[Induction of Th1 immune response against tumor by genetically engineered fusion of tumor cells and dendritic cells].
1
Citation
0
Reference
10
Related Paper
Abstract:
To study the antitumor activity of engineered fusion of tumor cell and dendritic cells (DC).J558 tumor cells were transfected with mouse IL-12 (mIL-12) gene and then fused with DCs to develop a hybrid-engineered tumor vaccine. BALB/c mice were challenged with wild-type J558 tumor cells 14 days after vaccinated with hybrid-engineered J558.mIL-12 was detected at (870 +/- 60) pg.(10(5) cells)(-1).ml(-1) in the culture supernatants and the cell-fusion rate was about 30% by co-focal microscopy. In addition, the lymphocytes from popliteal nodes and groin nodes of these mice vaccinated with hybrid-engineered J558 secreted higher levels of IFN-gamma than that of other control mice, and vaccination of mice with the fusion vaccine induced more efficient tumor-specific CTL cytotoxicity against wild-type tumor cells in vitro and with efficient antitumor immunity in vivo.It suggested that vaccination of mice with the fusion vaccine induced stronger Th1-dominant responses and this approach could perhaps be applied to clinical settings of DCs-based cancer vaccines.Keywords:
CTL*
Cell fusion
Cite
Obese people are at high risk of developing infections, including COVID-19, and are prone to a more severe course and a poorer prognosis of diseases. The review summarizes information on the post-vaccination immune response in obesity in children and adults with infections. The SARS-COV-2 pandemic further exacerbates this problem of the adequacy of the immune response to vaccination of obese people. The purpose of this review is to present and summarize information on the changes in various links of cellular and humoral immunity in the experiment and in the clinic during the immune response to vaccination in obesity. Results. The mechanisms of action of obesity and associated chronic inflammation and metabolic dysregulation on the post-vaccination immune response in various infections are discussed. The systemic inflammatory response that occurs in obesity represents a barrier to the induction of a sustained immune response. In obese individuals, innate and adaptive immune responses are slowed down and diminished, contributing to the spread of infections. Conclusion. In obesity, the differentiation and proliferation of cells of the immune system is impaired, and the immune response to vaccination changes. Further research is needed to study post-vaccination immunity in obesity, taking into account the effect on the vaccination of the microbiota of a particular person, the presence of possible comorbid conditions.
Pandemic
Cite
Citations (0)
Lipofectamine
Cationic polymerization
Ethyleneimine
Cite
Citations (22)
这研究的目的是学习人的乳突淋瘤的效果的目的在 vitro 并且在 vivo 的 RMA 房间的生长上的病毒(HPV ) 类型 16 E7 蛋白质表示。带野类型 HPV 16 E7 的再结合向量 pcDNA3.1-E7 被定序识别的方法。再结合向量 pcDNA3.1-E7 是进由 liposome 的老鼠淋巴腺瘤房间线 RMA 的 transfected，并且 monoclonal 房间 transfected 被抗菌素 G418 sieving 和限制冲淡试金稳定地获得。RT-PCR 方法被用来在 RMA-E7 房间检测 HPV 16 E7 mRNA 的表示。RMA 细胞和在 vitro 有教养的 RMA-E7 细胞的生长被细胞计数 Kit-8 测试。RMA-E7 房间和 RMA 房间皮下地分别地在 syngeneic 老鼠被接种，肿瘤尺寸被一个星期，和 E7 蛋白质表示在老鼠的肿瘤织物滑动 caliper 两次测量被西方的污点在肿瘤以后检测形成。在忍受肿瘤的老鼠的 E7 特定的 T 房间的 cytolytic 活动的动力学被 LDH 工具包测量。再结合向量定序的结果证明被插入到 recombinant 的目标基因是正确的，并且稳定地表示 E7 蛋白质的 RMA-E7 房间被有限冲淡试金获得。在 vitro 在在 RMA 细胞和 RMA-E7 细胞之间的 morphous 和生长速度没有明显的差别。RMA-E7 房间在 syngeneic 老鼠成长了比 RMA 房间显著地慢。E7 蛋白质被表示在在 vivo 的 RMA-E7 房间更强壮比在 vitro。E7 特定的 CTL 的 cytolytic 能力在早阶段被激活，在中间的阶段到达了最大值，并且在结束阶段输了。从忍受肿瘤的老鼠孤立的 RMA-E7 房间对比在 vitro 有教养的 RMA-E7 房间杀死的 E7 特定的 CTL 更抵抗。E7 蛋白质表示没在 vitro 在 RMA-E7 房间的生长上有明显的影响的结论，和罐头在 vivo 压制 RMA-E7 房间的生长。E7 特定的 CTL 的活动曲线近似介绍鈥渂e ll 鈥 ? 形状。RMA-E7 房间在有的 vivo 成长了高表情 E7 铺平蛋白质，和对比在 vitro 有教养的那些杀死的 E7 特定的 CTL 抵抗的更多。在 vivo 的 E7 蛋白质表示不仅开始有免疫力的激活，而
CTL*
Cite
Citations (0)
