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

Eye disease animal models

[0001] The present disclosure relates to an animal model for eye diseases, particularly an animal model for meibomian gland dysfunction, an animal model for dry eye, and techniques related thereto.

[0002] Dry eye is classified into aqueous-deficient and evaporative dry eye, but a combination of these two types also occurs. Causes of evaporative dry eye include meibomian gland dysfunction and dyslipidemia. It is believed that 89% of dry eye patients also have meibomian gland dysfunction. However, clinically, there are no dry eye treatments that target the improvement of meibomian gland dysfunction. There is a need for therapeutic and preventative agents for dry eye that is complicated by meibomian gland dysfunction, including evaporative dry eye.

[0003] The development of therapeutic and preventive agents requires screening using model animals that can evaluate their therapeutic and preventive effects. However, there are few model animals that can evaluate the therapeutic and preventive effects of candidate compounds on meibomian gland dysfunction. Furthermore, there have been no model animals that can confirm the effects on dry eye symptoms in addition to the effects on meibomian gland dysfunction.

[0004] The present inventors have unexpectedly discovered that by administering cells from an allogeneic animal to an immunodeficient animal, the immunodeficient animal unexpectedly has meibomian gland dysfunction, and furthermore, has both meibomian gland dysfunction and dry eye. Thus, in one aspect, the present disclosure provides a model animal with meibomian gland dysfunction and a method for producing such a model animal, or a dry eye model animal with meibomian gland dysfunction and a method for producing such a dry eye model animal. In another aspect, the present disclosure provides a method for evaluating the efficacy of a therapeutic and / or preventive agent for dry eye or meibomian gland dysfunction using such a model animal, and a method for screening a therapeutic and / or preventive agent for dry eye or meibomian gland dysfunction.

