脓毒症免疫抑制监测与免疫治疗的研究进展

脓毒症是宿主感染病原微生物后导致免疫反应失调而出现危及生命的器官功能障碍综合征,是重症监护病房常见的致死性疾病之一。近年来,脓毒症患者的免疫功能监测与免疫治疗成为研究的热点,但尚未形成统一共识。本文就脓毒症免疫抑制监测方法与治疗的研究进展进行综述。

参龟培元颗粒对实验性自身免疫性重症肌无力的治疗作用

目的探讨参龟培元颗粒对重症肌无力(myastheniagravis,MG)的治疗作用。方法采用合成电鳗乙酰胆碱受体α亚单位125-147段多肽介导的实验性自身免疫性重症肌无力(experimentalautoimmunemyastheniagravis,EAMG)大鼠模型,研究参龟培元颗粒对造模大鼠临床症状、肌电与收缩力、血清AchR-Ab滴度及细胞因子TNF-α、IFN-γ含量的影响。结果参龟培元颗粒(16.0,8.0g/kg)可明显改善大鼠的肌无力症状,降低血清AchR-Ab滴度及TNF-α、IFN-γ含量,且对低频刺激引起的造模大鼠肌电衰减和肌肉收缩力的衰减具有显著的改善作用。结论参龟培元颗粒对大鼠EAMG有一定的治疗作用。

顺铂加免疫核糖核酸序贯法治疗恶性胸水的临床观察

目的:观察顺铂加免疫核糖核酸对20例恶性胸水的临床疗效及毒副作用。方法:采对肺癌晚期伴发恶性胸水的患者分别采用定期常规胸腔穿刺术(A组n=7)及CVP导管引流(B组n=13)尽量排尽胸水后,分3次注入顺铂加免疫核糖核酸。结果A组有效率为25.5%,B组有效率分别为78.5%;B组的毒副作用主要是一过性胃肠道反应(20.0%)和WBC下降(8.0%)。结论:胸腔内置CVP管后顺铂加免疫核糖核酸序贯治疗能有效地控制癌性胸水,毒副反应小,可有效控制恶性胸水,提高晚期肺癌患者的生存质量。

高免疫原性胶质瘤细胞疫苗体外抗瘤时靶细胞超微结构变化

目的观察高免疫原性人多形性胶质母细胞瘤(GBM)U251细胞疫苗体外诱发产生效应细胞杀伤靶细胞的超微结构变化。方法以500U/mL干扰素-γ(IFN-γ)诱导48h+热43℃休克2h诱导膜型热休克蛋白70(HSP70)及主要组织相容性抗原复合体I类分子(MHC-I)双高表达,经丝裂霉素(MMC)灭活制成高免疫原性细胞疫苗。体外刺激健康捐献者外周血单个核细胞(PBMCs)作为效应细胞(MHC-HSP-CTL),进行肿瘤特异性杀伤试验;MTT比色法检测其杀伤活性;透射电镜观察靶细胞受攻击后情况。结果MHC-HSP-CTL对野生型U251细胞特异性的杀瘤活性明显高于HSP70分子单高表达的细胞疫苗刺激组(HSP-CTL)、MHC-I类分子单高表达的细胞疫苗刺激组(MHC-CTL)以及野生型对照组(W-CTL)(P<0.05),透射电镜显示靶细胞受到攻击后出现不同程度凋亡样坏死和胀亡样坏死,胀亡样坏死细胞在高免疫原性细胞疫苗刺激的效应细胞组较多。结论介导细胞胀亡是高免疫原性即膜型HSP70和MHC-I类分子双高表达疫苗的U251细胞疫苗一种重要主动特异性抗瘤机制。

南沙参多糖对小鼠免疫器官辐射损伤的防护

口服南沙参多糖1g／kg、0．5g／kg一周后能明显对抗60Co－γ线所致小鼠外周血白细胞总数降低和胸腺、脾脏重量减轻；使照射小鼠外周血酸性α-萘酚醋酸酯酶染色法（ANAE）显示淋巴细胞（ANAE－）比率升高，TH／TS比值升高，腹腔巨噬细胞吞噬功能明显增强；显著减轻小鼠胸腺、脾脏病理损伤，使修复加速，从而使6．0Gy照射小鼠30d存活率提高15％。

甲下黑色素瘤的治疗

目的探讨治疗甲下黑色素瘤的治疗方法。方法回顾性分析了自2002年以来,采用跨跖趾(掌指)关节截趾(指)术加免疫治疗的6例甲下黑色素瘤患者的临床资料与随访结果。结果所有患者随访4年,1例死亡,5例存活。结论采用跨跖趾(掌指)关节截趾(指)术加免疫治疗对治疗甲下黑色素瘤有较好的疗效,但本组病例数较少,术后观察时间较短,远期效果尚有待于进一步观察。

