CN101972476A - DNA vaccine adjuvant using Micro RNA-155 and construction method thereof - Google Patents
DNA vaccine adjuvant using Micro RNA-155 and construction method thereof Download PDFInfo
- Publication number
- CN101972476A CN101972476A CN201010280671XA CN201010280671A CN101972476A CN 101972476 A CN101972476 A CN 101972476A CN 201010280671X A CN201010280671X A CN 201010280671XA CN 201010280671 A CN201010280671 A CN 201010280671A CN 101972476 A CN101972476 A CN 101972476A
- Authority
- CN
- China
- Prior art keywords
- plasmid
- nucleic acid
- microrna
- acid vaccine
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention relates to the technical field of medical biology. Micro RNA-155 is micro ribonucleic acid (RNA) for regulating the maturation and natural immunoreaction of T cells and B cells. The invention provides a micro RNA-155-based plasmid for enhancing the immune effect of deoxyribonucleic acid (DNA) vaccine cells. A plasmid pcDNA 3.1-pre-miR-155 is obtained by inserting an expression sequence of a micro RNA-155 precursor molecule into a pcDNA-3.1 plasmid, and the micro RNA-155 precursor molecules can be expressed efficiently without influencing the expression capability of the co-transfected DNA vaccine. In an in-vivo test, the plasmid and the DNA vaccine are injected at the same time and have the capability of effectively introducing cellular immunoreaction, and such capability is represented by the increase of antigenically specific spleen IFN-gamma positive cells and enhancement of target killing capability of spleen CD8 positive cells. The plasmid of the invention can be added into the conventional DNA vaccine preparation and injected together so as to obviously promote the cellular immunoreaction induced by the DNA vaccine, and serves as a novel cell immune adjuvant.
Description
Technical field
The present invention relates to the medicine bioengineering field that learns a skill, relate in particular to the immunocompetence enhancement techniques field of nucleic acid vaccine.
Background technology
MicroRNAs is a kind of abundant endogenic noncoding RNAs, matches the expression of regulating targeting mRNAs in 3 ' UTR district by the defective base complementrity in the process of transcribing, thus regulating mRNA division and transcribe inhibition.(Kato, M.and Slack, F.J. (2008) microRNAs:small molecules with big roles-C.elegans to human cancer.Biol.Cell, 100,71-81.) MicroRNAs mainly regulates a lot of biological processess, be included in the natural immunity reply in the differentiation and the activation of many cells.(Garzon, R.and Croce, C.M. (2008) MicroRNAs in normal and malignant hematopoiesis.Curr.Opin.Hematol., 15,352-358.) wherein a kind of typical MicroRNA:MicroRNA-155 is the microRNA of a kind of T of adjusting cell and B cell maturation and natural immunity reaction.(Rodriguez, A., Vigorito, E., Clare, S., Warren, M.V., Couttet, P., Soond, D.R., van, D.S., Grocock, R.J., Das, P.P., Miska, E.A.et al.2007) Requirement ofbic/microRNA-155for normal immune function.Science, 316,608-611.) it is a kind of immune negative regulation molecule, it mainly acts on is the release of restriction inflammatory mediator, excite granulocyte and monocytic increment to strengthen the natural immunity, make the IgG1 antibody of B cell generation high-affinity simultaneously, weakening Th 1 cell and Th2 cell differentiation, the positive aspects such as making the antigen attenuating of regulating of T cell strengthens the acquired immune response.(346-348) under physiological conditions, this molecule is to immunoreation in the body for Mark A.Lindsay (2008) microRNAs andthe immune response, cell, and especially the natural attenuation process of cell immune response is significant.There are some researches show that MicroRNA-155 can make the Th2 cell produce a large amount of various types of cells factors in knock out mice, as IL-4, IL-5, IL-10 etc.(608-611) these cytokines are and B cellular immunization related cytokine for Rodriguez, A.et al. (2007) Requirement ofbic/microRNA-155 for normal immune function.Science 316.There are some researches show that simultaneously MicroRNA-155 can reduce the T lymphocyte immunity and reply, mainly show as the minimizing of IFN-(IFN-γ) and interleukin-22 (IL-2).(Thai，T.H.et?al.(2007)Regulation?of?the?germinal?center?response?by?microRNA-155.Science?316，604-608)。These find prompting, and therefore microRNA-155 may have very big research and using value in the vaccine field to the important regulating action of establishing of antigen specific immune response.
