Application of C-type lectin-like receptor-1 as therapeutic marker of fungal keratitis

A fungal keratitis, lectin technology, applied in biological testing, biomaterial analysis, disease diagnosis, etc., can solve the problems of difficult to restore transparency of the cornea, visual impairment of patients, and corneal tissue damage.

Pending Publication Date: 2022-05-24
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, an overly strong immune response can cause damage to the corneal tissue while controlling external infections, making it difficult to restore the transparency of the cornea, causing serious damage to the patient's vision

Method used

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  • Application of C-type lectin-like receptor-1 as therapeutic marker of fungal keratitis
  • Application of C-type lectin-like receptor-1 as therapeutic marker of fungal keratitis
  • Application of C-type lectin-like receptor-1 as therapeutic marker of fungal keratitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 The expression of CLEC-1 is increased in the cornea of ​​patients with Aspergillus fumigatus keratitis

[0054] 1. Experimental materials

[0055] 1.1 Collection of human corneal specimens

[0056] The corneas of 6 patients with Aspergillus fumigatus keratitis and 6 healthy patients were collected, and the specimens were confirmed by fungal culture and morphology. The purpose and methods of the study were explained to the patients in detail, and after informed consent was obtained, the samples were collected. Patients with any form of immunosuppression or topical steroid therapy or acute or chronic systemic disease were excluded.

[0057] 1.2 Experimental fungi

[0058] 3.0772 strains of Aspergillus fumigatus (China General Microorganism Culture Collection and Management Center)

[0059] 2. Experimental method

[0060] The corneas of 6 patients with Aspergillus fumigatitis and 6 healthy patients were collected for immunofluorescence and qRT-PCR detection....

Embodiment 2

[0064] Example 2 The expression of CLEC-1 decreased in the acute inflammatory phase of Aspergillus fumigatitis and increased in the recovery phase

[0065] 1. Experimental materials

[0066] 1.1 Experimental animals

[0067] C57BL / 6 mice (purchased from Jinan Pengyue Laboratory Animal Breeding Co., Ltd.)

[0068] 1.2 Experimental fungi

[0069] 3.0772 strains of Aspergillus fumigatus (China General Microorganism Culture Collection and Management Center)

[0070] 2. Experimental method

[0071] 2.1 Fungal Preparation

[0072] The Aspergillus fumigatus strain was inoculated on Sabouraud's medium, and cultured in a constant temperature incubator at 37°C for 2-3 days. The hyphae and conidia were scraped and put into 5ml sterile PBS to form a mixed solution, and the hyphae were removed by filtration with sterile gauze. Centrifuge at 4500 rpm at 4°C for 10 minutes, discard the supernatant, add 5 ml of bacterial PBS to the pellet to resuspend to form a conidia suspension. Adju...

Embodiment 3

[0087] Example 3 CLEC-1 overexpression reduces inflammatory response

[0088] 1. Experimental materials

[0089] 1.1 Experimental supplies

[0090] CLEC-1 adenovirus (AAV) vector (Genechem)

[0091] CLEC-1 lentiviral vector (Genechem)

[0092] 1.2 Experimental cells

[0093] THP-1 macrophages (Wuhan, China)

[0094] 1.3 Experimental fungi

[0095] 3.0772 strains of Aspergillus fumigatus (China General Microorganism Culture Collection and Management Center)

[0096] 2. Experimental method

[0097] 2.1 In vivo experiments

[0098] One eye from each mouse was randomly selected for subconjunctival injection (5 μl) of 1011 vg AAV vector. Two weeks later, enhanced green fluorescent protein (EGFP) was detected with a fluorescence stereo microscope to determine the uniform distribution of the virus in the corneal stroma. The overexpression of CLEC-1 was detected by qRT-PCR and western blot. Corneas were harvested 1 day after infection for MPO detection, immunofluorescence st...

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Abstract

The invention discloses an application of a C-type lectin-like receptor-1 as a therapeutic marker in fungal keratitis, and provides a kit for evaluating the medication opportunity of glucocorticoid drugs in the process of treating fungal keratitis. The C-type lectin-like receptor-1 can be used as a biomarker for simply, conveniently and safely judging the use opportunity of glucocorticoid drugs during fungal keratitis, and has important significance in the aspect of guiding clinical decision.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to the application of C-type lectin-like receptor-1 (C-type lectin-like receptor-1, CLEC-1) as a treatment marker for fungal keratitis. Background technique [0002] Fungal keratitis (FK) is a serious infectious eye disease worldwide. At present, due to the lack of effective clinical treatment methods, it can often lead to serious eye complications, and severe cases can eventually lead to Permanent vision loss. my country is an agricultural developing country, and it is a high-incidence area of ​​fungal keratitis, especially vegetative corneal trauma during farming is the main cause of the disease. Although many fungal keratitis drugs have been used clinically in recent years, the efficacy of these drugs is poor, and fungal resistance has gradually emerged, especially resistance to azoles. Due to the lack of effective treatment, many patients require corneal transplantation or even en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53
CPCG01N33/6893G01N33/53G01N2333/705G01N2800/16
Inventor 车成业张倩吕乐雨
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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