Symposia Report Immunoglobulin G for the Treatment of Chronic Pain:Report of an Expert Workshop

背景:慢性疼痛的治疗效果仍不理想.尽管现在治疗慢性疼痛的药物种类较多,但许多患者对疗效仍不满意或诉药物的副作用太大.越来越多的证据表明,免疫系统参与了伤害性和神经病理性慢性疼痛的病理过程.设计:在英国利物浦的专题会议上,专家们出示了免疫系统参与慢性疼痛的证据.近来的研究表明,静脉(IVIg)或皮下(SCIg)注射免疫调节药物——多价免疫球蛋白(IgG)可缓解外周神经病理性疼痛和其他疼痛性疾病.专题会议讨论了IVIg和SCIg治疗的适应证、效价比及其副作用.结果:IgG可缓解某些伤害性和神经病理性慢性疼痛,如糖尿病、干燥综合征、纤维肌痛症、复杂性区域疼痛综合征、小儿麻痹后遗症和继发于病理性自身抗体的疼痛.结论:IgG对某些慢性疼痛具有一定的治疗前景.IgG是一种相对安全的治疗方法,副作用少而轻,但价格较贵.今后有必要对IgG治疗顽固性疼痛进行随机对照研究和预测性临床试验.

AIM To study the effect of immune treatment on gastric cancer.METHODS Thirteen patients with advanced gastric cancer were given TIL adoptive immunotherapy in thisstudy. Histological findings showed that 13 patients had gastric adenocarcinoma. Patients received operationson their primary tumor, which could not be resected. Small tumor tissue and metastic lymph nodes were gotduring the operation for TIL preparation. Ten patients were treated as control group. During TILtreatment, the patients did not received any other treatment. Surgical speciments (metastatic lymph nodes )with pathological diagnosis were obtained from operating room. The lymph nodes were minced anddissociated in RPMI 1640 with 0.03% hyaluronidase type V (1500U/g), 0.05% collagenase type Ⅳ(200 U/g), and 0.008% deoxyribonuclease type I (100 U/g) (Sigma, USA) at 37℃ for 12 h. The cellmixture was then filtered through 4-layer gauge, washed twice in Hank's and separated on Ficoll-Hypague(Shanghai ist Chemical Reagent Factory) at 900×g for 20min. Finally, the cells were harvested andcounted. Cells suspension containing TIL and tumor cells were extensively washed and resuspended at a finalconcentration of 106 lymphocytes/mL in complete medium containing 15% human AB serum, 100 U/mLpenicillin and 100 μg/mL streptomycin in RPMI 1640 (Gibco). The final concentration of rIL-2 (Military Medical Institute, Nanjing ) was 500 U/mL. Cultured after 3 d-4 d, lymphoid cells were counted andculture was separated into more flasks when the concentration of lymmphoid cells reached or exceeded 2×106/mL until the total amount reached about 5×109/mL cells. Cytotoxic activities of TIL were tested by 6 h51Cr-release assay. Target cells (5×10s/mL) (human gastric adenocarcinoma) in 1 mL of culture mediumwere labelled with 100 μci of Na251CrO4(Beijing Atomic Energy Research Institute, Beijing), washed andadjusted to 105cells/mL. Labelled cells (E/T: 50: 1, 25: 1 and 12.5: 1) were seeded in round-bottom microtest plates (Corning, Japan) at 104cells/well. Isotope release was measured in a gamma counter (Packard,USA). The percentage of cytotoxicity was calculated according to the following formula: Cytotoxicity% = (Experimental- Spontaneous)/ (Maximum-Spontaneous)×100. Target cells without effective cells weremixed with 0.1 mL of culture media to obtain spontaneous release, and with 0.1 mL of 0.1 mol/L HCl toobtain maximum 51Cr release. TIL cells so induced were counted, washed twice, resupended in 100 mL 0.9%NaC1 solution and intravenouslly transferred. The number of total autologous TIL cells injected was morethan 5×109 cells for one patient and usually separated into 2 - 3 injection during the treatment, rIL-25000 U/d (Nanjing Military Medical institute, Nanjing) in 2 mL of 0.9% NaCl solution was intramuscularlyinjected starting from 5 days before TIL cells transfer to 5 days after transfer of TIL cells. All patients weregiven scheduled gastric roentgenograms. CT scanning, B type ultrasound, histological examination andimmune function were used to observe the changes before, during and after treatment. The curative effectswere judged by the standard of WHO. The methods of the assays of SIL-2R, NK cytotoxicity and CD4/CD8were carried out respectively according to the references.RESULTS The Nk cytotoxicity and CD4/CD8 were significantly increased (P<0.01) after 3- 6 monthstreatment. The soluble IL-2 receptor in sera of patients was significantly decreased (P<0.01) after 3- 6months treatment. There were no significantly differences in the test of CD4/CD8, the cytotoxicity of Nkcells and the soluble IL-2 receptor in serum between the group before treated by TIL and the control group(P >0.05). The NK cytotoxicity and CD4/CD8 in patients treated by TIL were significantly more than thosein the control group. On the contrary, the soluble IL-2 receptor in serum of patients treated by TIL wassignificantly less than those in serum of the control group. The patients of control group survived from 4.5months to 9 months (less than one year) after operation. However seven of the thirteen patients treated byTIL after operation survived over one year. Appetite was improved, sinew enhanced, weight increased andpain relieved in most of patients treated by TIL. On the contrary, the symptoms and signs of patients ofcontrol group were not improved. According to the standard of WHO, there were significantly differences ofPD(Disease Progress), MR (Minor remission), and PR(partial remission) between TIL group and controlgroup. The results indicated that tumor focus completely disapeared in 1(80%) of 13 patients, significantlydecreased in 4 (30%) of 13 patients and slightly decreased in 7 (53%) of 13 patients, suggesting that thetreatment of TIL in the patients with advanced cancer was effective. No side effects were found except fortransient fever in 2 patients.CONCLUSION TIL should be one of the fundamental therapies for the advanced gastric cancer, it canregulate the balance of immunity, relieve pain, improve symptoms and signs and prolong survival time.

