Association of Vav1 with Activity of Infiltrating T Cells and Local Expression of IDO in Tumor
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摘要:
目的 通过对Vav1与肿瘤浸润T淋巴细胞（tumor infiltrating T lymphocytes，TIL-T）活性关系的研究，提出T细胞失能的可能分子机制；初步探讨TIL-T中Vav1的表达情况及其与肿瘤局部微环境中吲哚胺2，3双加氧酶（indoleamine-2，3-dioxygen? ase，IDO）表达的相关性。 方法 收集天津医科大学附属肿瘤医院肺外科手术切除的新鲜肺癌标本、癌旁正常肺组织及蜡块40例，通过实时定量RT-PCR检测TIL-T中Vav1 mRNA表达变化；免疫印迹及免疫沉淀技术检测Vav1蛋白表达及磷酸化活性。BrdU法检测T细胞增殖活性。此外，运用Real time-PCR法检测肺癌组织中IDO mRNA表达水平，免疫印迹及免疫组化检测IDO蛋白表达情况。 结果 部分肺癌局部浸润的T细胞处于功能抑制状态，这种抑制状态可能与细胞内重要的信号传导蛋白Vav1的表达量及活性相关。IDO表达阳性组肺癌标本局部TIL-T中Vav1 mRNA水平及Vav1蛋白的表达和磷酸化水平明显低于IDO表达阴性组（P < 0.05）。 结论 Vav1的表达和活化在TIL-T功能中具有重要的作用。肿瘤局部微环境中的IDO蛋白可能是影响TIL-T中Vav1表达和活化的重要因素之一。IDO可能通过抑制Vav1的表达及磷酸化活化过程，使TIL-T的主动免疫受损，从而降低宿主的抗肿瘤免疫效应。 -
关键词:
- Vav1 /
- 吲哚胺2，3双加氧酶 /
- 肿瘤浸润T淋巴细胞 /
- 磷酸化 /
- 增殖
Abstract:Objective This study explored the relationship between Vav1 and tumor-infiltrating T lymphocytes (TIL - T), which provide a molecular mechanism inducing a state of T cell anergy. The expression of Vav1 in TIL-T and indoleamine 2, 3-dioxygenase (IDO) from the tumor microenvironment was invetigated in order to present a possible molecular mechanism for tumor-induced T cell immune tolerance. Methods A total of 40 lung cancer patients who had undergone surgery in the Tianjin Medical University Cancer Institute and Hospital were involved. The expression levels of Vav1 mRNA in TIL-T were detected by real-time PCR, while the expression and activation of Vav1 were determined using Western blot and immunoprecipitation. T cell proliferation was detected using the BrdU method. The levels of IDO expression in lung cancer and corresponding normal lung tissues were determined using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. The SPSS17.0 software was used for statistical analysis. Results TIL-Ts are a part of cancer tissues and are in a state of functional suppression. This suppressive condition may involve the key signal transducer Vav1. Both mRNA and protein levels of Vav1 in T cells were significantly decreased in lung cancer tissues with IDO-positive expression compared with those with IDO-negative expression. In addition, the levels of Vav1 phosphorylation was decreased (P < 0.05). Conclusion The expression and activation of Vav1 play critical roles in the function of TIL-T. IDO, secreted by the tumor cells themselves or antigen-presenting cells, has an important impact on the expression and activation of Vav1 in TIL-T. IDO may suppress the expression and phosphorylation of Vav1, leading to inhibition of the T cell active immune response, which may consequently reduce the anti-tumor defenses of the host. -
Key words:
- Vav1 /
- Indoleamine 2, 3-dioxygenase /
- Tumor infiltrating T lymphocytes /
- Phosphorylation /
- Proliferation
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图 1 Vav1高表达组及低表达组TIL-T的增殖情况
Figure 1. Proliferation of TIL-T in the group with high and low expression of Vav1
图 2 IDO在肺癌组织中的表达（SP× 400）
Figure 2. Expression of indoleamine 2, 3-dioxygenase in lung cancer tissues
图 3 肺癌组织、癌旁正常肺组织中IDO基因及蛋白的表达
A：IDO阳性表达的肺癌组织及其相应癌旁正常肺组织；M：Marker；1、2：肺癌组织中β-actin和IDO；3、4：正常肺组织中β-actin和IDO；B：IDO阴性表达的肺癌组织及其相应癌旁正常肺组织；M：Marker；1、2：肺癌组织中β-actin和IDO；3、4：正常肺组织中β-actin和IDO；C：1、2：肺癌组织；3：癌旁正常肺组织
Figure 3. Expression of IDO mRNA and protein in lung cancer and normal lung tissues
图 4 肺癌组织TIL-T中Vav1 mRNA、蛋白表达及磷酸化情况
A：IDO阳性组及阴性组中Vav1 mRNA的表达；B：IDO阳性组及阴性组中Vav1蛋白及磷酸化情况
Figure 4. Expression of Vav1 mRNA, protein, and phosphorylation in TIL-T derived from lung cancer tissues
表 1 IDO在不同组肺癌组织中的表达
例 Table 1. Expression of indoleamine 2, 3-dioxygenase in different groups of lung cancer tissues