Objective To investigate the feasibility and the efficacy of gene transfection to cultured cells and mouse muscles, mediated by ultrasound combined with a new ultrasound contrast agent SonoVue. The efficiency of gene expression with regard to different parameters of ultrasound and different DNA dosages was also studied. Methods Human liver cancer cell line HepG2 was used in the in vitro gene transfection experiment. While intramuscular injection of plasmid DNA combined with SonoVue solution was used in the in vivo experiment. Results pEGFP-C1 plasmids could be readily transfected to HepG2 cells in vitro and to mouse tibialis anterior muscles in vivo. One MHz ultrasound was ideal both in vitro and in vivo, but a more powerful ultrasound was needed in vivo (3 W/cm 2) than in vitro (1.5 W/cm 2). Conclusion Ultrasound combined with SonoVue could promote gene transfection to cultured cells and mouse muscles. In both experiments 1 MHz was an ideal ultrasound frequency.
Sonoporation
Cite
Citations (0)
CTL*
Cite
Citations (16)
Beside lymphocytes, HTLV-1 can infect monocytes, blood- or monocytes-derived dendritic cells (DCs) and plasmacytoid DCs in vitro. Considering that DCs might be the first cells to be infected during primo infection, we hypothesize that they may constitute viral reservoirs and eventually spread the virus to surrounding lymphocytes. Interestingly, ATL and HAM/TSP diseases display opposite immunological features, i.e. an impaired CTL response versus a chronic inflammation respectively. Since DCs are major effectors of both innate and adaptative immune responses, infection of specific DC subsets and the subsequent alteration of their functions might also lead to the orientation of the adaptive immune response towards either viral tolerance associated with impaired CTL responses or chronic inflammation, and thus directly participate to the determination of the infection outcome. Using various cytokines cocktails, we therefore generated distinct monocytes-derived DC (MDDC) subsets and infected these cells with HTLV-1 biofilm. We first show that the different MDDC subsets are not equally susceptible to HTLV-1 infection, as measured by FACS analyses and real time PCR assays. We then show that following infection, DC activation or IFN alpha production are not affected by infection in the MDDC subtypes tested. However, while DC maturation alters their susceptibility to the virus, we demonstrate that IFN alpha treatment does not. Finally, the ability of MDDC subsets to transmit HTLV-1 to T-cells will be discussed. Taken together, our results suggest that differential susceptibility of various DC subsets to HTLV-1 infection could differently shape immune responses and therefore affect viral pathogenesis.
CTL*
Cite
Citations (2)
Genetic immunization with a single injection of dendritic cells (DCs) expressing a model melanoma antigen generates antigen-specific, MHC-restricted, protective immune responses. After initiating the immune response, additional vaccinations may increase the protection or conversely downregulate the immune response. Groups of mice were vaccinated several times with DCs transduced with the MART-1 gene, and the anti-tumor protection was compared with that of mice receiving a single vaccination. C3H mice had poorer protection from a syngeneic MART-1-expressing tumor challenge with multiple vaccinations. This was accompanied by lower levels of splenic CTL effectors and a shift from a type 1 to a type 2 cytokine profile. On the contrary, multiple vaccinations in C57BL/6 mice generated greater in vivo antitumor protection with no decrease in splenic CTLs and no cytokine shift. Antiadenoviral humoral or cellular immune responses did not seem to contribute to these effects. When studies were performed in Fas receptor-negative C3H.(lpr) mice, the adverse effect of multiple vaccinations disappeared, and there was no cytokine shift pattern. In conclusion, C3H mice but not C57BL/6 mice receiving multiple vaccinations with DCs expressing the MART-1 tumor antigen show decreased protection associated with deviation from a type 1 to a type 2 cytokine response attributable to a Fas-receptor mediated clearance of antigen-specific IFN-gamma-producing cells.
