CN111407883B - New use of IFN-lambda 3 in toxoplasma gondii infection - Google Patents
New use of IFN-lambda 3 in toxoplasma gondii infection Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/21—Interferons [IFN]
- A61K38/217—IFN-gamma
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
Abstract
The invention belongs to the technical field of biomedical treatment, and relates to a novel application of IFN-lambda 3 in toxoplasma gondii infection. The invention discloses an application of IFN-lambda 3 in preparing a medicament for treating or preventing toxoplasma gondii infection. The invention also discloses a composition for preventing or treating toxoplasma gondii infection, which is characterized in that the active ingredient of the composition is IFN-lambda 3. The invention also provides a use of a composition for the preparation of a medicament for the prophylaxis or treatment of toxoplasma gondii infection, wherein the composition comprises IFN-lambda 3 and one or more pharmaceutically acceptable carriers. The invention provides a new way and a new method for treating toxoplasma gondii infection, and solves the problem that bad pregnancy caused by toxoplasma gondii infection cannot be treated in the prior art.
Description
Technical Field
The invention belongs to the technical field of biomedical treatment, and relates to a novel application of IFN-lambda 3 in toxoplasma gondii infection, in particular to an application of IFN-lambda 3 in preparing a medicament for treating or preventing toxoplasma gondii infection, a composition for preventing or treating toxoplasma gondii infection, and an application of the composition in preparing a medicament for preventing or treating toxoplasma gondii infection.
Background
Toxoplasma gondii (Toxoplasma gondii) is an important opportunistic pathogenic protozoan. Individuals with normal immunity often present with a recessive infection after infection with toxoplasma. For immunocompromised or immunodeficiency patients, such as aids, organ transplants, and malignant patients, toxoplasma infection is a significant cause of death. Infection of pregnant women can affect fetal development, resulting in congenital toxoplasmosis such as abortion, teratogenesis, stillbirth, premature birth, birth defects, etc. In recent years, as urban population increases sharply, the number of pets (particularly cats) kept is increasing, and more people are exposed to the risk of infection of toxoplasma due to bad eating habits such as wild animals. About 10 million people worldwide are infected with toxoplasma, wherein the infection rate of pregnant women is 10% -27.5%, and about 9 tens of thousands of newborns are threatened by toxoplasma infection every year. China, a large population, is a great challenge to us how to effectively prevent and treat congenital toxoplasmosis.
Interferons (IFNs) are largely classified as type 3: type I interferons (IFN- α and IFN- β), type II interferons (IFN- γ), and type III interferons (IFN- λ). Type I IFNs (IFN-alpha and IFN-beta) mainly have antiviral effects by inducing expression of various interferon activating genes (ISGs) through related signal paths. Type II interferon (IFN- γ) is an important cytokine that inhibits toxoplasma proliferation. After the toxoplasma tachyzoite invades the body, macrophages are stimulated to produce IL-12, so that NK cells and T cells are activated to secrete IFN-gamma. IFN-gamma then induces IFN-gamma-inducible genes expression, which in turn directly kills tachyzoites that are parasitic in the host cell. However, during pregnancy, IFN-gamma production and excessive secretion are the main factors in the occurrence of bad pregnancy. IFN-gamma is recruited by CD49b + NK, while regulating expression of NK cell Ly-49 receptor, leads to occurrence of abortion. In the rat abortion model, IFN-gamma upregulates tumor necrosis factor-alpha (TNF-alpha) while downregulating Matrix metalloproteinases-2 and-9 (MMP-2 and MMP-9) expression, further exacerbating abortion. In human pregnancy, the effect of IFN-gamma leading to poor pregnancy was also demonstrated, and T cells of fetal origin promote uterine contractions by secreting IFN-gamma and TNF-alpha, leading to the occurrence of premature labor. Thus, IFN-gamma does not protect the mother against toxoplasma infection during pregnancy, maintaining normal pregnancy.
