Recombinant E.coli LLO/OVA Vaccination Effectively Inhibits Murine Melanoma Metastasis to Lung by CD8^+T Cells Immunity

Objective： To construct recombinant E.coli LLO/OVA and investigate its tumor metastatic inhibition effect in B16 OVA melanoma challenged mice. Methods： Recombinant E.coli LLO/OVA was constructed and the expression of listeriolysin O （LLO） and ovalbumin （OVA） of the vaccine was determined by coomassie brilliant blue staining and western blotting, After 3 subcutaneous injections of E.coli LLO/OVA, the percentages of CD3^＋CD4^＋T, CD4^＋CD25^＋T, CD3^CD8^＋T and OVA257-264 SIINFEKL specific CD8^＋T cells were determined by flow cytomytry, and the tumor metastatic inhibition effect in B16 OVA melanoma challenged mice was observed. Results： Recombinant E.coli LLO/OVA was successfully constructed, and the expression of LLO and OVA of the vaccine was confirmed. After 3 subcutaneous injections of E.coli LLO/OVA and E.coli OVA in mice, the percentages of CD3^＋CD4^＋T, CD4^＋CD25^＋T and CD3^＋CD8^＋T cells were equivalent in the two groups of mice. However, there were significantly more OVA257-264 SIINFEKL specific CD8^＋T cells in E.coli LLO/OVA vaccinated mice than that in E.coli OVA vaccinated mice. The prophylactic E.coli LLO/OVA vaccination effectively prevented the tumor metastasis to lungs in B16 OVA melanoma challenged mice. Depletion of CD8^＋T cells significantly impaired the tumor inhibition effect of the vaccine in B16 OVA challenged mice. The therapeutic vaccination of E.coli LLO/OVA significantly prevented melanoma metastasis to lungs in B I6 OVA challenged mice too. Conclusion： Recombinant E.coli LLO/OVA vaccination is highly effective in inhibiting murine malignant melanoma metastasis by promoting CD8^＋T cell immunity.

Inoculation with IL-2 and B7-1 co-transfected G422 murine glioblastoma cells resulting in enhanced antitumor immunity in vivo

It’s well known that interleukin-2 (IL-2) plays animportant role in eliciting antitumor immunityparticularly mediated by T cells. In addition, theexpression of MHC can be enhanced by IL-2 genetransfection in tumor cells. Recent studies have indicatedthat at least two signals are required for the activation ofnaive T cells by antigen-bearing target cells: an antigen-sopific signal and a crystimulatory signal. B7-l is

Role of cellular immunity in halothane hepatitis: an in vitro study

Objective: To explore the effect of cellular immunity in halothane hepatitis. Methods: Hepatotoxicity model was established by exposing male Hartley guinea pigs to 1% halothane via inspiration for 4 h each time for 1 or 3 times within a 42-day interval. Then their hepatocytes and lymphocytes were collected and divided into 2 parts for different cultures. Hepatocytes were cultivated with or without 1% halothane for 4 h and lymphocytes were cultivated with or without 12.5 μg/ml trifluoroacetylated guinea pig serum albumin (TFA-GSA). Then the 2 kinds of hepatocytes were co-cultivated with lymphocytes (1:100) with or without TFA-GSA induction respectively and the supernatant fluid was taken after 24, 48 and 72 h to determine the concentration of alanine aminotransferase (ALT). The halothane cultivated hepatocytes were co-cultivated with various proportion of TFA-GSA antigen induced lymphocytes and ALT was determined after 48 h to determine the proper proportion of hepatocytes and lymphocyte. Results: Lymphocytes of 3 times halothane induced guinea pigs caused a significant increase of ALT in hepatocytes with or without halothane induction. But the lymphocytes of 1 time halothane induced guinea pigs only caused a significant increase of ALT in hepatocytes with induction of halothane. The increase of ALT was only seen after 48- and 72-hour co-culture. The proper proportion of hepatocytes and lymphocytes was 1:100 for lymphocytes cytotoxicity. Conclusion: Lymphocytes is sensitized after inhalation of halothane and generates cytotoxicity to hepatocytes. The immune response of lymphocytes to hepatocytes will be enhanced by repeated inhalation of halothane. The cellular immunity may be one of the mechanisms of halothane induced hepatotoxicity.

