Eye disease model animal

Abstract

The present disclosure provides an eye disease model animal, a method for producing the eye disease model animal, and a method for screening therapeutic agents and / or prophylactic agents for dry eye. One aspect of the present disclosure provides a dry eye model animal that has a characteristic associated with meibomian gland dysfunction. Another aspect of the present disclosure provides a method for producing the dry eye model animal, the method including a step for transplanting cells from an animal that is allogeneic to the dry eye model animal into an animal that has reduced or deficient immune function.

Description

Technical Field

[0001] The present disclosure relates to model animals for eye diseases, particularly meibomian gland dysfunction model animals, dry eye model animals, and related technologies thereof.

Background Art

[0002] Dry eye is classified into a tear-deficient type and an evaporative type, and there are also combined types thereof. The evaporative type includes meibomian gland dysfunction and lipid abnormalities as factors. It is considered that 89% of dry eye patients have concurrent meibomian gland dysfunction. However, clinically, there is no dry eye therapeutic agent targeted at improving meibomian gland dysfunction. There is a need for therapeutic and prophylactic agents for dry eye with concurrent meibomian gland dysfunction, including evaporative dry eye.

[0003] For the development of therapeutic and prophylactic agents, screening using model animals capable of evaluating therapeutic and prophylactic effects is essential. However, there are few model animals capable of evaluating the therapeutic and prophylactic effects of candidate compounds against meibomian gland dysfunction. Furthermore, there has been no model animal capable of concurrently confirming the effect on dry eye symptoms in addition to the effect on meibomian gland dysfunction.

Summary of the Invention

Means for Solving the Problems

[0004] By administering cells of an allogeneic animal to an immunodeficient animal, the inventors have newly found that the immunodeficient animal unexpectedly has meibomian gland dysfunction, and further has both meibomian gland dysfunction and dry eye. Accordingly, in one aspect, the present disclosure provides a model animal having meibomian gland dysfunction and a method for producing such a model animal, or a dry eye model animal having meibomian gland dysfunction and a method for producing such a dry eye model animal. In another aspect, a method for evaluating the efficacy of a therapeutic and / or prophylactic agent for dry eye or meibomian gland dysfunction using such a model animal, and a method for screening a therapeutic and / or prophylactic agent for dry eye or meibomian gland dysfunction are provided.