IFN-lambda (type III interferon) has a relatively independent and specific ability to resist pathogen infection. The IFN-lambda receptor consists of Interleukin 28 receptor alpha (Interleukin-28 receptor alpha,IL-28 Ralpha) and Interleukin 10 receptor beta (Interleukin-10 Receptor Beta,IL-10 Rbeta), wherein the IL-10 Rbeta is widely existing in cells and tissues, and the IL-28 Ralpha is mainly expressed on the surfaces of epithelial cells, neutrophils and liver cells. The limitations in receptor expression mean that IFN-lambda acts as a relatively independent and specific pathogen-resistant agent in certain tissues. The human IFN-lambda family consists of IFN-lambda 1, IFN-lambda 2, IFN-lambda 3 and IFN-lambda 4, whereas mice have only two functional IFN-lambda, IFN-lambda 2 and IFN-lambda 3. In rotavirus studiesThe researcher finds that IFN-lambda 2 can obviously inhibit rotavirus replication in intestinal tracts of young mice, and IFN-lambda receptor knockout promotes rotavirus proliferation in mice. Mu ir et al have proposed for the first time that IFN-. Lamda.1a inhibits the proliferation of hepatitis C virus (hepatitis C virus, HCV). IFN-. Lambda.1a can inhibit HCV replication rapidly (within 12 hours) and achieve a similar effect to IFN-. Alpha.within 24 hours. However, the patient has significantly decreased the proportion of complications such as thrombocytopenia and neutropenia. Therefore, IFN- λ1a is expected to be clinically used for the treatment of HCV in place of IFN- α. Rebecca L et al model Aspergillus fumigatus (Aspergillus fumigatus, af) for the investigation of anti-fungal immune responses to find CCR2 + Depletion of monocytes reduces the ability of neutrophils to inhibit the growth of invasive fungi. IFN-lambda 2/3 acts directly on neutrophils to activate their antifungal response, whereas neutrophil-specific mice lacking the IFN-lambda receptor die from invasive aspergillosis. Transfer of CCR2 by adoptive transfer + Either monocytes or IFN- λ2/3 are effective in treating mice depleted of neutrophil CCR 2. Therefore, IFN- λ2/3 is a key regulator of neutrophils, exerting antifungal effect. Cryptosporidium parvum (Cryptosporidium parvum, C.parvum) acts as an important opportunistic pathogenic protozoa, causing a clinical manifestation with diarrhea as a major component. Exogenous IFN- λ3 can reduce the number of insect charges in intestinal epithelial cells (Intestinal Epithelial Cells, IECs), restore transmembrane resistance (Transepithelial electrical resistance, TEER), and resist C.parvum invasion.
Although a large amount of experimental data support the ability of IFN-. Lamda.2/3 to inhibit proliferation of viruses, fungi and Cryptosporidium parvum, whether IFN-. Lamda.3 can inhibit proliferation of Toxoplasma gondii was yet to be studied further.
Disclosure of Invention
The invention aims to provide an application of IFN-lambda 3 in preparing a medicament for treating or preventing toxoplasma gondii infection, a composition for preventing or treating toxoplasma gondii infection and an application of the composition in preparing a medicament for preventing or treating toxoplasma gondii infection, so as to solve the problems in the prior art.
In view of the above objects, embodiments of the present invention provide the use of IFN- λ3 for the manufacture of a medicament for the treatment or prevention of Toxoplasma gondii infection.
Further, the IFN-lambda 3 intervenes in up-regulating expression of CK functional molecules in placenta and inhibiting proliferation of toxoplasma gondii.
Embodiments of the present invention also provide a composition for preventing or treating toxoplasma gondii infection, wherein the active ingredient of the composition is IFN-lambda 3.
Further, the composition includes IFN- λ3 and one or more pharmaceutically acceptable carriers.
Wherein the carrier comprises pharmaceutically acceptable diluents, excipients, fillers, binders, absorption promoters, surfactants and synergists.
Embodiments of the invention additionally provide for the use of a composition for the manufacture of a medicament for the prevention or treatment of toxoplasma gondii infection, wherein the composition comprises IFN-lambda 3 and one or more pharmaceutically acceptable carriers.
Further, the carrier includes pharmaceutically acceptable diluents, excipients, fillers, binders, absorption promoters, surfactants and synergists.
The technical scheme of the invention has the following beneficial effects:
(1) According to the embodiment of the invention, a plurality of groups of pregnant mouse models are prepared, and experiments of the pregnant mouse models prove that IFN-lambda 3 intervention can obviously reduce the flow rate caused by toxoplasma infection, reduce the damage of mouse placenta caused by toxoplasma infection and up-regulate the expression of functional molecules such as CK in placenta; meanwhile, after the toxoplasma gondii and JEG-3 cells are co-cultured, the expression of toxoplasma gondii SAG1 in each cell culture model group is detected by PCR, and the IFN-lambda 3 intervention can inhibit proliferation of toxoplasma gondii, so that the embodiment of the invention verifies that the IFN-lambda 3 intervention can treat bad pregnancy caused by toxoplasma gondii and inhibit proliferation of toxoplasma gondii, and provides a theoretical basis for treating toxoplasma gondii infection.
