Current status of interleukin-2 therapy in cancer.

In vitro studies and animal experiments showed the existence of a physiological immune response against tumors. Interleukin-2 was the first immunological agent which demonstrated an anti-tumor effect by activating immune effectors. In vitro IL2 may generate Lymphokine Activated Killer (LAK) cells from peripheral blood lymphocytes or Tumor Infiltrating Lymphocytes (TIL) expanded from tumor. In melanoma and renal cell carcinoma, IL2 alone or associated with LAK cells or TIL, mediated clinical responses. However, their clinical efficacy was associated with some toxicity related to a capillary leak syndrome. This implies an improvement in the selection of patients and in the understanding of IL2 action. Future directions in immunotherapy included combination IL2 with other cytokines or monoclonal antibodies or chemotherapy. Lymphokine gene therapy is designed to introduce IL2 or other cytokine genes into tumor infiltrating lymphocytes or directly into tumors to reduce systemic toxicity and to achieve high local cytokine concentration. Animal models and the first human trials make this approach promising.     

The effects of interleukin-2 therapy on the viral reservoir in HIV+ patients.

The initial idea that potent antiretroviral therapies could eradicate HIV infection within a few years of treatment has been recently challenged by the demonstration that the viral reservoir persists in the peripheral blood and in the lymphoid tissue. For this reason, an alternative approach based on the use of interleukin-2 has been developed. This cytokine, in fact, may be able to activate infected cells, promoting viral integration and replication, making HIV susceptible to antiretroviral treatments; this fact may ultimately contribute to the eradication of the virus itself. The measurement of the viral reservoir appears therefore essential to monitor the effects of combination therapies. We summarize here the technical approaches that have been used to quantitatively assess the HIV reservoir. We also show that the prolonged use of IL-2 in association with antiretroviral drugs promotes a reduction of the viral reservoir, but is unable to eradicate HIV, even after two years of therapy. The available in vitro and in vivo data do not exclude the fact that IL-2 may have a future in the treatment of HIV infection, though new therapeutic approaches using different strategies are required to clarify this issue.     

Intravesical liposome-mediated interleukin-2 gene therapy in orthotopic murine bladder cancer model

Using a novel orthotopic MBT-2 murine bladder tumor model, we evaluated the feasibility of intravesical gene therapy utilizing a cationic liposome, DMRIE/DOPE. Superficial bladder tumors were consistently established by intravesical instillation of 5x10(5) MBT-2 cells in syngeneic C3H female mice. In situ gene transfer to bladder tumors was accomplished via intravesical instillation of plasmid DNA/DMRIE/DOPE lipoplex. Beta-Galactosidase (beta-gal) gene expression was preferentially evident in bladder tumors and was present for at least 7 days after a single 30 min in situ transfection. Murine interleukin-2 (IL-2) gene was used for treatment of 3-day-old pre-established bladder tumors. Forty percent of animals treated with IL-2 gene were completely free of tumors by 60 days following the initial tumor implantation, while all control groups treated with beta-gal gene died. Those animals initially cured of pre-established tumors were completely resistant to a subsequent tumor re-challenge and their splenocyte-derived cytotoxic T lymphocytes were shown to be specific to MBT-2 cells, indicating that immunological memory against MBT-2 tumors was elicited by the treatment. These results demonstrate the possibility of an effective clinical application of this in situ intravesical IL-2 gene delivery system to high-risk superficial bladder tumors, obviating a need for tumor procurement and ex vivo gene transfer.     

Adenovirus-mediated interleukin-2 gene therapy of nociception

The effect of adenovirus-mediated interleukin-2 (IL-2) gene on rat basal nociceptive response and chronic neuropathic pain was explored. The paw withdrawal latency induced by radiant heat was used to evaluate the antinociceptive effect of adenovirus type 5 (Ad5) and Ad5-IL-2. The results showed that intrathecal delivery of Ad5-IL-2 exhibited obvious antinociceptive effects on basal nociceptive response and chronic neuropathic pain, which were maintained for 3 and 4 weeks, respectively. This suggested that the antinociceptive effect of Ad5-IL-2 on chronic neuropathic pain was greater than its effect on basal nociceptive response. Human IL-2 mRNA was detected by in situ hybridization in the spinal pia mater and parenchyma of the lumbar, sacral, thoracic and cervical regions, and gray matter had higher level of IL-2 expression than white matter. These data demonstrated that the IL-2 gene was transfected into spinal cord regions relevant to pain modulation. The expressed IL-2 protein profile in spinal cord detected by enzyme-linked immunosorbent assay coincided almost exactly with its antinociceptive effect. This supported the hypothesis that the therapeutic effect of IL-2 gene was related to IL-2 protein expression. The study indicates that intrathecal delivery of adenovirus-mediated IL-2 gene has a relatively long antinociceptive effect.     