[0005] The present disclosure provides, for example, the following items. (Item 1) A dry eye model animal having characteristics related to meibomian gland dysfunction. (Item 2) The model animal according to the above item, wherein the model animal is an animal with reduced or impaired immune function transplanted with cells of an allogeneic animal. (Item 3) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 4) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse with a major histocompatibility complex (MHC) type of H-2D d The model animal according to any one of the above items, which is a SCID mouse or a NOD-SCID mouse with a major histocompatibility complex (MHC) type of H-2D (Item 5) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 6) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 7) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen. (Item 8) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having a type of MHC class I different from that of the model animal. (Item 9) The allogeneic animal is a mouse having an MHC type of H-2D b The model animal according to any one of the above items. (Item 10) The model animal according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 11) The model animal according to any one of the above items, wherein the feature related to meibomian gland dysfunction is at least one selected from the group consisting of occlusion of the meibomian gland opening, atrophy of the meibomian gland, and eyelid margin irregularity. (Item 12) The model animal according to any one of the above items, wherein the feature related to meibomian gland dysfunction is occlusion of the meibomian gland opening. (Item 13) The model animal according to any one of the above items, further having a feature related to dry eye selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. (Item 14) The model animal according to any one of the above items, further having a feature related to dry eye selected from the group consisting of abnormal tear film break-up time and abnormal corneal staining score. (Item 15) A dry eye model animal, wherein the dry eye model animal is a dry eye model animal in which the immune function is reduced or impaired by transplantation of immune cells of an allogeneic animal. An animal having at least one feature selected from the features associated with the following meibomian gland dysfunction and at least one feature selected from the features associated with the following dry eye, The feature associated with the meibomian gland dysfunction is selected from occlusion of the meibomian gland opening, atrophy of the meibomian gland, and eyelid margin irregularity, The feature associated with the dry eye is selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate, Dry eye model animal. (Item 15A) The dry eye model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 15B) The dry eye model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse having a major histocompatibility complex (MHC) type of H-2D d as described above. (Item 15C) The dry eye model animal according to any one of the above items, wherein the allogeneic animal cells are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 15D) The dry eye model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC class I type different from that of the model animal. (Item 15E) The dry eye model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type of H-2D b as described above. (Item 15F) The dry eye model animal according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 16) A dry eye model animal, The dry eye model animal is an animal with reduced or impaired immune function due to transplantation of cells from an allogeneic animal of the same species, The type of the major histocompatibility complex (MHC) of the dry eye model animal is H-2D d and The type of the MHC of the allogeneic animal is H-2D b and having obstruction of the meibomian gland orifice, abnormal tear film break-up time, and abnormal corneal staining score, dry eye model animal. (Item 16A) The dry eye model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 16B) The dry eye model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 16C) The dry eye model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 16D) The dry eye model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen. (Item 16E) The dry eye model animal according to any one of the above items, wherein the allogeneic animal is a mouse having a type of MHC class I different from that of the model animal. (Item 16F) The dry eye model animal according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 17) A dry eye model animal characterized in that features related to meibomian gland dysfunction develop by 40 weeks of age. (Item 17A) The dry eye model animal according to any one of the above items, wherein the features related to meibomian gland dysfunction are at least one selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item 18) A model animal characterized by containing spleen-derived cells in the meibomian gland. (Item 18A) The model animal according to any one of the above items, wherein the cells are cells of an animal that is allogeneic to the model animal. (Item 19) The model animal according to any one of the above items, wherein the model animal is a dry eye model animal. (Item 20) The model animal according to any one of the above items, wherein the model animal is a meibomian gland dysfunction (MGD) model animal. (Item 21) A dry eye model animal containing at least one type of cell selected from the group consisting of T cells, B cells, macrophages, dendritic cells, sinusoidal endothelial cells, and NK cells in the meibomian gland. (Item 22) A dry eye model animal, which is an animal in which cells of an animal that is allogeneic to the model animal have been transplanted and whose immune function is reduced or impaired. (Item 23) A meibomian gland dysfunction (MGD) model animal, which is an animal in which cells of an animal that is allogeneic to the model animal have been transplanted and whose immune function is reduced or impaired. (Item 23A) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 23B) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse with a major histocompatibility complex (MHC) type of H-2D d as described above. (Item 23C) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 23D) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 23E) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC class I type different from that of the model animal. (Item 23F) The allogeneic animal is a mouse having an MHC type of H-2D b The model animal according to any one of the above items. (Item 23G) The model animal according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 23H) The model animal according to any one of the above items, wherein the model animal has at least one feature related to meibomian gland dysfunction selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item 23I) The model animal according to any one of the above items, wherein the model animal further has a feature related to dry eye selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. (Item 24) A meibomian gland dysfunction (MGD) model animal having occlusion of the meibomian gland orifice, wherein cells of an animal allogeneic to the model animal are transplanted, and the animal has reduced or defective immune function. (Item 24A) The MGD model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 24B) The model animal is a SCID mouse or a NOD-SCID mouse with the type of major histocompatibility complex (MHC) being H-2D d The MGD model animal according to any one of the above items, which is a SCID mouse or a NOD-SCID mouse with the type of major histocompatibility complex (MHC) being H-2D d . (Item 24C) The MGD model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 24D) The MGD model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 24E) The MGD model animal according to any one of the above items, wherein the allogeneic animal is a mouse having a type of MHC class I different from that of the model animal. (Item 24F) The allogeneic animal is a mouse with the type of MHC being H-2D b The MGD model animal according to any one of the above items, which is a mouse with the type of MHC being H-2D b . (Item 24G) The MGD model animal according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 25) A method for producing a dry eye model animal, the method comprising the step of transplanting cells of an allogeneic animal, which is of the same species as the dry eye model animal, into an animal with reduced or impaired immune function. (Item 26) A method for producing an MGD model animal, the method comprising the step of transplanting cells of an allogeneic animal, which is of the same species as the MGD model animal, into an animal with reduced or impaired immune function. (Item 26A) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 26B) The model animal has the type of major histocompatibility complex (MHC) being H-2D dThe method according to any one of the above items, wherein the animal is a SCID mouse or a NOD-SCID mouse. (Item 26C) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 26D) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 26E) The method according to any one of the above items, wherein the allogeneic animal is a mouse having a type of MHC class I different from that of the model animal. (Item 26F) The allogeneic animal has an MHC type of H-2D b The method according to any one of the above items, wherein the animal is a mouse. (Item 26G) The method according to any one of the above items, wherein the allogeneic animal is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 26H) The method according to any one of the above items, wherein the model animal has at least one characteristic related to meibomian gland dysfunction selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item 26I) The method according to any one of the above items, wherein the model animal further has a characteristic related to dry eye selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. (Item 27) A method for evaluating the efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) administering the test substance to an eye disease model animal; and (2) evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye in the eye disease model animal. The animal model of eye disease is an animal with reduced or impaired immune function in which cells of allogeneic animals are transplanted. Method. (Item 28) The method according to any one of the above items, wherein the eye disease is dry eye, and the method includes a step of evaluating characteristics related to dry eye in the step (2). (Item 29) The method according to any one of the above items, wherein the eye disease is meibomian gland dysfunction, and the method includes a step of evaluating characteristics related to meibomian gland dysfunction in the step (2). (Item 30) In the step (2), the characteristics related to meibomian gland dysfunction include at least one selected from occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity, and the characteristics related to dry eye include at least one selected from abnormal tear film break-up time, abnormal corneal staining score, decrease in tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The method according to any one of the above items. (Item 31) In the step (2), the characteristics related to meibomian gland dysfunction include at least occlusion of the meibomian gland orifice. The method according to any one of the above items. (Item 32) The method according to any one of the above items, wherein the step (2) includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction and at least one of the characteristics related to dry eye. (Item 33) In the step (2), the method includes a step of evaluating at least occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate. The method according to any one of the above items. (Item 34) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 35) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 36) The method according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item 37) The method according to any one of the above items, wherein the model animal has the type d of H-2D, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item 38) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 39) The method according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item 40) The method according to any one of the above items, wherein the allogeneic animal is a mouse having the type b of H-2D, which is a subclass constituting the MHC class I. (Item 41) A method for evaluating the efficacy of a test substance as a therapeutic agent and / or prophylactic agent for dry eye, the method comprising: (1) administering the test substance to a dry eye model animal; and (2) evaluating, in the dry eye model animal, the occlusion of the meibomian gland orifice, the abnormality of the tear film break-up time, and the abnormality of the blink rate wherein the dry eye model animal is an animal in which the immune function is reduced or impaired by transplantation of cells of an allogeneic animal, the major histocompatibility complex (MHC) type of the dry eye model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b and a method for evaluating the efficacy. (Item 41A) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 41B) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 41C) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 42) A method for screening a therapeutic agent and / or a prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an eye disease model animal; (2) evaluating, in the eye disease model animal, a feature related to meibomian gland dysfunction or a feature related to dry eye; and (3) identifying the test substance as a therapeutic agent and / or a prophylactic agent for the eye disease when the evaluated feature is improved or the progression is suppressed. comprising the eye disease model animal being an animal in which the immune function has been reduced or impaired by transplantation of cells of an allogeneic animal. method. (Item 42A) The method according to any one of the above items, wherein the eye disease is dry eye, and the step (2) includes the step of evaluating a feature related to dry eye. (Item 42B) The method according to any one of the above items, wherein the eye disease is meibomian gland dysfunction, and the step (2) includes the step of evaluating a feature related to meibomian gland dysfunction. (Item 42C) In the step (2), the features related to meibomian gland dysfunction include at least one selected from occlusion of the meibomian gland opening, atrophy of the meibomian gland, and eyelid margin irregularity, and the features related to dry eye include at least one feature selected from abnormal tear film break-up time, abnormal corneal staining score, decrease in tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The method according to any one of the above items. (Item 42D) The method according to any one of the above items, wherein in the step (2), the features related to meibomian gland dysfunction include at least the occlusion of the meibomian gland opening. (Item 42E) The method according to any one of the above items, including a step of evaluating at least one feature related to meibomian gland dysfunction and at least one feature related to dry eye in the step (2). (Item 42F) The method according to any one of the above items, wherein in the step (2), at least the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blink rate are evaluated. (Item 42G) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 42H) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item 42I) The method according to any one of the above items, wherein the model animal has the H-2D type of d, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item 42J) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 42K) The method according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item 42L) The method according to any one of the above items, wherein the allogeneic animal is a mouse having the H-2D type of b, which is a subclass constituting MHC class I. (Item 43) A method for screening a therapeutic and / or prophylactic agent for dry eye, the method comprising: (1) a step of administering a test substance to a dry eye model animal; (2) evaluating, in the dry eye model animal, the occlusion of the meibomian gland orifice, the abnormality of the tear film break-up time, and the abnormality of the blink rate; and (3) identifying, in the dry eye model animal, the test substance as a therapeutic agent and / or a prophylactic agent for dry eye when the occlusion of the meibomian gland orifice, the abnormality of the tear film break-up time, and the abnormality of the blink rate are improved or the progression is suppressed comprising the dry eye model animal being an animal in which the immune function is reduced or impaired due to transplantation of cells of an allogeneic animal the type of the major histocompatibility complex (MHC) of the dry eye model animal being H-2D d and the type of the MHC of the allogeneic animal being H-2D b and a screening method. (Item 43A) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 43B) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item 43C) The method according to any one of the above items, wherein the model animal has a type d of H-2D, a subclass constituting the major histocompatibility complex (MHC) class I. (Item 43D) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 43E) The method according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item 43F) The method according to any one of the above items, wherein the allogeneic animal is a mouse having a type b of H-2D, a subclass constituting the MHC class I. (Item A1) A method for evaluating the medicinal efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) administering the test substance to an animal model of an eye disease; and (2) evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye in the animal model of an eye disease, wherein the animal model of an eye disease is an animal with reduced or impaired immune function into which cells of allogeneic animals have been transplanted. Method. (Item A2) The method according to any one of the above items, wherein in the step (2), the method includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction. (Item A3) The method according to any one of the above items, wherein in the step (2), the characteristics related to meibomian gland dysfunction include at least one selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item A4) The method according to any one of the above items, wherein in the step (2), the characteristics related to meibomian gland dysfunction include at least occlusion of the meibomian gland orifice. (Item A5) The method according to any one of the above items, wherein in the step (2), the method includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction and at least one of the characteristics related to dry eye. (Item A6) The method according to any one of the above items, wherein in the step (2), the method includes a step of evaluating at least occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate. (Item A7) The method according to any one of the above items, wherein the eye disease is meibomian gland dysfunction. (Item A8) The method according to any one of the above items, wherein the eye disease is dry eye. (Item A9) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item A10) The method according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item A11) The method according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item A12) The method according to any one of the above items, wherein the model animal has the type d of H-2D, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item A13) The method according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item A14) The method according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item A15) The method according to any one of the above items, wherein the allogeneic animal is a mouse having the type b of H-2D, which is a subclass constituting the MHC class I. (Item A16) A method for screening a therapeutic agent and / or a prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an eye disease model animal; (2) evaluating, in the eye disease model animal, a feature related to meibomian gland dysfunction or a feature related to dry eye; and (3) when the evaluated feature is improved or the progression is suppressed, identifying the test substance as a therapeutic agent and / or a prophylactic agent for the eye disease. comprising the eye disease model animal is an animal in which the immune function has been reduced or impaired by transplantation of cells of an allogeneic animal. method. (Item A17) A method for screening a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction, the method comprising: (1) administering a test substance to a meibomian gland dysfunction model animal; (2) evaluating the occlusion of the meibomian gland orifice in the meibomian gland dysfunction model animal; and (3) identifying the test substance as a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction when the occlusion of the meibomian gland orifice is improved and / or the progression of the occlusion of the meibomian gland orifice is suppressed in the meibomian gland dysfunction model animal, wherein the meibomian gland dysfunction model animal is an animal with reduced or impaired immune function due to transplantation of cells from an allogeneic animal, the major histocompatibility complex (MHC) type of the meibomian gland dysfunction model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b such screening method. (Item A18) A method for screening a therapeutic agent and / or prophylactic agent for dry eye, the method comprising: (1) administering a test substance to a dry eye model animal; (2) evaluating the occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate in the dry eye model animal; and (3) identifying the test substance as a therapeutic agent and / or prophylactic agent for dry eye when the occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate are improved or the progression is suppressed in the dry eye model animal including wherein the dry eye model animal is an animal with reduced or impaired immune function due to transplantation of cells from an allogeneic animal, the major histocompatibility complex (MHC) type of the dry eye model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b such screening method.​ Screening method. (Item B1) Use of an animal model for eye diseases in a method for evaluating the medicinal effect of a test substance as a therapeutic and / or prophylactic agent for eye diseases, wherein the method comprises: (1) a step of administering the test substance to the animal model for eye diseases; and (2) a step of evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye in the animal model for eye diseases, wherein the animal model for eye diseases is an animal with reduced or impaired immune function into which cells of an allogeneic animal have been transplanted. Use. (Item B2) The use according to any one of the above items, wherein the eye disease is dry eye and the step (2) includes a step of evaluating the characteristics related to dry eye. (Item B3) The use according to any one of the above items, wherein the eye disease is meibomian gland dysfunction and the step (2) includes a step of evaluating the characteristics related to meibomian gland dysfunction. (Item B4) In the step (2), the characteristics related to meibomian gland dysfunction include at least one selected from the closure of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity, and the characteristics related to dry eye include at least one selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The use according to any one of the above items. (Item B5) The use according to any one of the above items, wherein in the step (2), the characteristics related to meibomian gland dysfunction include at least the closure of the meibomian gland orifice. (Item B6) The use according to any one of the above items, wherein the step (2) includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction and at least one of the characteristics related to dry eye. (Item B7) In the above step (2), the use according to any one of the above items, including at least the steps of evaluating the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blink rate. (Item B8) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item B9) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item B10) The use according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item B11) The use according to any one of the above items, wherein the model animal has the H-2D of the subclass constituting the major histocompatibility complex (MHC) class I of type d. (Item B12) The use according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item B13) The use according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item B14) The use according to any one of the above items, wherein the allogeneic animal is a mouse having the H-2D of the subclass constituting the MHC class I of type b. (Item B15) The use of a dry eye model animal in a method for evaluating the efficacy of a test substance as a therapeutic and / or prophylactic agent for dry eye, the method comprising: (1) administering the test substance to the dry eye model animal; and (2) evaluating, in the dry eye model animal, the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blink rate wherein the dry eye model animal is an animal in which the immune function is reduced or defective due to the transplantation of cells of an allogeneic animal. The type of the major histocompatibility complex (MHC) of the dry eye model animal is H-2D d and the type of the MHC of the allogeneic animal is H-2D b and Use (Item B16) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells (Item B17) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes (Item B18) The use according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse (Item B19) Use of an animal model for dry eye in a method for screening a therapeutic and / or prophylactic agent for an eye disease, the method comprising (1) a step of administering a test substance to the animal model for dry eye (2) a step of evaluating a feature related to meibomian gland dysfunction or a feature related to dry eye in the animal model for dry eye; and (3) a step of identifying the test substance as a therapeutic and / or prophylactic agent for the eye disease when the evaluated feature is improved or the progression is suppressed comprising the animal model for dry eye is an animal in which the immune function is reduced or defective due to transplantation of cells of an allogeneic animal Use (Item B20) The use according to any one of the above items, wherein the eye disease is dry eye and the step (2) includes a step of evaluating a feature related to dry eye (Item B21) The use according to any one of the above items, wherein the eye disease is meibomian gland dysfunction and the step (2) includes a step of evaluating a feature related to meibomian gland dysfunction (Item B21) In the step (2), the features related to meibomian gland dysfunction include at least one selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity, and the features related to dry eye include at least one selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The use according to any one of the above items. (Item B22) In the step (2), the features related to meibomian gland dysfunction include at least occlusion of the meibomian gland orifice. The use according to any one of the above items. (Item B23) In the step (2), the use according to any one of the above items includes a step of evaluating at least one feature related to meibomian gland dysfunction and at least one feature related to dry eye. (Item B24) In the step (2), the use according to any one of the above items includes a step of evaluating at least occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate. (Item B25) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item B26) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item B27) The use according to any one of the above items, wherein the model animal has an H-2D type of d, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item B28) The use according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item B29) The use according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item B30) The use according to any one of the above items, wherein the allogeneic animal is a mouse in which the type of H-2D, a subclass constituting MHC class I, is b. (Item B31) Use of a dry eye model animal in a method for screening a therapeutic and / or prophylactic agent for dry eye, the method comprising: (1) administering a test substance to the dry eye model animal; (2) evaluating occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate; and (3) in the dry eye model animal, when occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate are improved or the progression is suppressed, identifying the test substance as a therapeutic and / or prophylactic agent for dry eye comprising: the dry eye model animal being an animal in which the immune function is reduced or defective due to transplantation of cells of an allogeneic animal; the major histocompatibility complex (MHC) type of the dry eye model animal being H-2D d and the MHC type of the allogeneic animal being H-2D b and use. (Item B32) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item B33) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item B34) The use according to any one of the above items, wherein the model animal has a type of H-2D, a subclass constituting the major histocompatibility complex (MHC) class I, of d. (Item B35) The use according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item B36) The use according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item B37) The use according to any one of the above items, wherein the allogeneic animal is a mouse having an H-2D type of subclass constituting MHC class I of b. (Item C1) Use of an animal model for eye diseases in a method for evaluating the efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) administering the test substance to the animal model for eye diseases; and (2) evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye in the animal model for eye diseases, wherein the animal model for eye diseases is an animal in which the immune function is reduced or impaired by transplantation of cells of an allogeneic animal. Use. (Item C2) The use according to any one of the above items, wherein the step (2) includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction. (Item C3) The use according to any one of the above items, wherein the characteristics related to meibomian gland dysfunction in the step (2) include at least one selected from occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item C4) The use according to any one of the above items, wherein the characteristics related to meibomian gland dysfunction in the step (2) include at least occlusion of the meibomian gland orifice. (Item C5) The use according to any one of the above items, wherein the step (2) includes a step of evaluating at least one of the characteristics related to meibomian gland dysfunction and at least one of the characteristics related to dry eye. (Item C6) In the step (2), the use according to any one of the above items, including at least the steps of evaluating the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blink rate. (Item C7) The use according to any one of the above items, wherein the eye disease is meibomian gland dysfunction. (Item C8) The use according to any one of the above items, wherein the eye disease is dry eye. (Item C9) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item C10) The use according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item C11) The use according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item C12) The use according to any one of the above items, wherein the model animal has the H-2D type of d, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item C13) The use according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item C14) The use according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item C15) The use according to any one of the above items, wherein the allogeneic animal is a mouse having the H-2D type of b, which is a subclass constituting the MHC class I. (Item C16) Use of an animal model for an eye disease in a method for screening a therapeutic and / or prophylactic agent for an eye disease, the method comprising: (1) A step of administering a test substance to the animal model for an eye disease (2) Evaluating features related to meibomian gland dysfunction or features related to dry eye in an animal model of eye disease; and (3) Identifying the test substance as a therapeutic agent and / or prophylactic agent for eye disease when the evaluated features are improved or the progression is suppressed comprising the animal model of eye disease is an animal with reduced or defective immune function in which cells of an allogeneic animal have been transplanted Use. (Item C17) Use of an animal model of meibomian gland dysfunction in a method for screening a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction, the method comprising (1) Administering a test substance to an animal model of meibomian gland dysfunction, (2) Evaluating occlusion of the meibomian gland orifice in the animal model of meibomian gland dysfunction; and (3) Identifying the test substance as a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction when occlusion of the meibomian gland orifice is improved and / or progression of occlusion of the meibomian gland orifice is suppressed in the animal model of meibomian gland dysfunction, the animal model of meibomian gland dysfunction is an animal with reduced or defective immune function in which cells of an allogeneic animal have been transplanted, the major histocompatibility complex (MHC) type of the animal model of meibomian gland dysfunction is H-2D d and the MHC type of the allogeneic animal is H-2D b and Use. (Item C18) Use of an animal model of dry eye in a method for screening a therapeutic agent and / or prophylactic agent for dry eye, the method comprising (1) Administering a test substance to an animal model of dry eye, (2) Evaluating occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate in the animal model of dry eye; and (3) In the dry eye model animal, when occlusion of the meibomian gland opening, abnormal break-up time of the tear film, and abnormal blink rate are improved or the progression is suppressed, identifying the test substance as a therapeutic agent and / or prophylactic agent for dry eye comprising the dry eye model animal is an animal with reduced or impaired immune function in which cells of an allogeneic animal have been transplanted, the type of major histocompatibility complex (MHC) of the dry eye model animal is H-2D d and the type of MHC of the allogeneic animal is H-2D b and Use. (Item D1) An eye disease model animal for use in a method for evaluating the medicinal effect of a test substance as a therapeutic agent and / or prophylactic agent for an eye disease, the method comprising (1) administering the test substance to the eye disease model animal; and (2) in the eye disease model animal, evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye, wherein the eye disease model animal is an animal with reduced or impaired immune function in which cells of an allogeneic animal have been transplanted, Model animal. (Item D2) The eye disease is dry eye, and the method according to any one of the above items, including the step of evaluating the characteristics related to dry eye in the step (2). (Item D3) The eye disease is meibomian gland dysfunction, and the method according to any one of the above items, including the step of evaluating the characteristics related to meibomian gland dysfunction in the step (2). (Item D4) In the step (2), the features related to meibomian gland dysfunction include at least one selected from meibomian gland orifice occlusion, meibomian gland atrophy, and eyelid margin irregularity, and the features related to dry eye include at least one selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The model animal according to any one of the above items. (Item D5) In the step (2), the features related to meibomian gland dysfunction include at least meibomian gland orifice occlusion. The model animal according to any one of the above items. (Item D6) In the step (2), the model animal according to any one of the above items includes a step of evaluating at least one feature related to meibomian gland dysfunction and at least one feature related to dry eye. (Item D7) In the step (2), the model animal for eye diseases according to any one of the above items includes a step of evaluating at least meibomian gland orifice occlusion, abnormal tear film break-up time, and abnormal blink rate. (Item D8) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item D9) The model animal for eye diseases according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item D10) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item D11) The model animal for eye diseases according to any one of the above items, wherein the model animal has the type d of H-2D, a subclass constituting the major histocompatibility complex (MHC) class I. (Item D12) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item D13) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item D14) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an H-2D type of subclass constituting MHC class I as b. (Item D15) A dry eye model animal for use in a method for evaluating the efficacy of a test substance as a therapeutic agent and / or prophylactic agent for dry eye, the method comprising: (1) administering the test substance to the dry eye model animal; and (2) evaluating, in the dry eye model animal, occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate The dry eye model animal is an animal in which the immune function is reduced or impaired by transplantation of cells of an allogeneic animal, the major histocompatibility complex (MHC) type of the dry eye model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b is model animal. (Item D16) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item D17) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item D18) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item D19) An eye disease model animal for use in a method for screening a therapeutic agent and / or prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to the eye disease model animal (2) evaluating, in an animal model of an eye disease, features associated with meibomian gland dysfunction or features associated with dry eye; and (3) when the evaluated features are improved or the progression is suppressed, identifying the test substance as a therapeutic agent and / or a prophylactic agent for the eye disease comprising: wherein the animal model of the eye disease is an animal with reduced or impaired immune function in which cells of an allogeneic animal have been transplanted; model animal. (Item D20) The model animal according to any one of the above items, wherein the eye disease is dry eye and the step (2) includes evaluating features associated with dry eye. (Item B21) The model animal according to any one of the above items, wherein the eye disease is meibomian gland dysfunction and the step (2) includes evaluating features associated with meibomian gland dysfunction. (Item D21) In the step (2), the features associated with meibomian gland dysfunction include at least one selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity, and the features associated with dry eye include at least one feature selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. The model animal according to any one of the above items. (Item D22) The model animal according to any one of the above items, wherein in the step (2), the features associated with meibomian gland dysfunction include at least occlusion of the meibomian gland orifice. (Item D23) The model animal according to any one of the above items, wherein the step (2) includes evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item D24) In the step (2), the model animal according to any one of the above items, including at least the step of evaluating the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blinking frequency. (Item D25) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item D26) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item D27) The model animal according to any one of the above items, wherein the model animal has an H-2D type of d, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item D28) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item D29) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item D30) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an H-2D type of b, which is a subclass constituting MHC class I. (Item D31) A dry eye model animal for use in a method of screening a therapeutic agent and / or a prophylactic agent for dry eye, the method comprising: (1) a step of administering a test substance to the dry eye model animal; (2) a step of evaluating the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blinking frequency in the dry eye model animal; and (3) a step of identifying the test substance as a therapeutic agent and / or a prophylactic agent for dry eye when the occlusion of the meibomian gland opening, the abnormality of the tear film break-up time, and the abnormality of the blinking frequency are improved or the progression is suppressed in the dry eye model animal comprising: The dry eye model animal is an animal with reduced or impaired immune function into which cells of an allogeneic animal have been transplanted, The type of the major histocompatibility complex (MHC) of the dry eye model animal is H-2D d and the type of the MHC of the allogeneic animal is H-2D b is model animal. (Item D32) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item D33) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item D34) The model animal according to any one of the above items, wherein the model animal has the type d of H-2D, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item D35) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item D36) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item D37) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having the type b of H-2D, which is a subclass constituting the MHC class I. (Item E1) An eye disease model animal for use in a method for evaluating the medicinal efficacy of a test substance as a therapeutic agent and / or a prophylactic agent for eye diseases, the method comprising: (1) administering the test substance to the eye disease model animal; and (2) evaluating at least one of the characteristics related to meibomian gland dysfunction or the characteristics related to dry eye in the eye disease model animal. The animal model for eye diseases is an animal with reduced or impaired immune function due to transplantation of cells from allogeneic animals. Model animal. (Item E2) The model animal according to any one of the above items, wherein in the step (2), the method includes evaluating at least one feature related to meibomian gland dysfunction. (Item E3) The model animal according to any one of the above items, wherein in the step (2), the features related to meibomian gland dysfunction include at least one selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item E4) The model animal according to any one of the above items, wherein in the step (2), the features related to meibomian gland dysfunction include at least occlusion of the meibomian gland orifice. (Item E5) The model animal according to any one of the above items, wherein in the step (2), the method includes evaluating at least one feature related to meibomian gland dysfunction and at least one feature related to dry eye. (Item E6) The model animal according to any one of the above items, wherein in the step (2), the method includes evaluating at least occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate. (Item E7) The model animal according to any one of the above items, wherein the eye disease is meibomian gland dysfunction. (Item E8) The model animal according to any one of the above items, wherein the eye disease is dry eye. (Item E9) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item E10) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item E11) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are cells derived from the spleen. (Item E12) The model animal according to any one of the above items, wherein the model animal has an H-2D type of d, which is a subclass constituting the major histocompatibility complex (MHC) class I. (Item E13) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item E14) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an MHC type different from that of the model animal. (Item E15) The model animal according to any one of the above items, wherein the allogeneic animal is a mouse having an H-2D type of b, which is a subclass constituting MHC class I. (Item E16) An eye disease model animal for use in a method of screening for a therapeutic agent and / or prophylactic agent for an eye disease, the method comprising: (1) A step of administering a test substance to the eye disease model animal (2) A step of evaluating, in the eye disease model animal, a feature related to meibomian gland dysfunction or a feature related to dry eye; and (3) A step of identifying the test substance as a therapeutic agent and / or prophylactic agent for the eye disease when the evaluated feature is improved or the progression is suppressed comprising: The eye disease model animal is an animal in which the immune function is reduced or impaired by transplantation of cells of an allogeneic animal. Model animal. (Item E17) A meibomian gland dysfunction model animal for use in a method of screening for a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction, the method comprising: (1) A step of administering a test substance to the meibomian gland dysfunction model animal, (2) Evaluating the occlusion of the meibomian gland orifice in a meibomian gland dysfunction model animal; and (3) Identifying the test substance as a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction when the occlusion of the meibomian gland orifice is improved and / or the progression of the occlusion of the meibomian gland orifice is suppressed in the meibomian gland dysfunction model animal, wherein the meibomian gland dysfunction model animal is an animal with reduced or impaired immune function due to transplantation of cells from an allogeneic animal, the major histocompatibility complex (MHC) type of the meibomian gland dysfunction model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b and the model animal. (Item E18) A dry eye model animal for use in screening a therapeutic agent and / or prophylactic agent for dry eye, wherein the method comprises: (1) Administering a test substance to the dry eye model animal, (2) Evaluating the occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate in the dry eye model animal; and (3) Identifying the test substance as a therapeutic agent and / or prophylactic agent for dry eye when the occlusion of the meibomian gland orifice, abnormal tear film break-up time, and abnormal blink rate are improved or the progression is suppressed in the dry eye model animal comprising: wherein the dry eye model animal is an animal with reduced or impaired immune function due to transplantation of cells from an allogeneic animal, the major histocompatibility complex (MHC) type of the dry eye model animal is H-2D d and the MHC type of the allogeneic animal is H-2D b and the model animal. (Item F1) A dry eye model animal produced by transplanting cells of an animal of the same species but different strain as the dry eye model animal into an animal with reduced or defective immune function. (Item F2) An MGD model animal produced by transplanting cells of an animal of the same species but different strain as the MGD model animal into an animal with reduced or defective immune function. (Item F3) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item F4) The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse with the type of major histocompatibility complex (MHC) being H-2D d The model animal according to any one of the above items, wherein the model animal is a SCID mouse or a NOD-SCID mouse with the type of major histocompatibility complex (MHC) being H-2D (Item F5) The model animal according to any one of the above items, wherein the cells of the animal of the same species but different strain are immune cells. (Item F6) The model animal according to any one of the above items, wherein the cells of the animal of the same species but different strain are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. (Item F7) The model animal according to any one of the above items, wherein the animal of the same species but different strain is a mouse having a type of MHC class I different from that of the model animal. (Item F8) The method according to any one of the above items, wherein the animal of the same species but different strain is a mouse with the type of MHC being H-2D b The method according to any one of the above items, wherein the animal of the same species but different strain is a mouse with the type of MHC being H-2D (Item F9) The model animal according to any one of the above items, wherein the animal of the same species but different strain is a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse. (Item 26H) The model animal according to any one of the above items, having at least one feature related to meibomian gland dysfunction selected from closure of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (Item F10) The model animal according to any one of the above items, further having a feature related to dry eye selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate.