Nucleic acid vaccine is a kind of important new generation vaccine.(Donnelly, J.J., J.B.Ulmer, J.W.Shiver, and M.A.Liu.DNA vaccines.Annu.Rev.Immunol., 1997,15:617-648) because the immunoreactive specific ability of its inducing cell, it is at some viral infection, and bacterium infects in the born of the same parents, and special meaning is arranged on the treatment of diseases such as tumor.Behind the nucleic acid vaccine immunity, produce and immunoreactively comprise mainly that the natural immunity is replied with acquired immunity and reply.But nucleic acid vaccine also exists tangible deficiency, and promptly nucleic acid vaccine stimulates a little less than the immunoreation that ability that body produces immunne response often causes than the conventional vaccine inoculation, and this has just proposed new challenge to Study on DNA Vaccine Against.Therefore, the nucleic acid vaccine adjuvant has become the research focus that receives much attention now.Adjuvant as nucleic acid vaccine mainly contains cytokine, costimulatory molecules, CpG DNA and liposome (Donnelly, JJ, Wahren at present, B, and Liu, MA.DNA vaccines:progress andchallenges.J Immunol., 2005,175:633-639.).
But, in the existing nucleic acid vaccine adjuvant, do not utilize the research report and relevant patent of MicroRNA-155 plasmid vector as vaccine adjuvant.
Summary of the invention
The object of the present invention is to provide a kind of nucleic acid vaccine adjuvant and construction method thereof of utilizing the MicroRNA-155 plasmid vector with cellular immunization enhanced activity.
The present invention has important adjusting meaning in view of MicroRNA-155 at B cell and T cellullar immunologic response, strengthens the expression of MicroRNA-155 in the nucleic acid vaccine immunity local cells, and may strengthen vaccine-induced specific immunity has very big meaning.
The invention provides a kind of nucleic acid vaccine adjuvant with cellular immunization enhanced activity, it is characterized in that this nucleic acid vaccine adjuvant is the plasmid that contains DNA sequence that can effective expression MicroRNA-155 precursor molecule, DNA sequence wherein is shown in SEQ ID NO:4.
Above-mentioned plasmid is pcDNA.3.1 (+), and the present invention also can substitute with other eukaryon expression plasmids such as pVAX1 or pEZX.Plasmid also can substitute with adenovirus, slow virus etc.
The present invention also provides the construction method of above-mentioned nucleic acid vaccine adjuvant, may further comprise the steps: A) design PCR primer is shown in SEQ ID NO:2 or 3, adopt the extractive genomic DNA of human peripheral blood mononuclear cell to carry out PCR, the DNA sequence of the MicroRNA-155 precursor molecule that obtains encoding is shown in SEQ ID NO:4;
B) DNA sequence with above-mentioned coding MicroRNA-155 precursor molecule is cloned into plasmid.
Particular content of the present invention is as follows:
1, at first according to the internet database information searching human microRNA-155 precursor molecule sequence (GENBANK NR 030784.1, shown in SEQ ID NO:1), at its flanking gene group coding sequential design PCR primer (upstream and downstream is added BamH I and EcoR I restriction enzyme site respectively), adopt the extractive genomic DNA of human peripheral blood mononuclear cell to carry out PCR, the clone is as follows with the PCR primer:
Forward primer: 3 ' GGATCCGGTGGCACAAACCAGGAA 5 ' (shown in SEQ ID NO:2)
Downstream primer: 5 ' GAATTCTCTAAGTTTATCCAGCAGGGTG 3 ' (shown in SEQ ID NO:3)
Thereby obtained comprise the microRNA-155 precursor molecule dna fragmentation (shown in SEQ IDNO:4,247bp).
2, be connected into above-mentioned clone's the dna fragmentation that comprises the pre-miR-155 coded sequence (as shown in Figure 1) in polyclone zone, adopt BamH I and EcoR I restriction enzyme site available from the commercialization plasmid pcDNA.3.1 (+) of American I nvitrogene company.The polyclone enzyme action site of this insertion sequence flank still keeps, and can be used to insert antigen sequence does not influence its effective expression.Described pre-miR-155 expression vector: the pcDNA.3.1-pre-miR-155 that is.
