Suppression of hepatocellular carcinoma by transplantation of ex-vivo immune-modulated NKT lymphocytes
Suppression of hepatocellular carcinoma by transplantation of ex-vivo immune-modulated NKT lymphocytes
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DOI：
10.1002/ijc.20889
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发表时间：
2005-06-20
影响因子：
6.4
通讯作者：
Ilan, Y
中科院分区：
文献类型：
--
作者：
Margalit, M;Shibolet, O;Ilan, Y
NKT细胞是T淋巴细胞的调节亚群，具有免疫调节作用，在抗肿瘤免疫中发挥重要作用。最近已经证明了NKT细胞的“离体教育”的可行性。评价体外免疫调节的NKT淋巴细胞在小鼠肝癌模型中的抗肿瘤作用。对无胸腺Balb/C小鼠进行亚致死剂量照射并移植人Hep 3B HCC。将从免疫活性BalB/C小鼠制备的NKT细胞用HCC衍生抗原（A组）、Hep 3 B细胞（B组）或BSA（C组）离体脉冲，并过继转移到携带HCC的小鼠中（每只小鼠1 × 106个NKT细胞）。D组小鼠不进行NKT细胞移植。E组小鼠移植1 × 106个来自HBV免疫供体的NKT细胞。跟踪小鼠的肿瘤大小和重量。为了确定抗肿瘤作用的机制，通过FACS分析脾内淋巴细胞群体的NKT、CD 4+和CD 8+淋巴细胞亚群;通过Western印迹评估脾细胞中STAT 1、4和6的表达，并通过ELISA测量血清细胞因子水平。连续转移用HCC衍生抗原脉冲的NKT细胞（A组）和来自免疫供体的NKT细胞（E组）导致肿瘤在4周内完全消失，并且减轻体重减轻（A组和E组分别为6.5%和7%）。相比之下，B、C和D组中的小鼠出现大的坏死性肿瘤和严重的体重减轻（B、C和D组中的体重减轻分别为21%、17%和23%）。A、E组NKT/CD_4、CD_8/CD_4比值明显升高（NKT/CD_4比值：A、D组分别为12.3、17.6，B、C、D组分别为6.4、4.8、5.6;对于CD 8/CD 4比率，A组和E组分别为41和19.8，而B、C和D组分别为6.5、11.8和3.2）。转录因子STAT 4在A组中表达明显，而在B-D组中无表达。A、E组血清IFN γ、IL 12、IL 4水平升高。NKT淋巴细胞暴露于负载于树突状细胞上的HCC衍生抗原和来自免疫供体的NKT细胞的体外连续转移导致小鼠中HCC的抑制。NKT介导的抗肿瘤活性与增加的NKT和CD 8 + T淋巴细胞数量、增加的STAT 4（IL-12活性的标志物）表达以及升高的促炎细胞因子IFN γ和IL 12以及IL 4的血清水平相关。NKT淋巴细胞的离体调节有望成为HCC免疫治疗的新模式。(c)2005 Wiley-Liss，Inc.
NKT cells are a regulatory subset of T lymphocytes with immune modulatory effects and an important role in anti-tumor immunity. The feasibility of "ex-vivo education" of NKT cells has recently been demonstrated. To evaluate the anti-tumor effect of ex-vivo immune-modulated NKT lymphocytes in a murine model of hepatocellular carcinoma. Athymic Balb/C mice were sublethally irradiated and transplanted with human Hep3B HCC. NKT cells prepared from immunocompetent Balb/C mice were pulsed ex vivo with HCC-derived antigens (Group A), Hep3B cells (group B) or BSA (group C, and adoptively transferred into HCC harboring mice (I X 106 NKT cells per mouse). Group D mice did not undergo NKT cell transplantation. Group E mice were transplanted with I X 106 NKT cells from HBV-immunized donors. Mice were followed for tumor size and weight. To determine the mechanism of the anti-tumor effect, intrasplenic lymphocyte populations were analyzed by FACS for NKT, CD4+ and CD8+ lymphocyte subpopulations; STAT 1, 4 and 6 expression in splenocytes was assessed by Western blot, and serum cytokine levels were measured by ELISA. Adoptive transfer of NKT cells pulsed with HCC-derived antigens (group A) and NKT cells from immunized donors (group E) resulted in complete disappearance of tumors within 4 weeks and attenuated weight loss (6.5% and 7% in groups A and E, respectively). In contrast, mice in groups B, C, and D developed large, necrotic tumors and severe weight loss (21%, 17% and 23% weight loss in groups B, C, and D, respectively). NKT/CD4 and CD8/CD4 ratios were significantly increased in groups A and E (12.3 and 17.6 in groups A and D, respectively, compared to 6.4, 4.8 and 5.6 in groups B, C and D, respectively, for the NKT/CD4 ratio; 41 and 19.8 in groups A and E, respectively, compared to 6.5, 11.8 and 3.2 in groups B, C, and D, respectively, for the CD8/CD4 ratio). Expression of the transcription factor STAT4 was evident in group A, but not in groups B-D. Serum IFN gamma, IL12 and IL4 levels were increased in groups A and E. Adoptive transfer of NKT lymphocytes exposed ex vivo by HCC-derived antigens loaded on dendritic cells and NKT cells from immunized donors led to suppression of HCC in mice. NKT-mediated anti-tumor activity was associated increased NKT and CD8+ T lymphocyte numbers, increased expression of STAT4, a marker for IL-12 activity and elevated serum levels of the proinflammatory cytokines IFN gamma and IL12, and of IL4. Ex-vivo modulation of NKT lymphocytes holds promise as a novel mode of immune therapy for HCC. (c) 2005 Wiley-Liss, Inc.