances were almost normal at just one month postoperatively.