Mouse immune state monitoring reagent and its preparing process

Abstract

This invention provides a kind of mouse immunity state test reagent, this reagent is PBS suspension of mouse spleen cell in two inbreeding system which is stained by CFSE, the total concentration of the two kind of mouse spleen cell is 0.5-1 multiply 10 to the power 8 per ml, the proportion is 1:1, fluorescence tinction differential concentration is 1:20. Mainline the reagent in the model mouse; it can judge the immunity state of the model mouse by the change of the proportion of the two cell in this reagent when letting blood to test.

Description

technical field [0001] The present invention relates to medical preparations, in particular to medical preparations derived from animal materials. Background technique [0002] Transplantation rejection is the loss of graft function or damage to the recipient's body caused by the immune response induced by transplantation antigens. It is the main reason for clinical transplantation failure and the key problem that transplantation immunology is trying to overcome. In allografts, even if the surgery is successful, the graft will not survive long-term due to rejection. Therefore, transplant rejection has always been the focus of research in the medical field. In the research process of transplant rejection, the detection of the recipient's immune status is an essential link. However, there is still no reliable reagent / method to detect the immune status of the receptor. The existing detection of the immune status of the receptor mainly reflects the immune status indirectly by d...

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Problems solved by technology

[0009] 6. In the induction of immune tolerance, chimera status or gene therapy may become an indicator of immune detection in the future

51Cr release test is still the most commonly used standard test method for detection of specific T lymphocyte toxicity. The results of this method are accurate and reproducible; but there are also the following shortcomings: ① The use of radioactive 51Cr has the risk of r-ray radiation, which is not conducive to safe operation and waste disposal, and special measuring instruments are required; ②The spontaneous release rate of 51Cr is high, and some cell labeling problems and low sensitivity often affect the result judgment due to the large difference in the labeling efficiency of different target cells; ③The half-life of 51Cr (27.8 days), cannot be used Suitable for animal experiments that require multiple determinations; ④The co-incubation time of cells is short and the test operation steps are many, so it cannot be measured at the level of a single cell

Moreover, the 51Cr release test is an in vitro test. The test itself only measures the activity of CTL and NK cells, which can only reflect the status of cellular immunity to a certain extent, and cannot fully reflect the complex immune environment in the body.

[0014] 2. Using the ELISA method to use the cytokine concentration in the recipient serum as a monitoring index, and the effectiveness of using the RT-PCR method to detect the cytokine mRNA in the graft biopsy specimens have been recognized by some scholars, but cytokines are a Complex network, single cytokine cannot comprehensively reflect immune status

But graft biopsy pathological examination also has its limitation: one. Biopsy tissue detection is a kind of destructive examination method, has certain influence to the normal survival of graft, can not take sample repeatedly, in addition, takes too few samples and will Affect the correctness of pathological slice reading

Second, biopsy pathological examination is a non-quantitative index, which affects the accuracy of clinical diagnosis. Pathological examination is easily affected by the subjective factors and technical level of the reader, resulting in bias and lack of objectivity

3. Pathological biopsy cannot judge the function of infiltrating lymphocytes, which is not conducive to the early diagnosis of rejection. In the early stage of rejection or recipients with graft failure, even clinically stable recipients , pathological biopsy sections can see a large number of T lymphocyte infiltration, or only dotted T cell infiltration

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