[0005] The present disclosure provides, for example, the following items: (Item 1) A dry eye model animal having characteristics associated with meibomian gland dysfunction. (Item 2) The model animal according to the above items, wherein the model animal is an animal with reduced or impaired immune function to which cells from an animal allogeneic to the model animal have been transplanted. (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 is an animal with a major histocompatibility complex (MHC) type of H-2D ｄ (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 node. (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 different MHC class I type from that of the model animal. (Item 9) The allogeneic animal is an animal whose MHC type is H-2D ｂThe model animal according to any one of the above items, wherein the animal is a mouse having the above structure. (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 characteristic associated with meibomian gland dysfunction is at least one selected from the group consisting of obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. (Item 12) The model animal according to any one of the above items, wherein the characteristic associated with meibomian gland dysfunction is obstruction of meibomian gland orifices. (Item 13) The model animal according to any one of the above items, wherein the model animal further has a characteristic associated with dry eye selected from the group consisting of an abnormal tear film breakup time, an abnormal corneal staining score, a decreased tear volume, an abnormal tear film optical coherence score, and an abnormal blink rate. (Item 14) The model animal according to any one of the above items, wherein the model animal further has a dry eye-related characteristic selected from the group consisting of an abnormal tear film breakup time and an abnormal corneal staining score. (Item 15) A dry eye model animal, wherein the dry eye model animal is a dry eye model animal having reduced or insufficient immune function to which immune cells from an allogeneic animal have been transplanted, the animal having at least one characteristic selected from the following characteristics associated with meibomian gland dysfunction and at least one characteristic selected from the following characteristics associated with dry eye, wherein the characteristic associated with meibomian gland dysfunction is selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and the characteristic associated with dry eye is selected from abnormal tear film breakup time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical interference score, and abnormal blink rate. (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 model animal is a mouse with a major histocompatibility complex (MHC) type of H-2D ｄ(Item 15C) The dry eye model animal according to any one of the preceding items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item 15D) The dry eye model animal according to any one of the preceding items, wherein the allogeneic animal is a mouse having an MHC class I type different from that of the model animal. (Item 15E) The allogeneic animal is a mouse having an MHC class I type H-2D ｂ (Item 15F) The dry eye model animal according to any one of the preceding 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, wherein the dry eye model animal is an animal with reduced or impaired immune function to which cells from an allogeneic animal have been transplanted, and wherein 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 ｂA dry eye model animal having obstruction of meibomian gland orifices, an abnormal tear film breakup time, and an abnormal corneal staining score. (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 node. (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 different MHC class I type 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) The dry eye model animal, characterized in that it develops features associated with meibomian gland dysfunction by 40 weeks of age. (Item 17A) The dry eye model animal according to any one of the above items, wherein the feature associated with meibomian gland dysfunction is at least one selected from the group consisting of obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. (Item 18) The model animal, characterized in that the meibomian glands contain spleen-derived cells. (Item 18A) The model animal according to any one of the above items, wherein the cells are from an animal 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) The dry eye model animal comprises at least one cell type selected from the group consisting of T cells, B cells, macrophages, dendritic cells, venous sinus endothelial cells, and NK cells in the meibomian gland.(Item 22) A dry eye model animal, wherein the model animal is an animal with reduced or insufficient immune function and into which cells from an allogeneic animal have been transplanted. (Item 23) A meibomian gland dysfunction (MGD) model animal, wherein the model animal is an animal with reduced or insufficient immune function and into which cells from an allogeneic animal have been transplanted. (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 has a major histocompatibility complex (MHC) type of H-2D. ｄ (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 node. (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 an animal in which the MHC type is H-2D ｂThe model animal according to any one of the preceding items, wherein the animal is a mouse comprising the formula: (Item 23G) The model animal according to any one of the preceding 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 preceding items, wherein the model animal has at least one characteristic associated with meibomian gland dysfunction selected from the group consisting of obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. (Item 23I) The model animal according to any one of the preceding items, wherein the model animal further has a characteristic associated with dry eye selected from the group consisting of an abnormal tear film breakup time, an abnormal corneal staining score, reduced tear volume, an abnormal tear film optical coherence score, and an abnormal blink rate. (Item 24) An animal model for meibomian gland dysfunction (MGD) having obstruction of the orifices of the meibomian glands, the animal being an animal with reduced or insufficient immune function and into which cells from an allogeneic animal have been transplanted. (Item 24A) The MGD model animal according to any one of the above items, wherein the animal model is a SCID mouse or a NOD-SCID mouse. (Item 24B) The animal model for MGD has a major histocompatibility complex (MHC) type of H-2D. ｄ (Item 24C) The MGD model animal of any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item 24D) The MGD model animal of 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 24E) The MGD model animal of 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 24F) The allogeneic animal is an MGD model animal of any one of the above items, wherein the MHC type is H-2D ｂ(Item 24G) The MGD model animal according to any one of the preceding 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 transplanting cells from an animal allogeneic to 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 transplanting cells from an animal allogeneic to the MGD model animal into an animal with reduced or impaired immune function. (Item 26A) The method according to any one of the preceding items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 26B) The model animal is a mouse with a major histocompatibility complex (MHC) type of H-2D ｄ (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 node. (Item 26E) The method 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 26F) The allogeneic animal is a mouse having an MHC class I type different from that of the model animal. ｂThe method of any one of the above items, wherein the model animal is a mouse comprising the formula: (Item 26G) The method of 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 of any one of the above items, wherein the model animal has at least one characteristic associated with meibomian gland dysfunction selected from the group consisting of obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. (Item 26I) The method of any one of the above items, wherein the model animal further has a characteristic associated with dry eye selected from the group consisting of abnormal tear film breakup time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item 27) A method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for an eye disease, the method comprising: (1) a step of administering the test substance to an eye disease model animal; and (2) a step of evaluating at least one of a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item 28) The method of any one of the above items, wherein the eye disease is dry eye, and wherein step (2) comprises a step of evaluating a characteristic associated with dry eye. (Item 29) The method of any one of the above items, wherein the eye disease is meibomian gland dysfunction, and wherein step (2) comprises a step of evaluating a characteristic associated with meibomian gland dysfunction. (Item 30) The method according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least one selected from obstruction of the meibomian gland orifice, meibomian gland atrophy, and eyelid margin irregularity, and the feature associated with dry eye includes at least one selected from abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item 31) The method according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least obstruction of the meibomian gland orifice.(Item 32) The method of any one of the above items, wherein step (2) comprises evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item 33) The method of any one of the above items, wherein step (2) comprises evaluating at least obstruction of the meibomian gland orifice, abnormalities in tear film break-up time, and abnormalities in blink frequency. (Item 34) The method of any one of the above items, wherein the allogeneic animal cells are immune cells. (Item 35) The method of any one of the above items, wherein the allogeneic animal cells are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item 36) The method of any one of the above items, wherein the allogeneic animal cells are cells derived from the spleen. (Item 37) The method of any one of the above items, wherein the model animal is of the H-2D type of the major histocompatibility complex (MHC) class I subclass d. (Item 38) The method according to any one of the preceding items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 39) The method according to any one of the preceding items, wherein the allogeneic animal is a mouse having a different MHC type from that of the model animal. (Item 40) The method according to any one of the preceding items, wherein the allogeneic animal is a mouse having an H-2D type b, which is a subclass that constitutes MHC class I. (Item 41) A method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for dry eye, the method comprising: (1) administering the test substance to a dry eye model animal; and (2) evaluating the dry eye model animal for obstruction of meibomian gland orifices, abnormalities in tear film breakup time, and abnormalities in the number of blinks, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(Item 41A) The method according to any one of the preceding items, wherein the cells of the allogeneic animal are immune cells. (Item 41B) The method according to any one of the preceding items, wherein the cells of the allogeneic animal are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph node. (Item 41C) The method according to any one of the preceding items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item 42) A method for screening a therapeutic and / or preventive agent for an ocular disease, comprising: (1) administering a test substance to an ocular disease model animal; (2) evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the ocular disease model animal; and (3) identifying the test substance as a therapeutic and / or preventive agent for an ocular disease when the evaluated characteristic improves or suppresses progression, wherein the ocular disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item 42A) The method of any one of the above items, wherein the ocular disease is dry eye and step (2) comprises evaluating features associated with dry eye. (Item 42B) The method of any one of the above items, wherein the ocular disease is meibomian gland dysfunction and step (2) comprises evaluating features associated with meibomian gland dysfunction. (Item 42C) The method of any one of the above items, wherein in step (2), the features associated with meibomian gland dysfunction include at least one selected from blocked meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and the features associated with dry eye include at least one selected from abnormal tear film breakup time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item 42D) The method of any one of the above items, wherein in step (2), the features associated with meibomian gland dysfunction include at least blocked meibomian gland orifices. (Item 42E) The method according to any one of the preceding items, wherein step (2) includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction and at least one characteristic associated with dry eye.(Item 42F) The method according to any one of the above items, wherein in step (2), at least obstruction of the meibomian gland orifice, abnormalities in tear film break-up time, and abnormalities in blink frequency 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 is of the H-2D type, a subclass that constitutes 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 preceding items, wherein the allogeneic animal is a mouse whose H-2D type is b, a subclass that constitutes MHC class I. (Item 43) A method for screening a therapeutic and / or preventive agent for dry eye, the method comprising: (1) administering a test substance to a dry eye model animal, (2) evaluating the dry eye model animal for obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink rate; and (3) identifying the test substance as a therapeutic and / or preventive agent for dry eye when the obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink rate improve or inhibit the progression of the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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 ｂA screening method comprising: (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 node; (Item 43C) The method according to any one of the above items, wherein the model animal has an H-2D type d of the subclass that constitutes 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 a different MHC type 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 an H-2D type b of the subclass that constitutes MHC class I. (Item A1) A method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for eye disease, the method comprising: (1) administering the test substance to an eye disease model animal; and (2) evaluating at least one of a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function to which cells from an allogeneic animal have been transplanted. (Item A2) The method according to any one of the above items, wherein in step (2), the characteristic associated with meibomian gland dysfunction includes at least one selected from meibomian gland orifice obstruction, meibomian gland atrophy, and eyelid margin irregularity. (Item A4) The method according to any one of the above items, wherein in step (2), the characteristic associated with meibomian gland dysfunction includes at least meibomian gland orifice obstruction. (Item A5) The method according to any one of the preceding items, wherein step (2) includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction and at least one characteristic associated with dry eye.(Item A6) The method according to any one of the above items, wherein step (2) comprises at least evaluating blockage of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in blink frequency. (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 allogeneic animal cells are immune cells. (Item A10) The method 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 node. (Item A11) The method according to any one of the above items, wherein the allogeneic animal cells are cells derived from the spleen. (Item A12) The method according to any one of the above items, wherein the model animal has a major histocompatibility complex (MHC) class I H-2D subclass type d. (Item A13) The method according to any one of the preceding items, wherein the model animal is a SCID mouse or a NOD-SCID mouse. (Item A14) The method according to any one of the preceding items, wherein the allogeneic animal is a mouse having a different MHC type from that of the model animal. (Item A15) The method according to any one of the preceding items, wherein the allogeneic animal is a mouse having a b type H-2D subclass that constitutes MHC class I. (Item A16) A method for screening a therapeutic and / or preventive agent for an eye disease, comprising: (1) administering a test substance to an eye disease model animal; (2) evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) identifying the test substance as a therapeutic and / or preventive agent for an eye disease when the evaluated characteristic improves or suppresses progression, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted.(Item A17) A method for screening for a therapeutic and / or preventive agent for meibomian gland dysfunction, the method comprising: (1) administering a test substance to a meibomian gland dysfunction animal model; (2) evaluating blockage of the meibomian gland orifice in the meibomian gland dysfunction animal model; and (3) identifying the test substance as a therapeutic and / or preventive agent for meibomian gland dysfunction when blockage of the meibomian gland orifice improves and / or the progression of blockage of the meibomian gland orifice is inhibited in the meibomian gland dysfunction animal model, wherein the meibomian gland dysfunction animal model is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and the major histocompatibility complex (MHC) type of the meibomian gland dysfunction animal model is H-2D. ｄ and the MHC type of the allogeneic animal is H-2D ｂ (Item A18) A method for screening therapeutic and / or preventive agents for dry eye, the method comprising: (1) a step of administering a test substance to a dry eye model animal, (2) a step of evaluating in the dry eye model animal for obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for dry eye when obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency are improved or suppressed from progressing in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(Item B1) Use of an eye disease model animal in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for eye disease, the method comprising: (1) a step of administering the test substance to the eye disease model animal; and (2) a step of evaluating at least one of a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item B2) The use according to any one of the above items, wherein the eye disease is dry eye, and wherein step (2) comprises a step of evaluating a characteristic associated with dry eye. (Item B3) The use according to any one of the above items, wherein the eye disease is meibomian gland dysfunction, and wherein step (2) comprises a step of evaluating a characteristic associated with meibomian gland dysfunction. (Item B4) The use according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least one selected from blocked meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularities, and the feature associated with dry eye includes at least one selected from abnormal tear film break-up time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item B5) The use according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least blocked meibomian gland orifices. (Item B6) The use according to any one of the above items, wherein step (2) includes a step of evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item B7) The use according to any one of the above items, wherein the step (2) comprises at least a step of evaluating blockage of the meibomian gland orifice, an abnormality in tear film breakup time, and an abnormality in the blink frequency. (Item B8) The use according to any one of the above items, wherein the allogeneic animal cells are immune cells. (Item B9) The use 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 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 an H-2D type d of the subclass that constitutes major histocompatibility complex (MHC) class I. (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 an H-2D type b of the subclass that constitutes MHC class I. (Item B15) Use of a dry eye model animal in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for dry eye, the method comprising: (1) a step of administering the test substance to the dry eye model animal; and (2) a step of evaluating in the dry eye model animal obstruction of meibomian gland orifices, abnormalities in tear film breakup time, and abnormalities in the number of blinks, wherein the dry eye model animal is an animal with reduced or impaired immune function to which cells from an allogeneic animal have been transplanted, and 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(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 node. (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 eye disease model animal in a method for screening for therapeutic and / or preventive agents for eye diseases, the method comprising: (1) a step of administering a test substance to the eye disease model animal; (2) a step of evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for eye diseases when the evaluated characteristic improves or suppresses progression, wherein the eye disease model animal is an animal with reduced or compromised immune function into which cells from an allogeneic animal have been transplanted. (Item B20) The use according to any one of the above items, wherein the eye disease is dry eye and step (2) comprises a step of evaluating a characteristic associated with dry eye. (Item B21) The use according to any one of the above items, wherein the eye disease is meibomian gland dysfunction and step (2) comprises a step of evaluating a characteristic associated with meibomian gland dysfunction. (Item B21) The use according to any one of the above items, wherein in step (2), the features associated with meibomian gland dysfunction include at least one selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularities, and the features associated with dry eye include at least one selected from abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item B22) The use according to any one of the above items, wherein in step (2), the features associated with meibomian gland dysfunction include at least obstruction of meibomian gland orifices.(Item B23) The use according to any one of the above items, wherein step (2) comprises evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item B24) The use according to any one of the above items, wherein step (2) comprises evaluating at least blockage of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in blink frequency. (Item B25) The use according to any one of the above items, wherein the allogeneic animal cells are immune cells. (Item B26) The use 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 node. (Item B27) The use according to any one of the above items, wherein the model animal is of the H-2D type of the major histocompatibility complex (MHC) class I subclass d. (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 having an H-2D type b of the subclass that constitutes MHC class I. (Item B31) Use of a dry eye model animal in a method for screening therapeutic and / or preventive agents 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 blockage of the meibomian gland orifice, abnormality in tear film breakup time, and abnormality in the blink frequency; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for dry eye when blockage of the meibomian gland orifice, abnormality in tear film breakup time, and abnormality in the blink frequency are improved or the progression of the test substance is suppressed in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(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 node. (Item B34) The use according to any one of the above items, wherein the model animal has an H-2D type d of the subclass that constitutes major histocompatibility complex (MHC) class I. (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 a different MHC type 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 b of the subclass that constitutes MHC class I. (Item C1) Use of an eye disease model animal in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for eye disease, the method comprising: (1) a step of administering the test substance to the eye disease model animal; and (2) a step of evaluating at least one characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function to which cells from an allogeneic animal have been transplanted. (Item C2) The use according to any one of the above items, wherein step (2) includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction. (Item C3) The use according to any one of the above items, wherein in step (2), the characteristic associated with meibomian gland dysfunction includes at least one selected from meibomian gland orifice obstruction, meibomian gland atrophy, and eyelid margin irregularity. (Item C4) The use according to any one of the preceding items, wherein in step (2), the features associated with meibomian gland dysfunction include at least blockage of a meibomian gland orifice. (Item C5) The use according to any one of the preceding items, wherein in step (2), the features associated with meibomian gland dysfunction include at least one blockage of a meibomian gland orifice. (Item C6) The use according to any one of the preceding items, wherein in step (2), the features associated with meibomian gland dysfunction and at least one feature associated with dry eye are evaluated.(Item C6) The use according to any one of the above items, wherein the step (2) comprises at least a step of evaluating blockage of the meibomian gland orifice, an abnormality in tear film breakup time, and an abnormality in blink frequency. (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 allogeneic animal cells are immune cells. (Item C10) The use 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 node. (Item C11) The use according to any one of the above items, wherein the allogeneic animal cells are cells derived from the spleen. (Item C12) The use according to any one of the above items, wherein the model animal has a major histocompatibility complex (MHC) class I H-2D subclass type d. (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 an H-2D type b of the subclass that constitutes MHC class I. (Item C16) Use of an eye disease model animal in a method for screening for a therapeutic and / or preventive 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 a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for an eye disease when the evaluated characteristic improves or suppresses progression, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted.(Item C17) Use of a meibomian gland dysfunction model animal in a method for screening for a therapeutic and / or preventive agent for meibomian gland dysfunction, the method comprising: (1) a step of administering a test substance to the meibomian gland dysfunction model animal; (2) a step of evaluating blockage of the meibomian gland orifice in the meibomian gland dysfunction model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for meibomian gland dysfunction when blockage of the meibomian gland orifice improves and / or the progression of blockage of the meibomian gland orifice is inhibited in the meibomian gland dysfunction model animal, wherein the meibomian gland dysfunction model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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 (Item C18) Use of a dry eye model animal in a method for screening therapeutic and / or preventive agents 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 in the dry eye model animal for obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for dry eye when obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency are improved or suppressed from progressing in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(Item D1) An eye disease model animal for use in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for eye disease, the method comprising: (1) a step of administering the test substance to the eye disease model animal; and (2) a step of evaluating at least one of a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item D2) The model animal according to any one of the above items, wherein the eye disease is dry eye, and wherein step (2) comprises a step of evaluating a characteristic associated with dry eye. (Item D3) The model animal according to any one of the above items, wherein the eye disease is meibomian gland dysfunction, and wherein step (2) comprises a step of evaluating a characteristic associated with meibomian gland dysfunction. (Item D4) The model animal according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least one selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and the feature associated with dry eye includes at least one selected from abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item D5) The model animal according to any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least obstruction of meibomian gland orifices. (Item D6) The model animal according to any one of the above items, wherein step (2) includes a step of evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item D7) The eye disease model animal according to any one of the above items, wherein the step (2) comprises at least a step of evaluating blockage of the meibomian gland orifice, an abnormality in tear film breakup time, and an abnormality in the blink frequency. (Item D8) The eye disease model animal according to any one of the above items, wherein the allogeneic animal cells are immune cells.(Item D9) The eye disease 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 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 eye disease model animal according to any one of the above items, wherein the model animal has H-2D type d of the subclass that constitutes 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 a different MHC type 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 H-2D type b of the subclass that constitutes MHC class I. (Item D15) A dry eye model animal for use in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for dry eye, the method comprising: (1) a step of administering the test substance to the dry eye model animal; and (2) a step of evaluating in the dry eye model animal obstruction of meibomian gland orifices, abnormalities in tear film breakup time, and abnormalities in the number of blinks, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(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 node. (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 for a therapeutic and / or preventive 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 a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for an eye disease when the evaluated characteristic improves or suppresses progression, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item D20) The model animal according to any one of the above items, wherein the eye disease is dry eye and the step (2) comprises a step of evaluating a characteristic 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) comprises a step of evaluating a characteristic associated with meibomian gland dysfunction. (Item D21) The model animal of any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least one selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and the feature associated with dry eye includes at least one selected from abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical interference score, and abnormal blink rate. (Item D22) The model animal of any one of the above items, wherein in step (2), the feature associated with meibomian gland dysfunction includes at least obstruction of meibomian gland orifices.(Item D23) The model animal according to any one of the above items, wherein the step (2) comprises a step of evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item D24) The model animal according to any one of the above items, wherein the step (2) comprises a step of evaluating at least blockage of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink 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 H-2D type of the major histocompatibility complex (MHC) class I subclass is d. (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 b of the subclass that constitutes MHC class I. (Item D31) A dry eye model animal for use in a method for screening therapeutic and / or preventive agents 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 in the dry eye model animal for obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for dry eye when obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink frequency improve or inhibit progression in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and the major histocompatibility complex (MHC) type of the dry eye model animal is H-2D. ｄand the MHC type of the allogeneic animal is H-2D ｂ(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 node. (Item D34) The model animal according to any one of the above items, wherein the H-2D type of the major histocompatibility complex (MHC) class I subclass is d. (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 a different MHC type 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 a H-2D type of the MHC class I subclass. (Item E1) An eye disease model animal for use in a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for eye disease, the method comprising: (1) a step of administering the test substance to the eye disease model animal; and (2) a step of evaluating at least one characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item E2) The model animal according to any one of the above items, wherein step (2) includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction. (Item E3) The model animal according to any one of the above items, wherein in step (2), the characteristic associated with meibomian gland dysfunction includes at least one selected from meibomian gland orifice obstruction, meibomian gland atrophy, and eyelid margin irregularity. (Item E4) The model animal according to any one of the preceding items, wherein in step (2), the characteristics associated with meibomian gland dysfunction include at least blockage of the meibomian gland orifice.(Item E5) The model animal according to any one of the above items, wherein the step (2) comprises a step of evaluating at least one feature associated with meibomian gland dysfunction and at least one feature associated with dry eye. (Item E6) The model animal according to any one of the above items, wherein the step (2) comprises a step of evaluating at least blockage of meibomian gland orifices, abnormalities in tear film breakup time, and abnormalities in blink frequency. (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 node. (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 a subclass that constitutes major histocompatibility complex (MHC) class I of d type. (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 a subclass that constitutes MHC class I of b type.(Item E16) An eye disease model animal for use in a method for screening for therapeutic and / or preventive agents for eye disease, the method comprising: (1) a step of administering a test substance to the eye disease model animal; (2) a step of evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for eye disease when the evaluated characteristic improves or suppresses progression, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. (Item E17) A meibomian gland dysfunction model animal for use in a method for screening for a therapeutic and / or preventive agent for meibomian gland dysfunction, the method comprising: (1) administering a test substance to the meibomian gland dysfunction model animal; (2) evaluating blockage of the meibomian gland orifice in the meibomian gland dysfunction model animal; and (3) identifying the test substance as a therapeutic and / or preventive agent for meibomian gland dysfunction when blockage of the meibomian gland orifice improves and / or the progression of blockage of the meibomian gland orifice is inhibited in the meibomian gland dysfunction model animal, wherein the meibomian gland dysfunction model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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(Item E18) A dry eye model animal for use in a method for screening therapeutic and / or preventive agents 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 in the dry eye model animal for obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink rate; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for dry eye when obstruction of the meibomian gland orifice, abnormalities in tear film breakup time, and abnormalities in the blink rate are improved or suppressed from progressing in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted, and 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 (Item F1) A dry eye model animal, which is prepared by transplanting cells from an animal allogeneic to the dry eye model animal into an animal with reduced or insufficient immune function. (Item F2) An MGD model animal, which is prepared by transplanting cells from an animal allogeneic to the MGD model animal into an animal with reduced or insufficient 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 is prepared by transplanting cells from an animal with a major histocompatibility complex (MHC) type of H-2D ｄ(Item F5) The model animal according to any one of the above items, wherein the cells of the allogeneic animal are immune cells. (Item F6) 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 F7) 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 F8) The allogeneic animal is an animal in which the MHC type is H-2D ｂ (Item F9) The model animal of 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 model animal of any one of the above items, wherein the model animal has at least one characteristic associated with meibomian gland dysfunction selected from the group consisting of obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. (Item F10) The model animal of any one of the above items, wherein the model animal further has a characteristic associated with dry eye selected from the group consisting of abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical coherence score, and abnormal blink rate.