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[1] Katzav S. Vav1: an oncogene that regulates specific transcriptional activation of T cells[J]. Blood, 2004, 103(7): 2443-2451. doi: 10.1182/blood-2003-08-2834 [2] Koneru M, Schaer D, Monu N, et al. Defective proximal TCR signaling inhibits CD8+tumor-infiltrating lymphocyte lytic function[J]. J Immunol, 2005, 174(4): 1830-1840. doi: 10.4049/jimmunol.174.4.1830 [3] Miller CH, Graham L, Bear HD. Phenotype, functions and fate of adoptively transferred tumor draining lymphocytes activated ex vivo in mice with an aggressive weakly immunogenic mammary carcinoma[J]. BMC Immunol, 2010, 11(4): 54. http://pubmedcentralcanada.ca/pmcc/articles/PMC2998465/ [4] Wei J, Barr J, Kong LY, et al. Glioblastoma cancer-initiating cells inhibit T-cell proliferation and effector responses by the signal transducers and activators of transcription 3 pathway[J]. Mol Cancer Ther, 2010, 9(1): 67-78. doi: 10.1158/1535-7163.MCT-09-0734 [5] Sucher R, Kurz K, Weiss G, et al. IDO-Mediated Tryptophan Degradation in the Pathogenesis of Malignant Tumor Disease[J]. Int J Tryptophan Res, 2010, 3: 113-120. http://pubmedcentralcanada.ca/pmcc/articles/PMC3195236/ [6] Whiteside TL. Immune responses to malignancies[J]. J Allergy Clin Immunol, 2010, 125(2): S272-S283. doi: 10.1016/j.jaci.2009.09.045 [7] Corzo CA, Condamine T, Lu L, et al. HIF-1α regulates function and differentiation of myeloid-derived suppressor cells in the tumor microenvironment[J]. J Exp Med, 2010, 207(11): 2439-2453. doi: 10.1084/jem.20100587 [8] Kudoh S, Wang Q, Hidalgo OF, et al. Responses to T cell receptor/ CD3 and interleukin-2 receptor stimulation are altered in T cells from B cell non-Hodgkin's lymphomas[J]. Cancer Immunol Immunother, 1995, 41(3): 175-184. http://www.ncbi.nlm.nih.gov/pubmed/7553687 [9] Turner M, Billadeau DD. Vav proteins as signal integrators for multisubunit immune-recognition receptors[J]. Nat Rev Immunol, 2002, 2 (7): 476-486. doi: 10.1038/nri840 [10] Ruiz S, Santos E, Bustelo XR. The use of knockout mice reveals a synergistic role of the Vav1 and Rasgrf2 gene deficiencies in lymphomagenesis and metastasis[J]. PLOS One, 2009, 4(12): e8229. doi: 10.1371/journal.pone.0008229 [11] Thornton MV, Kudo D, Rayman P, et al. Degradation of NF-kappa B in T cells by gangliosides expressed on renal cell carcinomas [J]. J Immunol, 2004, 172(6): 3480-3490. doi: 10.4049/jimmunol.172.6.3480 [12] Yu J, Sun J, Wang SE, et al. Upregulated expression of indoleamine 2, 3-dioxygenase in primary breast cancer correlates with increase of infiltrated regulatory T cells in situ and lymph node metastasis [J]. Clin Dev Immunol, 2011, 2011: 469135. http://pubmedcentralcanada.ca/pmcc/articles/PMC3202140/ [13] Inaba T, Ino K, Kajiyama H, et al. Indoleamine 2, 3-dioxygenase expression predicts impaired survival of invasive cervical cancer patients treated with radical hysterectomy[J]. Gynecol Oncol, 2010, 117(3): 423-428. doi: 10.1016/j.ygyno.2010.02.028 [14] Nakamura T, Shima T, Saeki A, et al. Expression of indoleamine 2, 3-dioxygenase and the recruitment of Foxp3-expressing regulatory T cells in the development and progression of uterine cervical cancer[J]. Cancer Sci, 2007, 98(6): 874-881. doi: 10.1111/j.1349-7006.2007.00470.x [15] Runmei L, Feng W, Jinpu Y, et al. IDO inhibits T-cell function through suppressing Vav1 expression and activation[J]. Cancer Biol Ther, 2009, 8(14): 1402-1408. doi: 10.4161/cbt.8.14.8882 [16] Lee GK, Park HJ, Macleod M, et al. Tryptophan deprivation sensitizes activated T cells to apoptosis prior to cell division[J]. Immunology, 2002, 107(4): 452-460. doi: 10.1046/j.1365-2567.2002.01526.x [17] Terness P, Chuang JJ, Opelz G. The immunoregulatory role of IDO-producing human dendritic cells revisited[J]. Trends Immunol, 2006, 27(2): 68-73. doi: 10.1016/j.it.2005.12.006 [18] Suzuki J, Ricordi C, Chen Z. Immune tolerance induction by integrating innate and adaptive immune regulators[J]. Cell Transplant, 2010, 19(3): 253-268. doi: 10.3727/096368909X480314 -
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