[0006] In the present disclosure, it is intended that the above one or more features can be provided in combination in addition to the explicitly stated combinations. Further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading the following detailed description as necessary.

Advantages of the Invention

[0007] The present disclosure provides an animal model for eye diseases having meibomian gland dysfunction, particularly a dry eye model animal having meibomian gland dysfunction. According to the present disclosure, it is possible to develop therapeutic and / or prophylactic agents for dry eye related to meibomian gland dysfunction and / or therapeutic and / or prophylactic agents for meibomian gland dysfunction, which have been difficult to develop heretofore.

Brief Description of the Drawings

[0008]

Figure 1

Figure 2

Figure 3

Mode for Carrying Out the Invention

[0009] Hereinafter, the present disclosure will be described. Throughout this specification, it should be understood that singular expressions include the concepts of their plural forms unless otherwise specifically mentioned. Therefore, singular articles (for example, in English, "a", "an", "the", etc.) should be understood to include the concepts of their plural forms unless otherwise specifically mentioned. Also, the terms used in this specification should be understood to be used in the meaning commonly used in the art unless otherwise specifically mentioned. Therefore, unless otherwise defined, all technical terms and scientific and technical terms used in this specification have the same meaning as generally understood by those skilled in the art to which the present disclosure pertains. In case of contradiction, this specification (including definitions) shall prevail. In this specification, "about" means ±10% of the value that follows.

[0010] (Definition) In this specification, "eye disease" means a disease that exhibits symptoms in the eye. It is a disease that shows subjective and objective symptoms in the eye and includes dry eye and meibomian gland dysfunction.

[0011] In this specification, "dry eye" refers to a disease diagnosed as "dry eye" in accordance with clinical diagnostic criteria, and is defined as "a multi-factorial disease that is a disease of the tear fluid and ocular surface caused by various factors, and is accompanied by eye discomfort, visual function abnormalities, destabilization of the tear film, or damage to the ocular surface". Dry eye is mainly classified into "aqueous tear-deficient dry eye" and "evaporative dry eye". In recent years, a classification called "dry eye with shortened tear film break-up time (BUT)" (hereinafter also referred to as "BUT-shortened dry eye") is also used.

[0012] In this specification, "aqueous tear-deficient dry eye" refers to dry eye that develops due to tissue destruction of the lacrimal gland or obstruction of tear drainage from the lacrimal gland to the ocular surface.

[0013] In this specification, "evaporative dry eye" refers to dry eye that develops when, although the tear secretion function is normal, water is excessively lost from the ocular surface due to various causes. Causes of evaporative dry eye include meibomian gland dysfunction (MGD) and abnormal lipids in the tear fluid, and one or both of them are present.