[0006] It is contemplated that the present disclosure may provide one or more of the above-described features in combinations other than those explicitly stated. Still further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading and understanding the following detailed description, if necessary.

[0007] The present disclosure provides an eye disease model animal with meibomian gland dysfunction, particularly a dry eye model animal with meibomian gland dysfunction. The present disclosure enables the development of therapeutic and / or preventive agents for dry eye associated with meibomian gland dysfunction and meibomian gland dysfunction, which have been difficult to develop until now.

[0008] Figure 1 shows photographs of eyelid sections stained with Oil Red O and hematoxylin. The dotted line indicates the boundary between the meibomian gland and other tissues. Figure 2 shows an image of an irregular eyelid margin. Figure 3 shows fluorescent photographs of the eyelid taken two days after administration of fluorescently labeled spleen-derived cells. Photographs were taken of a mouse administered PKH26 (excitation wavelength 551 nm, fluorescence wavelength 567 nm)-labeled spleen-derived cells with eyelid opening and eyelid eversion, and a mouse administered PKH67 (excitation wavelength 490 nm, fluorescence wavelength 502 nm)-labeled spleen-derived cells with eyelid opening and eyelid eversion. Observations were performed with an excitation wavelength bandpass of 460-480 nm and a fluorescence wavelength bandpass of 495-540 nm.

[0009] The present disclosure will be described below. Throughout this specification, singular expressions should be understood to include the plural concept unless otherwise specified. Therefore, singular articles (e.g., "a," "an," "the," etc. in English) should be understood to include the plural concept unless otherwise specified. Furthermore, it should be understood that terms used in this specification are used in the sense commonly used in the art unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. In the event of conflict, the present specification (including definitions) will prevail. In this specification, "about" means ±10% of the following value.

[0010] (Definition) In this specification, the term "ocular disease" refers to a disease that presents symptoms in the eyes, including dry eye and meibomian gland dysfunction, and is a disease that presents subjective and objective symptoms in the eyes.

[0011] As used herein, "dry eye" refers to a disease diagnosed as "dry eye" according to clinical diagnostic criteria, and is defined as "a multifactorial disease of the tears and ocular surface caused by various factors, and accompanied by ocular discomfort, abnormal visual function, tear film instability, or ocular surface damage." Dry eye is broadly classified into "aqueous tear-deficient" dry eye and "evaporative" dry eye. In recent years, a classification called short tear film break-up time (BUT) dry eye (hereinafter also referred to as "short BUT dry eye") has also been used.

[0012] As used herein, "aqueous-deficient dry eye" refers to dry eye that develops due to tissue destruction of the lacrimal gland or impaired tear conduction from the lacrimal gland to the ocular surface.

[0013] As used herein, "evaporative dry eye" refers to dry eye that develops when tear secretion is normal but excessive water loss from the ocular surface occurs due to various causes. Causes of evaporative dry eye include meibomian gland dysfunction (MGD) and tear lipid abnormalities, and the presence of either or both of these.

[0014] In this specification, "short BUT dry eye" refers to dry eye in which the tear breakup time (BUT) is short and there are subjective symptoms of dry eye, but tear secretion and the corneal and conjunctival epithelium are almost normal. Some believe that a shortened BUT is associated with meibomian gland dysfunction, which is thought to be one of the causes of evaporative dry eye.

[0015] As used herein, "meibomian glands" refer to sebaceous glands located within the tarsal plate and having openings at the upper and lower eyelid margins. Meibum (a lipid) is secreted from the meibomian glands. The presence of lipids in tears exerts effects such as inhibiting tear evaporation, promoting tear stability, and promoting the spreading of tears over the ocular surface. Although the number of meibomian glands varies from person to person, humans have approximately 50 meibomian glands in the upper eyelid and approximately 25 in the lower eyelid, while mice have approximately 11 meibomian glands in both the upper and lower eyelids.

