RU2504019C1 - Method for simulating experimental adenoviral uveitis complicated by optic neuritis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: cultural fluid 0.1 ml containing type 6 adenovirus adapted to a pig's finite embryo renal cell line in a dose of 10000 TCD₅₀ is introduced through a flat portion of a ciliary body into a chinchilla's vitreous body with a needle 33 G. That is followed by the analysis of the removed eyeball starting from the 7^th post-infection day.

EFFECT: higher rate and accuracy of the reproduction of adenoviral uveitis complicated by optic neuritis.

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Description

The invention relates to medicine, in particular to ophthalmology, and can be used to create disease models in experimental ophthalmology.

With the introduction of adenovirus under the conjunctiva of the rabbit's eyeball for 3-5 days, adenoviral conjunctivitis developed, limited by the place of administration of the virus suspension (MT Aznabaev, VB Malkhanov, Adenoviral and chlamydial eye diseases, 1995, pp. 54-59) .

When a suspension containing type 6 or 8 adenovirus (infectious titers of 10 ⁴ and 10 ² TDC _{50 / 0.1} ml, respectively) was introduced into the anterior chamber of the rabbit's eye, adapted to the transplanted cell line HE _p -2 or HELA, serous iritis developed histologically , which took place by 28 days of the experiment (MT Aznabaev, VB Malkhanov, Adenoviral and chlamydial eye diseases, 1995, pp. 54-59).

The development of experimental adenovirus uveitis of guinea pigs with the introduction of type 6 adenovirus into the anterior chamber of the eye is also shown. Histologically revealed lesions of the cornea in the area of the limbus, iris, flat part of the ciliary body. Inflammatory changes in the choroid were found only on the 7th day after infection (MT Aznabaev, VB Malkhanov, Adenoviral and chlamydial eye diseases, 1995, pp. 54-59).

In all of the above experimental works, damage to the optic nerve during experimental uveitis did not occur.

The closest analogue of the invention is a method for modeling uveitis of adenoviral etiology, which consists in intraocular infection of an experimental animal, characterized in that an adenovirus is introduced into the anterior chamber of the eye of the experimental animal with an insulin needle at a dose equal to or greater than 10 ³ TDC _{50 /} 0.1 ml for transplanted culture HEP-2 followed by examination of the choroid from the 14th to the 28th day after infection (application RU 97109970, published on 05.27.1999.).

When modeling the adenovirus infection of the eyes of experimental animals, the smaller the size of the eyeball, the more often traumatic damage to the membranes of the eyeball develops. Moreover, in all experimental studies, a large-sized insulin needle (at least 22G) was used to inject the herpes simplex virus, which injured the lining of the eye and caused a nonspecific inflammatory response.

In addition, when modeling an adenovirus infection of the eyeball, type 6 adenovirus was adapted to cells of transplantable HE _p- 2, HELA cells and was introduced only under the conjunctiva, on the cornea or in the anterior chamber of the eye. These methods did not lead to damage to the optic nerve.

The objective of the invention is to create a model of experimental adenoviral uveitis complicated by optic neuritis with a high reproduction frequency that is closest to the specified disease in humans and to reduce the likelihood of injury to the eye membranes by a needle.

The technical result when using the invention is to increase the frequency and accuracy of modeling adenoviral uveitis complicated by optic neuritis, preventing the development of an undesirable concomitant traumatic injury to the membranes of the eye.

The proposed method for modeling adenoviral uveitis complicated by optic neuritis is as follows. With a 33 G needle, chinchilla rabbits are injected into the vitreous body of the eye through a flat part of the ciliary body with a culture fluid containing type 6 adenovirus adapted to the transplanted line of pig kidney kidney cells at a dose of 10,000 TCST ₅₀ . Histological studies of the removed eyeball are performed, starting from the 7th day after infection, which allows us to study the course of this disease.