Objective: To investigate the antitumor mechanisms of the streptococcal preparation OK-432. Methods: Using C57BL/6 mouse bearing B16 melanoma, we observed the antitumor activity of OK-432 and investigated the effect of OK-432 on multi-cytokine (IL-2, IL-4, IL-6, IL-10, IL-12, IFN-() production of mouse splenocyte both in vitro and in vivo. Results: As compared with control, OK-432 significantly inhibited B16 melanoma growth and lengthened mice survival time (P<0.05). In vitro OK-432 could stimulate splenocyte from tumor bearing mice to secrete IL-6, IL-12, IFN-( and IL-10 remarkably (P<0.01). In vivo OK-432 led to the increased production of IL-2, IL-12 and IFN-( but decreased production of IL-10 (P<0.05). When the splenocytes harvested from OK-432 treated mice were stimulated with OK-432 again in vitro, the production of IFN-( increased and IL-10 decreased significantly (P<0.05). Conclusion: OK-432 could boost multiple cytokines production, especially IL-12 which skewed T cells in a Th1 dominant state and enhanced the host antitumor activities.

Screening humanized antibodies from a human Fab phage display library is an effective and quick method to obtain beta-amyloid oligomers. Thus, the present study prepared amyloid-beta 42 oli-gomers and constructed a na?ve human Fab phage display library based on blood samples from six healthy people. After three rounds of biopanning in vitro, a human single-domain antibody that spe-cifical y recognized amyloid-beta 42 oligomers was identified. Western blot and enzyme-linked immunosorbent assay demonstrated this antibody bound specifical y to human amyloid-beta 42 te-tramer and nonamer, but not the monomer or high molecular weight oligomers. This study suc-cessful y constructed a human phage display library and screened a single-domain antibody that specifical y recognized amyloid-beta 42 oligomers.

Three-month-old Alzheimer’s disease model transgenic mice were immunized with Aβ1-42, Plp-Adenovirus [Ad]-X-CMV-(Aβ3-10)10-CpG [AdCpG-(Aβ3-10)10] or AdCpG virus lfuid via na-sal mucosal inhalation, respectively. ELISA analysis of serum showed Aβ42 antibody titers were signiifcantly increased in mice immunized with Aβ1-42 and AdCpG-(Aβ3-10)10. Concanavalin A and AdCpG-(Aβ3-10)10 stimulation signiifcantly increased the number of proliferating spleen cells cultured from AdCpG(Aβ3-10)10 and Aβ42 groups compared with the control group. In the AdCp-G(Aβ3-10)10 group, levels of interleukin (IL)-4 and IL-10 were increased, while those of IL-2 and interferon-γwere decreased. In the Aβ42 group, levels of IL-4, IL-10, IL-2 and interferon-γwere all increased. Experimental ifndings indicate that AdCpG-(Aβ3-10)10 vaccine can produce strong T helper 2 (hT2) humoral immune responses in addition to the production of Aβ42 antibody. hTe cellular immunologic response was weak and avoided Aβ1-42-mediated cytotoxicity.