[0014] In this specification, "BUT-shortened dry eye" refers to dry eye with a short tear film break-up time (BUT) and having subjective symptoms of dry eye, but with almost normal tear secretion and corneal and conjunctival epithelium. There is also an idea of interpreting the shortening of BUT in connection with meibomian gland dysfunction, which is one of the causes of evaporative dry eye.

[0015] In this specification, "meibomian gland" refers to sebaceous glands that are located within the tarsal plate and have openings at the upper and lower eyelid margins. Meibum (lipid) is secreted from the meibomian gland. The presence of lipids in the tear fluid exerts effects such as suppressing tear evaporation, promoting tear stability, and promoting the spreading of the tear fluid on the ocular surface. The number of meibomian glands varies among individuals. In humans, there are approximately 50 in the upper eyelid and approximately 25 in the lower eyelid, and in mice, there are approximately 11 in both the upper and lower eyelids.

[0016] In this specification, "Meibomian gland dysfunction (MGD)" refers to a disease defined as "a state in which the function of the Meibomian glands is diffusely abnormal due to various causes and is accompanied by chronic eye discomfort." Meibomian gland dysfunction is also widely recognized as one of the causes of dry eye. Meibomian gland dysfunction (MGD) is broadly classified into a secretion-decrease type in which the secretion of sebum from the Meibomian glands is decreased and a secretion-increase type in which the secretion of sebum from the Meibomian glands is excessive, but the secretion-decrease type is overwhelmingly more common (Definition and diagnostic criteria of Meibomian gland dysfunction, New Ophthalmology 27(5):627-631, 2010). The secretion-decrease type of MGD leads to evaporative dry eye due to a decrease in the lipid phase of the tear film. The secretion-decrease type of MGD has subjective symptoms including eye discomfort and foreign body sensation, and abnormal findings around the Meibomian gland openings including eyelid margin irregularities, and findings of occlusion of the Meibomian gland openings (Table 4 of New Ophthalmology 27(5):627-631, 2010).

[0017] In this specification, "features related to Meibomian gland dysfunction" refers to features in the Meibomian glands associated with Meibomian gland dysfunction. Such features may be symptoms of Meibomian gland dysfunction or may cause the onset of Meibomian gland dysfunction.

[0018] In this specification, "occlusion of the Meibomian gland openings" refers to a state in which meibum or Meibomian gland components remain at the Meibomian gland openings without being secreted or discharged. "Occlusion of the Meibomian gland openings" refers to a state in which, when the eyelid is inverted under stereomicroscopic observation, white to yellow protrusions are observed at the Meibomian gland openings, or columnar or rice grain-shaped masses along the Meibomian gland ducts are observed inside the openings, or the Meibomian gland openings are expanded by the contents.

[0019] In this specification, "atrophy of the Meibomian glands" refers to a state in which the glandular acini of the Meibomian glands are reduced and the meibum stored in the Meibomian glands is decreased. "Atrophy of the Meibomian glands" is a state in which the white reflection of visible light decreases or the lipid reflected light of infrared light decreases when the eyelid is inverted under stereomicroscopy.

[0020] As used herein, "eyelid margin irregularity" refers to a line where the line (hereinafter referred to as the "eyelid margin") in contact with the cornea of the upper eyelid or lower eyelid is depressed in places. "Eyelid margin irregularity" is observed visually or under a stereomicroscope without treatment, or after instillation or injection of fluorescein.

[0021] As used herein, "break-up time of the tear film (BUT)" is an index of the stability of the tear film covering the cornea. "BUT" refers to the time until the "break-up of the tear film" is observed, where the tear film on the cornea is disrupted during forced eyelid opening for a certain period of time (10 seconds) while suppressing eye blinking, resulting in the appearance of a dry spot that gradually expands. It can be measured as the eyelid opening time from the state where the reflected light, interference light of the tear, or fluorescence of the fluorescein molecules contained in the tear after administration of fluorescein is uniformly observed on the cornea until a region where the reflected light or fluorescence disappears occurs.

[0022] As used herein, "blink rate" refers to the number of blinks in a certain period of time. "Blink rate" is one of the characteristics related to dry eye, which is the number of blinks in a certain period of time (for example, 1 minute) and can be observed visually.

[0023] As used herein, "corneal staining score" refers to the scoring of staining spots seen when the corneal surface layer is damaged in a punctate manner. The punctate damage of the corneal surface layer can be visualized as staining spots when pigments (such as fluorescein) are taken up by corneal epithelial cells or when pigments (such as fluorescein) are retained in the parts where corneal epithelial cells have fallen off. Corneal staining spots are observed, for example, under a stereomicroscope using an excitation wavelength of 400 - 540 nm and a fluorescence wavelength of 480 - 680 nm after instillation or injection of fluorescein. Also, the corneal staining score is obtained by dividing the cornea into three quadrants: upper, central, and lower. For each quadrant, it is determined whether there are no (0 points), sparse (1 point), intermediate between sparse and dense (2 points), or dense (3 points) punctate corneal staining spots, and is evaluated on a scale of up to 9 points in total. The corneal staining score may also be referred to as the superficial punctate keratopathy (SPK) score.

[0024] As used herein, the "tear volume" means the amount of liquid secreted from the lacrimal gland, and refers to the mixed amount of the tears stored in the meniscus (the accumulation of tears above the lower eyelid) and the tears secreted during the measurement time. One end of a Schirmer test strip or cotton thread can be inserted into the conjunctival fornix at the upper and lower or inner and outer corners so as not to stimulate the cornea and conjunctiva, and after a certain period of time, it can be removed and measured as the wet length of the Schirmer test strip or cotton thread. As an example, the said certain period of time is 15 seconds in the case of a mouse.

[0025] As used herein, the "tear film optical coherence score" is an index for evaluating tear abnormalities (for example, lipid abnormalities in tears). "Tear film optical coherence" refers to a phenomenon in which the phase of the light reflected from water and the light reflected from oil changes due to differences in the refractive index, content rate, and / or mixing rate of water and oil, which are the components of the tear film (composed of three layers: the lipid layer (oil layer), the aqueous layer, and the mucin layer), and interference fringes of rainbow colors are observed or parts with less reflected light are observed by enhancing a specific wavelength. The "tear film optical coherence score" is scored from 1 to 5 points based on the presence or absence of interference and the amount of reflected light observed using a stereomicroscope. The presence or absence of interference is visualized by the amount of rainbow-colored striped patterns, and the amount of reflected light is visualized by the brightness of the light reflection from the entire tear film. The evaluation can be determined, for example, as uniform white to silver reflection (1 point), non-uniform white to silver reflection (2 points), recognition of 2 to 3 color striped patterns (3 points), recognition of rainbow-colored striped patterns (4 points), and recognition of the exposed part of the cornea (5 points).

[0026] As used herein, the "subject" refers to the administration target of the medicament or method for treatment and prevention according to the present disclosure. Examples of the subject include mammals (for example, humans, mice, rats, hamsters, rabbits, cats, dogs, cows, horses, sheep, monkeys, etc.), and primates are preferred, and humans are particularly preferred.

[0027] As used herein, "treatment" means the cure, improvement, or suppression or alleviation of a disease or symptom. "Treating an eye disease" includes treating objective symptoms or subjective symptoms.

[0028] As used herein, "prophylaxis" means preventing the onset of a disease or symptom, and this concept includes minimizing the onset of a disease or symptom by delaying the onset of the disease or symptom or treating it before the onset. "Preventing and treating an eye disease" includes preventing objective or subjective symptoms.

[0029] As used herein, "subjective symptom" refers to a symptom among the symptoms of a disease that can be perceived by a patient suffering from the disease.

[0030] As used herein, "objective symptom" refers to a symptom among the symptoms of a disease that can be objectively proven by findings such as imaging findings and numerical values of examination results (objective findings).

[0031] As used herein, "animal" means an animal used as a model animal, and examples include mammals (e.g., mice, rats, hamsters, rabbits, cats, dogs, cows, horses, sheep, monkeys, etc.).

[0032] As used herein, "model animal" refers to an animal created so as to exhibit symptoms similar to those of human diseases, and is used for elucidating mechanisms, developing preventive or therapeutic agents, or evaluating drug efficacy. If an animal develops the target disease and exhibits the characteristics of the disease, but it is impossible to elucidate the mechanism, develop a preventive or therapeutic agent, or evaluate drug efficacy due to the irreversibility of the characteristics, or the need for a considerable amount of time until the onset and manifestation of the characteristics of the disease (e.g., 50 weeks of age or older), such an animal is not a model animal.

[0033] As used herein, "allogeneic" refers to individuals that are of the same species but genetically different.

[0034] As used herein, "reduction or deficiency of immune function" is characterized by a lack of function of immune cells, and refers to a state in which the immune response to allogeneic cells is reduced or not induced.

[0035] As used herein, "the major histocompatibility complex (MHC) type is H-2D" d means that H-2D of the subclass constituting MHC class I is a homozygous form of d. "The MHC type is H-2D" b means that H-2D of the subclass constituting MHC class I is a homozygous form of b.

[0036] As used herein, "immune cell" refers to cells constituting the immune system, including T cells, B cells, dendritic cells, macrophages, neutrophils, mast cells, eosinophils, basophils, and natural killer cells.

[0037] As used herein, "cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes" refers to cells prepared and / or isolated from the spleen, blood, bone marrow, thymus, or lymph nodes, or cultured cells obtained by culturing the prepared and / or isolated cells.

[0038] As used herein, "containing cells in the meibomian gland" means containing external cells not derived from the model animal itself in the meibomian gland. The cells contained in the meibomian gland are confirmed by detecting the presence of external cells through pathological examination of the meibomian gland, or by using an antibody against cell surface membrane proteins of external cells, etc.

[0039] As used herein, "developing characteristics related to meibomian gland dysfunction by 40 weeks of age" means developing characteristics related to meibomian gland dysfunction at any time before reaching 40 weeks of age and becoming a state available as a model animal.

[0040] As used herein, "test substance" is a substance used for evaluating drug efficacy and screening using a model animal, and can be any substance such as a low-molecular compound, polysaccharide, protein, peptide, nucleic acid, or their fusion.

[0041] As used herein, "evaluation" means confirming the state of characteristics related to meibomian gland dysfunction and / or characteristics related to dry eye in a model animal, and also includes observing or measuring the characteristics as necessary, thereby determining whether the test substance has the intended properties or effects, or the degree of the intended properties or effects of the test substance.

[0042] (Preferred Embodiment) The following describes preferred embodiments. It should be understood that these embodiments are illustrative of the present invention and that the scope of the present invention is not limited to such preferred embodiments. Those skilled in the art should also understand that modifications and changes within the scope of the present disclosure can be easily made with reference to the following preferred examples. Regarding these embodiments, those skilled in the art can appropriately combine any embodiments.

[0043] (Model Animal) In one aspect, the present disclosure provides an eye disease model animal having characteristics related to meibomian gland dysfunction, particularly a meibomian gland dysfunction model animal or a dry eye model animal. Most dry eye patients have concurrent meibomian gland dysfunction. Therefore, in the treatment of dry eye, treatment for both dry eye symptoms and meibomian gland dysfunction is desired. However, there was no model animal with concurrent dry eye symptoms and meibomian gland dysfunction, making it difficult to develop therapeutic and prophylactic agents. There was also a problem that there were few meibomian gland dysfunction model animals that could be used for the development of therapeutic and prophylactic agents for meibomian gland dysfunction. The present inventors newly found a model animal having characteristics related to meibomian gland dysfunction, and further a model animal having concurrent characteristics related to dry eye and meibomian gland dysfunction.

[0044] In some embodiments, the model animal of the present disclosure is a non-human animal, preferably a non-human mammal, more preferably a rodent, such as a mouse, a rat, or a hamster, and most preferably a mouse.

[0045] In some embodiments, the model animal of the present disclosure can be an animal with reduced or impaired immune function into which cells of an allogeneic animal of the same species as the model animal have been transplanted. Examples of cells of an allogeneic animal of the same species as the model animal include, but are not limited to, T cells, B cells, macrophages, dendritic cells, sinusoidal endothelial cells, and NK cells. In some embodiments, the cells of an allogeneic animal of the same species as the model animal can be immune cells, for example, immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. The cells of the allogeneic animal are preferably spleen cells.

[0046] The model animal with reduced or impaired immune function may lack T cells and / or B cells, and preferably may lack both T cells and B cells. The reduction or impairment of immune function is preferably a state that occurs congenitally, that is, a state that occurs due to a gene mutation.