[0016] As used herein, "meibomian gland dysfunction (MGD)" refers to a disease defined as "a condition in which the function of the meibomian glands is diffusely abnormal due to various causes, accompanied by chronic ocular discomfort." Meibomian gland dysfunction is also widely recognized as one of the causes of dry eye. Meibomian gland dysfunction (MGD) is broadly classified into hyposecretory type, in which oil secretion from the meibomian glands is reduced, and hypersecretory type, in which oil secretion from the meibomian glands is excessive. However, the hyposecretory type is overwhelmingly more common (Definition and Diagnostic Criteria of Meibomian Gland Dysfunction, Atarashii Ophthalmology 27(5):627-631, 2010). Hyposecretory MGD leads to evaporative dry eye due to a decrease in the tear oil phase. Hyposecretory MGD is characterized by subjective symptoms such as ocular discomfort and foreign body sensation, abnormal findings around the meibomian gland orifices including irregular eyelid margins, and findings of meibomian gland orifice obstruction (Table 4 in Atarashii Ophthalmology 27(5):627-631, 2010).

[0017] As used herein, the term "characteristics associated with meibomian gland dysfunction" refers to characteristics in the meibomian glands associated with meibomian gland dysfunction. The characteristics may be symptoms of meibomian gland dysfunction or may cause the onset of meibomian gland dysfunction.

[0018] As used herein, "obstruction of the meibomian gland orifice" refers to a state in which meibum or components of the meibomian glands remain in the orifice of the meibomian gland without being secreted or excreted. "Obstruction of the meibomian gland orifice" refers to a state in which, when the eyelid is everted under a stereomicroscope, a white to yellow protrusion is observed at the orifice of the meibomian gland, a cylindrical or rice-grain-shaped mass is observed within the orifice along the meibomian gland duct, or the orifice of the meibomian gland is pushed open by the contents.

[0019] As used herein, "atrophy of the meibomian glands" refers to a state in which the acini of the meibomian glands shrink and the amount of meibum stored in the meibomian glands decreases. "Atrophy of the meibomian glands" refers to a state in which the white reflex of visible light or the oily reflection of infrared light is reduced when the eyelid is everted under a stereomicroscope.

[0020] As used herein, "irregular eyelid margin" refers to a condition in which the line where the upper or lower eyelid meets the cornea (hereinafter referred to as "eyelid margin") is concave in places. "Irregular eyelid margin" is observed visually or under a stereomicroscope without treatment or after instillation of fluorescein.

[0021] As used herein, "tear film breakup time (BUT)" is an index of the stability of the tear film covering the cornea. "BUT" refers to the time required for the tear film to break down on the cornea, resulting in the appearance of a dry spot and gradually expanding tear film breakup, observed when blinking is suppressed and the eyelids are forcibly opened for a certain period of time (10 seconds). BUT can be measured as the eyelid-opening time from when reflected light from the tear film, interference light, or fluorescence from fluorescein molecules contained in the tear film after administration of fluorescein is uniformly observed on the cornea until an area where the reflected light or fluorescence disappears appears.

[0022] In this specification, the term "number of blinks" refers to the number of blinks in a certain period of time. The "number of blinks" is one of the characteristics related to dry eye, and is the number of blinks in a certain period of time (e.g., one minute), which can be observed visually.

[0023] As used herein, the term "corneal mottle score" refers to a score for mottles observed when the corneal surface is damaged in a punctate pattern. Punctate damage to the corneal surface can be visualized as mottles by the uptake of a dye (e.g., fluorescein) into corneal epithelial cells or by the accumulation of a dye (e.g., fluorescein) in areas where corneal epithelial cells have fallen off. For example, corneal mottles can be observed under a stereomicroscope after instillation or instillation of fluorescein, using a fluorescence wavelength of 480-680 nm with an excitation wavelength of 400-540 nm. The corneal mottle score is calculated by dividing the cornea into three quadrants: upper, middle, and lower. The punctate corneal mottle score is determined for each quadrant as absent (0 points), sparse (1 point), intermediate between sparse and dense (2 points), and dense (3 points), resulting in a total score of 9. The corneal mottle score is also sometimes referred to as the superficial punctate keratopathy (SPK) score.

[0024] As used herein, "tear volume" refers to the amount of fluid secreted from the lacrimal gland, and refers to the combined volume of tears stored in the meniscus (the tear reservoir above the lower eyelid) and tears secreted during the measurement period. One end of a Schirmer test strip or cotton thread is inserted into the conjunctival fornix of the upper or lower or medial or lateral cornea without irritating the cornea or conjunctiva, and after a certain period of time, it is removed and measured as the wet length of the Schirmer test strip or cotton thread. For example, the certain period of time is 15 seconds in mice.

[0025] As used herein, the term "tear film optical interference score" refers to an index for evaluating tear abnormalities (e.g., tear lipid abnormalities). The term "tear film optical interference" refers to a phenomenon in which the phase of light reflected from water and light reflected from oil changes due to differences in the refractive index, content, and / or mixing ratio of water and oil, which are components of tears (composed of three layers: lipid layer (oil layer), aqueous layer, and mucin layer), causing specific wavelengths to intensify, resulting in the observation of rainbow-colored interference fringes or areas with little reflected light. The "tear film optical interference score" is obtained by observation using a stereomicroscope and is scored from 1 to 5 based on the presence or absence of interference and the amount of reflected light. The presence or absence of interference is visualized by the number of rainbow-colored fringes, and the amount of reflected light is visualized by the brightness of light reflected from the entire tear film. The evaluation can be made, for example, as follows: uniform white to silvery reflection (1 point), non-uniform white to silvery reflection (2 points), two to three colored stripes (3 points), rainbow-colored stripes (4 points), or exposed areas of the cornea (5 points).

[0026] As used herein, the term "subject" refers to the recipient of the therapeutic and preventive medicine or method of the present disclosure, and examples of subjects include mammals (e.g., humans, mice, rats, hamsters, rabbits, cats, dogs, cows, horses, sheep, monkeys, etc.), with primates being preferred, and humans being particularly preferred.

[0027] As used herein, "treatment" means curing, improving, or suppressing or alleviating a disease or condition. "Treating an ocular disease" includes treating objective or subjective symptoms.

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

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

[0030] As used herein, the term "objective symptom" refers to a symptom of a disease that can be objectively proven by findings (objective findings) such as imaging findings or numerical values ​​from test results.

[0031] As used herein, the term "animal" refers to animals used as model animals, including mammals (e.g., mice, rats, hamsters, rabbits, cats, dogs, cows, horses, sheep, monkeys, etc.).

[0032] As used herein, the term "model animal" refers to an animal that has been engineered to produce symptoms similar to those of a human disease, and is used to elucidate mechanisms, develop preventive or therapeutic agents, or evaluate pharmacological efficacy. An animal develops the disease of interest and exhibits characteristics of the disease, but if the characteristics are irreversible, or it takes a considerable amount of time for the disease to develop or the characteristics to appear (e.g., approximately 50 weeks of age or older), and therefore it is not possible to elucidate mechanisms, develop preventive or therapeutic agents, or evaluate pharmacological efficacy, such an animal is not a model animal.

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

[0034] As used herein, "reduced or impaired immune function" refers to a condition characterized by a deficiency in immune cell function, resulting in a reduced or inability to elicit an immune response against allogeneic cells.

[0035] As used herein, "major histocompatibility complex (MHC) type H-2D" refers to ｄ"is" refers to the fact that the subclass H-2D constituting MHC class I is homozygous for d. ｂ The phrase "is" refers to the fact that the H-2D subclass constituting MHC class I is homozygous b.

[0036] As used herein, "immune cells" refers to cells that constitute the immune system, and includes 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 node" refers to cells prepared and / or isolated from the spleen, blood, bone marrow, thymus, or lymph node, or cultured cells obtained by culturing prepared and / or isolated cells.

[0038] As used herein, the phrase "containing cells in the meibomian gland" refers to the presence of foreign cells not derived from the model animal of the present disclosure in the meibomian gland. The presence of cells in the meibomian gland can be confirmed by detecting the presence of foreign cells through a pathological examination of the meibomian gland, or by using an antibody against a cell surface membrane protein or the like of the foreign cells.

[0039] As used herein, the phrase "developing characteristics associated with meibomian gland dysfunction by 40 weeks of age" refers to the development of characteristics associated with meibomian gland dysfunction at any time before the age of 40 weeks, and becoming usable as a model animal.

[0040] As used herein, the term "test substance" refers to a substance that is subjected to efficacy evaluation or screening using a model animal, and may be any substance such as a low molecular weight compound, polysaccharide, protein, peptide, nucleic acid, or fusion product thereof.

[0041] As used herein, the term "evaluation" refers to confirming the state of characteristics related to meibomian gland dysfunction and / or characteristics related to dry eye in a model animal, and may also include observing or measuring the characteristics as necessary, thereby determining whether or not a test substance has a desired property or effect, or the degree to which the test substance has a desired property or effect.

[0042] (Preferred Embodiments) The following describes preferred embodiments, but it should be understood that these embodiments are merely examples of the present invention and that the scope of the present invention is not limited to such preferred embodiments. It should also be understood that those skilled in the art can easily make modifications, changes, etc. within the scope of the present disclosure by referring to the following preferred examples. Those skilled in the art can combine any of these embodiments as appropriate.

[0043] (Model Animal) In one aspect, the present disclosure provides an ophthalmic disease model animal, particularly a meibomian gland dysfunction model animal or a dry eye model animal, having characteristics associated with meibomian gland dysfunction. Because most dry eye patients also suffer from meibomian gland dysfunction, treatment of both dry eye symptoms and meibomian gland dysfunction is desirable in dry eye treatment. However, there are no model animals that simultaneously suffer from both dry eye symptoms and meibomian gland dysfunction, making it difficult to develop therapeutic or preventive agents. Furthermore, there is a problem in that there are few meibomian gland dysfunction model animals that can be used to develop therapeutic or preventive agents for meibomian gland dysfunction. The present inventors have newly discovered a model animal having characteristics associated with meibomian gland dysfunction, and further a model animal that simultaneously suffers from characteristics associated with both dry eye and meibomian gland dysfunction.