A kind of plasmid pcDNA.3.1-pre-miR-155 that contains hsa-miR-155 precursor coded sequence of the present invention, this plasmid can efficiently express human MicroRNA-155 precursor molecule (pre-miR-155) in vivo and in vitro, and, can impel the ripe bulk concentration of cell MicroRNA-155 to improve more than 1000 times in the in vitro tests through the ripe body of shear action formation MicroRNA-155.Through being total to the injecting immune mice with the hepatitis B surface antigen nucleic acid vaccine, we find to improve especially to nucleic acid vaccine immunity effectiveness by this plasmid, and the enhancing of cell immune response plays an important role, therefore, described plasmid and nucleic acid vaccine are injected altogether and be can be used as a kind of new nucleic acid vaccine immunity effectiveness Enhancement Method.
Plasmid of the present invention is through checking, and have following attribute: (1) this plasmid can efficiently increase intracellular MicroRNA-155 developed by molecule level.(2) this plasmid can be total to immunity and not influence latter's antigen expressed with various nucleic acid vaccines.(3) this plasmid and hepatitis B surface antigen nucleic acid vaccine are injected mice altogether, can induce very strong antigenic specificity cell immune response, more traditional nucleic acid vaccine is significantly increased.But the influence of antagonist titre is not remarkable.(4) in the experiment of heavy dose of this carrier of immunity repeatedly, any unusual allergy or autoimmune phenomena do not appear in laboratory animal.
Description of drawings
Fig. 1: pcDNA3.1-pre-miR-155 plasmid ideograph
Disturb Expression element to pass through the polyclone zone of BamH I and EcoR I site insertion pcDNA3.1 (+) carrier pre-microRNA-155, the carrier of has-MicroRNA-155 is expressed in acquisition.Green area inserts the site for the pre-microRNA-155 expressed sequence among the figure, and the multiple clone site beyond the green area still keeps, and can be used for the antigen encoding sequence and inserts.This plasmid contains ammonia benzyl mycin resistant gene.
Fig. 2: the vivoexpression checking of pcDNA3.1-pre-miR-155 plasmid
Wherein Fig. 2 A is the expression efficiency checking of MicroRNA-155 behind the independent transfection HEK293 cell of pcDNA3.1-pre-miR-155 plasmid (pMiR-155), and pcDNA3.1 empty plasmid (pcDNA) transfectional cell and untreated cell are in contrast; After Fig. 2 B is the common transfection HEK293 cell of pcDNA3.1-pre-miR-155 plasmid (pMiR-155) and nucleic acid vaccine pVAX 1-HBsAg (pHBs), verify the pHBs expression efficiency by HBsAg expression in the ELISA detection supernatant, common transfectional cell of pcDNA3.1 empty plasmid and pHBs (pcDNA+pHBs) and the independent transfectional cell of pHBs are in contrast; Fig. 2 C is that pcDNA3.1-pre-miR-155 plasmid (pMiR-155) is verified the expression efficiency of MicroRNA-155 behind the transfection HEK293 cell jointly with nucleic acid vaccine pVAX1-HBsAg (pHBs), and common transfectional cell of pcDNA3.1 empty plasmid and pHBs (pcDNA+pHBs) and the independent transfectional cell of pHBs are in contrast; All experiments all repeat 3 times, and what show among the figure is meansigma methods+standard deviation.
Fig. 3: the pcDNA3.1-pre-miR-155 plasmid is to the adjuvant effect checking of pVAX1-HBsAg nucleic acid vaccine
To the mix preparation of C57/BL mouse inoculation pcDNA3.1-pre-miR-155 plasmid and nucleic acid vaccine pVAX1-HBsAg (pHBs+pMiR-155), and with Mock plasmid pcDNA3.1 and nucleic acid vaccine pVAX1-HBsAg combined immunization (pHBs+Mock); Separately two groups of injection PBS in contrast.Immunity 3 times detects its immune indexes.Wherein Fig. 3 A is the change curve of respectively organizing in the mouse immune process with immunity time serum hepatitis B specific antibody level.Carry out the OD450 numerical value that the HBsAg specific ELISA detects after being expressed as serum dilution in 1: 20 among the figure, repeat 3 times and average.Fig. 3 B is 1 week after the last immunity, gets spleen lymphocyte behind the execution mice and with the hepatitis B surface antigen stimulation, adopts the detection of ELISPOT method respectively to organize the IL-4 and the IFN-γ secretion situation of mouse spleen T cell.3 meansigma methods+standard deviations that mice detects in having shown every group among the figure; Fig. 3 C after mouse spleen lymphocyte process antigen induction and cd4 cell, cd8 cell are rejected respectively, detects the ability that it kills and wounds the hepatitis B antigen positive target cell.Shown among the figure at the different effect cell: under the target cell ratio, the killing-efficiency curve of spleen lymphocyte.Numerical value is 3 meansigma methods+standard deviations that mice detects in every group among the figure.