[0047] The model animal with reduced or impaired immune function may be in a state where the function of the protein Prkdc (Protein kinase, DNA activated, catalytic polypeptide) is deficient due to a mutation in the Prkdc gene, and preferably may have a mutation denoted as Prkdc scid Since the function of Prkdc is deficient, gene rearrangement of B cells and T cells cannot occur, resulting in the deficiency of mature B cells and T cells.

[0048] In some embodiments, the model animal of the present disclosure can be a SCID mouse or a NOD-SCID mouse.

[0049] In some embodiments, the model animal of the present disclosure can be CB17 / Icr-Prkdc scid / CrlCrlj.

[0050] In some embodiments, the allogeneic animal may have a major histocompatibility complex (MHC) type different from that of the model animal. The type of MHC class I of the model animal of the present disclosure is d, and the type of MHC class I of the allogeneic animal may be a, b, k, q, s other than d, but may preferably be b. In certain embodiments, H-2D, a subclass that constitutes MHC class I of the model animal of the present disclosure, is d, and H-2D, a subclass that constitutes MHC class I of the allogeneic animal, may be a, b, k, q, s other than d, but may preferably be b. The subclass that constitutes MHC class I may be homozygous or heterozygous. The H-2D of the model animal of the present disclosure is a homozygous form of d (H-2D d ), and the H-2D of the allogeneic animal may be a homozygous form of b (H-2D b ).

[0051] In some embodiments, the model animal of the present disclosure may be a SCID mouse or a NOD-SCID mouse whose major histocompatibility complex (MHC) type is H-2D d . In a further embodiment, the model animal of the present disclosure is a SCID mouse or a NOD-SCID mouse whose major histocompatibility complex (MHC) type is H-2D d , and the allogeneic animal may be a mouse whose MHC type is H-2D b .

[0052] In some embodiments, the allogeneic animal may be a C57BL / 6 mouse, a C57BL / 10 mouse, a C57L / J mouse, or a BXSB / Mp mouse.

[0053] In some embodiments, the model animal of the present disclosure may have at least one, at least two, or all of the features associated with meibomian gland dysfunction selected from the group consisting of occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity.

[0054] Preferably, in the model animals of the present disclosure, the number of blocked meibomian gland orifices is increased compared to healthy animals that have not developed an eye disease (for example, animals to which allogeneic cells of the same species as the model animals have not been administered) (hereinafter referred to as "non-model animals"). For example, the evaluation of "blockage of meibomian gland orifices" can be performed using the number of blocked meibomian gland orifices per individual.

[0055] Preferably, in the model animals of the present disclosure, atrophy of the meibomian gland can be confirmed, whereas atrophy of the meibomian gland is not observed in non-model animals. For example, the evaluation of "atrophy of the meibomian gland" can be performed using the number of individuals showing atrophy of the meibomian gland (that is, the ratio of the number of individuals showing atrophy of the meibomian gland) to the total number of individuals in the model animal group under the same conditions.

[0056] Preferably, in the model animals of the present specification, the eyelid margin is irregular, that is, the eyelid margin is indented in places, whereas non-model animals show a smooth eyelid margin line. For example, the evaluation of "irregular eyelid margin" can be performed using the number of individuals showing an irregular eyelid margin (that is, the ratio of the number of individuals showing an irregular eyelid margin) to the total number of individuals in the model animal group under the same conditions. The model animals of the present disclosure are advantageous as model animals because they develop characteristics related to meibomian gland dysfunction and / or characteristics of dry eye relatively early after administration of allogeneic cells. In some embodiments, it may develop on the 3rd day, 4th day, 5th day, 6th day, 7th day, 8th day, 9th day, 10th day, 11th day, 12th day, 13th day, 14th day, or at the latest on the 15th day to 31st day after administration of allogeneic cells. In some embodiments, the model animals of the present disclosure may be characterized in that they develop characteristics related to meibomian gland dysfunction by 40 weeks of age, 35 weeks of age, 25 weeks of age, 20 weeks of age, 15 weeks of age, 14 weeks of age, 13 weeks of age, 12 weeks of age, 11 weeks of age, 10 weeks of age, 9 weeks of age, 8 weeks of age, or 7 weeks of age.

[0057] In a further embodiment, the model animal of the present disclosure may have at least one, at least two, at least three, at least four or all of the features related to dry eye selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate.

[0058] Preferably, in the model animals of the present specification, the BUT is decreased as compared with non-model animals. For example, the evaluation of "BUT" can be performed using the BUT per individual in a group of model animals under the same conditions.

[0059] Preferably, in the model animals of the present specification, the blink rate is increased as compared with non-model animals. For example, the evaluation of "blink rate" can be performed using the blink rate per individual in a group of model animals under the same conditions.

[0060] Preferably, in the model animals of the present specification, the corneal staining score is increased as compared with non-model animals. For example, the evaluation of "corneal staining score" can be performed using the corneal staining score per individual in a group of model animals under the same conditions.

[0061] Preferably, in the model animals of the present specification, the tear volume is decreased as compared with non-model animals. For example, the evaluation of "tear volume" can be performed using the tear volume per individual in a group of model animals under the same conditions.

[0062] Preferably, in the model animals of the present specification, the tear film optical coherence score is increased as compared with non-model animals. For example, the evaluation of "tear film optical coherence score" can be performed using the tear film optical coherence score per individual in a group of model animals under the same conditions.

[0063] In a specific embodiment, the model animal of the present disclosure may have at least the features related to dry eye of abnormal tear film break-up time and / or abnormal corneal staining score as symptoms of dry eye.

[0064] In a further aspect, the present disclosure provides a meibomian gland dysfunction model animal, which is an immunodeficient animal with reduced or defective immune function, and is a meibomian gland dysfunction model animal in which immune cells of an allogeneic animal of the same species as the meibomian gland dysfunction model animal are transplanted, and is an animal having at least one characteristic selected from the characteristics associated with meibomian gland dysfunction, wherein the characteristic associated with meibomian gland dysfunction is selected from occlusion of the meibomian gland opening, atrophy of the meibomian gland, and eyelid margin irregularity. In a specific embodiment, the meibomian gland dysfunction model animal is an animal with reduced or defective immune function into which cells of an allogeneic animal have been transplanted, and the major histocompatibility complex (MHC) type is H-2D d and the allogeneic animal can be a mouse with an MHC type of H-2D b . In a further embodiment, the above-mentioned meibomian gland dysfunction model animal is an animal having at least one characteristic selected from the characteristics associated with dry eye, wherein the characteristic associated with dry eye is selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. In a further embodiment, the characteristic associated with meibomian gland dysfunction in the above-mentioned meibomian gland dysfunction model animal is occlusion of the meibomian gland opening, and the characteristic associated with dry eye can be abnormal tear film break-up time and / or abnormal corneal staining score.

[0065] In a further aspect, the present disclosure provides a dry eye model animal, which is an immunodeficient animal with reduced or impaired immune function, and is a dry eye model animal into which immune cells of an allogeneic animal of the same species as the dry eye model animal have been transplanted, and has at least one feature selected from the features related to meibomian gland dysfunction in the following (1) and at least one feature selected from the features related to dry eye in the following (2). (1) The features related to meibomian gland dysfunction are selected from the closure of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity. (2) The features related to dry eye are selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate. In a specific embodiment, the dry eye model animal is an animal with reduced or impaired immune function into which cells of an allogeneic animal have been transplanted, and the major histocompatibility complex (MHC) type is H-2D d and the allogeneic animal can be a mouse with an MHC type of H-2D b . In a further embodiment, the feature related to meibomian gland dysfunction in the dry eye model animal is closure of the meibomian gland orifice, and the features related to dry eye can be abnormal tear film break-up time and / or abnormal corneal staining score.

[0066] In a further aspect, the present disclosure provides a model animal characterized in that the features related to meibomian gland dysfunction develop by 40 weeks of age, 35 weeks of age, 25 weeks of age, 20 weeks of age, 15 weeks of age, 10 weeks of age, or 7 weeks of age.

[0067] In some embodiments, the model animal of the present disclosure may develop the features related to meibomian gland dysfunction after 4 weeks of age, 5 weeks of age, or 6 weeks of age.

[0068] In a further aspect, the present disclosure provides a model animal, characterized in that the meibomian gland contains cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes, preferably cells derived from the spleen. The model animal can be a dry eye model animal or a meibomian gland dysfunction (MGD) model animal, or both.

[0069] In a further aspect, the present disclosure provides a model animal, wherein the meibomian gland contains at least one cell selected from the group consisting of T cells, B cells, macrophages, dendritic cells, sinusoidal endothelial cells, and NK cells. The cells may be allogeneic cells.

[0070] The above cells contained in the meibomian gland can be confirmed as follows. (1) When the model animal is an immunodeficient mouse lacking immune cells, a pathological examination of the meibomian gland is performed using a marker (such as an antibody) specific for the lacking immune cells. If the lacking immune cells are detected in the meibomian gland, it can be confirmed that external cells have reached the meibomian gland of the model animal. Since immunodeficient mice such as SCID mice lack T cells and B cells, if T cells or B cells are present in the meibomian gland by pathological examination, it can be confirmed that external cells have reached the meibomian gland. (2) A pathological examination of the meibomian gland is performed using a marker (such as an antibody) specific for an MHC type different from that of the model animal. If an MHC type different from that of the model animal is detected in the meibomian gland, it can be confirmed that external cells have reached the meibomian gland of the model animal. In this case, it is desirable to confirm that the cells derived from the spleen have an MHC type different from that of the model animal.

[0071] In a further aspect, the present disclosure provides a dry eye model animal, which is an animal (preferably congenital) with reduced or defective immune function, into which cells of an animal of the same species but different strain as the model animal have been transplanted.

[0072] In a further aspect, the present disclosure provides a meibomian gland dysfunction (MGD) model animal, which is an animal having reduced or defective immune function (preferably congenital) and into which cells of an animal of the same species but different strain as the model animal have been transplanted.

[0073] In a further aspect, the present disclosure provides a meibomian gland dysfunction (MGD) model animal having an occlusion at the opening of the meibomian gland, which is an animal having reduced or defective immune function and into which cells of an animal of the same species but different strain as the model animal have been transplanted.

[0074] (Method for producing a model animal) In another aspect, the present disclosure provides a method for producing a dry eye model animal, the method including a step of transplanting cells of an animal of the same species but different strain as the dry eye model animal into an animal having reduced or defective immune function.

[0075] In a further aspect, the present disclosure provides a method for producing an MGD model animal, the method including a step of transplanting cells of an animal of the same species but different strain as the MGD model animal into an animal having reduced or defective immune function.

[0076] The production method of the present disclosure may include a step of breeding for 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or at least 15 days to 31 days after transplanting cells into an animal having reduced or defective immune function. The breeding can be carried out by general methods, but it is preferable to breed a plurality of mice in one space (e.g., in a cage, etc.) with a density of 1 mouse / 50 - 200 cm 2 1 mouse / 130 - 180 cm 2 and preferably breed them in an environment with such a density.

[0077] Embodiments such as model animals, animals of the same species but different strain, and cells of animals of the same species but different strain are described above.

[0078] Meibomian gland dysfunction clinically typically presents symptoms such as abnormal findings around the meibomian gland orifices, meibomian gland orifice occlusion, and meibomian gland atrophy. Abnormal findings around the meibomian gland orifices include eyelid margin irregularities. This model animal has "occlusion of the meibomian gland orifices", "atrophy of the meibomian glands", or "eyelid margin irregularities" as characteristics related to meibomian gland dysfunction. The said characteristics in this model animal are reversible, and this model animal can be used for the development of therapeutic and preventive agents for eye diseases, particularly meibomian gland dysfunction.

[0079] Dry eye clinically typically presents an objective sign, the tear breakup time (BUT), and subjective symptoms (eye discomfort or visual function abnormalities). BUT is an essential test item in the diagnosis of dry eye in Japan. As a criterion for the development of dry eye therapeutic drugs by the US Food and Drug Administration (FDA), it is essential that at least one of the objective findings is significantly improved, and the objective findings include BUT. Also, subjective symptoms (such as eye discomfort and visual function abnormalities) are essential test items in the diagnosis of dry eye in Japan and are clinically obtained by interviewing the patient himself / herself and scored. The subjective symptoms in human dry eye can be evaluated in model animals by substituting the blink rate, and an increase in the blink rate means worsening of the subjective symptoms. This model animal has "BUT" or "blink rate" as characteristics related to dry eye. The said characteristics in this model animal are reversible, and this model animal can be used for the development of therapeutic and preventive agents for eye diseases.