[0044] In some embodiments, the model animal of the present disclosure can be a non-human animal, preferably a non-human mammal, more preferably a rodent, such as a mouse, rat, or 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 transplanted with cells from an animal allogeneic to the model animal. Cells from animals allogeneic to the model animal can include, but are not limited to, T cells, B cells, macrophages, dendritic cells, sinus endothelial cells, and NK cells. In some embodiments, the cells from animals allogeneic to the model animal can be immune cells, such as immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes. The cells from the allogeneic animal can preferably be spleen cells.

[0046] A model animal with reduced or impaired immune function may be deficient in T cells and / or B cells, preferably in both T cells and B cells. The reduced or impaired immune function is preferably a congenital condition, i.e., may be a condition caused by a genetic mutation.

[0047] The model animal with reduced or impaired immune function may be in a state in which the function of the protein Prkdc (protein kinase, DNA activated, catalytic polypeptide) is deficient due to a mutation in the Prkdc gene, and preferably in a state in which the function of the protein Prkdc is deficient due to a mutation in the Prkdc gene. ｓｃｉｄ The lack of Prkdc function results in a failure of gene rearrangement in B and T cells, resulting in a deficiency in mature B 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 animals of the present disclosure are 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 MHC class I type of the model animal of the present disclosure is d, and the MHC class I type of the allogeneic animal may be a, b, k, q, or s other than d, but is preferably b. In a specific embodiment, the H-2D subclass constituting the MHC class I of the model animal of the present disclosure is d, and the H-2D subclass constituting the MHC class I of the allogeneic animal may be a, b, k, q, or s other than d, but is preferably b. The subclass constituting the MHC class I may be homozygous or heterozygous. The H-2D of the model animal of the present disclosure is homozygous for d (H-2D ｄ ), and H-2D in allogeneic animals is homozygous for b (H-2D ｂ ) can be.

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

[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 animals of the present disclosure may have at least one, at least two, or all of the characteristics associated with meibomian gland dysfunction selected from the group consisting of obstructed meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularities.

[0054] Preferably, the model animal of the present disclosure has an increased number of blocked meibomian gland orifices compared to a healthy animal that does not develop an ocular disease (e.g., an animal that has not been administered with allogeneic cells from the model animal) (hereinafter referred to as a "non-model animal"). For example, the "blockage of meibomian gland orifices" can be evaluated using the number of blocked meibomian gland orifices per individual.

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

[0056] Preferably, the model animals of the present disclosure have concave eyelid margins, i.e., irregular eyelid margins, whereas non-model animals have smooth eyelid margins. For example, evaluation of "irregular eyelid margins" can be performed using the number of individuals exhibiting irregular eyelid margins (i.e., the ratio of individuals exhibiting irregular eyelid margins) to the total number of individuals in a group of model animals under the same conditions. The model animals of the present disclosure are advantageous as model animals because they develop characteristics associated with meibomian gland dysfunction and / or dry eye characteristics relatively quickly after administration of allogeneic cells. In some embodiments, the onset of symptoms may occur on days 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14, or at the latest between days 15 and 31, after administration of allogeneic cells. In some embodiments, the model animals of the present disclosure may be characterized by the onset of features associated with 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 further embodiments, 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 characteristics associated with dry eye selected from the group consisting of an abnormal tear film breakup time, an abnormal corneal staining score, reduced tear volume, an abnormal tear film optical interference score, and an abnormal blink rate.

[0058] Preferably, the model animal of the present specification has a reduced BUT compared to a non-model animal. For example, the evaluation of "BUT" can be performed using the BUT per individual of a group of model animals under the same conditions.

[0059] Preferably, the model animals of the present specification have an increased number of blinks compared to non-model animals. For example, the "number of blinks" can be evaluated using the number of blinks per individual of a group of model animals under the same conditions.

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

[0061] Preferably, the model animal of the present specification has a reduced tear volume compared to a non-model animal. For example, the "tear volume" can be evaluated using the tear volume per individual of a model animal group under the same conditions.

[0062] Preferably, the tear film light interference score is increased in the model animal of the present specification compared to non-model animals. For example, the "tear film light interference score" can be evaluated using the tear film light interference score per individual of a group of model animals under the same conditions.

[0063] In certain embodiments, the model animal of the present disclosure may have at least dry eye-related characteristics, such as abnormal tear film breakup 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 insufficient immune function, into which immune cells from an allogeneic animal have been transplanted, the meibomian gland dysfunction model animal being an animal with at least one characteristic selected from characteristics associated with meibomian gland dysfunction, the characteristic associated with meibomian gland dysfunction being selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity. In a specific embodiment, the meibomian gland dysfunction model animal is an immunodeficient animal with reduced or insufficient immune function into which immune cells from an allogeneic animal have been transplanted, the major histocompatibility complex (MHC) type being H-2D ｄ and allogeneic animals have an MHC type of H-2D. ｂ In a further embodiment, the meibomian gland dysfunction animal model may be an animal further having at least one characteristic selected from 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 interference score, and abnormal blink rate. In a further embodiment, the characteristic associated with meibomian gland dysfunction in the meibomian gland dysfunction animal model may be obstruction of the meibomian gland orifice, and the characteristic associated with dry eye may 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, the dry eye model animal being an immunodeficient animal with reduced or insufficient immune function, into which immune cells from an allogeneic animal have been transplanted, the dry eye model animal having the following: (1) at least one characteristic selected from characteristics associated with meibomian gland dysfunction; and (2) at least one characteristic selected from characteristics associated with dry eye, wherein (1) the characteristic associated with meibomian gland dysfunction is selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and (2) the characteristic associated with dry eye is selected from abnormal tear film breakup time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical coherence score, and abnormal blink rate. In a specific embodiment, the dry eye model animal is an immunodeficient animal with reduced or insufficient immune function into which cells from an allogeneic animal have been transplanted, the major histocompatibility complex (MHC) type being H-2D ｄ and allogeneic animals have an MHC type of H-2D. ｂ In a further embodiment, the feature associated with meibomian gland dysfunction in the dry eye model animal may be obstruction of meibomian gland orifices, and the feature associated with dry eye may be an abnormality in tear film breakup time and / or an abnormality in corneal staining score.

[0066] In a further aspect, the present disclosure provides a model animal characterized by the onset of features associated with meibomian gland dysfunction 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 animals of the present disclosure may develop features associated with meibomian gland dysfunction after 4, 5, or 6 weeks of age.

[0068] In a further aspect, the present disclosure provides an animal model comprising meibomian glands containing cells derived from the spleen, blood, bone marrow, thymus, or lymph node, preferably cells derived from the spleen, which may 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 comprising at least one cell selected from the group consisting of T cells, B cells, macrophages, dendritic cells, venous sinus endothelial cells, and NK cells in the meibomian gland. The cell may be an allogeneic cell.

[0070] The presence of the above-mentioned cells in the meibomian glands can be confirmed as follows. (1) When the model animal is an immunodeficient mouse lacking immune cells, a pathological examination of the meibomian glands is performed using a marker (e.g., an antibody) specific to the lacking immune cells. If the lacking immune cells are detected in the meibomian glands, it can be confirmed that foreign cells have reached the meibomian glands of the model animal. Since immunodeficient mice, such as SCID mice, lack T cells and B cells, the presence of T cells or B cells in the meibomian glands can be confirmed as having reached the meibomian glands. (2) A pathological examination of the meibomian glands is performed using a marker (e.g., an antibody) specific to 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 glands, it can be confirmed that foreign cells have reached the meibomian glands of the model animal. In this case, it is desirable to confirm that the spleen-derived cells 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 with (preferably congenitally) reduced or deficient immune function into which cells from an animal allogeneic to the model animal are transplanted.

[0072] In a further aspect, the present disclosure provides a meibomian gland dysfunction (MGD) model animal, the MGD model animal being an animal with (preferably congenitally) reduced or deficient immune function, into which cells from an animal allogeneic to the model animal are transplanted.

[0073] In a further aspect, the present disclosure provides a meibomian gland dysfunction (MGD) model animal having obstruction of the orifices of the meibomian glands, the MGD model animal being an animal with reduced or impaired immune function and into which cells from an allogeneic animal have been transplanted.

[0074] (Method for producing a model animal) In a different aspect, the present disclosure provides a method for producing a dry eye model animal, the method comprising the step of transplanting cells from an animal allogeneic to the dry eye model animal into an animal with reduced or impaired immune function.

[0075] In a further aspect, the present disclosure provides a method for producing an MGD model animal, the method comprising transplanting cells from an animal allogeneic to the MGD model animal into an animal with reduced or impaired immune function.

[0076] The production method of the present disclosure may include a step of transplanting cells into an animal with reduced or insufficient immune function, followed by rearing the animals for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days, or at the latest 15 to 31 days. Rearing can be performed by a general method, but it is also possible to rear multiple mice in one space (e.g., in a cage) and place the multiple mice one per 50 to 200 cm. ２ , 1 animal / 130-180cm ２ It is preferable to raise them in an environment where

[0077] Embodiments of model animals, allogeneic animals, cells of allogeneic animals, etc. are described above.

[0078] In clinical practice, meibomian gland dysfunction typically presents with symptoms such as abnormal findings around the meibomian gland orifice, obstructed meibomian gland orifice, and meibomian gland atrophy. Abnormal findings around the meibomian gland orifice include irregular eyelid margins. This model animal has the characteristics associated with meibomian gland dysfunction, such as "obstructed meibomian gland orifice," "meibomian gland atrophy," or "irregular eyelid margins." These characteristics in this model animal are reversible, and this model animal can be used to develop therapeutic and preventive agents for eye diseases, particularly meibomian gland dysfunction.

[0079] In clinical settings, dry eye typically presents with objective symptoms such as tear film breakup time (BUT) and subjective symptoms such as ocular discomfort or visual dysfunction. BUT is an essential test item for diagnosing dry eye in Japan. The U.S. Food and Drug Administration (FDA) requires significant improvement in at least one objective finding for the development of dry eye treatments, including BUT. Subjective symptoms (e.g., ocular discomfort, visual dysfunction) are also essential test items for diagnosing dry eye in Japan, and are assessed clinically by patient self-assessment and scoring. Subjective symptoms of human dry eye can be assessed in animal models using blink rate, with an increase in blink rate indicating worsening of subjective symptoms. This animal model possesses "BUT" or "blink rate" as a characteristic associated with dry eye. This characteristic in this animal model is reversible, making it useful for developing therapeutic and preventative agents for ocular diseases.