(2) The invention provides an application of IFN-lambda 3 in inhibiting toxoplasma gondii infection, a medicament for preventing or treating toxoplasma gondii infection, a novel approach and a novel method for treating toxoplasma gondii infection are provided, and the problem that bad pregnancy caused by toxoplasma gondii infection cannot be treated in the prior art is solved.
Drawings
FIG. 1 is a diagram showing abortion in a group of Toxoplasma gondii infected with the same species as those of the pregnant mice in example 1 of the present invention, after the pregnant mice were pretreated with IFN-. Lambda.3 for 24 hours;
FIG. 2 is a graph showing the damage of the placenta of mice infected with Toxoplasma gondii in example 2 of the present invention, after the mice were not infected with Toxoplasma gondii, the mice were infected with Toxoplasma gondii, and the mice were pretreated with IFN-. Lambda.3 for 24 hours;
FIG. 3 is a graph showing the expression of placenta CK in the toxoplasma gondii infected group after IFN-. Lambda.3 pretreatment for 24 hours in the pregnant mice, the toxoplasma gondii infected group and the pregnant mice infected group in example 3 of the present invention;
FIG. 4 is a graph showing the expression of toxoplasma SAG1 in toxoplasma gondii infected groups, in which the cells were not infected with toxoplasma gondii, in which the cells were subjected to co-stimulation with toxoplasma gondii and IFN-. Lamda.3, in which the cells were subjected to pretreatment with IFN-. Lamda.324 h, in which the toxoplasma gondii, in which the cells were subjected to co-treatment with toxoplasma gondii and IFN-. Gamma.24 h, and in which the cells were subjected to pretreatment with IFN-. Gamma.24 h, in example 4 of the present invention.
Detailed Description
In order to make the technical problems, technical solutions and advantages to be solved by the present invention more apparent, the following detailed description will be made with reference to specific embodiments.
Experimental materials
1.1, JEG-3 cell line was purchased from Thermo Fisher Scientific.
1.2 Trizol reagent was purchased from Invitrogen, inc. of America.
1.3, reverse transcription reagents were purchased from Bio-Rad, inc. of America.
1.4, SYBR Green Master Mix, taqMan Universal Master Mix are available from U.S. Thermo Fisher Scientific.
1.5 primers were synthesized by Sigma, america.
1.6, fetal bovine serum was purchased from Hyclone Inc.
1.7, pancreatin and MEM were purchased from Thermo Fisher Scientific company in the united states.
Preparation of pregnant mouse experiment
At least 24 mice and 48 mice were divided into 24 groups, each group consisting of 2 mice and one mouse, 2 mice and 1 mouse were caged at 5 pm in the morning and white pessaries were detected at 7 pm. If a vaginal suppository is detected, the pregnancy of the mouse is determined to be E0.5 (electronic day 0.5).
The 24 groups of pregnant mice were grouped and prepared into model groups of different groups, the specific groupings were as follows:
(1) Pregnant mice were not infected with toxoplasma gondii group (normal pregnant group);
(2) A pregnant mouse infects toxoplasma gondii group;
(3) After the pregnant mice are pretreated by IFN-lambda 3 for 24 hours, the pregnant mice are infected with toxoplasma gondii;
examples 1 to 3 of the present invention three groups of pregnant mice, at least four in each group (the pregnancy of each pregnant mouse is the same), were prepared and the experiments of examples 1 to 3 below were performed.
Example 1 IFN- λ3 intervention significantly reduces abortion rate due to toxoplasma infection
Pregnant mice were injected with IFN-. Lamda.3 (2. Mu.g/ml) on day 9.5 (E9.5) of gestation, and on day E10.5, pregnant mice were infected with Toxoplasma gondii. Subsequently, pregnant mice were injected daily with IFN- λ3 until 18.5 days of gestation. The change in weight of pregnant mice was monitored, and the fluid productivity of pregnant mice, the weight of the fetal mice, and the size of the fetal mice were observed. In the middle gestation period, the pregnant mice are infected with toxoplasma gondii, and the weight of the pregnant mice is reduced compared with the weight of the pregnant mice. Killing by cutting at E18.5 days, passing through the length of the top buttocks (mm) x the occipital frontal diameter (mm) 2 ) To measure the size of the fetal mice while observing the flow rate. And collecting each tissue sample in the female mouse and the fetal mouse, and carrying out subsequent experiments.
As a result, as shown in FIG. 1, infection with Toxoplasma gondii in the pregnant mice infected with Toxoplasma gondii group resulted in an abortion rate of about 58.9% compared with the group (normal pregnant group) in which the pregnant mice were not infected with Toxoplasma gondii. And after IFN-lambda 3 is dried, after pregnant mice are pretreated by IFN-lambda 3 for 24 hours, the abortion rate of the pregnant mice in the toxoplasma gondii infection group is obviously reduced.