Effectiveness of colonoscopy,immune fecal occult blood testing,and risk-graded screening strategies in colorectal cancer screening

BACKGROUND Colorectal cancer(CRC)is one of the most common malignant tumors,and early screening is crucial to improving the survival rate of patients.The combination of colonoscopy and immune fecal occult blood detection has garnered significant attention as a novel method for CRC screening.Colonoscopy and fecal occult blood tests,when combined,can improve screening accuracy and early detection rates,thereby facilitating early intervention and treatment.However,certain risks and costs accompany it,making the establishment of a risk classification model crucial for accurate classification and management of screened subjects.AIM To evaluate the feasibility and effectiveness of colonoscopy,immune fecal occult blood test(FIT),and risk-graded screening strategies in CRC screening.METHODS Based on the randomized controlled trial of CRC screening in the population conducted by our hospital May 2020 to May 2023,participants who met the requirements were randomly assigned to a colonoscopy group,an FIT group,or a graded screening group at a ratio of 1:2:2(after risk assessment,the high-risk group received colonoscopy,the low-risk group received an FIT test,and the FITpositive group received colonoscopy).The three groups received CRC screening with different protocols,among which the colonoscopy group only received baseline screening,and the FIT group and the graded screening group received annual follow-up screening based on baseline screening.The primary outcome was the detection rate of advanced tumors,including CRC and advanced adenoma.The population participation rate,advanced tumor detection rate,and colonoscopy load of the three screening programs were compared.RESULTS A total of 19373 subjects who met the inclusion and exclusion criteria were enrolled,including 8082 males(41.7%)and 11291 females(58.3%).The mean age was 60.05±6.5 years.Among them,3883 patients were enrolled in the colonoscopy group,7793 in the FIT group,and 7697 in the graded screening group.Two rounds of follow-up screening were completed in the FIT group and the graded screening group.The graded screening group(89.2%)and the colonoscopy group(42.3%)had the lowest overall screening participation rates,while the FIT group had the highest(99.3%).The results of the intentional analysis showed that the detection rate of advanced tumors in the colonoscopy group was greater than that of the FIT group[2.76%vs 2.17%,odds ratio(OR)=1.30,95%confidence interval(CI):1.01-1.65,P=0.037].There was no significant difference in the detection rate of advanced tumors between the colonoscopy group and the graded screening group(2.76%vs 2.35%,OR=1.9,95%CI:0.93-1.51,P=0.156),as well as between the graded screening group and the FIT group(2.35%vs 2.17%,OR=1.09%,95%CI:0.88-1.34,P=0.440).The number of colonoscopy examinations required for each patient with advanced tumors was used as an index to evaluate the colonoscopy load during population screening.The graded screening group had the highest colonoscopy load(15.4 times),followed by the colonoscopy group(10.2 times),and the FIT group had the lowest(7.8 times).CONCLUSION A hierarchical screening strategy based on CRC risk assessment is feasible for screening for CRC in the population.It can be used as an effective supplement to traditional colonoscopy and FIT screening programs.

Dendritic Cell-Derived Exosomes Stimulate Stronger CD8^+ CTL Responses and Antitumor Immunity than TVimor Cell-Derived Exosomes

Exosomes （EXO） derived from dendritic cells （DC） and tumor cells have been used to stimulate antitumor immune responses in animal models and in clinical trials. However, there has been no side-by-side comparison of the stimulatory efficiency of the antitumor immune responses induced by these two commonly used EXO vaccines. In this study, we selected to study the phenotype characteristics of EXO derived from a transfected EG7 tumor cells expressing ovalbumin （OVA） and OVA-pulsed DC by flow cytometry. We compared the stimulatory effect in induction of OVA-specific immune responses between these two types of EXO. We found that OVA protein-pulsed DCOVA-derived EXO （EXODC） can more efficiently stimulate naive OVA-specific CD8^＋ T cell proliferation and differentiation into cytotoxic T lymphocytes in vivo, and induce more efficient antitumor immunity than EG7 tumor cell-derived EXO （EXOEG7）. In addition, we elucidated the important role of the host DC in EXO vaccines that the stimulatory effect of EXO is delivered to T cell responses by the host DC. Therefore, DC-derived EXO may represent a more effective EXO-based vaccine in induction of antitumor immunity. Cellular ＆ Molecular Immunology. 2006;3（3）：205-211.

