Novel strategy of sirolimus plus thymalfasin and huaier granule on tumor recurrence of hepatocellular carcinoma beyond the UCSF criteria following liver transplantation: A single center experience.

Although liver transplantation (LT) lengthens the survival time of patients with hepatocellular carcinoma (HCC), LT patients exhibit a high recurrence rate; particularly those that had advanced HCC associated with the tumor biological characteristics and long-term application of immunosuppressants. A consensus on optimal prophylaxis and treatment for recurrent HCC following LT does not currently exist. The present study retrospectively analyzed data from 36 non-University of California at San Francisco criteria-eligible patients with advanced HCC who underwent LT, and then treated them with sirolimus (SRL)-based therapy with thymalfasin and huaier granules (SRL+, n=18), or with tacrolimus-based therapy (controls; n=18). The SRL+ group had significantly longer recurrence times (P=0.008) and survival times (P<0.0001) (OS, 1-year: 100%, 3-year: 94.4%, 5-year: 77.8%; DFS, 1-year: 88.9%, 3-year: 55.6%, 5-year: 50.0%). Furthermore, compared with pre-LT values and the control group, the SRL+ group had significantly lower serum α-fetoprotein (AFP) levels (both P<0.0001) and percentage of Forkhead box P3 (FoxP3)+ Treg lymphocytes (P<0.001) during the first year. In the SRL+ group, FoxP3+/cluster of differentiation (CD)8+ Treg lymphocyte percentages decreased significantly following LT (P<0.001); however, CD8+/CD3+ T-cells significantly increased (P<0.001). Levels of serum AFP and FoxP3+ Treg cells increased when tumors relapsed, and decreased to near-normal when relapse foci were cured or stabilized. SRL+ therapy may decrease AFP and Treg levels, while increasing CD8+ T cells, indicating an associated mechanism among them. In conclusion, SRL+ therapy appears to be safe and effective in preventing HCC recurrence following LT with no significant adverse events, and warrants further investigation.

[Immature dendritic cells phagocytosing human spleen cells treated by PUVA present the characteristics of regulatory dendritic cells].

Objective To investigate the effect of psoralen combined with A-band ultraviolet (PUVA)-treated human spleen lymphocytes on the phenotype and function of immature dendritic cells (imDCs). Methods Human peripheral blood mononuclear cells (PBMCs) were isolated and induced to produce DCs by interleukin-4 (IL-4) and recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF). On the sixth day, the imDCs were obtained and stimulated by lipopolysaccharide (LPS). One day later, mature DCs were harvested. Human spleen cells (SPs) were isolated and treated with PUVA to prepare apoptotic PUVA-SPs. Co-culture of imDCs with PUVA-SPs resulted in extracorporeal photochemotheraputic DCs (ecpDCs). Co-culture of imDCs with SPs resulted in SP-DCs. The expressions of CD11c, CD83 and CD86 were detected by flow cytometry. The levels of IL-10 and IL-12 in the supernatants of the above cells were determined by ELISA. Results The early apoptosis rate of PUVA-SPs was (94.21±3.75)%. There was no significant difference in the expressions of CD83 and CD86 between imDCs and ecpDCs. But the positive rates of CD83 and CD86 in ecpDCs were lower than those in DCs. However, the positive rates of CD83 and CD86 in SP-DCs were significantly higher than those of the imDCs. Conclusion The imDCs phagocytosing apoptotic human SPs present phenotype and function of regulatory DCs.

[Immature dendritic cells phagocytosing apoptotic human spleen cells treated with PUVA inhibits the maturation induced by LPS].

