Application of salivaricin in the preparation of medicines for preventing and/or treating autoimmune diseases

A technology of streptococcus salivarius and streptococcus, which is applied in the field of treatment of autoimmune diseases, can solve problems such as concomitant infection, and achieve the effect of regulating immune balance and promoting secretion

Active Publication Date: 2022-03-04
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, excessive use of hormones, immunosuppressants, and biological agents to suppress the immune response cannot achieve the purpose of effectively alleviating the disease, and is usually accompanied by adverse reactions such as infection

Method used

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  • Application of salivaricin in the preparation of medicines for preventing and/or treating autoimmune diseases
  • Application of salivaricin in the preparation of medicines for preventing and/or treating autoimmune diseases
  • Application of salivaricin in the preparation of medicines for preventing and/or treating autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The gene encoding the synthesis of salivaricin is significantly reduced in the tonsillar microbiota of patients with rheumatoid arthritis

[0027] experimental method

[0028] (1) Collection of tonsil flora samples from patients with rheumatoid arthritis RA and healthy subjects: 1) Instruct the subjects to rinse their mouth gently and avoid touching the throat; 2) Use a moist sterile cotton swab to dip Take the surface of the tonsils of the subjects, and pay attention to avoid touching the saliva, the back of the tongue, the palate, the inner wall of the oral cavity and other parts to prevent bacterial contamination; 3) The bacterial flora samples are temporarily stored in a -80°C low-temperature refrigerator for future use.

[0029] (2) Next-generation sequencing analysis of tonsil flora: 1) Use DNA extraction kit (Catalog No. 12888-100, MO BIO Laboratories, Carlsbad, CA 92010, USA) to extract the genomic DNA of all flora in the sample; 2) use The Illumina library const...

Embodiment 2

[0037] Isolation and purification of salivaricin A2 and B from Streptococcus salivarius

[0038] experimental method:

[0039] (1) Expanded culture of Streptococcus salivarius:

[0040] In a sterile workbench, use a 20 μL pipette to draw the bacterial solution into the M17 agar medium, spread it evenly on the flat medium with a coating stick, and place it at a temperature of 37°C and CO 2 Cultivate for 24 hours at a concentration of 5%; in a sterile workbench, take 5 1.5ml centrifuge tubes and add 1ml of liquid medium respectively, and use a sterile probe to take the plaques on the petri dish that are full, smooth, and independently distributed Transfer them into centrifuge tubes respectively, shake to dissolve them fully, add THB liquid medium to culture for 18 hours, OD 600 The value is between 20 and 40.

[0041] (2) Separation and purification of salivaricin:

[0042] ①Spread the above bacterial solution evenly in M17 medium (each 1L of M17 medium also includes 5g sucr...

Embodiment 3

[0051] Salivaricin A2 and B regulate the proportion of immune cells in human peripheral blood

[0052] experimental method:

[0053] (1) Isolation and culture of human peripheral blood mononuclear cells (PBMC): add 4ml of whole blood to 8ml of lymphocyte separation medium, and centrifuge at 1800rpm for 20min; 10ml sterile normal saline), this layer is the mononuclear cell layer, centrifuged at 1800rpm for 10min, discarded the supernatant; adjusted to 2×10 with 1640 complete medium 6 cell / ml single cell suspension, inoculated to 48-well plate, 1ml per well. The cells were pretreated with salivaricin A2 and B for 1 hour respectively, then co-stimulated with phorbol ester and ionomycin, and cultured in a 37°C incubator for 72 hours. The doses of salivaricin A2 and B were set at three levels, 50 μg / mL, 100 μg / mL, and 200 μg / mL, respectively. The negative control group did not receive any stimulation, and the drug control group only added phorbol ester and ionomycin.

[0054] (...

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Abstract

The invention provides the application of salivaricin in the preparation of medicines for preventing and / or treating autoimmune diseases, and belongs to the technical field of autoimmune diseases treatment. Streptococcus salivarius is an antimicrobial peptide synthesized and secreted by Streptococcus salivarius, which plays an important role in maintaining the local microecological balance of the mucosa. Salivaricin can significantly promote the secretion of anti-inflammatory cytokine IL-10, and inhibit the expression of pro-inflammatory cytokines IL-17, IL-21 and TNF-α. Simultaneously, salivaricin upregulates the proportion of anti-inflammatory regulatory T (Treg) cells, and downregulates the proportion of pro-inflammatory follicular helper T cells and helper T‑17 cells among immune cell subsets. Salivaricin can improve immune imbalance by inhibiting immune response, rebuild immune balance, and provide a new treatment method for autoimmune diseases such as rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the technical field of autoimmune diseases, and in particular relates to the application of salivaricin in the preparation of medicaments for preventing and / or treating autoimmune diseases. Background technique [0002] Autoimmune diseases, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS), are characterized by immune inflammation, and a variety of pathogenic autoantibodies and multiple pathogenic autoantibodies appear in serum. Systemic involvement is the main clinical feature of this disease. The lesions of rheumatoid arthritis mainly invade the joints, and the symptoms of the joints generally recur, and the damage of the joints becomes more and more serious, eventually leading to varying degrees of dysfunction and deformity. In addition to joints, skin rheumatoid nodules, arteritis, pericarditis, scleritis, lymphadenitis, hepatosplenomegaly, and neuropathy are not uncommon. T...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61P37/02
CPCA61K38/164A61P37/02
Inventor 栗占国李静何菁靳家扬覃俊杰李胜辉
Owner PEOPLES HOSPITAL PEKING UNIV
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