Cytotoxicity Research of Recombinant Human Lactoferrin on Primary Hepatocyte and Nephrocyte Cell

[Objective] The aim was to study whether recombinant Human Lactoferrin has toxic effect on Primary Hepatocyte and Nephrocyte Cell of rat to provide reference for further safety evaluation.[Method] Recombinant Human Lactoferrin and its digested products were taken as tested compound,cow Lactoferrin was used for contrast.Primary Hepatocyte and Nephrocyte Cell of rat were cultured and IC50 values were tested by MTT,and cytotoxic dose-response relationship was tested.[Result]Target toxicity was not found from recombinant Human Lactoferrin on hepatocytes and nephrocytes,in accordance with sub-chronic toxicity test.[Conclusion] This study is of reference value for further safety evaluation of recombinant Human Lactoferrin and safety of evaluation method of GM food.

Characterization of SARS-CoV-2-specific humoral immunity and its potential applications and therapeutic prospects

Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is an ongoing pandemic that poses a great threat to human health worldwide.As the humoral immune response plays essential roles in disease occurrence and development,understanding the dynamics and characteristics of virus-specific humoral immunity in SARS-CoV-2-infected patients is of great importance for controlling this disease.In this review,we summarize the characteristics of the humoral immune response after SARS-CoV-2 infection and further emphasize the potential applications and therapeutic prospects of SARSCoV-2-specific humoral immunity and the critical role of this immunity in vaccine development.Notably,serological antibody testing based on the humoral immune response can guide public health measures and control strategies;however,it is not recommended for population surveys in areas with very low prevalence.Existing evidence suggests that asymptomatic individuals have a weaker immune response to SARS-CoV-2 infection,whereas SARS-CoV-2-infected children have a more effective humoral immune response than adults.The correlations between antibody(especially neutralizing antibody)titers and protection against SARS-CoV-2 reinfection should be further examined.In addition,the emergence of cross-reactions among different coronavirus antigens in the development of screening technology and the risk of antibody-dependent enhancement related to SARS-CoV-2 vaccination should be given further attention.

Detecting SARS-CoV-2 neutralizing immunity: highlighting the potential of split nanoluciferase technology

The coronavirus disease 2019(covID-19)pandemic has progressed over 2 years since its onset causing significant health concerns all over the world and is currently curtailed by mass vaccination.mmunity acquired against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)can be following either infection or vaccination.However,one can never be sure whether the acquired immunity is adequate to protect the individual from subsequent infection because of three important factors:individual variations in humoral response dynamics,waning of protective antibodies over time,and the emergence of immune escape mutants.Therefore,a test that can accurately dfferentiate the protected from the vulnerable is the need of the hour.The plaque reduction neutralization assay is the conventional gold standard test for estimating the titers of neutralizing antibodies that confer protection.However,it has got several drawbacks,which hinder the practical application of this test for wide-scale usage.Hence,various tests have been developed to detect protective immunity against SARS-CoV-2 that directly or indirectly assess the presence of neutralizing antibodies to SARS-Cov-2 in a lower biosafety setting.In this review,the pros and cons of the currently available assays are elaborated in detail and special focus is put on the scope of the novel split nanoluciferase technology for detecting SARS-CovV-2 neutralizing antibodies.

Boosting T cell immunity against cytomegalovirus:a potential strategy combating human aging and age-related diseases

In a recent publication in Cell,Hasegawa and colleagues reveal that cytotoxic CD4-positive cytotoxic T lymphocytes(CD4+CTLs)eliminate senescent fibroblasts with a surface expression of the immunologically important human leukocyte antigen class Il(HLA-)and the human cytomegalovirus glycoprotein B(HCMV-gB).1 This implies that a virome-immune cell interaction controls the fate of human skin cells that are linked to the aging process and associated diseases.

