Recombinant adenovirus and application thereof

A technology of recombinant adenovirus and recombinant vector, which is applied in the field of genetic engineering and can solve problems such as insensitivity to treatment

Inactive Publication Date: 2011-08-31
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the focus is on inhibiting the activity of pro-inflammatory factors TNF-α and IL-1β, and the use of antibodies against these two pro-inflammatory factors and their receptor fusion proteins has also achieved certain results in clinical applications. However, there are still some Patients are insensitive to treatments that block both factors

Method used

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  • Recombinant adenovirus and application thereof
  • Recombinant adenovirus and application thereof
  • Recombinant adenovirus and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1, the construction of recombinant adenovirus

[0031] 1. Formation of human interleukin-17 receptor D extracellular domain Fc fusion protein gene

[0032] Artificially synthesized sequence 1 was used as a template, and Sense primer: 5'-ATAAAGCTTATGGCCCCGTGGCTGC-3'Antisense primer: 5'-ATTCTCGAGTCATTTACCCGGGAGACAG-3' was used as a primer, and it was digested with Hind III and EcoRV, and the 1536bp target band was recovered from the gel. The recovered fragment was ligated into Ad-Track-CMV (purchased from Stratagene, catalog #240009) double-digested with the same restriction endonuclease, and the bacteria Top10 (invitrogen Catalog #C404003) was transformed to obtain a transformant. Extract the plasmid from the transformant and send it for sequencing. The result is that the nucleotide (the nucleotide of hIL-17RD-ECD-Fc) shown in sequence 1 in the sequence listing is a positive plasmid, and the plasmid is the sequence in the sequence listing. 1 was inserted into...

Embodiment 2

[0046] Embodiment 2, the application of recombinant adenovirus

[0047] 1. Construction of CIA mice

[0048] Take 40 male DBA-1 mice (DBA-1 mice, Shanghai Experimental Animal Center, Chinese Academy of Sciences) (10-12 weeks old) according to Erik Lubberts (IL-1-Independent Role of IL-17 in Synovial Inflamation and Joint Destruction During Collagen- Induced Arthritis, J.Immunol.2001, 167:1004-1013) and other methods for sensitization: Antigen Bovine type II collagen (CII) (bovine type II collagen SIGMA C7806-10MG) was diluted with 0.05M acetic acid to a concentration of 2mg / ml, and then emulsify this concentration of CII with an equal volume of complete Freund's adjuvant (CFA). After emulsification, 100 ul of CII / mouse was subcutaneously injected into the tail of the mouse (the body weight of each mouse was about 25 g.) to obtain allergic mice. Arthritis was observed as early as 21 days after injection, with significant redness or swelling in the knee and ankle joints of th...

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Abstract

The invention discloses a recombinant adenovirus and application thereof. The recombinant adenovirus is obtained by transferring an encoding gene of an interleukin 17 receptor D extracellular region Fc fusion protein to a host cell, wherein the amino acid sequence of the interleukin 17 receptor D extracellular region Fc fusion protein is a sequence 2 in a sequence table; and through a recombinant vector, the interleukin 17 receptor D extracellular region Fc fusion protein is transferred to the host cell. The experiment in the invention proves that a CIA (collagen in-duced arthritis) mice model is constructed by using an adenovirus vector of constructed and purified Ad / Hil-17RD-ECD-Fc, the prevention and treatment on the CIA mice are achieved by using the Ad / Hil-17RD-ECD-Fc, and finally, the alleviation degrees of CIA mice arthritis and bone injury are estimated and analyzed through He dyeing. Thus, a novel and effective method is provided for treating rheumatoid arthritis.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a recombinant adenovirus and its application. Background technique [0002] Rheumatoid arthritis (RA) is a chronic autoimmune system disorder, and its clinical manifestations are mainly symmetrical joint swelling and pain, erosive synovitis, and even progressive joint damage, deformity, disability, sometimes involving Eyes, lungs and other extra-articular organs. This disease afflicts 1-2% of the population worldwide, most of whom are middle-aged women, and most patients have a chronic relapsing course. The morbidity rate of rheumatoid arthritis is very high, and its treatment has always been a focus of attention. At present, the treatment of rheumatoid arthritis mainly includes various methods such as non-steroidal anti-inflammatory drugs, slow-acting anti-rheumatic drugs, glucocorticoids, biological agents, autologous hematopoietic stem cell transplantation, and gene therap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/62C12N15/861C07K19/00A61K48/00A61P19/02A61P29/00A61P37/02
Inventor 常智杰孙小军杨世高王银银任芳丽夏永静
Owner TSINGHUA UNIV
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