临床肿瘤热疗的几点思考

肿瘤热疗已经从热对药物增敏,热对放射增敏的研究走出。近年来全身热疗引起机体免疫的研究逐渐深入,热休克蛋白-肿瘤抗原肽疫苗的开发和初步应用,对临床热疗的方案设计将会有很大的改变。作者对肿瘤热疗特别是全身热疗的发展,提出个人见解。

Clinical observation of 125 I labeled anti alpha fetoprotein antibody radioimmunotherapy in hepatocellular carcinoma *

AIM To observe the therapeutic effects and toxic side reactions of 125 I labeled hourse anti human AFP polyclonal antibodies in immuno targeting therapy against hepatocellular carcinoma (HCC).

Immunotherapy of EGFR-positive Tumor Based on Recombinant EGFR Phage Vaccine

Objective： To construct the recombined phage which was inserted with the gene of extracellular domain of chicken EGFR as a new type vaccine and to evaluate the efficiency of the phage vaccine against the EGFR positive tumor. Methods： The T7 phage display system was used to display 5 fragments of the extracellular domain of chicken EGFR. The EGFR was expressed as a fused protein on the surface of the T7 phage 10B capsid protein. The EGFR expression of the phage vaccine was verified by the Western blot analysis. The anti-EGFR antibody was detected by ELISA. The splenic lymphocytes of the immunized mice were separated and used to determine the cellular immunotoxic effect against A431 cells. The phage vaccines were injected into the C57 mice 4 times before lewis lung cancer cells were inoculated. The tumor volume was recorded to evaluate the anti-tumor effect of each vaccine. Results： Five phage vaccines inserted with the chicken EGFR gene were constructed successfully. Western blot assay showed that the extracellular domain of chicken EGFR proteins was displayed on the surface of the phage. The specific antibody was induced in the immunized mice compared with the control group. The splenic lymphocytes of the immunized mice were shown to be immunotoxic against A431 cells. The killing rates of the experimental groups were higher than in the control group （P〈0.001, by t test）. The highest killing rate was （45.74±7.21）%. The tumor growth was inhibited in the experimental groups as compared with the control group （P〈0.05 in C1, C2, C3 and C4 groups, P〉0.05 in C5 group）. Conclusion： The phage vaccine could induce both the protective and therapeutic antitumor immunity against EGFR-positive tumor.

Fixed-Tumor Vaccine: A Practical Formulation with Cytokine-Microspheres for Protective and Therapeutic Antitumor Immunity

Objective: To study the protective and therapeutic antitumor immunity against hepatocellular carcinoma (HCC) with the fixed-tumor vaccine.Methods: A tumor vaccine consisting of fixed tumor cells or fixed tumor fragments combined with sustained-releasers of cytokines and a non-toxic adjuvant was developed. C57BL/6J mice were immunized intra-dermally with the vaccine on day 0 and 7, followed by intrahepatic challenge with live Hepa 1–6 cells.Results: All of 15 nonimmunized control mice developed the hepatoma. Protection of mice immunized with fixed Hepa 1–6 cells and both of IL-2/GM-CSF microspheres or further mixed with TiterMax Gold reached 80% and 87%, respectively. Mass growth of the established tumors, vaccinated twice at 5 mm in diameter, the tumor of control animals continued to grow. However, 7–10 days after the second injection of the tumor vaccine, the tumor growth was suppressed in 9 of 10 mice and then markedly reduced. Complete tumor regression was observed in 60% (6/10) of mice. Splenocytes from the control mice were not able to lyse target Hepa 1–6 cells and other tumor cells. In contrast splenocytes from the vaccinated mice exhibited a 41% lytic activity against the Hepa 1–6 cells tested at an effector/target (E/T) ratio of 5, whereas they did not exhibited such activity against the melanoma cells (B16-F1), Lewis lung carcinoma cells (LLC), renal carcinoma cells (Renca), and bladder carcinoma cells (MBT-2). The cytotoxic activity was inhibited by the treatment with anti-CD3, anti-CD8, and anti-MHC-class I monoclonal antibodies but not with anti-CD4 and anti-MHC-class II antibodies. In the Phase-I clinical trial, vaccination of HCC patients with the autologous vaccine is a well-tolerated treatment and induces fixed tumor fragment-specific immunity.Conclusion: Fixed HCC vaccination elicited protective and therapeutic antitumor immunity against HCC. The tumor vaccine elicited antigen specific CTL response lysis of the target HCC was mediated by the typical MHC-class I restricted CD8+ T cells. Key words cancer vaccine - cytotoxic T lymphocyte - immunotherapy - hepatoma