Mannose-6-phosphate receptor: a novel regulator of T cell immunity

CD8^(+) cytotoxic T cells(CTLs)protect against pathogens and cancer.1,2,3,4 Over the past few years,CTL responses have been extensively studied.1,2,3,4 In response to an infection,the CTL response develops in three phases:expansion,contraction,and memory.2,3,In the expansion phase,pathogen-derived antigens(Ags)trigger activation and proliferation of Ag-specific CTLs,leading to the generation of effector CTLs(eCTLs)with cytotoxic functions.2,5,Later,in the contraction phase,90–95%of eCTLs die as a result of activation-induced cell apoptosis.

The role of a novel protein ligand hMSH 2 in Vγ9δ2 T cell-mediated anti-cervix cancer immunity

Objective To explore the role of hMSH2,a novel endogenous tumor-associated protein ligand recognized by Vγ9δ2 T cells,in innate anti-cervix cancer immunity.Methods hMSH2 that expressed on the surface of cervical cancer cell line HeLa cells was blocked by specific antibody.Then the differences in their effects on Vγ9δ2T cells before and after antibody blockage were evaluated by cytotoxicity of Vγ9δ2 T cells and cytokines secretion.

In Vitro Cytotoxicity of Polyphosphoester as a Novel Injectable Alveolar Replacement Material

The aim of this study was to investigate the in vitro cytotoxicity of polyphosphoester polymer used as a novel injectable alveolar bone substitutes for controlled delivery of tetracycline. Cell culture medium was exposed to the polymer （0.01-10 mg/mL） for 24 h. The L-929 mouse fibro- blasts were then exposed to the treated cell culture medium for 24 h. Finally, cell viability and growth were assessed by using MTT assay and Alamar Blue assay. No significant cytotoxicity of the polyphosphoester against L-929 mouse fibroblasts was observed at a concentration up to 10 mg/mL （P〉0.05）. The two evaluation methods showed no significant differences （P〉0.05）. This study suggests that polyphosphoester does not demonstrate any significant toxic effects to cells in vitro and has the potential to be used both as a medical device and as scaffolds in tissue engineering applications.

Immune checkpoint inhibitor-mediated colitis in gastrointestinal malignancies and inflammatory bowel disease

Immune checkpoint inhibitors(ICI)have markedly changed the landscape of cancer therapy.By re-invigorating the immune system against tumors,ICI provide novel therapeutic options for a broad variety of malignancies,including many gastrointestinal(GI)cancers.However,these therapies can also induce autoimmune-like side effects in healthy tissue across the body.One of the most common of these side effects is ICI-mediated colitis and diarrhea(IMC).Here,we review the incidence and risk of IMC in ICI therapy,with a focus on what is known regarding IMC in patients with GI malignancies.We also discuss data available on the use of ICI and risk of IMC in patients with pre-existing inflammatory bowel disease,as these patients may have increased risk of IMC due to their underlying intestinal pathology.

Clinical characteristics of gastrointestinal immune-related adverse events of immune checkpoint inhibitors and their association with survival

BACKGROUND Despite the popularity of immune checkpoint inhibitors(ICIs)in the treatment of advanced cancer,patients often develop gastrointestinal(GI)and non-GI immune-related adverse events(irAEs).The clinical characteristics and survival outcomes of GI-irAEs have not been fully elucidated in previous reports.This necessitates the evaluation of the impact of GI-irAEs on patients receiving ICI treatment.AIM To evaluate the clinical characteristics of GI-irAEs and their impact on survival in patients treated with ICIs.METHODS In this single-center,retrospective,observational study,we reviewed the records of 661 patients who received ICIs for various cancers at Nagoya University Hospital from September 2014 to August 2020.We analyzed the clinical characteristics of patients who received ICI treatment.We also evaluated the correlation between GI-irAE development and prognosis in non-small cell lung cancer(LC)and malignant melanoma(MM).Kaplan-Meier analysis was used to compare the median overall survival(OS).Multivariate Cox proportional hazards models were used to identify prognostic factors.A P value<0.05 was considered statistically significant.RESULTS GI-irAEs occurred in 34 of 605 patients(5.6%)treated with an anti-programmed cell death-1/programmed death-ligand 1(anti-PD-1/PD-L1)antibody alone and in nine of 56 patients(16.1%)treated with an anti-cytotoxic T-lymphocyte antigen 4(CTLA-4)antibody alone or a combination of anti-PD-1 and anti-CTLA-4 antibodies.The cumulative incidence and median daily diarrhea frequency were significantly higher in patients receiving anti-CTLA-4 antibodies(P<0.05).In 130 patients with MM,OS was significantly prolonged in the group that continued ICI treatment despite the development of GI-irAEs compared to the group that did not experience GI-irAEs(P=0.035).In contrast,in 209 patients with non-small cell LC,there was no significant difference in OS between the groups.The multivariate analyses showed that a performance status of 2-3(hazard ratio:2.406;95%confidence interval:1.125–5.147;P=0.024)was an independent predictive factor for OS in patients with MM.CONCLUSION Patients receiving anti-CTLA-4 antibodies develop GI-irAEs more frequently and with higher severity than those receiving anti-PD-1/PD-L1 antibodies.Continuing ICI treatment in patients with MM with GI-irAEs have better OS.