These results suggest: IFN- λ3 can improve the poor pregnancy outcome caused by toxoplasma infection.
EXAMPLE 2 IFN- λ3 reduces injury to the mouse placenta from toxoplasma infection
After infection of pregnant mice with toxoplasma on day E10.5, the mouse placenta was collected on day E18.5 and HE stained with formalin to observe the structural changes of the placenta.
The method of HE staining is as follows: the slide was rinsed with tap water for 3min. Distilled water was rinsed for 1min. The slide was permeabilized in a pre-chilled 3% Triton X-100 solution at 4℃for 5min. Hematoxylin staining for 10min. The tap water was rinsed for 30s. Differentiation solution (2% ethanol hydrochloride) for 20s. And washing with tap water to blu for 5-10min. Eosin staining for 30s-2min. And (3) carrying out gradient dehydration on 50%, 70%, 80%, 95% absolute ethyl alcohol for 1-3min. Xylene I, II is transparent for 1-3min. And (3) sealing the sheet by neutral resin, and shooting images under a microscope.
As can be seen from the HE staining results, the normal structure of the placenta of normal mice was divided into Decidua (DE), junction Zone (JZ) and Labyrinth Zone (Labyrinth Zone, LZ). In comparison with the group of Toxoplasma gondii not infected with the pregnant mice (normal gestation group), the infection of Toxoplasma gondii in the group of Toxoplasma gondii destroys the normal structure of placenta, which is manifested by thinning of decidua layer and junction region, accompanied by a significant decrease in the number of cells in the labyrinthine region of placenta, as shown in FIG. 2, suggesting a decrease in placenta nutrient supply and gas exchange disorder. After IFN-lambda 3 intervention, pregnant mice are pretreated for 24 hours by IFN-lambda 3, and in the toxoplasma gondii infection group, the thickness of decidua layers and connecting areas is close to that of normal mouse placenta, and meanwhile, the cell number of the labyrinth area is obviously increased.
These results indicate that toxoplasma infection disrupts the normal structure of the mouse placenta, while IFN- λ3 significantly reduces the damage to the placenta.
Example 3 IFN- λ3 intervention up-regulates expression of functional molecules such as CK in placenta
Cytokeratin (CK) is a marker protein of mouse placental trophoblast cells.
The method for detecting the cell immunofluorescence comprises the following steps: the placenta tissue of the mice is collected and is prepared into 6-8 micron slices. When the slices are dyed, the slices are dried for 15 minutes at room temperature. Then soaking in PBS for 10min to remove OCT;0.5% Triton X-100 (PBS) was allowed to permeate for 20min at room temperature; sections were blocked with PBS containing 10% normal goat serum for 1h at room temperature; the diluted primary antibody was dropped onto a slide and incubated overnight at 4 ℃. After 3 washes with PBS, the diluted fluorescent secondary antibodies were dropped onto the slide and incubated at room temperature for 90min in the dark. After 3 washes in PBS, hoechst stain was dropped onto the slide and incubated at room temperature in the dark for 15min. After 3 washes with PBS, 50% glycerol lock up. And shooting images by using laser confocal.
In normal gestational groups (pregnant mice were not infected with toxoplasma gondii), a large amount of CK expression was observed in the mouse placenta, whereas in pregnant mice infected with toxoplasma gondii, the number of CK in the mouse placenta was significantly reduced, indicating that toxoplasma gondii was able to reduce the number of mouse placental trophoblast cells. As shown in FIG. 3, after pregnant mice were pretreated with IFN- λ3 for 24 hours, the infection of toxoplasma gondii group, i.e. after IFN- λ3 intervention, the CK expression was significantly increased, suggesting that IFN- λ3 can alleviate placenta injury and improve placenta function.
EXAMPLE 4 IFN- λ3 intervention to inhibit proliferation of toxoplasma gondii
In this example, in vitro experiments using cell culture were validatedIFN-lambda 3 intervention inhibits toxoplasma gondii proliferation Performing reproduction; the cell model groups used in this example can be divided into the following six groups:the cells are not infected with toxoplasma gondii, the cells are subjected to the common stimulation of toxoplasma gondii and IFN-lambda 3, and after the cells are subjected to the pretreatment of IFN-lambda 3 for 24 hours, the toxoplasma gondii, the cells, the toxoplasma gondii and IFN-gamma common treatment, and the cells are subjected to the pretreatment of IFN-gamma for 24 hours, so that the toxoplasma gondii is infected.