Effect of Leukemia Vaccine on the Macrophage of Mice

Objective: To evaluate the e?ect of the prepared leukemia vaccine on the cytotoxicity of macrophage of C57BL/6 mice. Methods: The model of mice bearing leukemia was established and three types of leukemia vaccine were prepared before they were administered on the mice respectively. The cy- totxicity of M? derived from the mice was measured after the active immunotherapy or prevention for 2–4 weeks later by using MTT colorimetric method and compared with the control group. Results: (1) With the growth of leukemia cells in the mice, the cytotoxicity of M? was seriously depressed; (2) The administration of leukemia vaccine prepared from inactivated leukemic cells, incomplete Freund’s adjuvant (IFA) and cytokines, such as rGM-CSF, rIL-2 and rIL-6, promoted the cellular immunity of mice bearing leukemia, more e?cient than leukemia vaccine from either inactivated leukemic cells and IFA or inactive leukemic cells per se. Conclusion: The leukemia vaccine prepared with inactive leukemic cells, IFA and cytokines could activate the non-speci?c cellular immunity such as M?, as well as, antigen present, immune surveillance and so on, and this activation constructed a base for further activate T lymphocyte. Naturally, this will certainly have a promising future in the therapy against hematopietic tumor.

Expression and Mutation of MAGE-A3 mRNA in Lung Cancer Tissues

Objective： To inwvetigate the expression of MAGE-A3 mRNA in tissue samples derived from lung cancers and to discuss the possibility of using MAGE-A3 antigens as a new peptide vaccine for inunotherapy for lung cancers. Methods： Tumor tissue samples of lung cancers and paired non-tumor tissues of the lung were obtaimed from 31 lung cancer patients. Total RNA was extracted and cDNA was synthesized. Nested polymernse chain reaction amplification using MAGE-A3 specific primer was performed to detect the expression of MAGE-A3. The 10 clones of 5 samples of MAGE-A3 mRNA positive PCR products were DNA sequenced by using DNAs sequencer （PE-377）. Results： Of 31 lung cancers, 26 （83.9%） expressed MACE-A3 mRNA. The expression of MAGE-A3 gene was not detectable in the adjacent lung tissues. The DNA sequencing confirmed that the target gene fragment in all 5 samples of PCR products was MACE-A3 cDNA. Point nmtations occurred in 4 samples （8 clones） detected （C^2773→T^2773; G^2807→A^2807） resulting in alternation of amino acid residue in one position （E^143→K）. Conclusion： （1） The MAGE-A3 gene was expressed exclusively in tumor tissues of the patients with lung cancer in China. This tumor rejection antigen may have potential to be used as a new peptide vaccine for immunotherapy for lung eancers. （2） There are two point mutations of MAGE-A3 gene sequence in some Chinese lung cancer patients.

Expression of HMGB1 Protein in Human Cervical Squamous Epithelium Carcinoma

OBJECTIVE To investigate the expression of the high mobility group boxl（HMGB1） in human cervical squamous epithelial carcinoma （CSEC） and to explore the relationship of HMGB1 expression to the differentiation degree, size, invasion and metastasis of CSEC. METHODS Immunohistochemical staining of tissue microarrays and Western blot analysis were conducted to detect the expression of HMGB1 in the following tissue samples： 30 carcinoma in situ, 90 invasive CSEC without metastasis, 30 invasive CSEC with metastasis, 30 cases of normal cervical squamous epithelia. RESULTS The positive-expression rate of HMGB1 was 58.7% （88/150） in CSEC, showing a significant difference compared to normal cervical squamous epithelia. The expression of HMGB1 was correlated with tumor size, invasion and metastasis of CSEC （respectively, P〈0.01）, but had no relationship with the degree of differentiation （P〉0.05）. CONCLUSION The over-expression of HMGB1 in CSEC might be a useful parameter as an indication of tumor invasion, metastasis, prognosis and overall biological behavior of human CSEC, as well as a noval target site for gene therapy.