Objective To investigate whether lipopolysaccharide (LPS) can induce the maturation of immature dendritic cells (imDCs) which phagocytose apoptotic spleen lymphocytes. Methods Human peripheral blood mononuclear cells (PBMCs) were induced to produce DCs by interleukin 4 (IL-4) and recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF). Human spleen cells (hSPs) were isolated and treated with psoralen combined with ultraviolet A(PUVA) to obtain apoptotic PUVA-hSPs. Co-culture of imDCs with PUVA-hSPs resulted in extracorporeal photochemotherapeutic dendritic cells (ecpDCs). The imDCs and ecpDCs were collected and stimulated by 10 ng/mL LPS for 1 day. The expressions of CD11c, CD83 and CD86 were detected by flow cytometry. The level of IL-10 in the supernatants of the above cells was detected by ELISA. Results There was no significant difference in the expressions of CD83 and CD86 between ImDCs and ecpDCs. However, the positive rates of CD83 and CD86 in the imDCs stimulated by LPS were significantly higher than those in the ecpDCs treated by LPS. The level of IL-10 in imDCs culture supernatant was lower than that in ecpDCs. The level of IL-10 in LPS-stimulated imDCs was lower than that in LPS-stimulated ecpDCs. Conclusion Both imDCs and ecpDCs showed immature phenotype, but ecpDCs can inhibit the maturation of DC induced by LPS.

Sirolimus treatment for cirrhosis or hepatocellular carcinoma patients accompanied by psoriasis after liver transplantation: A single center experience.

There is currently no consensus on the most suitable therapeutic approach for psoriasis (PS) co-existing with posthepatic cirrhosis (PCs) and hepatocellular carcinoma (HCC) following liver transplantation (LT). The present study provides an analysis of the therapeutic experience of such patients. Five LT recipients (two with PC and three with HCC) with accompanying PS were included. The induction program consisted of methylprednisolone plus basiliximab treatment. The initial postoperative treatment scheme consisted of tacrolimus (FK506) plus mycophenolate mofetil (MMF) and hormone; the latter was withdrawn 1 week after LT. The patients with PC had been using FK506 with or without a postoperative MMF program; the patients with HCC and recurrence of PS had been switched to a sirolimus (SRL)-based replacement therapy. Furthermore, all patients received anti-hepatitis B virus (HBV) therapy. The patients were followed up after 8.3±1.5 years. There was a positive correlation between HBV-DNA copy numbers, and psoriatic area and severity index (PASI) scores (r=0.97; P=0.006). The PASI scores were decreased significantly at 6 months following surgery compared with pre-transplantation (P<0.05). The patients who had received the FK506-based treatment experienced PS recurrence two years post-transplantation. The PASI scores increased significantly (P<0.05) and then declined gradually, maintaining a stable level (P<0.05) by 1 year after switching to the SRL-based treatment. The patients who had received the SRL-based treatment exhibited no recurrence of PS. The results of the present study suggest that SRL therapy provides a promising novel treatment method for patients with PS following LT that may be superior to tacrolimus treatment. When co-existing HBV is present pre-transplantation, regular injection of human hepatitis B immunoglobulin should be used to prevent the HBV from relapsing or aggravating the PS.

The role of regulatory B cells (Bregs) in the Tregs-amplifying effect of Sirolimus.

Sirolimus can significantly amplify regulatory T cells (Tregs) in vivo and in vitro, but the specific mechanism of this has not been well documented. The role of regulatory B cells (Bregs) in the Tregs-amplifying effect of Sirolimus was investigated in peripheral blood mononuclear cells (PBMCs) in vitro in this study. The results showed that the percentages of both CD19+CD24+CD38+TGF-ß1+ Bregs and CD19+CD24+CD38+IL-10+ Bregs to B cells were elevated by Sirolimus in PBMCs including B cells. Sirolimus significantly enhances the cytokine production of transforming growth factor-ß1 (TGF-ß1) and interleukin-10 (IL-10) in PBMCs with B cells, and the enhancement significantly decreased in PBMCs without B cells. The percentage of CD4+CD25+Foxp3+ Tregs to T cells was also elevated by Sirolimus in PBMCs including B cells. The elevation of Tregs percentage decreased in PBMCs without B cells and recovered when additional TGF-ß1 and IL-10 were added. The amplification of Tregs by Sirolimus was partially inhibited when either TGF-ß1 or IL-10 was neutralized, and it even disappeared when these two cytokines were both neutralized. These results indicate that Sirolimus can amplify Bregs and Tregs in PBMCs in vitro, and Bregs may be the why Sirolimus amplifies Tregs.