To ten rabbits (20 eyes) of chinchilla breed, 0.1 ml of culture fluid containing 10,000 TCD _{50 of} type 6 adenovirus adapted to the transplanted line of pig fetal kidney cells obtained using standard pigs was introduced into the vitreous body of both eyes through the flat part of the ciliary body into the vitreous body of both eyes technique [Virology. Methods: Per. with English // Under. ed. B. Meikh. - M .: Mir, 1988. - S.254-269.]. On days 7 and 21, animals were slaughtered and histological studies of distant eyeballs were performed. On the 7th day, an acute inflammatory process was detected in the ciliary body and optic nerve. In the ciliary body, pronounced edema, diffuse inflammatory infiltration, represented by neutrophilic leukocytes, eosinophils, monocytes and lymphocytes, were observed, and in some places, the formation of abscesses was observed. The epithelium with signs of vacuolization of the cytoplasm, in places desquamated. At the same time, in the thickness of the optic nerve, between the nerve fibers, an accumulation of edematous fluid, foci of lymphocytic infiltration with an admixture of single neutrophils, as well as perivascular infiltration by lymphocytes and single leukocytes were detected. In the collateral vessels of the ophthalmic artery and optic vein, an increase in the permeability of their walls, exudation of blood plasma into the perivascular zone and tissue structures were noted. On the 21st day, an increase in the inflammatory response in the ciliary body, iris and optic nerve was revealed in histological sections. The degree of infiltration by inflammatory cells of the ciliary body has increased. Vascular congestion increased, hemorrhage areas were determined. Diffuse tissue infiltration by lymphocytes, monocytes, single neutrophils was observed in the ciliary body, and plasmocytic cells appeared in the area of inflammation. Epithelial cells looked swollen, with signs of vacuolization, in places they were desquamated. The epithelial lining is destroyed, homogenized. In the flat part of the ciliary body, ulceration of the inner retinal layer was determined, and infiltration by inflammatory cells was noticed. An optic nerve tissue destruction center with massive hemorrhagic impregnation, cystic enlargement, lymphocytic macrophage infiltration, and dystrophically altered neurocytes was revealed in the optic nerve.

Example. A rabbit No. 9 of chinchilla breed weighing 2.15 kg through the flat part of the ciliary body into the vitreous body of both eyes with a 33 G needle was injected with 0.1 ml of culture fluid containing 10,000 TCD _{50 of} type 6 adenovirus adapted to the transplanted line of pig kidney cells. On days 7 and 21, animals were slaughtered and histological studies of distant eyeballs were performed. On the 7th day, an acute inflammatory process was detected in the ciliary body and optic nerve. In the ciliary body, pronounced edema, diffuse inflammatory infiltration, represented by neutrophilic leukocytes, eosinophils, monocytes and lymphocytes, were observed, and in some places, the formation of abscesses was observed. The epithelium with signs of vacuolization of the cytoplasm, in places desquamated. At the same time, in the thickness of the optic nerve, between the nerve fibers, an accumulation of edematous fluid, foci of lymphocytic infiltration with an admixture of single neutrophils, as well as perivascular infiltration by lymphocytes and single leukocytes were detected. In the collateral vessels of the ophthalmic artery and optic vein, an increase in the permeability of their walls, exudation of blood plasma into the perivascular zone and tissue structures were noted. On the 21st day, the following histological changes corresponded to the development of uveitis. The degree of infiltration by inflammatory cells of the ciliary body has increased. Vascular congestion increased, there were areas of hemorrhage. Diffuse tissue infiltration by inflammatory cells was observed in the ciliary body, and the appearance of plasmacytic cells was also observed. Epithelial cells looked swollen, with signs of vacuolization, sometimes desquamated. The epithelial lining is destroyed, homogenized. In the flat part of the ciliary body, ulceration of the inner retinal layer was determined, and infiltration by inflammatory cells was noticed. An optic nerve tissue destruction center with massive hemorrhagic impregnation, cystic enlargement, lymphocytic macrophage infiltration, and dystrophically altered neurocytes was revealed in the optic nerve.

Thus, the proposed method for modeling adenoviral uveitis, complicated by optic neuritis, allows you to create a model that is closest to this disease in humans. In addition, there is no traumatic damage to the membranes of the eyeball and lens.

Claims (1)

A method for modeling adenoviral uveitis complicated by optic neuritis, characterized in that 0.1 ml of a culture fluid containing type 6 adenovirus adapted to the transplanted line of pig kidney cells is injected into the vitreous body of a chinchilla rabbit eye with a 33 G needle through the flat part of the ciliary body , at a dose of 10,000 TCD ₅₀ , while conducting a histological examination of the removed eyeball, starting from 7 days after infection.