Colon cancer and the immune system:The role of tumor invading T cells

Colon cancer is still one of the leading causes of cancer death worldwide. Although the host immune system has been shown to react against tumor cells, mainly through tumor infi ltrating lymphocytes and NK cells, tumor cells may utilize different ways to escape anti-tumor immune response. Tumor infi ltration of CD8+ and CD4+ (T-bet+) effector T cells has been attributed to a beneficial outcome, and the enhancement of T cell activation through T cell receptor stimulation and co-stimulatory signals provides promising strategies for immunotherapy of colon cancer. Growing evidence supports a role for the Fas/FasL system in tumor immunology, although the mechanisms and consequences of FasL activation in colon cancer are not completely understood. In animal models, depletion of regulatory T cells (CD4+ CD25+ T cells) can enhance the anti-tumor immune response under certain conditions. Taken together, recent insights in the immune reaction against colon carcinoma have provided new approaches to immunotherapy, although much remains to be learned about the exact mechanisms.

Pharmacokinetics of radioimmunotherapeutic agent of direct labeling mAb ^(188)Re-HAb18

AIM:To labed Anti-hepatoma monoclonal antibody(mAb) fragment HAb18 F(ab')_2 was labeled with 188 Re for the pharmacokinetic model of ^(188)Re-HAb18 F(ab')_2 and to evaluate its pharmacokinetic parameters in hepatoma- bearing nude mice. METHODS:HAb18 F(ab')_2 was directly labeled with ^(188)Re using 2-mercaptoethanol(2-ME)as reducing agents. Labeling efficiency and immunoreactivity of ^(188)Re-HAb18 F (ab')_2 were evaluated by Whatman 3MM paper chromatography and live cell assay,respectively. Biodistribution analysis was also conducted in nude mice bearing human hepatoma in which animals were sacrificed at different time points(1,4,18,24 and 24h)after ^(188)Re-HAb18 F(ab')_2 was injected through tail-vein into hepatoma-bearing nude mice.The blood and radioactivity of organs and mass were measured.The concentrations of ^(188)Re-HAb18 F(ab')_2 were evaluated with a pharrnacokinetic 3P97 software. RESULTS:The optimum labeling efficiency and immunoreactive fraction were 91.7% and 0.78%, respectively.The parameters of ^(188)Re-HAb18 F(ab')_2 were: T_(1/2),2.29h;Vd,1.49×10^(-9)L·Bq^(-1);AUC,20.49×10~9Bq·h· L^(-1);CL,0.45×10^(-3)L·h^(-1).^(188)Re-HAb18 F(ab')_2 could locate specially in hepatoma with high selective reactivity of HAb18 F(ab')_2.^(188)Re-HAbl8 F(ab')_2 was mainly eliminated by kidney.The maximal tumor to blood ratio was at 48h,and maximal tumor to liver ratio was at 18h. CONCLUTION:The pharmacokinetics of ^(188)Re-HAb18 F(ab')_2 fit a I-compartment model.^(188)Re-HAb18 F(ab')_2 can be uptaken selectively at the hepatoma site.

Food allergy in gastroenterologic diseases:Review of literature

Food allergy is a common and increasing problem worldwide. The newly-found knowledge might provide novel experimental strategies, especially for laboratory diagnosis. Approximately 20% of the population alters their diet for a perceived adverse reaction to food, but the application of double-blind placebo-controlled oral food challenge, the ＂gold standard＂ for diagnosis of food allergy, shows that questionnaire-based studies overestimate the prevalence of food allergies. The clinical disorders determined by adverse reactions to food can be dassified on the basis of immunologic or nonimmunologic mechanisms and the organ system or systems affected. Diagnosis of food allergy is based on clinical history, skin prick tests, and laboratory tests to detect serum-food specific IgE, elimination diets and challenges. The primary therapy for food allergy is to avoid the responsible food. Antihistamines might partially relieve oral allergy syndrome and IgE-mediated skin symptoms, but they do not block systemic reactions. Systemic corticosteroids are generally effective in treating chronic IgE-mediated disorders. Epinephrine is the mainstay of treatment for anaphylaxis. Experimental therapies for IgE-mediated food allergy have been evaluated, such as humanized IgG anti-IgE antibodies and allergen specific immunotherapy.

Overview of immunosuppression in liver transplantation

Continued advances in surgical techniques and immunosuppressive therapy have allowed liver transplantation to become an extremely successful treatment option for patients with end-stage liver disease.Beginning with the revolutionary discovery of cyclosporine in the 1970s,immunosuppressive regimens have evolved greatly and current statistics confirm one-year graft survival rates in excess of 80%. Immunosuppressive regimens include calcineurin inhibitors,anti-metabolites,mTOR inhibitors,steroids and antibody-based therapies.These agents target different sites in the T cell activation cascade,usually by inhibiting T cell activation or via T cell depletion.They are used as induction therapy in the immediate periand post-operative period,as long-term maintenance medications to preserve graft function and as salvage therapy for acute rejection in liver transplant recipients. This review will focus on existing immunosuppressive agents for liver transplantation and consider newer medications on the horizon.

