Method for simulating experimental adenoviral uveitis complicated by optic neuritis. RU patent 2504019.
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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 7th post-infection day.
EFFECT: higher rate and accuracy of the reproduction of adenoviral uveitis complicated by optic neuritis.
1 ex
Invention refers to medicine, in particular to ophthalmology, and can be used to create models of the disease in experimental ophthalmology.
With the introduction of adenovirus under the conjunctiva eyeball rabbit on 3-5 day there was development of an adenoviral conjunctivitis, limited place a suspension of the virus (M.T. Aznabaev, V.B. , Adenovirus and chlamydial diseases of the eye, 1995, P.54-59).
At introduction in front of the camera eye rabbit suspension containing adenovirus type 6 or 8 (infectious titles respectively 10 4 and 10 2 TDC 50/0,1 ml), adapted to the endothelial cell line NO R -2 or HELA histologically developed severe iritis, which took place on the 28th day experience (M.T. Aznabaev, V.B. , Adenovirus and chlamydial diseases of the eye, 1995, P.54-59).
Also shows the development of the pilot adenoviral uveitis Guinea pigs with the introduction of adenovirus type 6 in front of the camera eye. Histologically revealed lesions of the cornea in the zone of the limb, the iris, the flat part of the ciliary body. Inflammatory changes in were found only on 7 days after exposure (M.T. Aznabaev, V.B. , Adenovirus and chlamydial diseases of the eye, 1995, P.54-59).
In all the above experimental studies of the defeat of the optic nerve at the experimental uveitis occurred.
The closest analogue of the invention is a method of modelling of uveitis adenovirus etiology, which consists in infection of an experimental animal, characterized in that into the anterior chamber of the eye of an experimental animal insulin needle is injected adenovirus in the dose equal to or exceeding 10 3 TDC 50/ 0.1 ml for endothelial culture Hep-2 with the following research choroid from 14 to 28 days after infection (application EN 97109970, published 27.05.1999.).
When modeling adenovirus infections of the eye of experimental animals, the smaller the size of the eyeball, the more often develops traumatic injury of the shells of the eyeball. Moreover, in all experimental studies for the introduction of herpes simplex virus used insulin needle large size (at least 22G), which injure the eye and cause nonspecific inflammatory response.
In addition, during the simulation adenovirus infection eyeball adenovirus type 6 was adapted to the cells transplanted cells R -2, HELA and was administered only under the conjunctiva, cornea or in front of the camera eye. These methods do not lead to the defeat of the optic nerve.
Object of the present invention is to create a model of experimental adenoviral uveitis, complicated by the optic nerve neuritis, with a high frequency of play is closest to the specified disease in humans and to reduce the likelihood of injury to the eye membranes needle.
Technical result in the use of the invention is increasing the frequency and accuracy of modeling of adenoviral uveitis, complicated by the optic nerve neuritis, preventing the development of not desirable associated traumatic injury membranes in the eyes.
The proposed method of modelling of adenoviral uveitis, complicated by the optic nerve neuritis, as follows. Needle 33 G rabbits of chinchilla breed in the vitreous body of the eye through a flat part of the ciliary body introduce environmental liquid containing adenovirus type 6 adapted to the endothelial lines of embryonic pig kidney cells in a dose of 10,000 50 . Spend histological studies of remote eyeball, starting from 7 days after infection, which allow the study of the course of the disease.
Ten rabbits (20 eyes) of the chinchilla breed through the flat part of the ciliary body, vitreous both eyes needle 33 G was administered 0.1 ml of cultural liquid containing 10000 50 adenovirus type 6 adapted to endothelial lines of embryonic pig kidney cells, obtained by the standard technique [Virology. Methods: Per. s angl // Under. Ed. by B. . - M: Mir, 1988. - .254-269.]. On 7 and 21 days have made the slaughter of animals and histological studies conducted remote eyeballs. On the 7th day revealed an acute inflammatory process in the ciliary body and the optic nerve. In the ciliary body observed a marked swelling, a diffuse inflammatory infiltration provided leikotitami, , monocytes and lymphocytes, seats observed the formation of abscesses. Epithelium with signs cytoplasm, seats . Simultaneously in the thickness of the optic nerve, between nerve fibers revealed the accumulation of edematous fluid pockets of lymphocytic infiltration with admixture of single neutrophils, and infiltration of lymphocytes and isolated by leukocytes. The collateral vessels of the ophthalmic artery and vein of the optic nerve increased permeability of the walls, blood plasma exudation in zone and tissue structures. On the 21st day in histological sections identified the strengthening of the inflammatory reaction in the ciliary body, iris and the optic nerve. The degree of infiltration of inflammatory cells of the ciliary body has increased. Increased plethora of vessels, determined areas of hemorrhage. In the ciliary body observed diffuse infiltration tissue lymphocytes, monocytes, single neutrophils, in areas of inflammation appeared cells. Skin cells looked swollen, with signs of , places they were . Epithelial covering destroyed, homogenized. In the flat part of the ciliary body determined by pitting domestic retinal layer, was spotted infiltration cells inflammation. In the optic nerve identified hearth destruction of nerve tissue with a massive hemorrhagic drenching, cystic extensions, lymphocytic-macrophage infiltration, dystrophic modified .
Example. Rabbit №9 of the chinchilla breed mass 2.15 kg through a flat part of the ciliary body, vitreous both eyes needle 33 G was administered 0.1 ml of cultural liquid containing 10000 50 adenovirus type 6 adapted to endothelial lines of embryonic pig kidney cells. On 7 and 21 days have made the slaughter of animals and histological studies conducted remote eyeballs. On the 7th day revealed an acute inflammatory process in the ciliary body and the optic nerve. In the ciliary body observed a marked swelling, a diffuse inflammatory infiltration provided leikotitami, , monocytes and lymphocytes, seats observed the formation of abscesses. Epithelium with signs cytoplasm, seats . Simultaneously in the thickness of the optic nerve, between nerve fibers revealed the accumulation of edematous fluid pockets of lymphocytic infiltration with admixture of single neutrophils, and infiltration of lymphocytes and isolated by leukocytes. The collateral vessels of the ophthalmic artery and vein of the optic nerve increased permeability of the walls, blood plasma exudation in zone and tissue structures. On the 21st day development of uveitis meet the following histological changes. The degree of infiltration of inflammatory cells of the ciliary body has increased. Increased plethora of vessels, were the areas of hemorrhage. In the ciliary body observed diffuse infiltration tissue cells inflammation, also observed the emergence of cells. Epithelial cells looked swollen, with signs of , seats . Epithelial covering destroyed, homogenized. In the flat part of the ciliary body determined by pitting domestic retinal layer, was spotted infiltration cells inflammation. In the optic nerve identified hearth destruction of nerve tissue with a massive hemorrhagic drenching, cystic extensions, lymphocytic-macrophage infiltration, dystrophic modified .
Thus, the proposed method of modelling of adenoviral uveitis, complicated by the optic nerve neuritis, to create a model closest to the disease in humans. Furthermore, there is no traumatic injury of the shells of the eyeball and the lens.
A method of modelling of adenoviral uveitis, complicated neuritis of the optic nerve, characterized by the fact that through the flat part of the ciliary body of the vitreous body of the eye rabbit breed chinchilla needle 33 G impose 0.1 ml of cultural liquid containing adenovirus type 6 adapted to the endothelial lines of embryonic pig kidney cells, in the dose of 10,000 50 , in doing so, a histological study of the remote eyeball, starting from 7 days after infection.