[0080] Dry eye may also be accompanied by objective symptoms such as "corneal and conjunctival epithelial damage," "decreased tear volume," and "abnormal tear lipid layer." "Corneal and conjunctival epithelial damage" refers to damage to the cornea or conjunctiva and is clinically assessed by staining tests. Corneal and conjunctival epithelial damage includes superficial punctate keratopathy, which is a loss of the corneal epithelium down to the wing cells; corneal erosion, which is a loss of the entire epithelium, including the basal cells, but without damage to the basement membrane; and corneal ulcers, which extend beyond the basement membrane to the stroma. While corneal and conjunctival epithelial damage is not essential for the diagnosis of dry eye, it is desirable to demonstrate the efficacy of a drug for treating dry eye. "Corneal and conjunctival epithelial damage" is scored by examining slit-lamp images stained or unstained with a dye (e.g., fluorescein) using a staining test. In animal models, the "corneal staining spot score" corresponds to the scoring of the corneal and conjunctival epithelial damage. Furthermore, "decreased tear volume" is not essential for the diagnosis of dry eye, but it is desirable to demonstrate the efficacy of a drug for treating dry eye. Tear volume is one of the objective findings that must be improved in the FDA's standards for developing dry eye treatments. Clinically, tear volume can be measured using the Schirmer test, and similar methods can be used to measure it in model animals. In addition, while "tear lipid layer abnormalities" are not essential for diagnosing dry eye, it is desirable to confirm their improvement as an indicator for developing dry eye treatments. Tear lipid layer abnormalities are abnormalities in the quantity and quality of lipids and affect tear stability. Clinically, "tear lipid layer abnormalities" are detected by observing "tear film optical interference," but they can also be scored and measured using similar methods in model animals. This model animal has dry eye-related characteristics such as "abnormal corneal staining score," "decreased tear volume," or "abnormal tear film optical interference score," 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 ocular diseases.

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

[0082] (Method for Evaluating Pharmacological Efficacy) In a different aspect, the present disclosure provides a method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for an ocular disease, the method comprising: (1) administering the test substance to an ocular disease model animal; and (2) evaluating at least one of a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye, wherein the ocular disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted. In a specific embodiment, the ocular disease may be dry eye and / or meibomian gland dysfunction.

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

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

[0085] In some embodiments, in step (1), the method of administering the test substance is oral administration, eye drop administration, intraocular injection administration, etc., and the dosage form of the administered test substance may be a tablet, eye drop, injection, etc. In a preferred embodiment, the test substance may be administered by eye drop administration. For example, the test substance may be administered 1 to 3 days, preferably 1 to 2 days, after the onset of characteristics associated with meibomian gland dysfunction or characteristics associated with dry eye, or 6 to 9 days, preferably 7 to 8 days, after transplantation of cells from an animal allogeneic to the model animal into the model animal.

[0086] The method for evaluating medicinal efficacy in the present disclosure may include a step of evaluating the test substance as having medicinal efficacy when the test substance improves or inhibits the progression of features associated with meibomian gland dysfunction and / or features associated with dry eye in a model animal to which the test substance has been administered.

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

[0088] In some embodiments, in step (2), the feature associated with meibomian gland dysfunction may be at least one feature selected from obstruction of meibomian gland orifices, meibomian gland atrophy, and eyelid margin irregularity, and the feature associated with dry eye may be at least one feature selected from abnormal tear film breakup time, abnormal corneal staining score, decreased tear volume, abnormal tear film optical interference score, and abnormal blink rate.

[0089] In some embodiments, dry eye-related features and / or meibomian gland dysfunction-related features may be evaluated in step (2). In certain embodiments, at least one, at least two, or three meibomian gland dysfunction-related features and at least one, at least two, at least three, at least four, or five dry eye-related features may be evaluated in step (2).

[0090] Preferably, the obstruction of the meibomian gland orifices in the model animal can be determined to be an improvement in the characteristic, for example, when the number of obstructions is reduced compared to before administration of the test substance, or the progression of the characteristic can be determined to be suppressed when the number of obstructions is reduced when the test substance is administered compared to when a placebo is administered.

[0091] Preferably, the atrophy of the meibomian glands in the model animal can be determined to have improved, for example, if the size of the meibomian gland acini increases compared to before administration of the test substance, if the accumulation of meibum increases, or if the reflection of infrared light by the accumulated oil increases, or if the reflex that should be anatomically present is restored. Alternatively, the progression of the atrophy of the meibomian glands can be determined to have been inhibited, for example, if the meibomian gland acini are larger when the test substance is administered than when a placebo is administered, if the accumulation of meibum increases, or if the reflection of infrared light by the accumulated oil increases, or if the reflex that should be anatomically present is restored.

[0092] Preferably, the eyelid margin irregularity characteristic in a model animal can be determined to have improved, for example, when the eyelid margin line becomes less concave and smoother compared to before administration of the test substance; alternatively, the progression of the characteristic can be determined to have been inhibited, for example, when the eyelid margin line becomes less concave and smoother when the test substance is administered compared to when a placebo is administered.

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

[0094] Preferably, the characteristic can be determined to be improved when the number of blinks in the model animal is reduced compared to before administration of the test substance, or the progression of the characteristic can be determined to be inhibited when the number of blinks is reduced when the test substance is administered compared to when a placebo is administered.

[0095] Preferably, when the corneal staining score in the model animal is reduced compared to before administration of the test substance, it can be determined that the characteristic has improved; alternatively, when the score is reduced when the test substance is administered compared to when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

[0096] Preferably, when the tear volume in the model animal is increased compared to before administration of the test substance, it can be determined that the characteristic has improved. Alternatively, for example, when the tear volume in the model animal is increased when the test substance is administered compared to when a placebo is administered, it can be determined that the progression of the characteristic has been suppressed.

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

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

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

[0100] The significant difference can be confirmed by a general statistical method, and means that there is a statistically significant difference.

[0101] In a further embodiment, in step (2), at least the obstruction of the meibomian gland orifice, the tear film breakup time, and the blink frequency may be evaluated.

[0102] Embodiments of model animals, allogeneic animals, cells of allogeneic animals, etc. are described above.

[0103] (Screening Method) In a different aspect, the present disclosure provides a method for screening for therapeutic and / or preventive agents for dry eye, the method comprising the steps of: (1) administering a test substance to a dry eye model animal; (2) evaluating characteristics associated with meibomian gland dysfunction and characteristics associated with dry eye; and (3) identifying the test substance as a therapeutic and / or preventive agent for dry eye when the evaluated characteristics improve or the progression is inhibited in the dry eye model animal.

[0104] In a different aspect, the present disclosure provides a method for screening for therapeutic and / or preventive agents for meibomian gland dysfunction, the method comprising the steps of: (1) administering a test substance to an animal with meibomian gland dysfunction; (2) evaluating characteristics associated with meibomian gland dysfunction; and (3) identifying the test substance as a therapeutic and / or preventive agent for meibomian gland dysfunction if the evaluated characteristics improve or the progression of the evaluated characteristics is inhibited in the animal with meibomian gland dysfunction.

[0105] In a further aspect, the present disclosure provides a method for screening for therapeutic and / or preventive agents for ocular diseases, the method comprising: (1) a step of administering a test substance to an ocular disease model animal, wherein the ocular disease model animal is an immunodeficient animal transplanted with immune cells from an allogeneic animal; and (2) a step of evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for ocular diseases when the evaluated characteristic improves or progression is inhibited in the ocular disease model animal.

[0106] Embodiments of model animals, allogeneic animals, cells of allogeneic animals, endpoints, etc. are described above.

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

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

[0109] The present disclosure will be described in more detail below based on examples, but these are intended to provide a better understanding of the present disclosure and are not intended to limit the scope of the present disclosure. In addition, this example was conducted with the approval of the Animal Experiment Ethics Committee of Senju Pharmaceutical Co., Ltd.

[0110] (Example 1: Creation and evaluation of eye disease model mice) 1. Test method 1-1. Creation of model mice Five to seven-week-old female C.B17 ICR SCID mice (CB17 / Icr-Prkdc ｓｃｉｄ / CrlCrlj, H-2D ｄ (Purchased from Charles River Japan, Inc.) were divided into a sham group and a spleen-derived cell-administered group (six mice per group). In each group, five mice were used for time-course observation and one mouse for pathological observation.

[0111] The sham group received 200 μL / mouse of PBS (Thermo Fisher Scientific) administered intravenously to the tail vein. The spleen-derived cell administration group received 200 μL / mouse of a PBS suspension of spleen-derived cells (2.5 × 10 ７　 The spleen-derived cells were suspended in PBS (hereinafter referred to as "spleen-derived cell suspension") and administered into the tail vein of C57BL / 6 mice (H-2D ｂ After incision was made in the spleen, the cells were dispersed by pipetting several times and suspended in PBS. After administration of PBS or the spleen-derived cell suspension, the mice were housed in cages (dimensions: length 338 mm, width 225 mm, height 140 mm) with groups of five mice.

[0112] 1-2. Evaluation of Each Characteristic (a) Evaluation of Characteristics Related to Meibomian Gland Dysfunction (a1) Number of Blocked Meibomian Gland Orifices The day of administration of the spleen-derived cell suspension was designated as day 0, and the number of blocked meibomian glands in the right eyes of mice in each group was measured under anesthesia using a stereomicroscope on days 8, 15, and 21 after administration. The number of blocked meibomian glands was measured under observation under the stereomicroscope by gently everting the upper and lower eyelids to expose the meibomian gland orifices, and counting the number of orifices that were found to be blocked from the medial canthus toward the lateral canthus.

[0113] Obstruction of the meibomian gland orifice was determined when, under a stereomicroscope, white to yellow protrusions were observed at the orifice, the meibomian orifice was expanded by the contents, or a cylindrical or rice-grain-shaped mass was observed within the orifice along the meibomian gland duct. The total number of obstructions observed on the upper and lower eyelids was used as the individual value for the number of obstructed meibomian glands, and the average value for five mice was calculated.