Immune therapy including dendritic cell based therapy in chronic hepatitis B virus infection

Hepatitis B virus （HBV） infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of chronic HBV carriers suffer from progressive liver diseases. In addition, all HBV carriers are permanent source of this virus. There is no curative therapy for chronic HBV carriers. Antiviral drugs are recommended for about 10% patients, however, these drugs are costly, have limited efficacy, and possess considerable side effects. Recent studies have shown that immune responses of the host to the HBV are critically involved at every stage of chronic HBV infection： （1） These influence acquisition of chronic HBV carrier state, （2） They are important in the context of liver damages, （3） Recovery from chronic HBV-related liver diseases is dependent on nature and extent of HBV-specific immune responses. However, induction of adequate levels of HBV-specific immune responses in chronic HBV carriers is difficult. During the last one decade, hepatitis B vaccine has been administered to chronic HBV carriers as a therapeutic approach （vaccine therapy）. The present regimen of vaccine therapy is safe and cheap, but not so effective. A dendritic cell-based therapeutic vaccine has recently been developed for treating chronic HBV infection. In this review, we will discuss about the concept, scientific logics, strategies and techniques of development of HBV- specific immune therapies including vaccine therapy and dendritic cell-based vaccine therapy for treating chronic HBV infection.

PD-1/PD-L1 pathway blockade works as an effective and practical therapy for cancer immunotherapy

Cancer immunotherapy has greatly advanced in recent years,and PD-1/PD-L1 blocking therapy has become a major pillar of immunotherapy.Successful clinical trials of PD-1/PD-L1 blocking therapies in cancer treatments have benefited many patients,which promoted the Food and Drug Administration(FDA)approval of PD-1/PD-L1 blocking drugs.In this review,we provide a detailed introduction of five PD-1/PD-L1 blocking drugs,with indications and studies,as a valuable reference for doctors and medical investigators.Moreover,the characteristics of PD-1/PD-L1 blocking therapies,including their universality and sustainability,are discussed in this review.Furthermore,we also discuss and predict the possibility of PD-L1 as an indication marker of PD-1/PD-L1 blocking therapy for pan-cancer treatment,and the current status of combination therapies.

In vivo anti-tumor effect of hybrid vaccine of dendritic cells and esophageal carcinoma cells on esophageal carcinoma cell line 109 in mice with severe combined immune deficiency

AIM： To develop a fusion vaccine of esophageal carcinoma cells and dendritic cells （DC） and observe its protective and therapeutic effect against esophageal carcinoma cell line 109 （EC109）. METHODS： The fusion vaccine was produced by fusing traditional polyethyleneglycol （PEG）, inducing cytokine, sorting CD34＋ magnetic microbead marker and magnetic cell system （MACS）. The liver, spleen and lung were pathologically tested after injection of the fusion vaccine. To study the therapeutic and protective effect of the fusion vaccine against tumor EC109, mice were divided immune group and therapeutic group. The immune group was divided into P, E, D and ED subgroups, immunized by phosphate buffered solution （PBS）, inactivated EC109, DC and the fusion vaccine respectively, and attacked by EC109 cells. The tumor size, weight, latent period and mouse survival period were recorded and statistically analyzed. The therapeutic group was divided into four subgroups： P, inactivated EC109, D and ED subgroups, which were attacked by EC109 and then treated with PBS, inactivated EC109, DC, and EC109-DC respectively. Pathology and flow cytometry were also used to study the therapeutic effect of the fusion vaccine against EC109 cells.RESULTS： Flow cytometry showed that the expression of folate receptor （FR）, EC109 （C）, Des （D） in human nasopharyngeal carcinoma cell line （HNE1） （B） was 78.21%, 89.50%, and 0.18%, respectively. The fusion cells （C） were highly expressed. No tumor was found in the spleen, lung and liver after injection of the fusion vaccine. Human IgG was tested in peripheral blood lymphocytes （PBL）. In the immune group, the latent period was longer in EC109-DC subgroup than in other subgroups, while the tumor size and weight were also smaller than those in ED subgroup. In the therapeutic group, the tumor size and weight were smaller in ED subgroup than in P, inactivated EC109 and DC subgroups. CONCLUSION： Fusion cells are highly expressed not only in FR but also in CD80. The fusion vaccine has a distinctive protective effect against tumor EC109 and can inhibit the growth of tumor in mice, and its immune protection against tumor attack is more significant.