The method for obtaining the toxoplasma gondii infected JEG-3 cell strain is as follows: JEG-3 cell line was cultured in a cell culture flask using MEM complete medium (containing 100. Mu.g/mL streptomycin and 100U/mL penicillin) containing 10% fetal bovine serum, and placed in 5% CO at 37 ℃C 2 In a cell incubator of (C)Culturing. The cells are replaced every other day with culture medium, and when the cells reach about 80% of fusion degree, the cells are passaged or subjected to subsequent experiments. 2X 10 5 After plating the JEG-3 cells on 6-well plates and culturing in a cell culture incubator for 24 hours, toxoplasma gondii was infected with the JEG-3 cell line at a ratio of a multiplicity of infection (multiplicity ofinfection, MOI) of 2.
After IFN-. Lambda.3 pretreatment of JEG-3 cells, it was co-cultured with toxoplasma gondii, and real-time PCR was used to detect SAG1 expression. The method for detecting the number of toxoplasma gondii is as follows: tissue or cells were collected, and a suitable weight of tissue or cells was taken and added to Trizol for cleavage, RNA was extracted, and reverse transcribed into cDNA using Oligo (dT) 18 using a reverse transcription kit. Frozen at-80 ℃. After infection of JEG3 cells with toxoplasma gondii diluted at a 10-fold ratio, the extracted RNA is reverse transcribed into cDNA, and amplified on a fluorescent quantitative PCR instrument by using the cDNA as a template, and a standard curve is established. Reaction conditions: pre-denaturation at 95℃for 3 min; 40 cycles: 95 ℃ for 15s;60 ℃ for 1min. Corresponding to the standard curve, the calculated relative amount of toxoplasma gondii SAG 1.
After IFN-. Lambda.3 pretreatment of JEG-3 cells, it was co-cultured with toxoplasma gondii, and real-time PCR was used to detect SAG1 expression. The results were as follows: IFN-gamma and toxoplasma gondii co-act on JEG-3 cells in vitro, after co-culturing for 48 hours, cell culture supernatant is collected, and the expression of toxoplasma gondii SAG1 is detected by using a Real-time PCR method. As a result, it was found that the cells were treated with toxoplasma gondii and IFN-gamma as positive control groups, wherein IFN-gamma had the ability to inhibit toxoplasma gondii proliferation. And the proliferation of toxoplasma in the toxoplasma-infected group (another positive control group) was inhibited after the cells were pretreated with IFN-. Gamma.for 24 h. Similarly, IFN- λ3 in the co-stimulated group of cells via toxoplasma gondii and IFN- λ3 significantly reduced SAG1 expression. And after IFN- λ3 pretreatment, the expression of toxoplasma SAG1 in toxoplasma gondii infection group was further decreased after the cells were subjected to IFN- λ324h pretreatment as shown in FIG. 4.
These results indicate that IFN-. Lambda.3 inhibits toxoplasma gondii proliferation in vitro.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the present invention.
Claims (3)
- Use of ifn- λ3 for the preparation of a medicament for the treatment or prevention of toxoplasma gondii infection, characterized in that: the IFN-lambda 3 intervenes in up-regulating expression of CK functional molecules in placenta and inhibiting proliferation of toxoplasma gondii.
- 2. Use of a composition comprising IFN- λ3 and one or more pharmaceutically acceptable carriers for the manufacture of a medicament for the prevention or treatment of toxoplasma gondii infection.
- 3. Use of a composition according to claim 2 for the manufacture of a medicament for the prevention or treatment of toxoplasma gondii infection, wherein the carrier comprises pharmaceutically acceptable diluents, excipients, fillers, binders, absorption enhancing agents, surfactants and synergists.
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102512666A (en) * | 2011-12-15 | 2012-06-27 | 武汉大学 | Application of interferon lambda1 in preparation of anti-enterovirus 71 medicines |
| CN107412739A (en) * | 2017-08-09 | 2017-12-01 | 南通大学 | New applications of the IFN λ in Zika virus infection |
Non-Patent Citations (3)
| Title |
|---|
| Protective and Pathogenic Effects of Interferon Signaling During Pregnancy;Rebecca L.Casazza等;《Viral Immunology》;第第33卷卷(第第1期期);摘要，图1，第5-7页 * |
| 干扰素生物学特性及应用研究进展;王斌斌等;《广东畜牧兽医科技》;20070618(第03期);全文 * |
| 干扰素研究进展;刘运龙等;《动物医学进展》;20080220(第02期);第85页左栏最后1段至右栏第1段，第87页右栏最后1段 * |
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