[0114] (a2) Atrophy of Meibomian Glands As in the case of confirming the number of obstructed meibomian glands, the day of administration of the spleen-derived cell suspension was designated as day 0, and atrophy of the meibomian glands in the right eye was confirmed under anesthesia using a stereomicroscope on days 8, 15, and 21 after administration, and the number of mice with atrophy was counted.

[0115] Meibomian gland atrophy was assessed by observing the area of ​​the meibomian glands under a stereomicroscope by everting the upper and lower eyelids and observing the area of ​​the meibomian glands visible through the conjunctiva. Atrophy was determined when the area was smaller than the size of meibomian glands in a typical mouse, and the number of mice with one or more atrophied meibomian glands was counted.

[0116] Atrophy of the meibomian glands was also confirmed anatomically. Twenty-eight days after administration of the spleen-derived cell suspension, the mice were euthanized and their eyelids were collected. The collected eyelids were embedded and frozen in OCT compound (Sakura Finetech Japan), and 10-μm-thick sagittal frozen sections were prepared. The obtained sections were stained red for lipids in the meibomian glands using Oil Red O, and blue for cell nuclei using hematoxylin dye. After staining, the condition of the meibomian glands in the sections was observed under a microscope.

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

[0118] A fluorescein sodium solution (a solution prepared by dissolving uranine (Fujifilm Wako Pure Chemical Industries, Ltd.) in physiological saline and filtering the solution) was instilled into the right eye of anesthetized and immobilized mice, and the solution was mixed well with the tear fluid by forced blinking several times. The mice were then placed under fluorescent observation with a stereomicroscope (excitation wavelength bandpass 460-480 nm, fluorescence wavelength bandpass 495-540 nm), and fluorescent stereomicroscope images of the eyelid margin were taken and observed. A smooth curve of the eyelid edge visualized by fluorescein was considered normal, while an irregularity was visualized as an eyelid margin irregularity.

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

[0120] The tear film breakup time was measured using a fluorescein sodium solution, as in (a3). Similarly to (a3), the fluorescein sodium solution was instilled into the right eye of anesthetized, immobilized mice, and mixed thoroughly with the tear fluid by forced blinking several times. The mice were then placed under fluorescent observation with a stereomicroscope (excitation wavelength bandpass 460-480 nm, fluorescence wavelength bandpass 495-540 nm). The mixed solution of tear fluid and fluorescein covered the cornea, and tear film breakup could be evaluated by confirming the disappearance of fluorescein fluorescence. After placing the mice under fluorescent observation with a stereomicroscope, the right eyelids of the mice were forced open. The time from the moment of eyelid opening to the appearance of the area where the fluorescence disappeared was measured three times, and the average of the three measurements was used as the individual value. The average of the individual values ​​for five mice was calculated and used as the tear film breakup time (BUT).

[0121] (b2) Number of Blinks The day of administration of the spleen-derived cell suspension was designated as day 0. On days 4, 11, and 18 after administration, the mice were transferred to small observation cages and isolated individually, and the number of blinks of the left eye of each mouse was measured while it was moving freely in the cage. Measurements were performed 5 times per minute per mouse, and the total of the 5 measurements was used as the individual value, and the average value for the 5 mice was calculated. After measurement, the mice were returned to their breeding cages, and breeding continued as usual, with 5 mice placed in one cage.

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

[0123] After measuring the tear film breakup time (b1), the mice were washed with saline and observed for corneal staining. After washing the eyes, the mice were again placed under fluorescent observation with a stereomicroscope (excitation wavelength bandpass: 460-480 nm, fluorescence wavelength bandpass: 495-540 nm). Corneal staining was observed by confirming punctate fluorescence from fluorescein in the outermost layer of the cornea.

[0124] Observations were performed on five quadrants of the cornea: upper, right, lower, left, and center. The punctate fluorescence was evaluated as absent (0 points), sparse (1 point), slightly dense (2 points), or dense (3 points). The total score for the five quadrants was calculated as an individual value for the corneal staining plaque score (SPK score). The average value was calculated from the individual scores obtained.

[0125] (b4) Tear Volume The day of administration of the spleen-derived cell suspension was set as day 0, and tear volume in the right eye was measured under a stereomicroscope with the mouse awake and restrained on days 7, 14, and 21 after administration.

[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 a mouse, held there for 15 seconds, and then immediately removed. The length of discoloration of the phenol red contained in the cotton thread due to 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 Interference Score The day of administration of the spleen-derived cell suspension was set as day 0, and on day 31 after administration, tear film optical interference of the right eye was observed under anesthesia using a stereomicroscope.

[0128] After forcibly opening the right eyelid of the mouse, the focus of a stereomicroscope was adjusted to the tear film, and the tear film was scored from 1 to 5 based on the number of rainbow-colored stripes produced by optical interference and the brightness of the light reflection from the entire tear film. Scoring was performed according to the report by Yokoi et al., Am J Ophthalmol 1996 Dec;122(6):818-24. The average score was calculated from the obtained scores.

[0129] 2. Results (a1) Number of obstructed meibomian gland orifices The number of obstructed meibomian gland orifices was significantly increased in the spleen-derived cell-administered group compared to the sham group on days 8 and 15 after administration of the spleen-derived cell suspension, and this increasing trend continued until day 22 (Table 1).

[0130]

[0131] (a2) Atrophy of Meibomian Glands Meibomian gland atrophy developed in 4 out of 5 mice in the spleen-derived cell administration group from day 8 onwards after administration of the spleen-derived cell suspension (Table 2). Furthermore, pathological sections of the eyelids of the spleen-derived cell administration group collected 28 days after administration of the spleen-derived cell suspension showed severe atrophy of the meibomian glands (Figure 1).

[0132]

[0133] (a3) Irregular eyelid margins Irregular eyelid margins were observed from day 7 after administration of the spleen-derived cell suspension. On day 21 after administration, the eyelid edges in the sham group were visualized as smooth curves by fluorescein, whereas those in the spleen-derived cell-administered group were visualized as uneven, confirming the irregular eyelid margins ( FIG. 2 ).

[0134] (b1) Tear film breakup time (BUT) The tear film breakup time was significantly shorter in the spleen-derived cell-administered group compared to the sham group on days 7, 14, and 21 after administration of the spleen-derived cell suspension (Table 3).

[0135]

[0136] (b2) Number of Blinks The number of blinks tended to increase in the spleen-derived cell administration group on day 11 after administration of the spleen-derived cell suspension, and showed a significant increase compared to the sham group on day 18 after administration (Table 4).

[0137]

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

[0139]

[0140] (b4) Tear Volume Tear volume was significantly reduced in the spleen-derived cell administration group compared to the sham group on days 7 and 14 after administration of the spleen-derived cell suspension (Table 6).

[0141]

[0142] (b5) Tear Film Optical Interference Score The tear film optical interferometry score was significantly increased in the spleen-derived cell administration group compared to the sham group on day 31 after administration of the spleen-derived cell suspension (Table 7).

[0143]

[0144] These results demonstrated that C.B17 ICR SCID mice intravenously administered with a suspension of spleen-derived cells collected from C57BL / 6 mice exhibited symptoms of human meibomian gland dysfunction and human dry eye symptoms.

[0145] Example 2: Observation of localization of spleen-derived cells The purpose of this example was to explore the mechanisms by which meibomian gland dysfunction (MGD) and dry eye (DE) symptoms are exhibited in the model animal of the present disclosure. As in Example 1, donor C57BL / 6 mice (H-2D ｂ Spleen-derived cells isolated from C. B17 icr-SCID mice (CB17 / Icr-Prkdc) were fluorescently labeled using PKH reagent (Sigma-Aldrich) and then cultured in a 5% CO2-free medium. ｓｃｉｄ / CrlCrlj, H-2D ｄ(purchased from Charles River Japan, Inc.) was administered intravenously to the tail vein, and the anterior segment of the eye was observed over time.

[0146] Specifically, spleen-derived cell suspensions prepared as in Example 1 were mixed with a PKH reagent for labeling. The fluorescently labeled spleen-derived cell suspension (hereinafter referred to as "fluorescently labeled spleen-derived cells") was then administered via the tail vein to C.B17 icr-SCID mice. Two days after administration of the fluorescently labeled spleen-derived cells, the cells were observed under a stereomicroscope using an excitation wavelength bandpass of 460-480 nm and an emission wavelength bandpass of 495-540 nm. The PKH reagent binds to the lipid region of the cell membrane and is thought to label T cells, B cells, and other cells. The peak excitation wavelength of PKH26 is 551 nm and the peak emission wavelength is 567 nm, while the peak excitation wavelength of PKH67 is 490 nm and the peak emission wavelength is 502 nm. Therefore, at these observation wavelengths, PKH26 does not emit fluorescence, while PKH67 does. When PKH26 is used, only autofluorescence from the living body is observed, and therefore, the fluorescence observed only when PKH67 is used can be determined to be the localization of labeled cells derived from the spleen.

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

[0148] From these results, it is believed that the MGD and DE symptoms exhibited by the model animals of the present disclosure are the result of the administered spleen-derived cells themselves or components of the administered spleen-derived cells reaching tissues that contribute to tear homeostasis, such as the meibomian glands. Without wishing to be bound by theory, it is believed that when the spleen-derived cells themselves reach the meibomian glands, the spleen-derived cells recognize the host cells' meibomian glands as non-self, resulting in an immune reaction and damage to the meibomian glands. Without wishing to be bound by theory, it is believed that when the cell membrane components of the spleen-derived cells reach the meibomian glands, the cell membrane components are supplied to the meibomian glands via the blood and absorbed as a raw material for meibum. However, the properties of the synthesized meibum change, the openings become blocked, and the balance of differentiation from stem cells into ductal epithelial cells or acinar cells shifts toward differentiation into ductal epithelial cells, resulting in atrophy of the meibomian glands. The spleen-derived cells include the spleen itself and immune cells contained in the spleen, such as T cells and B cells.

[0149] (Example 3: Evaluation of efficacy of test substance in model animal) In this example, the efficacy of an azithromycin-containing eye drop as a test substance was evaluated using this model animal. Azithromycin is known to be effective against meibomian gland dysfunction. This example was conducted with the approval of the Animal Experiment Ethics Committee of Senju Pharmaceutical Co., Ltd.