AbstrAct Immunotherapy has become a key strategy for cancer treatment, and two immune checkpoints, namely, programmed cell death 1 (PD-1) and its ligand (PD-L1), have recently emerged as important targets. hTe interaction blockade of PD-1 and PD-L1 demonstrated promising activity and antitumor effcacy in early phase clinical trials for advanced solid tumors such as non-small cell lung cancer (NSCLC). Many cell types in multiple tissues express PD-L1 as well as several tumor types, thereby suggesting that the ligand may play important roles in inhibiting immune responses throughout the body. hTerefore, PD-L1 is a critical immunomodulating component within the lung microenvironment, but the correlation between PD-L1 expression and prognosis is controversial. More evidence is required to support the use of PD-L1 as a potential predictive biomarker. Clinical trials have measured PD-L1 in tumor tissues by immunohistochemistry (IHC) with different antibodies, but the assessment of PD-L1 is not yet standardized. Some commercial antibodies lack speciifcity and their reproducibility has not been fully evaluated. Further studies are required to clarify the optimal IHC assay as well as to predict and monitor the immune responses of the PD-1/PD-L1 pathway.

AbstrAct Survival rates for metastatic lung cancer, including non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), are poor with 5-year survivals of less than 5%. The immune system has an intricate and complex relationship with tumorigenesis;a groundswell of research on the immune system is leading to greater understanding of how cancer progresses and presenting new ways to halt disease progress. Due to the extraordinary power of the immune system—with its capacity for memory, exquisite speciifcity and central and universal role in human biology—immunotherapy has the potential to achieve complete, long-lasting remissions and cures, with few side effects for any cancer patient, regardless of cancer type. As a result, a range of cancer therapies are under development that work by turning our own immune cells against tumors. However deeper understanding of the complexity of immunomodulation by tumors is key to the development of effective immunotherapies, especially in lung cancer.

The last ten years have seen remarkable progress in cancer research. However, despite significant breakthroughs in the understanding, prevention, and treatment of cancer, the disease continues to affect millions of people worldwide. Cancer’s complexity compounded with ifnancial, policy and regulatory roadblocks has slowed the rate of progress being made against cancer. In this paper, we review a few of the most recent breakthroughs that are fueling medical advances and bringing new hope for patients affected by this devastating disease. We also address the challenges facing us and the opportunities to accelerate future progress against cancer. The efforts of the American Association for Cancer Research (AACR) to address the cancer burden already extend beyond the borders of the United States of America. hTe AACR is committed to increasing its efforts to stem the tide of cancer worldwide by promoting innovative programs, strategies, and initiatives for cancer researchers and all those engaged in cancer-related biomedical sciences around the world.

预防肿瘤术后复发和转移的新疗法 :热休克蛋白 /肽复合物疫苗的研究与应用

Deepened understanding of the mechanism involved in the activation of T cells and improved molecular biology techniques have brought a promising strategy to active a patient's immune system to prevent tumor recurrence and metastasis. Heat shock proteins (HSPs) are chaperones of peptides,It can elicit array of immune responses,such as:present tumor antigens to T cells, stimulate antigen presenting cells to secrete cytokines,mediate maturation of dentritic cells,active NK cells and/T cells.Extract HSP/peptides complex from tumor cells can be used as a polyvalent vaccine for treatment of cancers,The elicited antigen specific immune response is restricted to the tumor from which the HSPs are purified.HSP/peptides complex vaccine has been started in third clinical trials.The rationale,feasibility,advantages and safety of this new approach were discussed.

In recent years, immunotherapy has emerged as a viable and attractive strategy for the treatment of prostate cancer. While there are multiple ways to target the immune system, therapeutic cancer vaccines and immune checkpoint inhibitors have been most successful in late-stage clinical trials. The landmark Food and Drug Administration approval of sipuleucel-T for asymptomatic or minimally symptomatic metastatic prostate cancer set the stage for ongoing phase III trials with the cancer vaccine PSA-TRICOM and the immune checkpoint inhibitor ipilimumab. A common feature of these immune-based therapies is the appearance of improved overall survival without short-term changes in disease progression. This class effect appears to be due to modulation of tumor growth rate kinetics, in which the activated immune system exerts constant immunologic pressure that slows net tumor growth. Emerging data suggest that the ideal population for clinical trials of cancer vaccines is patients with lower tumor volume and less aggressive disease. Combination strategies that combine immunotherapy with standard therapies have been shown to augment both immune response and clinical beneift.

