Secondary T-cell-dependent antibody responses to detect immunosuppressive methods for exogenous compounds

An antibody reaction, exogenous technology, applied in the screening of compounds, detection of programmed cell death, biological testing, etc., can solve specific T cell-dependent antigen insensitivity, large amount of tested exogenous compounds, difficult early stage Drug screening and other issues, to achieve the effect of short immune response time, small inter-individual differences, and strong response

Active Publication Date: 2016-01-13
SHANGHAI INNOSTAR BIO TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to overcome the defects that the existing TDAR technology is not sensitive to specific T cell-dependent antigens, the experimental period is long, and the amount of exogenous compounds used is large, which makes it difficult to use in early drug screening. A Secondary T Cell Dependent Antibody Response Method for Detecting Immunosuppression by Exogenous Compounds

Method used

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  • Secondary T-cell-dependent antibody responses to detect immunosuppressive methods for exogenous compounds
  • Secondary T-cell-dependent antibody responses to detect immunosuppressive methods for exogenous compounds
  • Secondary T-cell-dependent antibody responses to detect immunosuppressive methods for exogenous compounds

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Experimental method: Select 20 SD female rats with good growth and the same size, and inject 300 μg / kg KLH into the tail vein; 21 days after the first injection of KLH, inject 300 μg / kg KLH intravenously again to stimulate secondary immunity Response, the rats to be tested were equally divided into 4 groups:

[0033] At the same time as the second injection of KLH, deionized water, 7.5, 15, and 30 mg / kg cyclosporine were given orally once respectively; peripheral blood was collected, serum was separated, and anti-KLHIgG and peripheral blood white blood cell count were detected by ELISA method 10 days after the last oral administration of deionized water and different concentrations of cyclosporine, the organs were dissected for histopathological examination.

[0034] Experimental results: peripheral blood leukocyte counts showed that secondary immune 15, 30mg / kg cyclosporine group appeared immune function suppression as shown in Table 1;

[0035] The effects of differe...

Embodiment 2

[0037] Experimental method: Select 15 SD female rats with good growth and the same size, and inject 300 μg / kg KLH into the tail vein; 21 days after the first injection of KLH, inject 300 μg / kg KLH intravenously again to stimulate secondary immunity Response, the rats to be tested were divided into 3 groups on average:

[0038] At the same time as the second injection of KLH, deionized water and cyclophosphamide at 5 and 20 mg / kg were given orally once respectively; peripheral blood was collected, serum was separated, and anti-KLHIgG and peripheral blood white blood cell count were detected by ELISA method. On the 10th day after oral administration of deionized water and cyclophosphamide of different concentrations, the organs were dissected for histopathological examination.

[0039] Experimental results: the peripheral blood leukocyte count shows that the secondary immune function of 20mg / kg cyclophosphamide is suppressed as shown in Table 1; it can be seen that after 3 days ...

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Abstract

The invention discloses a method for detecting exogenous compound immunosuppression through secondary TDAR (T cell-dependent antibody response). The method comprises the following steps of (1) injecting T cell-independent antigens into an animal; (2) injecting the same T cell-independent antigens again 15 to 25 days after the injection in the step (1) to excite secondary immune response, and starting to continuously feed exogenous compounds to the animal for 2 to 5 days through the mouth; and (3) after 6-10 days since the step (2) is ended, collecting peripheral blood, separating serum, and detecting a specific antibody of the antigens in the step (1). The method is short in immune response time and high in response degree; the number of the required exogenous compounds is small; furthermore, the method can be applied to early medicine screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting immunosuppression of exogenous compounds by secondary T cell-dependent antibody response. Background technique [0002] Immunotoxicity is any permanent or reversible change in the structure or function of the immune system caused by a foreign substance. Immunotoxic effects are mainly divided into 5 types: immunosuppression, immune enhancement, hypersensitivity, autoimmunity, and immunogenicity. Among them, immunosuppression is the most common and has been studied more in immunotoxicity. [0003] T cell-dependent antibody response (Tcell-dependent antibody response, TDAR) is currently used to detect exogenous compound immunosuppression gold standard. TDAR means that when animals are given exogenous compounds, they are attacked by T cell-dependent antigens at the same time. After continuing to administer exogenous compounds for a certain period of time, serum i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6803G01N2500/00
Inventor 马璟洪敏李华周国民程一帆林曼
Owner SHANGHAI INNOSTAR BIO TECH
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