In vitro corrosion behavior and cytotoxicity property of magnesium matrix composite with chitosan coating

Mg-6%Zn-10%β-Ca3(PO4)2 composite was prepared through powder metallurgy methods with different chitosan coatings on its surface. The properties of the chitosan coatings on the surface of Mg-6%Zn-10%β-Ca3(PO4)2 composite, such as the adhesion ability, the corrosion behavior and the cytotoxicity properties, were investigated, and the microstructure of the chitosan coating was observed by scanning electron microscope(SEM). The results show that chitosan coating improves the corrosion resistance of the magnesium composite specimens significantly. Mg-6%Zn-10%β-Ca3(PO4)2 composite specimens exhibit good corrosion resistance and low p H values in simulated body fluid(SBF) at 37 °C in the immersion test with 7-layer chitosan coating whose relative molecular mass is 30×104 Da. The cytotoxicity tests indicate that Mg-6%Zn-10%β-Ca3(PO4)2 with chitosan coating is nontoxic with a cytotoxicity grade of zero against L-929 cells, which is better than that of uncoated composites.

Cytotoxicity evaluation of extracts and fractions of five marine sponges from the Persian Gulf and HPLC fingerprint analysis of cytotoxic extracts

Objective: To screen the cytotoxic effects of some marine sponges extracts on HeLa and PC12 cells.Methods: Five marine sponges including Ircinia echinata(I. echinata), Dysidea avara,Axinella sinoxea, Haliclona tubifera and Haliclona violacea were collected from the Persian Gulf(Hengam Island). The cytotoxic effect of these sponges was evaluated by using MTT assay. The metabolic high performance liquid chromatography fingerprint of I. echinata was also carried out at two wavelengths(254 and 280 nm).Results: Among the sponges tested in this study, the extracts of I. echinata and Dysidea avara possessed the cytotoxic effect on HeLa and PC12 cells. The obtained fractions from high performance liquid chromatography were evaluated for their cytotoxic properties against the cell lines. The isolated fractions did not show significant cytotoxic properties.Conclusions: I. echinata could be considered as a potential extract for chemotherapy.Further investigation is needed to determine the accuracy of mechanism.

Immune blockade inhibitors and the radiation abscopal effect in gastrointestinal cancers

The field of tumor immunology has produced in the recent years a revolution in cancer therapeutics putting an end in the long lasting frustration of investigators in the area stemming from largely unsuccessful strides to develop cancer vaccines. This progress has come from the introduction of immune checkpoint inhibitors, monoclonal antibodies blocking ligand/receptor pairs with inhibitory effects for immune cells. Through this blockade immune checkpoint blockers are able to ac-tivate the immune system and create an anti-tumoral effect. A significant sub-set of patients with various types of cancers such as melanoma, lung carcinomas and urothelial cancers benefit from treatment with these drugs and survivals have improved in some ca-ses. However other cancers are primarily resistant to immune blockers and secondary resistance is also the norm. Radiation therapy is often used in the palliative treatment of patients with advanced cancers and, in addition to the local effect in the irradiated field, it may in rare cases produce a systemic antitumor effect, termed "abscopal". This effect has been suggested to be produced by immune mechanisms. Thus an opportunity presents for a synergistic effect of immune stimulation between radiation and immune blockade inhibitors. The therapeutic opportunities presented with the combination of radiation and these drugs for gastrointestinal cancers will be discussed in this editorial overview.