[0150] (Preparation and Evaluation of Eye Disease Model Mice) 1. Test Method 1-1. Preparation of Model Mice Model mice were prepared in the same manner as in Example 1. Five to seven-week-old female C.B17 ICR SCID mice (CB17 / Icr-Prkdc ｓｃｉｄ / CrlCrlj: purchased from Charles River Japan Co., Ltd.) was added with 200 μL of PBS suspension of spleen-derived cells per animal (2.5 × 10 ７　 A PBS suspension of spleen-derived cells (hereinafter referred to as "spleen-derived cell suspension") was prepared by making an incision in a spleen collected from a C57BL / 6 mouse, pipetting the cells several times, and dispersing them in PBS.

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

[0152] 1-2. Administration of Test Substances The day of administration of the spleen-derived cell suspension was designated as day 0. On day 10 after administration, the mice were divided into a control eye drop administration group and an azithromycin-containing eye drop administration group (1% azithromycin ophthalmic solution, manufactured by Senju Pharmaceutical Co., Ltd.) (8 mice per group). Starting on day 10 after administration, the control eye drop or azithromycin-containing eye drop was administered to both eyes at a dose of 2 μL twice daily for two days, and then starting on day 12 after administration, the control eye drop or azithromycin-containing eye drop was administered to both eyes at a dose of 2 μL once daily for 12 days. The control eye drop was an isotonic buffer solution. The day on which administration of the control eye drop or azithromycin-containing eye drop was initiated (day 10 after administration of the spleen-derived cell suspension) was counted as day 1 of eye drop administration.

[0153] 1-3. Confirmation of the number of blocked meibomian gland orifices, BUT, and blink count The number of blocked meibomian gland orifices in the right eyes of mice in each group was measured under anesthesia on days 10 (before the start of eye drop administration), 15 (sixth day after eye drop administration), and 22 (thirteenth day after eye drop administration) after cell suspension administration. The number of blocked meibomian gland orifices was determined by gently everting the upper and lower eyelids of the mice under a stereomicroscope to expose the meibomian gland orifices, and counting the number of blocked orifices from the medial canthus toward the lateral canthus. Blockage of the meibomian gland orifices was evaluated in the same manner as in Example 1.

[0154] The total number of occlusions observed on the upper and lower eyelids was taken as the individual value for the number of occlusions of the meibomian gland orifices, and the average of the individual values ​​was calculated. BUT and the number of blinks were also evaluated in the same manner as in Example 1.

[0155] 2. Results The mean number of blocked meibomian gland orifices in the control eye drop group increased from 10.8 before administration to 15.0 on Day 6 and 17.8 on Day 13, a change of +7.0. In the azithromycin-containing eye drop group, the mean number of blocked meibomian gland orifices increased from 10.5 before administration to 12.0 on Day 6 and 11.3 on Day 13, demonstrating a significant decrease in the number of blocked meibomian gland orifices in the azithromycin-containing eye drop group compared to the control eye drop group (p=0.00158, t-test, 2-sided).

[0156] The mean BUT in the control ophthalmic solution group changed from 0.59 seconds before administration to 0.7 seconds on day 6 and 0.38 seconds on day 13. In the azithromycin ophthalmic solution group, the mean BUT changed from 0.59 seconds before administration to 0.76 seconds on day 6 and 1.10 seconds on day 13. A significant prolongation of BUT was observed in the azithromycin ophthalmic solution group on day 13 compared with the control ophthalmic solution group (p=0.003122, t-test, 2-sided).

[0157] The mean number of blinks in the control ophthalmic solution group changed from 2.79 before administration to 2.25 on day 5 and 3.53 on day 12, a change of +0.74 blinks.In the azithromycin ophthalmic solution group, the mean number of blinks changed from 3.67 before administration to 4.00 on day 5 and 2.84 on day 12, a change of -0.83 blinks.

[0158] Therefore, it was found that in this model animal, the occlusion of the meibomian gland orifice, BUT, and blink frequency were reversible and could be used to evaluate drug efficacy.

[0159] Using the same method, we also confirmed that the tear film optical interference score in this animal model improved with the drug. We also found that the corneal staining score and tear volume were variable in this animal model. Therefore, these evaluation items are reversible and can be used to evaluate drug efficacy.

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

[0161] After euthanizing the mice, the eyelids are collected and embedded and frozen in OCT compound (Sakura Finetech Japan), and 10 μm-thick sagittal frozen sections are prepared.

[0162] 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 Class I H2-D ｄ Immunohistochemical staining is performed using antibodies (SCID mouse tissue markers).

[0163] If the mouse is a SCID mouse that is deficient in T cells and B cells, the presence or absence of CD3 or CD19 positive cells is determined, and if positive cells are present, it can be determined that cells from an allogeneic animal have been mixed in.

[0164] Macrophages, dendritic cells, NK cells, and endothelial cells are included in cells derived from the spleen, and these cells have MHC class I. Therefore, whether the mouse is a SCID mouse or not, the presence of spleen-derived cells in the eyelid of the mouse and whether the spleen-derived cells have a different MHC class I type from that of the mouse can be detected to confirm the contamination with cells from an allogeneic animal. When the mouse is a SCID mouse, the MHC class I H2-D ｄ The presence or absence of cells that are negative and positive for F4 / 80, CD11, NK1.1, CD31 or Claudin-5 is determined, and if cells that satisfy both criteria are present, it can be determined that cells from an allogeneic animal have been mixed in.

[0165] While the present disclosure has been illustrated using preferred embodiments thereof, it is understood that the scope of the present disclosure should be construed solely in terms of the claims. It is understood that the patents, patent applications, and literature cited herein are incorporated by reference in their entirety as if the contents themselves were specifically set forth herein.

Claims

1. A method for evaluating the efficacy of a test substance as a therapeutic and / or preventive agent for an eye disease, the method comprising: (1) a step of administering the test substance to an eye disease model animal; and (2) a step of evaluating at least one of characteristics associated with meibomian gland dysfunction or characteristics associated with dry eye in the eye disease model animal, wherein the eye disease model animal is an animal with reduced or impaired immune function into which cells from an allogeneic animal have been transplanted.

2. The method of claim 1, wherein step (2) includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction.

3. The method of claim 1 or 2, wherein in step (2), the characteristics associated with meibomian gland dysfunction include at least one selected from the group consisting of obstruction of meibomian gland orifices, atrophy of the meibomian glands, and eyelid margin irregularities, and the characteristics associated with dry eye include at least one selected from the group consisting of abnormal tear film break-up time, abnormal corneal staining score, reduced tear volume, abnormal tear film optical interference score, and abnormal blink rate.

4. The method according to any one of claims 1 to 3, wherein in step (2), the characteristics associated with meibomian gland dysfunction include at least blockage of the meibomian gland orifice.

5. The method according to any one of claims 1 to 4, wherein in step (2), the method includes a step of evaluating at least one characteristic associated with meibomian gland dysfunction and at least one characteristic associated with dry eye.

6. The method according to claim 5, wherein in step (2), the characteristics associated with meibomian gland dysfunction include at least blockage of the meibomian gland orifice, and the characteristics associated with dry eye include at least an abnormality in tear film breakup time and an abnormality in blink rate.

7. The method according to any one of claims 1 to 6, wherein the eye disease is meibomian gland dysfunction.

8. The method according to any one of claims 1 to 6, wherein the eye disease is dry eye.

9. The method of any one of claims 1 to 8, wherein the allogeneic animal cells are immune cells.

10. The method of claim 9, wherein the allogeneic animal cells are immune cells derived from the spleen, blood, bone marrow, thymus, or lymph nodes.

11. The method of claim 10, wherein the allogeneic animal cells are cells derived from the spleen.

12. The eye disease model animal has an MHC type of H-2D ｄ The method according to any one of claims 1 to 11, wherein the mouse is 13. The method according to any one of claims 1 to 12, 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 13, wherein the allogeneic animal is a mouse having an MHC type different from that of the eye disease model animal.

15. The allogeneic animal has an MHC type of H-2D ｂ The method according to any one of claims 1 to 14, wherein the mouse is 16. A method for screening therapeutic and / or preventive agents for eye diseases, comprising: (1) a step of administering a test substance to an eye disease model animal; (2) a step of evaluating a characteristic associated with meibomian gland dysfunction or a characteristic associated with dry eye in the eye disease model animal; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for eye diseases when the evaluated characteristic improves or inhibits progression, wherein the eye disease model animal is an animal with reduced or compromised immune function into which cells from an allogeneic animal have been transplanted.

17. A method for screening a therapeutic and / or preventive agent for meibomian gland dysfunction, comprising: (1) a step of administering a test substance to a model animal of meibomian gland dysfunction; (2) a step of evaluating blockage of the meibomian gland orifice in the model animal of meibomian gland dysfunction; and (3) a step of identifying the test substance as a therapeutic and / or preventive agent for meibomian gland dysfunction when blockage of the meibomian gland orifice is improved and / or progression of blockage of the meibomian gland orifice is inhibited in the model animal of meibomian gland dysfunction, wherein the model animal of meibomian gland dysfunction is an animal having reduced or insufficient immune function to which cells from an allogeneic animal have been transplanted, and the MHC type of the model animal of meibomian gland dysfunction is H-2D ｄ and the MHC type of the allogeneic animal is H-2D. ｂ A screening method.

18. A method for screening a therapeutic and / or preventive agent for dry eye, comprising: (1) administering a test substance to a dry eye model animal; (2) evaluating in the dry eye model animal blockage of the meibomian gland orifice, abnormality in tear film breakup time, and abnormality in the blink frequency; and (3) identifying the test substance as a therapeutic and / or preventive agent for dry eye when the blockage of the meibomian gland orifice, abnormality in tear film breakup time, and abnormality in the blink frequency are improved or inhibited from progressing in the dry eye model animal, wherein the dry eye model animal is an animal with reduced or insufficient immune function to which cells from an allogeneic animal have been transplanted, and the MHC type of the dry eye model animal is H-2D. ｄ and the MHC type of the allogeneic animal is H-2D. ｂ A screening method.

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