[0080] Furthermore, dry eye may be accompanied by objective symptoms such as "corneal and conjunctival epithelial disorders", "decrease in tear volume", and "abnormal tear lipid layer". "Corneal and conjunctival epithelial disorders" refer to the state in which the cornea and conjunctiva are damaged. Clinically, it is evaluated by staining tests. Corneal and conjunctival epithelial disorders include superficial punctate keratopathy, which is a defect up to the wing cells of the corneal epithelial layer, corneal erosion, which is a defect of the entire epithelial layer including basal cells but the basement membrane is not damaged, and corneal ulcer, in which the lesion extends to the substantia beyond the basement membrane. Although corneal and conjunctival epithelial disorders are not essential for the diagnosis of dry eye, it is desirable for them to show efficacy as a therapeutic agent for dry eye. "Corneal and conjunctival epithelial disorders" are scored by confirming the slit lamp microscopic images under pigment (e.g., fluorescein) staining / non-staining in a staining test. The "corneal staining score" in model animals corresponds to the scoring of such corneal and conjunctival epithelial disorders. Also, "decrease in tear volume" is not essential for the diagnosis of dry eye, but it is desirable for it to show efficacy as a therapeutic agent for dry eye. Tear volume is included as one of the objective findings that must be improved in the FDA criteria for the development of dry eye therapeutic agents. Clinically, tear volume can be measured by the Schirmer test, and the same means can be used to measure it in model animals. In addition, "abnormal tear lipid layer" is not essential for the diagnosis of dry eye, but it is desirable to confirm improvement as an index for the development of therapeutic agents for dry eye. Abnormal tear lipid layer is an abnormality in the amount and quality of lipids, which affects the stability of tears. Clinically, "abnormal tear lipid layer" is detected by observing "tear film interferometry", and it can also be scored and measured in model animals using the same means. This model animal has "abnormal corneal staining score", "decrease in tear volume", or "abnormal tear film interferometry score" as characteristics related to dry eye, and these characteristics in this model animal are reversible. Therefore, this model animal can be advantageously used in the development of therapeutic and preventive agents for eye diseases.

[0081] The model animals of the present disclosure can be used to develop therapeutic and / or prophylactic agents for treating meibomian gland dysfunction, develop therapeutic and / or prophylactic agents for treating dry eye associated with meibomian gland dysfunction, for example, develop therapeutic and / or prophylactic agents for evaporative dry eye.

[0082] (Method for evaluating drug efficacy) In different aspects, the present disclosure provides a method for evaluating the drug efficacy of a test substance as a therapeutic and / or prophylactic agent for an eye disease, the method comprising: (1) administering the test substance to an eye disease model animal; and (2) evaluating at least one of the characteristics associated with meibomian gland dysfunction or the characteristics associated with dry eye, wherein the eye disease model animal is an animal with reduced or impaired immune function transplanted with cells of allogeneic animals. In a particular embodiment, the eye disease can be dry eye and / or meibomian gland dysfunction.

[0083] In a further aspect, the present disclosure provides a method for evaluating the drug efficacy of a test substance as a therapeutic and / or prophylactic agent for meibomian gland dysfunction, the method comprising: (1) administering the test substance to a meibomian gland dysfunction model animal; and (2) evaluating the characteristics associated with meibomian gland dysfunction.

[0084] In a further aspect, the present disclosure provides a method for evaluating the drug efficacy of a test substance as a therapeutic and / or prophylactic agent for dry eye, the method comprising: (1) administering the test substance to a dry eye model animal; and (2) evaluating the characteristics associated with meibomian gland dysfunction and the characteristics associated with dry eye.

[0085] In some embodiments, in the step (1), the method of administering the test substance may be oral administration, eye drop administration, intravitreal injection, etc., and the dosage form of the test substance to be administered may be tablets, eye drops, injection solutions, etc. In a preferred embodiment, the test substance can be administered by eye drops. For example, the test substance can be administered 1 to 3 days, preferably 1 to 2 days, after the onset of characteristics related to meibomian gland dysfunction or characteristics related to dry eye, or can be administered 6 to 9 days, preferably 7 to 8 days, after transplanting cells of an animal of the same species but different strain as the model animal into the model animal.

[0086] The method for evaluating the drug efficacy in the present disclosure may include a step of evaluating that the test substance has drug efficacy when characteristics related to meibomian gland dysfunction and / or characteristics related to dry eye are improved or the progression is suppressed in a model animal administered with the test substance.

[0087] In some embodiments, the drug efficacy can be evaluated by changing the administration route of the test substance, the composition of the formulation, the dosage form and concentration of the formulation.

[0088] In some embodiments, in the step (2), the characteristics related to meibomian gland dysfunction are at least one characteristic selected from occlusion of the meibomian gland orifice, atrophy of the meibomian gland, and eyelid margin irregularity, and the characteristics related to dry eye may be at least one characteristic selected from abnormal tear film break-up time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate.

[0089] In some embodiments, in the step (2), the characteristics related to dry eye and / or the characteristics related to meibomian gland dysfunction can be evaluated. In a specific embodiment, in the step (2), at least one, at least two or three of the characteristics related to meibomian gland dysfunction and at least one, at least two, at least three, at least four or five of the characteristics related to dry eye can be evaluated.

[0090] Preferably, the occlusion of the meibomian gland orifice in the model animal can be determined to have improved when, for example, the number of occlusions has decreased compared to before administration of the test substance, or, for example, when the number of occlusions is lower when the test substance is administered than when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

[0091] Preferably, atrophy of the meibomian gland in the model animal can be determined to have improved when, for example, the size of the acini of the meibomian gland has increased, the retention of meibum has increased, or the reflection of infrared light by the retained sebum has increased, or when the reflection that should anatomically be present has recovered compared to before administration of the test substance, or, for example, when the acini of the meibomian gland are larger, the retention of meibum has increased, or the reflection of infrared light by the retained sebum has increased, or when the reflection that should anatomically be present has recovered when the test substance is administered than when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

[0092] Preferably, eyelid margin irregularity in the model animal can be determined to have improved when, for example, compared to before administration of the test substance, the indentation of the eyelid margin line has decreased and become a smooth line, or, for example, when the indentation of the eyelid margin line has decreased and become a smooth line when the test substance is administered than when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

[0093] Preferably, BUT in the model animal can be determined to have improved when, for example, it has increased compared to before administration of the test substance, or, for example, when it has increased when the test substance is administered than when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

[0094] Preferably, when the number of blinks in the model animal decreases, for example, compared to before administration of the test substance, it can be determined that the characteristics have improved. Alternatively, for example, when the test substance is administered, if the number of blinks is less than when a placebo is administered, it can be determined that the progression of the characteristics has been suppressed.

[0095] Preferably, when the corneal staining score in the model animal decreases, for example, compared to before administration of the test substance, it can be determined that the characteristics have improved. Alternatively, for example, when the test substance is administered, if the score is less than when a placebo is administered, it can be determined that the progression of the characteristics has been suppressed.

[0096] Preferably, when the amount of tear fluid in the model animal increases compared to before administration of the test substance, it can be determined that the characteristics have improved. Alternatively, for example, when the test substance is administered, if the amount of tear fluid in the model animal is greater than when a placebo is administered, it can be determined that the progression of the characteristics has been suppressed.

[0097] Preferably, when the score of the tear film optical interference in the model animal decreases compared to before administration of the test substance, it can be determined that the characteristics have improved. Alternatively, for the tear film optical interference in the model animal, for example, if it is less than when a placebo is administered, it can be determined that the progression of the characteristics has been suppressed.

[0098] Preferably, the improvement of each characteristic refers to the case where there is a significant difference in the measured value of the characteristic after administration of the test substance compared to before administration of the test substance, or a difference of 10% or more, 20% or more, 30% or more, 40% or more, 45% or more, 46% or more, 48% or more.

[0099] Preferably, the suppression of the progression of each feature refers to the case where there is a significant difference in the measured value of the feature when the test substance is administered compared to when a placebo is administered, or when there is a difference of 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more.

[0100] Note that the significant difference can be confirmed by general statistical methods and means that there is a statistically significant difference.

[0101] In a further embodiment, in step (2), at least the occlusion of the meibomian gland orifice, the tear film break-up time, and the blink rate can be evaluated.

[0102] Embodiments such as model animals, allogeneic animals, and cells of allogeneic animals are described above.

[0103] (Screening method) In different aspects, the present disclosure provides a method for screening a therapeutic agent and / or prophylactic agent for dry eye, the method comprising: (1) administering a test substance to a dry eye model animal; (2) evaluating features related to meibomian gland dysfunction and features related to dry eye; and (3) identifying the test substance as a therapeutic agent and / or prophylactic agent for dry eye when the evaluated features are improved or the progression is suppressed in the dry eye model animal.

[0104] In different aspects, the present disclosure provides a method for screening a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction, the method comprising: (1) administering a test substance to a meibomian gland dysfunction animal; (2) evaluating features related to meibomian gland dysfunction; and (3) identifying the test substance as a therapeutic agent and / or prophylactic agent for meibomian gland dysfunction when the evaluated features are improved or the progression is suppressed in the meibomian gland dysfunction animal.

[0105] In a further aspect, the present disclosure provides a method for screening a therapeutic agent and / or a prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an animal model of an eye disease, wherein the animal model of an eye disease is an immunodeficient animal transplanted with immune cells of an allogeneic animal; (2) evaluating a feature related to meibomian gland dysfunction or a feature related to dry eye; and (3) identifying the test substance as a therapeutic agent and / or a prophylactic agent for the eye disease when the evaluated feature is improved or the progression is suppressed in the animal model of the eye disease.

[0106] Embodiments such as the model animal, the allogeneic animal, the cells of the allogeneic animal, and the evaluation items are described above.

[0107] In a further aspect, in step (1) of the screening method, the test substance may be two or more candidate substances, and in step (3), a step of selecting a therapeutic agent and / or a prophylactic agent for the eye disease from among the plurality of candidate substances may be performed.

[0108] The present disclosure has been described above with reference to preferred embodiments for ease of understanding. Hereinafter, the present disclosure will be described based on examples. However, the above description and the following examples are provided for illustrative purposes only and are not provided for the purpose of limiting the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiments and examples specifically described herein, but is limited only by the claims.

Example

[0109] The present disclosure will be described in more detail below based on examples. These are for better understanding of the present disclosure and do not limit the scope of the present disclosure. In addition, these examples were carried out with the approval of the Animal Experiment Ethics Review Committee of Chisso Corporation.

[0110] (Example 1: Preparation and evaluation of an animal model of an eye disease) 1. Test method 1-1. Preparation of a model mouse Female C.B17 ICR SCID mice at 5 to 7 weeks of age (CB17 / Icr-Prkdc scid / CrlCrlj, H-2D d (purchased from Charles River Laboratories Japan, Inc.) were divided into a Sham group and a spleen-derived cell administration group (6 mice in each group). In each group, 5 mice were used for observation over time and 1 mouse was used for pathological observation.

[0111] In the Sham group, PBS (manufactured by Thermo Fisher Scientific) was administered into the tail vein at 200 μL / mouse. In the spleen-derived cell administration group, a PBS suspension of spleen-derived cells (2.5×10 7 cells / mL) at 200 μL / mouse was administered into the tail vein. The PBS suspension of spleen-derived cells (hereinafter referred to as the "spleen-derived cell suspension") was prepared by making incisions in the spleens collected from C57BL / 6 mice (H-2D b ), dispersing them by pipetting several times, and suspending them in PBS. After administration of PBS or the spleen-derived cell suspension, the mice were housed in breeding cages (size: 338 mm long, 225 mm wide, 140 mm high) in groups of 5.

[0112] 1-2. Evaluation of each characteristic (a) Evaluation of characteristics related to meibomian gland dysfunction (a1) Number of occlusions of meibomian gland openings Taking the day of administration of the spleen-derived cell suspension as day 0, on days 8, 15, and 21 after administration, the number of occlusions of the meibomian glands in the right eyes of the mice in each group was measured under anesthesia using a stereomicroscope. The number of occlusions of the meibomian glands was measured by gently inverting the upper and lower eyelids respectively under stereomicroscopic observation to expose the meibomian gland openings, and counting the number of openings with occlusions from the inner canthus side towards the outer canthus.

[0113] Under a stereomicroscope, when white to yellow protrusions were observed at the openings of the meibomian glands, when the meibomian gland openings were expanded by the contents, or when columnar or rice grain-shaped masses along the meibomian gland ducts were observed within the openings, it was determined that the meibomian gland openings were blocked. The total number of blockages observed in the upper and lower eyelids was taken as the individual value of the meibomian gland blockage number, and the average value of 5 animals was calculated.

