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A kind of method of constructing Treponema pallidum mouse model

A Treponema pallidum and mouse model technology, applied in the biological field, can solve problems such as not easy to obtain, expensive rabbits, difficult to enter the next step, etc.

Inactive Publication Date: 2021-03-30
NANHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The primary animal model for syphilis research is the rabbit, but inbred rabbits are expensive and not readily available
Candidate vaccines in the existing research results can inhibit the spread of Treponema pallidum to distant organs, but there is a lack of reagents for studying humoral immunity and cellular immunity in rabbits, and it is difficult to go to the next step in the study of this protective mechanism

Method used

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  • A kind of method of constructing Treponema pallidum mouse model
  • A kind of method of constructing Treponema pallidum mouse model
  • A kind of method of constructing Treponema pallidum mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Construction of Treponema pallidum mouse model and verification of blood diffusion

[0019] Treponema pallidum was propagated and cultured by inoculating adult New Zealand white rabbits into the testes, and mice were inoculated with Treponema pallidum intradermally (between the shoulder blades), intraperitoneally, rectally (gastrically) and in the cavernous body of the penis, each site 2.5×10 6 Treponema pallidum (a total of 1 × 10 7 indivual). The rectal inoculation is administered by gavage, and the other parts are inoculated by injection. Mice were assessed every other day for signs of skin or systemic infection. Mice were sacrificed on day 30 after inoculation. Testes, lymph nodes (groin / brachial / armpit), spleen, liver, blood and brain tissues were collected for RT-qPCR analysis.

[0020] Mice immunization: mice were randomly assigned, 9 in each group. Mice were immunized by intramuscular injection, and both pcDNA / CpG-FlaB3 and pcDNA3 / FlaB3 could inh...

Embodiment 2

[0022] Embodiment 2: Treponema pallidum mouse model immunohistochemical verification

[0023] Mouse model construction and immunization are the same as in Example 1

[0024] Biopsy punch samples (2 mm) were taken from the testis, liver and spleen of each mouse 30 days after infection and sent to South China University. Tissues were fixed in formalin and stained with hematoxylin and eosin or immunohistochemically stained with anti-T ( figure 2 ).

[0025] figure 2 The results showed that a large number of Treponema pallidum was found in the testis, liver and spleen of the control group, while the load of Treponema pallidum was significantly reduced in the testis, liver and spleen of the infected mice immunized with pcDNA3 / FlaB3 and pcDNA3 / CpG-FlaB3.

Embodiment 3

[0026] Example 3: Verification of lymph node spread in treponema pallidum mouse model

[0027] 1) After 30 days of Tp infection, aseptically isolate the inguinal, brachial plexus and axillary lymph nodes of C57BL / 6 mice (3 mice / group), place them in a sterile petri dish, and add 1 mL of normal saline;

[0028] 2) Fully grind the lymph nodes with the soft part of a medical syringe (5 mL), place the culture dish on a platform rotator for 10 minutes, remove the lymph node fragments, and absorb 2 mL of the bacterial solution;

[0029] 3) Infect three New Zealand rabbits with the Tp bacterial solution collected from each group of mice, and inject 1 mL of the bacterial solution after the testis was disinfected with povidone iodine;

[0030] 4) Observe the testicular inflammation of New Zealand rabbits every day, and perform serological detection (RPR and TPPA) every other week;

[0031] 5) After 9 weeks of infection, the New Zealand rabbits were anesthetized and sacrificed, the tes...

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Abstract

The invention relates to the technical field of biology, and discloses a method for constructing a treponema palladium mouse model. The method comprises the steps that a treponema pallidum is inoculated to a rabbit for breeding, then, treponema pallidum is inoculated into mouse skin, peritoneum, rectum and corpus cavernosum, and the treponema palladium mouse model is obtained. In order for solve the problems that an existing has low reactivity for treponema pallidum infection, and no treponema pallidum mouse model exists, mice of the specific strain are adopted for inoculating treponema pallidum into skin, peritoneum, rectum and corpus cavernosum, and the treponema pallidum mouse model is constructed, the mouse model can detect treponema pallidum in blood, the liver, the spleen and the testis, dissemination of blood and lymph glands of the treponema pallidum can be achieved, the inhibition effect on treponema pallidum DNA vaccines can be reflected, and the requirement for being used asa treponema pallidum animal model is met.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a Treponema pallidum mouse model. Background technique [0002] Syphilis is a chronic sexually transmitted disease caused by Treponema pallidum. Currently, it remains a globally prevalent disease with an estimated 36 million cases worldwide. The incidence of syphilis among gay men has risen dramatically in recent years. The incidence of congenital syphilis infection is also on the rise, with an estimated 1.36 million pregnant women infected each year worldwide, approximately 520,000 of which result in miscarriage or stillbirth, etc. In addition, syphilis increases the risk of HIV infection. Despite the fact that Treponema pallidum remains sensitive to penicillin therapy, the prevalence of syphilis continues to increase worldwide, thus creating an urgent need to develop effective syphilis vaccines to complement traditional screening and treatment approache...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/74A61K49/00
Inventor 吴移谋郑康徐嫚
Owner NANHUA UNIV
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