Adenovirus-mediated interleukin-12 gene therapy for prostate cancer

As a human prostate cancer gene therapy trial, we have performed Herpes Simplex Virus-thymidine kinase gene transduction and ganciclovir therapy in locally advanced prostate cancer after radiation therapy(USA) or endocrine therapy(Japan). Safety and clinical efficacy were confirmed. In our current studies, we demonstrated the efficacy and safety of gene therapy approaches using recombinant adenoviral vector that transduce interleukin-12 in an orthotopic mouse model of prostate cancer. These preclinical studies will soon be resulted in the initiation of clinical trials using this approach(USA: FDA approved, Japan: Under IRB review) in advanced prostate cancer patients with or without metastasis after endocrine therapy. In this chapter, preclinical data and clinical protocol are introduced.     

Late effects of cancer therapy on the central nervous system

The number of studies documenting the long-term morbidity of CNS treatment has increased dramatically in recent years. Cranial irradiation, in particular, has been associated with cognitive deficits and neuroanatomic pathology. Children who are treated at an early age and individuals who receive higher doses (2,400 cGy or greater) appear to be at greatest risk for these sequelae. Much more research on the pathogenesis of delayed injury following CNS treatment is needed. Although several compelling mechanisms have been proposed, little empirical evidence is available. This knowledge is essential to the identification of agents that may protect normal brain tissue from injury. Even less is known about the effects of age at time of treatment, type of treatment (radiation v chemotherapy), or dose of radiation in relation to delayed injury. Of utmost importance are studies that will establish the predictive relationship between brain injury and cognitive deficits. This would allow clinicians to predict patients who are at risk for cognitive impairment in order to institute appropriate preventive or remedial interventions.     

Late effects of childhood cancer therapy

One in every 900 young adults is a survivor of childhood cancer. Survivors may experience a wide variety of late effects stemming from the treatment they received. It is estimated that a significant portion of adult survivors of childhood cancer are not followed regularly in a center familiar with the late effects of their specific therapy. Therefore it is important for health care professionals in any setting to have an understanding of these possible late effects and encourage the survivor to seek appropriate follow-up. This article will provide a general overview of the potential late effects of childhood cancer therapy.     

The role of interleukin-2 in cancer immunotherapy

The arena of cellular immunotherapy is an emerging field of study. The field has strong foundations in numerous animal tumor models, which provide avenues of research for refinements in human immunotherapy. Experience from a number of research centers can be generalized: LAK cell trials in humans have demonstrated that PBMCs activated with IL-2 possess reproducible antitumor activity in vitro. Exogenous IL-2 administration to patients is required to maintain the biologic activity of the activated cells, although the requirement for LAK cells in tumors like melanoma is not clear-cut. Overall, the response rate to immunotherapy in humans is approximately 30%. The toxicity of immunotherapy, which is related to the dose of systemic IL-2, can be significantly reduced. The next generation of trials of cellular immunotherapy in humans will use TILs. Murine experiments indicate that this population of cells is more powerful and potentially more specific for tumor than LAK cells. Like LAK cells, TILs also require exogenous IL-2 for optimum performance in vivo. The difficulty in reproducible TIL growth can be overcome by the use of monoclonal antibody activation and IL-2. Improvements in TIL therapy are anticipated from the use of TILs combined with hybrid antibodies, the addition of other biologic response modifiers, and the implementation of genetically engineered cells and cell products.     

Preliminary report on 1592

A preliminary report on Glaxo-Wellcome's nucleoside analogue 1592 indicates that the drug is a powerful antiretroviral; however, cross-resistance has occurred in patients treated previously with AZT/3TC. The drug may be most effective in 3TC-naive patients. The dosage is 300 mg taken orally twice daily; however, optimal use of 1592 remains unknown. A telephone number is provided for further information about 1592.     