The specific embodiment
Below in conjunction with drawings and Examples the present invention is described in detail, but embodiments of the invention only are used to the present invention is described and are not used in restriction protection scope of the present invention.
The structure of embodiment 1:pcDNA3.1-pre-miR-155 plasmid
We have at first searched the human microRNA-155 precursor molecule (GENBANK:NR_030784 of coding according to internet data library information (http://www.mirbase.org), shown in SEQ IDNO:1) genome sequence, at its flanking sequence design PCR primer (upstream and downstream is added BamH I and EcoR I restriction enzyme site respectively), adopt the extractive genomic DNA of human peripheral blood mononuclear cell to carry out PCR, microRNA-155 precursor molecule sequence and clone use PCR primer such as following table.Thereby obtained the dna fragmentation that comprises microRNA-155 precursor molecule coded sequence.
We are connected into above-mentioned clone's the dna fragmentation that comprises the pre-miR-155 coded sequence (shown in Fig. 1 and following sequence) in the polyclone zone available from the commercialization plasmid pcDNA.3.1 (+) of American I nvitrogene company, adopt BamH I and EcoR I restriction enzyme site.The polyclone enzyme action site of this insertion sequence flank still keeps, and can be used to insert antigen sequence does not influence its effective expression.Described pre-miR-155 expression vector: the pcDNA.3.1-pre-miR-155 that is.
The vivoexpression checking of embodiment 2:pcDNA3.1-pre-miR-155 plasmid
Utilize 293T cell (available from U.S. ATCC company) as external model, utilize Lipofectamine2000 transfection reagent (American I nvitrogene company operates with reference to its description) with pcDNA3.1-pre-miR-155 plasmid (pMiR-155) transfectional cell.Set up pcDNA3.1 (pcDNA) empty plasmid transfection group and untreated fish group (Untreated) in contrast.Adopt Trizol reagent (American I nvitrogene company after 24 hours, operate with reference to its description) the extracting cell total rna, adopt the microRNA real-time quantitative detection kit of Guangzhou?FulenGen?Co., Ltd., with reference to the expression of its company's description with the ripe body of has-microRNA-155 in the realtime PCR method detection cell, the expression of the ripe body of has-microRNA-155 rises above 1000 times (shown in Fig. 2 A) behind the discovery pcDNA3.1-pre-miR-155 plasmid transfection.
Embodiment 3:pcDNA3.1-pre-miR-155 plasmid and pVAX1-HBsAg nucleic acid vaccine cotransfection situation lower body appearance Danone power detect.
Utilize the 293T cell as external model, utilize the following three groups of plasmids of Lipofectamine2000 transfection reagent z transfection, 1.pVAX1-HBsAg and pcDNA3.1 empty plasmid group (pHBs+pcDNA) transfection; 2.pcDNA3.1-pre-miR-155 plasmid and pVAX1-HBsAg transfection (pHBs+pMiR-155); 3.pVAX1-HBsAg transfection (pHBs) separately.Adopt double-antibodies sandwich ELISA to detect the antigenic expression of HBsAg in the cell culture medium supernatant after 24 hours, after finding pcDNA3.1-pre-miR-155 plasmid and the common transfection of pVAX1-HBsAg, pVAX1-HBsAg still can efficiently express HBsAg antigen, with pVAX1-HBsAg expression suitable (shown in Fig. 2 B).
Above-mentioned 3 groups of cells of handling are adopted American I nvitrogene company's T rizol reagent extracting cell total rna simultaneously, adopt the expression (experimental technique with reference to company test kit description) of the microRNA real-time quantitative detection kit of Guangzhou?FulenGen?Co., Ltd. with the ripe body of has-miR-155 in the realtime PCR method detection cell, the expression of the ripe body of has-miR-155 rises above 1000 times (shown in Fig. 2 C) behind the discovery pcDNA3.1-pre-miR-155 plasmid transfection.