[0114] (a2) Atrophy of the meibomian gland Similar to the confirmation of the meibomian gland blockage number, with the administration day of the spleen-derived cell suspension as day 0, on days 8, 15, and 21 after administration, the atrophy of the right eye's meibomian gland was confirmed under anesthesia using a stereomicroscope, and the number of individuals with developed atrophy was counted.

[0115] For the atrophy of the meibomian gland, the upper and lower eyelids were each flipped under stereomicroscopic observation, and the area of the meibomian gland that could be seen through the conjunctiva was observed. When the area was smaller than the size of the meibomian gland of a general mouse, it was determined to be atrophied, and the number of individuals in which one or more atrophied meibomian glands were observed was counted.

[0116] In addition, the atrophy of the meibomian gland was also anatomically confirmed. On day 28 after the administration of the spleen-derived cell suspension, the mice were euthanized and the eyelids were collected. The collected eyelids were embedded and frozen in OCT compound (manufactured by Sakura Finetek Japan Co., Ltd.), and 10-μm-thick sagittal frozen sections were prepared. The obtained sections were stained with Oil Red O to stain the lipids in the meibomian gland red and with hematoxylin pigment to stain the cell nuclei blue. After staining, the state of the meibomian gland in the sections was observed using a microscope.

[0117] (a3) Eyelid margin irregularity With the administration day of the spleen-derived cell suspension as day 0, on days 7, 14, and 21 after administration, the eyelid margin irregularity of the right eye was observed under anesthesia using a stereomicroscope.

[0118] A fluorescein sodium solution (a solution prepared by dissolving uranine (manufactured by Fujifilm Wako Pure Chemical Corporation) in physiological saline and filtering it through a filter) was instilled into the right eye of anesthetized mice, and thoroughly mixed with the tears by several forced blinks. Thereafter, under a stereomicroscope fluorescence observation (excitation wavelength bandpass 460 - 480 nm, fluorescence wavelength bandpass 495 - 540 nm), a fluorescence stereomicroscopic image of the eyelid margin was taken and observed. A case where the groove of the eyelid was depicted as a smooth curve by fluorescein was regarded as normal, and a case where irregularities were depicted was determined as eyelid margin irregularity.

[0119] (b) Evaluation of characteristics related to dry eye (b1) Tear film breakup time (BUT) Taking the administration day of the spleen-derived cell suspension as day 0, on days 7, 14, and 21 after administration, the tear film breakup time of the right eye was measured under anesthesia using a stereomicroscope.

[0120] The tear film breakup time was measured using a fluorescein sodium solution in the same manner as (a3). In the same manner as (a3), a fluorescein sodium solution was instilled into the right eye of anesthetized mice, and thoroughly mixed with the tears by several forced blinks. Thereafter, it was placed under a stereomicroscope fluorescence observation (excitation wavelength bandpass 460 - 480 nm, fluorescence wavelength bandpass 495 - 540 nm). The mixed solution of tears and fluorescein covered the cornea, and by confirming the disappearance of the fluorescence of fluorescein, the breakup of the tear film could be evaluated. After placing the mice under a stereomicroscope fluorescence observation, the right eye of the mice was forcibly opened, and the time from the moment of opening the eyelid until the site where the fluorescence disappeared appeared was measured 3 times, and the average value of the 3 times was taken as the individual value, and the average value was calculated from the individual values of 5 individuals and used as the tear film breakup time (BUT).

[0121] (b2) Blink frequency The administration date of the spleen-derived cell suspension was defined as day 0. On days 4, 11, and 18 after administration, the mice were transferred to small cages for observation and isolated individually. The blinking frequency of the left eye of the mice moving freely in the cage was measured. The measurement was performed 5 times for 1 minute per individual, and the total of the 5 times was used as the individual value, and the average value of 5 individuals was calculated. After the measurement, the mice were returned to the breeding cage, and 5 mice were placed in one cage as usual and breeding was continued.

[0122] (b3) Corneal staining spot score The administration date of the spleen-derived cell suspension was defined as day 0. On days 7, 14, and 21 after administration, the corneal staining spot score (SPK score) of the right eye was measured under anesthesia using a stereomicroscope.

[0123] After measuring the tear film break-up time in (b1), the eyes were washed with physiological saline for observation of corneal staining spots. After eye washing, the mice were placed under a stereomicroscope fluorescence observation again (excitation wavelength band pass 460 - 480 nm, fluorescence wavelength band pass 495 - 540 nm). Observation of corneal staining spots was performed by confirming the fluorescence of punctate fluorescein in the outermost layer of the cornea.

[0124] Observation was performed for 5 quadrants of the upper, right, lower, left, and central parts of the cornea. Each punctate fluorescence was evaluated as none (0 points), sparse (1 point), moderately dense (2 points), or dense (3 points), and the total score of the 5 quadrants was calculated and used as the individual value of the corneal staining spot score (SPK score). The average value was calculated from the obtained individual values.

[0125] (b4) Tear volume The administration date of the spleen-derived cell suspension was defined as day 0. On days 7, 14, and 21 after administration, the tear volume of the right eye was measured under wakeful restraint using a stereomicroscope.

[0126] One end of a cotton thread containing phenol red (Zone Quick, manufactured by Ayumi Pharmaceutical Co., Ltd.) was inserted into the conjunctival fornix of the right eye of the mouse and held for 15 seconds, then immediately removed. The length of the discolored phenol red contained in the cotton thread by tears was measured and used as an index of tear volume. The average value was calculated from the measured lengths.

[0127] (b5) Tear film optical coherence score The administration date of the cell suspension derived from the spleen was defined as day 0. On day 31 after administration, slit lamp microscopy was used to observe the optical coherence of the tear film in the right eye under anesthesia.

[0128] Observation was performed with the focus of the slit lamp microscope adjusted to the tear film after forced eyelid opening of the right eye of the mouse. Scoring was performed on a scale of 1 to 5 based on the number of iridescent stripe patterns generated by optical coherence and the brightness of light reflection from the entire tear film. Scoring was determined as uniform white-silver reflection (1 point), non-uniform white-silver reflection (2 points), recognition of 2-3 color stripe patterns (3 points), recognition of iridescent stripe patterns (4 points), and recognition of the exposed part of the cornea (5 points), and was performed according to the report by Yokoi et al. (Am J Ophthalmol 1996 Dec;122(6):818-24). The average value was calculated from the obtained scoring values.

[0129] 2. Results (a1) Number of blocked openings of Meibomian glands The number of blocked openings of Meibomian glands significantly increased in the spleen-derived cell administration group compared to the Sham group on days 8 and 15 after administration of the cell suspension derived from the spleen, and the increasing trend continued until day 22 (Table 1).

[0130]

Table 1

[0131] (a2) Atrophy of Meibomian glands Atrophy of Meibomian glands developed in 4 out of 5 mice in the spleen-derived cell administration group after day 8 of administration of the cell suspension derived from the spleen (Table 2). In addition, strong atrophy of Meibomian glands was observed in the eyelid pathological sections of the spleen-derived cell administration group collected on day 28 after administration of the cell suspension derived from the spleen (Figure 1).

[0132]

Table 2

[0133] (a3) Eyelid margin irregularity Eyelid margin irregularity was observed from the 7th day after administration of the spleen-derived cell suspension. On the 21st day after administration, while the sulcus of the eyelid in the Sham group was depicted as a smooth curve by fluorescein, concavities and convexities were depicted in the spleen-derived cell administration group, and eyelid margin irregularity was confirmed (Figure 2).

[0134] (b1) Break-up time of the tear film (BUT) The break-up time of the tear film was significantly shortened in the spleen-derived cell administration group compared to the Sham group on the 7th, 14th, and 21st days after administration of the spleen-derived cell suspension (Table 3).

[0135]

Table 3

[0136] (b2) Blink rate The blink rate showed an increasing trend in the spleen-derived cell administration group on the 11th day after administration of the spleen-derived cell suspension and showed a significant increase compared to the Sham group on the 18th day after administration (Table 4).

[0137]

Table 4

[0138] (b3) Corneal staining score The corneal staining score was significantly increased in the spleen-derived cell administration group compared to the Sham group on the 14th day after administration of the spleen-derived cell suspension (Table 5).

[0139]

Table 5

[0140] (b4) Tear volume The tear volume was significantly decreased in the spleen-derived cell administration group compared to the Sham group on the 7th and 14th days after administration of the spleen-derived cell suspension (Table 6).

[0141]

Table 6

[0142] (b5)Tear film optical coherence score The tear film optical coherence score was significantly increased in the spleen-derived cell administration group compared to the Sham group on the 31st day after administration of the spleen-derived cell suspension (Table 7).

[0143]

Table 7

[0144] From these results, it was shown that mice administered intravenously with a suspension of spleen-derived cells collected from C57BL / 6 mice in C.B17 ICR SCID mice presented with human meibomian gland dysfunction symptoms and human dry eye symptoms.

[0145] (Example 2: Observation of localization of spleen-derived cells) This example aimed to explore the mechanism by which the model animals of the present disclosure present with meibomian gland dysfunction (MGD) and dry eye (DE) symptoms. In the same manner as in Example 1, spleen-derived cells isolated from donor C57BL / 6 mice (H-2D b ) were fluorescently labeled using PKH reagent (Sigma-Aldrich), and after intravenous administration to C.B17 icr-SCID mice (CB17 / Icr-Prkdc scid / CrlCrlj, H-2D d : purchased from Charles River Laboratories Japan, Inc.), the anterior eye was observed over time.

[0146] Specifically, the PKH reagent was added to and mixed with the spleen-derived cell suspension prepared in the same manner as in Example 1 for labeling. Thereafter, the fluorescence-labeled spleen-derived cell suspension (hereinafter referred to as "fluorescence-labeled spleen-derived cells") was administered to C.B17 icr-SCID mice via the tail vein. Two days after the administration of the fluorescence-labeled spleen-derived cells, observation was performed under a stereomicroscope using an excitation wavelength band pass of 460 to 480 nm and an emission wavelength band pass of 495 to 540 nm. The PKH reagent binds to the lipid region of the cell membrane and is considered to label T cells, B cells, etc. Since the peak excitation wavelength of PKH26 is 551 nm and the peak emission wavelength is 567 nm, and the peak excitation wavelength of PKH67 is 490 nm and the peak emission wavelength is 502 nm, PKH26 does not show fluorescence and PKH67 shows fluorescence at the above observation wavelengths. When PKH26 is used, only autofluorescence by the living body is observed. Therefore, the fluorescence observed only when PKH67 is used can be determined to be the localization of the labeled spleen-derived cells.

[0147] Fig. 3 shows a photograph taken two days after the administration of the fluorescence-labeled spleen-derived cells. In the animals transplanted with the fluorescence-labeled spleen-derived cells of PKH67, fluorescence was observed in the myobom gland tissue and fluorescent secretions were observed from the myobom gland.

[0148] From these results, it was considered that the model animals of the present disclosure exhibited MGD and DE symptoms as a result of the administered spleen-derived cells themselves or the components of the administered spleen-derived cells reaching tissues that contribute to the homeostasis of tears, such as the meibomian gland. Without wishing to be bound by theory, when the spleen-derived cells themselves reach, it is thought that the spleen-derived cells recognize the host cell's meibomian gland as non-self, an immune reaction occurs, and the meibomian gland is damaged. Without wishing to be bound by theory, when the cell membrane components of the spleen-derived cells reach, the cell membrane components of the spleen-derived cells are supplied to the meibomian gland via the blood and taken up as a raw material for meibum. However, the properties of the synthesized meibum change, the opening is blocked, and the differentiation balance from stem cells to duct epithelial cells or acinar cells tilts toward the differentiation side into duct epithelial cells, and the meibomian gland atrophies. The spleen-derived cells include the spleen itself and immune cells such as T cells and B cells contained in the spleen.

[0149] (Example 3: Evaluation of the drug efficacy of a test substance in a model animal) In this example, using this model animal, the drug efficacy of azithromycin-containing eye drops as a test substance was evaluated. Azithromycin is known to exhibit drug efficacy against meibomian gland dysfunction. This example was carried out with the approval of the Animal Experiment Ethics Review Committee of Chisso Corporation.

[0150] (Preparation and evaluation of a mouse model of eye disease) 1. Test method 1-1. Preparation of model mice In the same manner as in Example 1, model mice were prepared. Female C.B17 ICR SCID mice (CB17 / Icr-Prkdc scid / CrlCrlj: purchased from Charles River Laboratories Japan, Inc.) were intravenously administered 200 μL / mouse of a PBS suspension of spleen-derived cells (2.5×10 7 cells / mL). The PBS suspension of spleen-derived cells (hereinafter referred to as the "spleen-derived cell suspension") was prepared by making incisions in the spleen collected from C57BL / 6 mice, pipetting several times, and dispersing it in PBS.