氯诺昔康超前镇痛对术后疼痛及病人血清白介素-2和白介素-6水平的影响

目的研究以氯诺昔康行超前镇痛对术后疼痛及病人围术期血清白细胞介素．2（IL-2）和自细胞介素-6（IL-6）水平的影响。方法60例ASAⅠ～Ⅱ级的子宫肌瘤患者，硬膜外麻醉下行经腹子宫全切术，随机分为三组：A组，不施行超前镇痛；B组，术前30min缓慢静注氯诺昔康16mg；C组，术毕时缓慢静注氯诺昔康16mg。三组术后均不行自控镇痛（PCIA），如果术后静息时患者主诉疼痛难忍，肌注曲马多镇痛（100ms／次）。分别于术前、术毕时、术后12h和术后24h抽取外周静脉血，采用放射免疫分析法测定患者血清中IL-2和IL-6的浓度，并于术后不同时间点观察视觉模拟评分（VAS）、术后应用镇痛药的比例和不良反应的发生率。结果B组的VAS评分在术后各时间点均显著低于A组，C组在术后4hVAS疼痛评分显著低于A组（P〈0．01）。A组术后24h内用镇痛药的患者占30％，显著高于B、C组（P〈0．05）。B组IL-2和IL-6的浓度波动均不明显；在A组和C组，术后IL-2的浓度显著降低而IL-6的浓度显著升高，与B组的差异具有显著性；C组IL-2和IL-6的浓度与A组比较差异无显著性。结论以氯诺昔康行超前镇痛能够减轻下腹部手术的术后疼痛，减少术后镇痛药的应用，能有效的抑制应激反应，减轻术后免疫抑制。     

Multicenter phase 2 study of interleukin-2 and 13-cis retinoic acid as maintenance therapy in advanced non-small-cell lung cancer

High serum levels of vascular endothelial growth factor (VEGF) are a poor prognostic factor for patients with advanced non-small-cell lung cancer (NSCLC). We have previously shown that low-dose interleukin (IL)-2 and 13-cis retinoic acid (RA) decreased VEGF and improved the immune function of patients with advanced tumors treated with chemotherapy. The primary end point of this study was to verify whether IL-2 and RA decreased serum VEGF in NSCLC patients showing a clinical benefit from chemotherapy. The secondary end point was the evaluation of clinical outcome. We treated 38 patients with advanced NSCLC who had a complete or partial response or disease stability to chemotherapy and had a median serum VEGF level of 508 ng/mL; as maintenance therapy, they received subcutaneous IL-2 (1.8 x 10(6) IU) and oral RA. Matched controls (n = 87) were selected from a large cohort of patients with a similar disease status, including clinical benefit from chemotherapy. The most common adverse events were mild cutaneous skin rash and fever. Serum VEGF decreased to a mean level of 152 ng/mL (P = 0.0002). A statistically significant improvement in immune function was observed (lymphocyte and natural killer cell numbers and CD4+/CD8+ ratio) with respect to baseline values and controls. An improvement in the clinical outcome was also observed compared with controls. These data show that the administration of low-dose subcutaneous IL-2 and oral RA to patients with advanced NSCLC showing a clinical benefit from chemotherapy is feasible with a low-toxicity profile, decreases VEGF, and seems to improve progression-free and overall survival.     

Intravesical interleukin-15 gene therapy in an orthotopic bladder cancer model

Interleukin-15 (IL-15) is known to stimulate the proliferation of CD8(+) T-cells and natural killer cells, and also to help to maintain memory CD8(+) T cells, suggesting that it may be of value in cytokine treatment of bladder cancer. In this experiment, we tested the efficiency of intravesical liposomal IL-15 gene delivery and its antitumor effect in a mouse orthotopic bladder cancer model. We established an orthotopic bladder cancer model by implanting 5×10(5) MBT-2 cells into female C3H/HeN mice through the urethra. The mice received repeated intravesical gene delivery injected with liposome-mediated plasmids (5?μg) transurethrally. On day 23, the bladder weights in the group receiving medium alone, the beta-galactosidase gene delivery control group, and the IL-15 gene therapy group were 196±36?mg, 201±35?mg, and 96±29?mg, respectively (p<0.05), demonstrating the antitumor effect of intravesical IL-15 gene therapy in this model. In the bladders treated with IL-15 gene plasmid instillation, histological analysis revealed that many inflammatory cells were induced around the tumors. Immunohistochemical analysis confirmed that there was predominant infiltration of CD8(+) T cells around the tumor nest. After the intravesical IL-15 gene therapy, the growth of rechallenged subcutaneous MBT-2 cells in surviving mice was inhibited again via tumor-specific cytotoxic T lymphocytes, although newly implanted FM3A cells in the same mice were not rejected. The present findings indicate that IL-15 gene therapy may be a promising new adjuvant therapy for bladder cancer.     