A REVIEW ON CANCER IMMUNOTHERAPY VIA DENDRITIC CELLS

Cancer is considered as a devastating disease and a leading cause of deaths around the world.According to WHO,1.4 million cancer cases are seen in 2012.Cancer is usually asymptomatic at its initial stages,but it shows very pathetic signs and symptoms at advanced stages.There are various ways to diagnose cancer as taking images of body parts,some blood tests and histological study of body cells.The advanced malagnancies are not sensitive to therapies including surgery,chemotherapy and radiotherapy,and also accompanied by adverse effects,and also do not give the promising results.These adverse effects are usually toxic.Recent studies in immune therapy has shown non-toxdc and effective results given by dendritic cells(DCs).DCs are the potent antigen presenting cells and bave ability to stimulate strong immune response in the body by activating T cells.DCs can be prepared in vitro,can be combined with cytokines,peptides and different molecules to develop cancer vaccines.These cancer vaccines offer low toxicity and have ability to induce anticancer immune response.Under the banner of immunotherapy,the aim of these vaccines is to increase body's immune response against cancers.This article reviews the basic biology of dendritic cells,and future direction of DCs based cancer vaccines.

In vitro amplified human leukocyte antigen (HLA)-haploidentical donor immune cellinfusion (HDICI) is not commonly used in children. Therefore, our study sought to evaluate its safety for treating childhood malignancies. Between September 2011 and September 2012, 12 patients with childhood malignancies underwent HDICI in Sun Yat-sen University Cancer Center. The median patient age was 5.1 years (range, 1.7-8.4 years). Of the 12 patients, 9 had high-risk neuroblastoma (NB) [7 showed complete response (CR), 1 showed partial response (PR), and 1 had progressive disease (PD) after multi-modal therapies], and 3 had Epstein-Barr virus (EBV)-positive lymphoproliferative disease (EBV-LPD). The 12 patients underwent a total of 92 HDICIs at a mean dose of 1.6×108 immune cells/kg body weight: 71 infusions with natural kil er (NK) cells, 8 with cytokine-induced kil er (CIK) cells, and 13 with cascade primed immune cells (CAPRIs); 83 infusions with immune cells from the mothers, whereas 9 with cells from the fathers. Twenty cases (21.7%) of fever, including 6 cases (6.5%) accompanied with chil s and 1 (1.1%) with febrile convulsion, occurred during infusions and were al eviated after symptomatic treatments. Five cases (5.4%) of mild emotion changes were reported. No other adverse events occurred during and after the completion of HDIDIs. Neither acute nor chronic graft versus host disease (GVHD) was observed following HDICIs. After a median of 5.0 months (range, 1.0-11.5 months) of follow-up, the 2 NB patients with PR and PD developed PD during HDICIs. Of the other 7 NB patients in CR, 2 relapsed in the sixth month of HDICIs, and 5 maintained CR with disease-free survival (DFS) ranging from 4.5 to 11.5 months (median, 7.2 months). One EBV-LPD patient achieved PR, whereas 2 had stable disease (SD). Our results show that HDICI is a safe immunotherapy for childhood malignancies, thus warranting further studies.

SA-IL-2锚定修饰治疗表浅膀胱癌的实验研究

Background and Objective: Intravesical administration of Bacillus Calmette-Guerin (BCG) after transurethral resection is by far the most effective local therapy for superficial bladder cancer, the fifth most common cancer in the world. However, approximately one-third of patients fail to respond and most patients eventually relapse. In addition, there are pronounced side effects of BCG therapy, such as BCG sepsis and a high frequency of BCG-induced cystitis. This study established a novel immunotherapy through immobilization of streptavidin-tagged human IL-2 (SA-hlL-2) on the biotinylated mucosal surface of bladder wall. Methods: A mouse orthotopic model of MB49 bladder cancer was established by perfusing MB49 cells into mouse bladders. The SA-hlL-2 fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall. Treatment began on day 1 after MB49 implantation, once every 3 days for 6 times. Immunohistochemical assay was performed to assess the persistence of SA-hlL-2 immobilized on the biotinylated mucosal surface of the bladder wall. The mice were monitored for tumor growth and survival. On day 60 after MB49 implantation, the SA-hlL-2-cured mice, which were found to have no hematuria or palpable tumors, were challenged with wild-type MB49 cells implanted into the pretreated bladder and monitored for survival. Results: SA-hlL-2 could be immobilized efficiently and durably on the bladder mucosal surface as long as 7 days. On day 60 after MB49 implantation, 9 out of 20 SA-hlL-2-treated mice survived, but all mice in PBS control group died. More importantly, 5 out of 9 tumor-free mice in the SA-hlL-2 group were protected against a second intravesical wild-type MB49 tumor challenge. Conclusions: SA-hlL-2 fusion protein could significantly inhibit tumor growth and extend the survival time in the orthotopic model of MB49 bladder cancer.

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor in adults. Standard therapeutic approaches provide modest improvement in the progression-free and overall survival, necessitating the investigation of novel therapies. We review the standard treatment options for GBM and evaluate the results obtained in clinical trials for promising novel approaches, including the inhibition of angiogenesis, targeted approaches against molecular pathways, immunotherapies, and local treatment with low voltage electric fields.