CTL*
Cite
Citations (39)
Cultured SV40-transformed fibroblasts from C3H mice (SV-C3H) were "adapted" to in vivo growth by serial passage through sublethally irradiated, syngeneic recipients. After four in vivo passages, a population of cells was obtained (V4) that was weakly oncogenic in nonirradiated mice. Cells isolated from large V4 tumors (V5) were found to be highly oncogenic, producing lethal tumors at doses of less than 10(3) cells. V5 is insensitive to SV40-specific transplantation immunity in syngeneic animals but can be rejected completely by H-2 allogeneic mice. In vitro studies revealed that although V4 and the parent SV-C3H cells can induce SV40-specific cytotoxic T cells (CTL) in vitro and are lysed by these CTL, V5 does neither. The failure of V5 to interact with CTL was traced to the loss of H-2Kk antigen expression on these cells. The correlation between H-2Kk loss and immunoresistance in vivo suggests a central role for the cytotoxic T cell in in vivo tumor elimination in this system.
CTL*
Cite
Citations (95)
HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP), is a debilitating neurodegenerative disease characterized by a robust immune response including the oligoclonal expansion of cytotoxic T lymphocytes (CTLs) specific for the viral oncoprotein Tax.However, the underlying mechanism resulting in the disease process is currently unknown.The CTL response is affected by many factors including the efficiency of epitope processing and presentation.In this respect, dendritic cells (DCs), the most potent antigen presenting cells, have long been recognized as key regulators of the immune system.We have previously demonstrated that DCs are capable of priming a pronounced Tax-specific CTL response in naïve PBLs and in HLA-A2 transgenic mice.Since DCs are such crucial cells of the immune system, an extensive assessment of their function and interaction with T cells in HAM/TSP is critical.Therefore, utilizing a newly standardized DC and pre-standardized T cell polychromatic antibody cocktails, we have investigated the immune activation of these cells in HTLV-1 infected samples from the Jamaican region including the seronegative controls, asymptomatic carriers (ACs), and HAM/TSP patients.The extensive immune cell profiling was compared to the matched proviral loads and Tax mRNA levels leading to the identification of unique signatures distinguishing ACs from HAM/TSP patients.Collectively, these studies possess great potential to enable immune cell monitoring and development of diagnostic and therapeutic strategies for the HTLV-associated neuroinflammatory disease.
Cite
Citations (1)
Balb/c (H-2d) thyroid lobes were cultured for 48 hours in 2.5 atmospheres of O/sub 2/ and inserted under the kidney capsule of C57B1/6 (H-2b) mice. 36 out of 52 grafts survived for more than 30 weeks. Unlike freshly grafted mice long term recipients failed to reject the original grafts or a new transplant following the injection of donor spleen cells. Cytotoxic T lymphocytes precursor (CTLp) frequency was found to be in the same range (1/500 to 1/1000 spleen cells) in tolerant and normal mice. Nevertheless CTL responses were reduced both in vivo and in vitro. CTL responses in vivo were measured by injecting live P815 tumor cells (H-2d) i.p. into primed tolerant and normal C57B1/6 mice. The peak of specific /sup 51/Cr release (E:T = 80:1) of 9 normal mice was 39 +/- 7% on day 3, while the peak response of 11 tolerant mice was 12 +/- 2% on day 5. CTL responess measured in vitro were less strinkingly reduced than the in vivo responses. However, the in vitro responses were reduced in different experiments by either (1) reduced specific /sup 51/Cr release at the same E:T ratio or (2) reduced cell number at the end of the culture.more » Further evidence that the in vitro CTL responses of tolerant mice were suppressed was found in their restoration to normal levels by the addition of ConA supernatant to the culture media.« less
CTL*
Ratón
Cite
Citations (0)