[0151] After administration of the spleen-derived cell suspension, the mice were housed in breeding cages (size: 338 mm in length, 225 mm in width, and 140 mm in height) in groups of five or six.

[0152] 1-2. Administration of the Test Substance Taking the day of administration of the spleen-derived cell suspension as day 0, on the 10th day after administration, the mice were divided into a control eye drop administration group and an azithromycin-containing eye drop (azithromycin eye drop 1%, manufactured by Chisso Pharmaceutical Co., Ltd.) administration group (8 mice in each group). From the 10th day after administration for 2 days, each group was administered control eye drops or azithromycin-containing eye drops into both eyes twice a day at a dose of 2 μL per time, and from the 12th day after administration for 12 days, each group was administered once a day at a dose of 2 μL per time. The control eye drops were an isotonic buffer solution. The day when the instillation administration of the control eye drops or azithromycin-containing eye drops was started (the 10th day after administration of the spleen-derived cell suspension) was counted as the first day of instillation administration.

[0153] 1-3. Confirmation of the Number of Closed Meibomian Gland Openings, BUT, and Blink Frequency On the 10th day (before the start of instillation administration), 15th day (6th day of instillation administration), and 22nd day (13th day of instillation administration) after administration of the cell suspension, the number of closed Meibomian gland openings in the right eyes of the mice in each group was measured under anesthesia. The number of closed Meibomian gland openings was measured by gently inverting the upper and lower eyelids of the mice to expose the Meibomian gland openings under a stereomicroscope and counting the number of openings with occlusion from the inner canthus side to the outer canthus. The occlusion of the Meibomian gland openings was evaluated in the same manner as in Example 1.

[0154] The total number of occlusions observed in the upper and lower eyelids was taken as the individual value of the number of closed Meibomian gland openings, and the average value of the individual values was calculated. BUT and blink frequency were also evaluated in the same manner as in Example 1.

[0155] 2. Results The average number of blocked meibomian gland orifices changed from 10.8 before the start of eye drops administration in the control eye drops administration group to 15.0 on the 6th day of eye drops administration and 17.8 on the 13th day of eye drops administration, and the amount of change was +7.0. In the azithromycin-containing eye drops administration group, it changed from 10.5 before the start of eye drops administration to 12.0 on the 6th day of eye drops administration and 11.3 on the 13th day of eye drops administration. In the azithromycin-containing eye drops administration group, a significant decrease in the number of blocked meibomian gland orifices was observed on the 13th day of eye drops administration compared to the control eye drops administration group (p = 0.00158, t-test, 2-sided).

[0156] The average value of BUT changed from 0.59 seconds before the start of eye drops administration in the control eye drops administration group to 0.7 seconds on the 6th day of eye drops administration and 0.38 seconds on the 13th day of eye drops administration. In the azithromycin eye drops administration group, it changed from 0.59 seconds before the start of eye drops administration to 0.76 seconds on the 6th day of eye drops administration and 1.10 seconds on the 13th day of eye drops administration. In the azithromycin eye drops administration group, a significant prolongation of BUT was observed on the 13th day of eye drops administration compared to the control eye drops administration group (p = 0.003122, t-test, 2-sided).

[0157] The average value of blink frequency changed from 2.79 times before the start of eye drops administration in the control eye drops instillation group to 2.25 times on the 5th day of eye drops administration and 3.53 times on the 12th day of eye drops administration, and the amount of change was +0.74 times. In the azithromycin eye drops administration group, it changed from 3.67 times before the start of eye drops administration to 4.00 times on the 5th day of eye drops administration and 2.84 times on the 12th day of eye drops administration, and the amount of change was -0.83 times.

[0158] Therefore, it was found that the blockage of meibomian gland orifices, BUT, and blink frequency in this model animal are reversible and can be used for the evaluation of drug efficacy.

[0159] Furthermore, by the same method, it was confirmed that the tear film optical coherence score in this model animal was improved by the drug. In this model animal, it was found that the corneal staining score and the amount of tear fluid could also vary. Therefore, these evaluation items are reversible and can be used for evaluating the drug efficacy.

[0160] (Example 4: Confirmation of the distribution of immune cells in the meibomian gland) That immune cells (for example, cells derived from the spleen) have reached the meibomian gland can be confirmed by the following method.

[0161] After euthanizing the mouse, collect the eyelids, embed and freeze them in OCT compound (manufactured by Sakura Finetek Japan Co., Ltd.), and prepare sagittal frozen sections with a thickness of 10 μm.

[0162] Perform immunohistochemical staining with anti-CD3 (T cell surface antigen) antibody, anti-CD19 (B cell surface antigen) antibody, anti-F4 / 80 (macrophage surface antigen) antibody, anti-CD11 (dendritic cell surface antigen) antibody, anti-NK1.1 (NK cell surface antigen) antibody, anti-CD31 (endothelial cell marker) or anti-Claudin-5 (endothelial cell marker) and anti-mouse MHC ClassI H2-D d antibody (SCID mouse tissue marker).

[0163] When the mouse is a SCID mouse lacking T cells and B cells, determine the presence or absence of CD3 or CD19 positive cells, and if positive cells are present, it can be determined that cells of allogeneic animals are contaminated.

[0164] Macrophages, dendritic cells, NK cells, and endothelial cells are contained in cells derived from the spleen, and these have MHC class I. Therefore, whether the mouse is a SCID mouse or not, detect whether cells derived from the spleen are present in the mouse eyelid and whether the MHC class I type of the cells derived from the spleen is different from that of the mouse, and confirm the contamination of cells of allogeneic animals. When the mouse is a SCID mouse, MHC ClassI H2-D dThe presence or absence of cells that are negative and positive for F4 / 80, CD11, NK1.1, CD31, or Claudin-5 is determined. If cells that satisfy both conditions are present, it can be determined that cells of an allogeneic animal are contaminated.

[0165] As described above, the present disclosure has been illustrated using preferred embodiments of the present disclosure. However, it is understood that the scope of the present disclosure should be interpreted only by the claims. It is understood that patents, patent applications, and documents cited herein should be incorporated by reference herein as if the content thereof were specifically set forth herein.

Claims

1. A method for evaluating the medicinal efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) a step of administering the test substance to an animal model of an eye disease; and (2) a step of evaluating at least one feature associated with meibomian gland dysfunction, wherein the animal model of the eye disease is a non-human animal in which the immune function is reduced or impaired by transplantation of cells from an allogeneic animal, the feature associated with the meibomian gland dysfunction includes at least occlusion of the meibomian gland orifice, the cells of the allogeneic animal are immune cells derived from the spleen, a method.

2. A method for evaluating the medicinal efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) a step of administering the test substance to an animal model of an eye disease; and (2) a step of measuring the number of occlusions of the meibomian gland orifice in the animal model of the eye disease, wherein the animal model of the eye disease is a non-human animal in which the immune function is reduced or impaired by transplantation of cells from an allogeneic animal, the cells of the allogeneic animal are immune cells derived from the spleen, a method.

3. A method for evaluating the medicinal efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) a step of administering the test substance to an animal model of an eye disease; and (2) a step of evaluating the occlusion of the meibomian gland orifice and the abnormality of the blinking frequency in the animal model of the eye disease, wherein the animal model of the eye disease is a non-human animal in which the immune function is reduced or impaired by transplantation of cells from an allogeneic animal, the cells of the allogeneic animal are immune cells derived from the spleen, a method.

4. A method for evaluating the medicinal efficacy of a test substance as a therapeutic and / or prophylactic agent for eye diseases, the method comprising: (1) a step of administering the test substance to an animal model of an eye disease; and (2) a step of evaluating the occlusion of the meibomian gland orifice and the abnormality of the tear film break-up time in the animal model of the eye disease, wherein the animal model of the eye disease is a non-human animal in which the immune function is reduced or impaired by transplantation of cells from an allogeneic animal, the cells of the allogeneic animal are immune cells derived from the spleen, a method.

5. In the step (2), the method according to claim 1 includes a step of evaluating at least one of the features related to meibomian gland dysfunction and at least one selected from an abnormal tear film break-up time, an abnormal corneal staining score, a decrease in tear volume, an abnormal tear film optical coherence score, and an abnormal blink rate.

6. In the step (2), the method according to claim 5 includes a step of evaluating an occlusion of the meibomian gland orifice, an abnormal tear film break-up time, and an abnormal blink rate.

7. The method according to any one of claims 1 to 4, wherein the eye disease is meibomian gland dysfunction.

8. The method according to any one of claims 1 to 4, wherein the eye disease is dry eye complicated with meibomian gland dysfunction.

9. The method according to any one of claims 1 to 4, wherein the allogeneic animal is a mouse having an H-2D type of a subclass constituting the major histocompatibility complex (MHC) class I different from that of the eye disease model animal.

10. The method according to any one of claims 1 to 4, wherein the allogeneic animal is a mouse having an H-2D type of b in a subclass constituting MHC class I.

11. The method according to any one of claims 1 to 4, wherein the allogeneic animal is a C57BL / 6 mouse.

12. The method according to any one of claims 1 to 4, wherein the eye disease model animal is a mouse having an H-2D type of d in a subclass constituting MHC class I.

13. The method according to any one of claims 1 to 4, wherein the eye disease model animal is a SCID mouse or a NOD-SCID mouse.

14. The method according to any one of claims 1 to 4, wherein the eye disease model animal is a CB17 / Icr-Prkdc scid / CrlCrlj mouse.

15. A method for screening a therapeutic agent and / or a prophylactic agent for an eye disease, the method comprising: (1) a step of administering a test substance to an eye disease model animal, (2) a step of evaluating at least one of the features related to meibomian gland dysfunction; and (3) a step of identifying the test substance as a therapeutic agent and / or a prophylactic agent for the eye disease when the evaluated feature is improved or the progression of the feature is suppressed and the eye disease model animal is a non-human animal in which the immune function is reduced or impaired by transplantation of cells of an allogeneic animal. The features related to the meibomian gland dysfunction include at least the occlusion of the meibomian gland orifice, the cells of the allogeneic animal are immune cells derived from the spleen, Method.

16. A method for screening a therapeutic and / or prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an eye disease model animal, (2) measuring the number of occlusions of the meibomian gland orifice of the eye disease model animal; and (3) when the number of occlusions of the meibomian gland orifice decreases and / or the increase in the number of occlusions of the meibomian gland orifice is suppressed, identifying the test substance as a therapeutic and / or prophylactic agent for the eye disease including, the eye disease model animal is a non-human animal with reduced or impaired immune function transplanted with cells of an allogeneic animal, the cells of the allogeneic animal are immune cells derived from the spleen, Method.

17. A method for screening a therapeutic and / or prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an eye disease model animal, (2) evaluating the occlusion of the meibomian gland orifice and the abnormality of the blink rate of the eye disease model animal; and (3) when the occlusion of the meibomian gland orifice is improved and / or the progression of the occlusion of the meibomian gland orifice is suppressed, and the blink rate decreases and / or the increase in the blink rate is suppressed, identifying the test substance as a therapeutic and / or prophylactic agent for the eye disease including, the eye disease model animal is a non-human animal with reduced or impaired immune function transplanted with cells of an allogeneic animal, the cells of the allogeneic animal are immune cells derived from the spleen, Method.

18. A method for screening a therapeutic and / or prophylactic agent for an eye disease, the method comprising: (1) administering a test substance to an eye disease model animal, (2) evaluating the occlusion of the meibomian gland orifice and the abnormality of the tear film break-up time of the eye disease model animal; and (3) when the occlusion of the meibomian gland orifice is improved and / or the progression of the occlusion of the meibomian gland orifice is suppressed, and the tear film break-up time is prolonged and / or the shortening of the tear film break-up time is suppressed, identifying the test substance as a therapeutic and / or prophylactic agent for the eye disease including, the eye disease model animal is a non-human animal with reduced or impaired immune function transplanted with cells of an allogeneic animal, The cells of the allogeneic animal are immune cells derived from the spleen, Method.

19. The method according to claim 15, wherein in the step (2), at least one of the features related to the meibomian gland dysfunction and at least one selected from an abnormality in the tear film break-up time, an abnormality in the corneal staining score, a decrease in the amount of tears, an abnormality in the tear film optical coherence score, and an abnormality in the blink rate are evaluated.

20. The animal model for an eye disease is a mouse of type d of H-2D, which is a subclass constituting MHC class I, The allogeneic animal is a mouse of type b of H-2D, which is a subclass constituting MHC class I, The method according to any one of claims 15 to 18.

21. The method according to any one of claims 15 to 18, wherein the eye disease is meibomian gland dysfunction.

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