Differential effects of interleukin-15 and interleukin-2 on differentiation of bipotential T/natural killer progenitor cells

Bipotential T/natural killer (NK) progenitor cells are destined to differentiate mainly into T cell receptor (TCR) alpha beta and TCR gamma delta cells in a thymic microenvironment, whereas extrathymically they selectively develop into NK cells. The exact environmental conditions that are required for differentiation into these three leukocyte populations are largely unknown. In this report, we have investigated and compared the effect of interleukin (IL)-15 and IL-2 in this process. The IL-15 receptor is composed of the gamma and beta chains of the IL-2 receptor (IL-2R gamma and IL-2R beta) and of a specific alpha chain (IL-15R alpha). Here, it is shown that IL-15 mRNA is mainly expressed in thymic epithelial stromal cells, whereas IL-2 mRNA is exclusively expressed in thymocytes. IL-2R beta-expressing cells were present in the fetal thymus with a CD25-CD44+Fc gamma R+HSA- /low TCR- phenotype, which is characteristic of progenitor cells. These cells also expressed IL-15R alpha messenger RNA. Sorted IL-2R beta + TCR- cells differentiated into TCR alpha beta and TCR gamma delta cells after transfer to alymphoid thymic lobes, whereas culture of the same sorted cells in cell suspension in the presence of IL-15 resulted in the generation of functional NK cells. This shows that IL-2R beta +TCR- cells of the fetal thymus contain bipotential T/NK progenitors. Addition of low concentrations of IL-15 to fetal thymic organ culture (FTOC) resulted in an increase of all T cell subpopulations. The largest expansion occurred in the TCR gamma delta compartment. In contrast, low concentrations of IL-2 did not result in a higher total cell number and did not induce outgrowth of TCR gamma delta cells. High concentrations of IL-15 blocked TCR alpha beta development and shifted differentiation towards NK cells. Differentiation towards TCR gamma delta cells still proceeded. High concentrations of IL-2 similarly induced development into NK cells, but the cell number was fourfold lower than in IL-15 cultures. Importantly, blocking of IL-2R alpha in IL-2-treated FTOC resulted in a drastic increase in cell number, indicating that IL-2R alpha negatively regulates cell expansion. Collectively, these experiments provide direct evidence that IL-15 and IL-2 differentially affect the differentiation of bipotential T/NK progenitors.     

中西医结合疗法治疗脊髓损伤幻觉痛初探

用耳压疗法、气功、电动按摩器和经皮神经电刺激仪相结合的中西医结合疗法治疗脊髓损伤幻觉痛１８，莫克吉尔疼痛答卷等方面于治疗前后对幻觉痛进行评测。结果表明，治疗后病人疼痛的６项参数值有非常显著性降低）Ｐ＜０．００５），四项参数值有显著性降低（Ｐ＜０．００５０，一项无显著性差异。证实这种疗法是切实有效的，并初步探讨了其作用的可能机制。     

The effects of calcitriol therapy on serum interleukin-1, interleukin-6 and tumour necrosis factor-alpha concentrations in post-menopausal patients with osteoporosis

Seventy post-menopausal women with osteoporosis were randomized into two groups: 40 patients received calcitriol (0.5 microg/day) and calcium (1000 mg/day); and 30 control patients received calcium (1000 mg/day) alone. Thirty healthy women formed the healthy control group. Bone mineral density (BMD) and serum interleukin (IL)-1, IL-6 and tumour necrosis factor-alpha (TNF-alpha) concentrations were measured at baseline and after 6 months of treatment. Calcitriol treatment for 6 months significantly increased BMD and reduced serum IL-1 and TNF-alpha concentrations compared with no significant changes in patients treated with calcium alone. Both treatments increased serum calcium and decreased parathyroid hormone concentrations. The healthy control group had a significantly lower IL-6 concentration than the post-menopausal women with osteoporosis. We have shown that calcitriol was an effective treatment for osteoporosis. Significant reductions in serum IL-1 and TNF-alpha concentrations suggest that, in addition to increasing the absorption of calcium, calcitriol may directly affect bone metabolism via cytokines.