Through above-mentioned design and verification experimental verification, we proves in experiment in vitro, and the pcDNA3.1-pre-miR-155 plasmid can effective expression MicroRNA-155 molecule, and the nucleic acid vaccine that does not influence cotransfection carries out the vaccine antigen expression.
Embodiment 4: immune efficacy in the body after pcDNA3.1-pre-miR-155 plasmid and the common immunity of pVAX1-HBsAg nucleic acid vaccine is detected.
This embodiment main reference document (Wang, Y., Li, D., He, Y.and et al.Proteomicanalysis of augmented immune responses in mouse by prime-and-boostimmunization strategy with DNA vaccine coding HBsAg and rHBsAgprotein.Vaccine, 2007,25:8146-8153.) (Yang, F., Yan, S., He, Y., and et al.Expression of HBV Proteins in Transgenic Mice Disturbs Liver LipidMetabolism and Induces Oxidative Stress.J.Hepatol., 2008,48:12-19.) method carry out, be summarized as follows:
With pcDNA3.1-pre-miR-155 with 0.1mg/ only with pVAX1-HBsAg with 0.1mg/ dosage immunity 6-8 C57BL/6J mice in age in week, immunity is 3 times altogether, at interval 2 weeks, mix incomplete Freund's adjuvant booster immunization once with HBsAg recombiant protein (10 μ g/ only) the 6th weekend.
Set up following three groups of contrasts: 1.PBS blank group (PBS); 2.pcDNA3.1-pre-miR-155 the common immune group of plasmid and pVAX1-HBsAg (pHBs+pMiR-155); 3.Mock plasmid pcDNA3.1 and pVAX1-HBsAg be immunity (pHBs+Mock) separately.
The humoral immunization aspect through different immune time point tail vein bloods, adopts ELISA to detect serum hepatitis B surface antigen specific antibody titre; The cellular immunization aspect, 1 week was put to death mice (3 every group) after the last immunity, aseptic separating spleen lymphocyte, and with hepatitis b surface antigen protein (10 μ g/ml) processing 5 days, adopt the ELISPOT method to detect the IL-4 and the IFN-γ secretion situation of spleen t-cell, adopt external CTL to kill the target experiment and detect antigenic specificity CD8
+The T cell quantity.
Found that, pcDNA3.1-pre-miR-155 with 0.1mg/ only with pVAX1-HBsAg with the only common immunity of 0.1mg/ after, the antibody titer peak value does not have significant change with respect to the pVAX1-HBsAg immune group, but antibody peak value time of occurrence (as Fig. 3 A) in advance.And in cellular immunization detects, pcDNA3.1-pre-miR-155 with 0.1mg/ only with pVAX1-HBsAg with the only common immunity of 0.1mg/ after effective inducing antigen-specific ctl response, IFN-γ secretion positive cell significantly increases (as Fig. 3 B), the target cell killing-efficiency significantly improves (as Fig. 3 C), and prompting antigenic specificity cellular immunization obtains obviously to strengthen.
Claims (4)
1. nucleic acid vaccine adjuvant with cellular immunization enhanced activity is characterized in that this nucleic acid vaccine adjuvant is the plasmid that contains DNA sequence that can effective expression MicroRNA-155 precursor molecule, and DNA sequence wherein is shown in SEQ ID NO:4.
2. the nucleic acid vaccine adjuvant with cellular immunization enhanced activity according to claim 1 is characterized in that plasmid wherein is pcDNA.3.1 (+).
3. construction method with nucleic acid vaccine adjuvant of cellular immunization enhanced activity as claimed in claim 1 or 2 is characterized in that this method may further comprise the steps:
A) design PCR primer adopts the extractive genomic DNA of human peripheral blood mononuclear cell to carry out PCR shown in SEQ ID NO:2 and 3, and the DNA sequence of the MicroRNA-155 precursor molecule that obtains encoding is shown in SEQ ID NO:4;
B) DNA sequence with above-mentioned coding MicroRNA-155 precursor molecule is cloned into plasmid.