Testing Point of Care Portable Data Capture in a Hospital HIV Transmission Prevention Programme and Experience from Computerized Patient Management in a Sub-Saharan Clinic Setting

Clinicians involved in HIV/AIDS （Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome） prevention of mother to child transmission （PMTCT） programme and research activities can benefit from the advantages that computerized systems add to medical practice even in resource constrained sub-Saharan clinic settings. Their continued use of paper based systems presents clinical data management and patient care challenges. A portable point of care data capture electronic system and a computerized clinic patient management system （CCPMS） were implemented to remedy these challenges. PMTCT report compilation was easier with the portable data collection system whose data were found to be more complete and accurate with a 0.83% error rate compared to a 4.1% error rate in the paper registers. A resounding majority of clinicians preferred using the new CCPMS with many of the view that it improved drug inventory and general clinic management with a positive effect on patient care.

骆驼布鲁氏菌活疫苗A19株免疫试验

为通过骆驼布鲁氏菌A19株疫苗免疫试验筛选出适宜的免疫方案,选择布鲁氏菌抗体检测阴性骆驼60头,按照不同剂量、不同年龄分4组颈部皮下注射免疫,并于免疫后不同时间采样检测免疫抗体。结果显示,1~3岁骆驼标准剂量(6×10^(10)CFU/头)免疫后23 d,免疫抗体阳性率为100%,至免疫12个月,免疫抗体阳性率0;1~3岁骆驼减低剂量(1×10^(9)CFU/头)免疫23 d,免疫抗体阳性率为94.1%,至免疫6个月,免疫抗体阳性率为0。3~12月龄骆驼标准剂量免疫38 d,免疫抗体阳性率为50%,免疫6个月,免疫抗体阳性率0。3~12月龄骆驼减低剂量免疫38 d,免疫抗体阳性率为16.7%,免疫6个月,免疫抗体阳性率为0。结果表明,1~3岁骆驼使用布氏菌活疫苗(A19株)标准剂量(6×10^(10)CFU/头)皮下注射免疫,效果最好。

基于小白鼠试验评价乳清醋增强免疫功能研究

用乳清醋饲喂小白鼠试验,分析检验其免疫学指标,评价乳清醋是否具有增强机体免疫的功能特性。试验鼠选用体质量(BW)为20 g的雄性小白鼠,共320只,从中每次随机取40只作为1个批次饲喂的试验鼠,分为4组,3个处理组和1个对照组,每组10只。实验室制备总酸为5.52 g/100 mL的乳清醋,乳清醋经稀释后的高、中、低浓度的剂量组,分别对处理组小鼠以剂量为10 mL/kg BW灌胃,对照组同步灌胃等量生理盐水,日饲1次,连续30 d为1个批次饲喂试验。研究以试验鼠的8项免疫学指标为检测目标:(a)免疫器官重量及胸腺脾脏指数,(b)迟发型超敏反应,(c)淋巴细胞转化率,(d)抗体生成细胞检测,(e)血清溶血素检测,(f)单核巨噬细胞攻击和吞噬能力,(g)碳粒廓清试验,(h)NK(Natural killer)细胞活性。每一批次饲喂试验结束后,针对其中1项免疫学指标要求对该批次的小鼠进行处置、解剖、检测。取平均数据为该批次各组鼠的该项免疫学指标数据。经8批次的饲喂试验,完成了试验鼠8项免疫学指标的检测。结果显示,处理组与对照组相比,除了(d)和(e)2项指标没有明显的统计学差异(P>0.05)外,中、高剂量组小鼠的其余6项免疫学指标均明显增强,均有统计学差异(P<0.05)。本研究发现,乳清醋饲喂白鼠后在促进和提高小鼠的免疫器官增重,细胞免疫功能,单核巨噬细胞吞噬功能和NK细胞活性等方面起了明显作用,可以判定乳清醋具有增强人体免疫力的功能。