4. the construction method with nucleic acid vaccine adjuvant of cellular immunization enhanced activity according to claim 3 is characterized in that plasmid wherein is pcDNA.3.1 (+).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010280671 CN101972476B (en) | 2010-09-14 | 2010-09-14 | DNA vaccine adjuvant using Micro RNA-155 and construction method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010280671 CN101972476B (en) | 2010-09-14 | 2010-09-14 | DNA vaccine adjuvant using Micro RNA-155 and construction method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101972476A true CN101972476A (en) | 2011-02-16 |
| CN101972476B CN101972476B (en) | 2012-12-19 |
Family
ID=43572409
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010280671 Expired - Fee Related CN101972476B (en) | 2010-09-14 | 2010-09-14 | DNA vaccine adjuvant using Micro RNA-155 and construction method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101972476B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102776178A (en) * | 2011-05-12 | 2012-11-14 | 华中农业大学 | MicroRNA molecule marker associated with pig blood parameter properties, and its application |
| KR20170082534A (en) * | 2014-11-14 | 2017-07-14 | 보이저 테라퓨틱스, 인크. | Modulatory polynucleotides |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007081680A2 (en) * | 2006-01-05 | 2007-07-19 | The Ohio State University Research Foundation | Microrna expression abnormalities in pancreatic endocrine and acinar tumors |
-
2010
- 2010-09-14 CN CN 201010280671 patent/CN101972476B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007081680A2 (en) * | 2006-01-05 | 2007-07-19 | The Ohio State University Research Foundation | Microrna expression abnormalities in pancreatic endocrine and acinar tumors |
Non-Patent Citations (9)
| Title |
|---|
| 《Biochimica et Biophysica Acta》 20090630 Isabella Faraoni et al. miR-155 gene: A typical multifunctional microRNA 497-505 1-4 第1792卷, 第6期 * |
| 《The Journal of Immunology》 20071015 Esmerina Tili et al. Modulation of miR-155 and miR-125b Levels following Lipopolysaccharide/TNF-alpha Stimulation and Their Possible Roles in Regulating the Response to Endotoxin Shock 5082-5089 1-4 第179卷, 第8期 * |
| 《The Journal of Immunology》 20071015 Esmerina Tili et al. Modulation of miR-155 and miR-125b Levels following Lipopolysaccharide/TNF-alpha Stimulation and Their Possible Roles in Regulating the Response to Endotoxin Shock 5082-5089 1-4 第179卷, 第8期 2 * |
| 《医学研究生学报》 20090331 肖斌等 微小RNA在免疫调节中的作用研究进展 303-305,310 1-4 第22卷, 第3期 * |
| 《生命科学》 20100228 李江,胡维新 microRNA-155 研究进展 139-142 1-4 第22卷, 第2期 * |
| ESMERINA TILI ET AL.: "Modulation of miR-155 and miR-125b Levels following Lipopolysaccharide/TNF-α Stimulation and Their Possible Roles in Regulating the Response to Endotoxin Shock", 《THE JOURNAL OF IMMUNOLOGY》, vol. 179, no. 8, 15 October 2007 (2007-10-15), pages 5082 - 5089, XP002716885 * |
| ISABELLA FARAONI ET AL.: "miR-155 gene: A typical multifunctional microRNA", 《BIOCHIMICA ET BIOPHYSICA ACTA》, vol. 1792, no. 6, 30 June 2009 (2009-06-30), pages 497 - 505, XP026150247, DOI: doi:10.1016/j.bbadis.2009.02.013 * |
| 李江,胡维新: "microRNA-155 研究进展", 《生命科学》, vol. 22, no. 2, 28 February 2010 (2010-02-28), pages 139 - 142 * |
| 肖斌等: "微小RNA在免疫调节中的作用研究进展", 《医学研究生学报》, vol. 22, no. 3, 31 March 2009 (2009-03-31) * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102776178A (en) * | 2011-05-12 | 2012-11-14 | 华中农业大学 | MicroRNA molecule marker associated with pig blood parameter properties, and its application |
| KR20170082534A (en) * | 2014-11-14 | 2017-07-14 | 보이저 테라퓨틱스, 인크. | Modulatory polynucleotides |
| JP2021100435A (en) * | 2014-11-14 | 2021-07-08 | ボイジャー セラピューティクス インコーポレイテッドＶｏｙａｇｅｒ Ｔｈｅｒａｐｅｕｔｉｃｓ，Ｉｎｃ． | Regulatory polynucleotide |
| EP3907287A1 (en) * | 2014-11-14 | 2021-11-10 | Voyager Therapeutics, Inc. | Modulatory polynucleotides |
| US11198873B2 (en) | 2014-11-14 | 2021-12-14 | Voyager Therapeutics, Inc. | Modulatory polynucleotides |
| JP7256223B2 (en) | 2014-11-14 | 2023-04-11 | ボイジャー セラピューティクス インコーポレイテッド | regulatory polynucleotide |
| KR102584655B1 (en) * | 2014-11-14 | 2023-10-06 | 보이저 테라퓨틱스, 인크. | Modulatory polynucleotides |
| JP7469538B2 (en) | 2014-11-14 | 2024-04-16 | ボイジャー セラピューティクス インコーポレイテッド | Regulatory Polynucleotides |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101972476B (en) | 2012-12-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20240091340A1 (en) | Pre-immunization and immunotherapy | |
| Vargas-Inchaustegui et al. | Distinct roles for MyD88 and Toll-like receptor 2 during Leishmania braziliensis infection in mice | |
| Bridle et al. | Oncolytic vesicular stomatitis virus quantitatively and qualitatively improves primary CD8+ T-cell responses to anticancer vaccines | |
| JP6118491B2 (en) | Methods for increasing immune response | |
| Salek-Ardakani et al. | Targeting OX40 promotes lung-resident memory CD8 T cell populations that protect against respiratory poxvirus infection | |
| Villarreal et al. | Ubiquitin-like molecule ISG15 acts as an immune adjuvant to enhance antigen-specific CD8 T-cell tumor immunity | |
| Coban et al. | Effect of plasmid backbone modification by different human CpG motifs on the immunogenicity of DNA vaccine vectors | |
| Bahadoran et al. | Induction of a robust immune response against avian influenza virus following transdermal inoculation with H5-DNA vaccine formulated in modified dendrimer-based delivery system in mouse model | |
| Miyahira et al. | Immune responses against a single CD8+-T-cell epitope induced by virus vector vaccination can successfully control Trypanosoma cruzi infection | |
| Soltani et al. | DNA vaccine: Methods and mechanisms | |
| CN103275971A (en) | RNA interference targets for hepatitis b virus (HBV) infection treatment | |
| CN101972476B (en) | DNA vaccine adjuvant using Micro RNA-155 and construction method thereof | |
| Wang et al. | The 3′ CCACCA sequence of tRNAAla (UGC) is the motif that is important in inducing Th1-like immune response, and this motif can be recognized by Toll-like receptor 3 | |
| CN101701218B (en) | Immune regulatory oligodeoxynucleotide for inhibiting differentiation of Th1 and Th17 and application thereof | |
| CN103215274B (en) | Epinephelus coioide interferon IFNgamma 2 and its preparation method and application | |
| CN102660579B (en) | HBx and human IL-12 double-gene recombinant vector and liver caner-resistant vaccine | |
| CN103243106B (en) | Epinephelus coioides interferon IFNgamma1 and preparation method and application thereof | |
| CN102000330B (en) | Nucleic acid vaccine adjuvant and construction method thereof | |
| Mao et al. | PD-1 blockade-driven anti-tumor CD8+ T cell immunity requires XCR1+ dendritic cells | |
| Coupet et al. | Intravenous injection of a novel viral immunotherapy encoding human interleukin-7 in nonhuman primates is safe and increases absolute lymphocyte count | |
| CN1324661A (en) | Human hepatitis B nucleic acid vaccine | |
| Zhu et al. | Plasmid vector-linked maturation of natural killer (NK) cells is coupled to antigen-dependent NK cell activation during DNA-based immunization in mice | |
| CN101629182B (en) | Eukaryotic expression plasmid of chicken defensin-1 gene and molecular adjuvants prepared by the gene and vaccine | |
| CN102038951B (en) | Nucleic acid vaccine adjuvant designed based on MCP-1 and construction method thereof | |
| CN103239717A (en) | Replicon DNA (Deoxyribonucleic Acid) vaccine for treating chronic hepatitis B |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20121219 Termination date: 20150914 |
|
| EXPY | Termination of patent right or utility model |