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ARTICLES
Year : 1972  |  Volume : 20  |  Issue : 2  |  Page : 63-69

Ocular response to uveal tissue


Department of Opthalmology, A.I.I.M.S., New Delhi, India

Correspondence Address:
L P Agarwal
Department of Opthalmology, A.I.I.M.S., New Delhi
India
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Source of Support: None, Conflict of Interest: None


PMID: 4668479

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How to cite this article:
Agarwal L P, Singh P, Khosla P K. Ocular response to uveal tissue. Indian J Ophthalmol 1972;20:63-9

How to cite this URL:
Agarwal L P, Singh P, Khosla P K. Ocular response to uveal tissue. Indian J Ophthalmol [serial online] 1972 [cited 2020 Oct 24];20:63-9. Available from: https://www.ijo.in/text.asp?1972/20/2/63/34666

Sympathetic ophthalmitis has been considered to be either of viral origin or a hypersenstive res­ponse to uveal tissue. Evidences in favour of each are equally balanced. Elschnig (1910a) first suggested that sympathetic ophth­almia is an allergic response to uveal tissue. In order that this could be accepted it had to be proved that uveal tissue has anti­genic properties and is organ specific. Elschnig (1910 b) showed by animal experiments that it is the uveal tissue pigment which is the fraction responsible for such a reaction and that this fraction is organ specific. His findings were supported by Kummel (1912 and 1913) who in addition demonstrat­ed the presence of antiuveal anti­bodies in 7 out of 13 cases `of sympathetic ophthalmia. Wissman, (1911) further strengthened this hypothesis by producing anaphy­laxis by injecting the emulsion of the diseased eye (sympathetic ophthalmitis) of the individual in guinea pigs. passively sensitized by the serum of the same patient.

Fuchs and Meller (1914) how­ever failed to demonstrate anti­uveal antibodies in the serum of these patients. Rados (1913) and von Szily (1916) failed to demon­strate the production of antiuveal antibodies in animal experiments.

This led to considerable criticism of Elschnig's (1910) allergy theory of sympathetic ophthalmia. Woods (1916) confirmed Elschnig's (1910) work and he attributed von Szily's (1916) failure to faulty technique of heating the antigen which re­sulted in denaturation of proteins. Woods (1921) also demonstrated complement fixing antiuveal anti­bodies in serum of patients with sympathetic ophthalmia.

Vannas, Wedman and Tier (1935) produced a histopathological picture resembling sympathetic ophthalmia following the intraper­itoneal implantation of the homo­logous eye. Collins (1949) and Aronson, Rogan and Zweigart (1963 a+b) produced in animals disease resembling sympathetic ophthalmia following repeated sys­temic injection of uveal tissue emulsion in combination with Freunds adjuvant (complete). But certain workers like Naquin (1955), failed to produce uveal hypersen­sitive reaction using a technique similar to that of Collins (1949).

A brief review of the sequence of events in this field indicates that nothing conclusive has been achi­eved so far. Allergic hypothesis needs more conclusive support to be finally accepted.


  Materials and Methods Top


Healthy pigmented guinea pigs weighing between 400 to 500 gms were taken for the study. Animals were kept under observation for 15 days to rule out the presence of any concurrent ocular disease. Whole uveal tissue was used as an­tigen, so as to include all the anti­genic components in their natural form.

The antigen was prepared from fresh enucleated guinea pig eyes. Uvea was dissected, pooled, weigh­ed and homogenised in known amount of saline using a glass homogenizer (Poter Eleveghan). The final mixture contained 50 gm (wet weight) of uvea per cc. All this procedure was carried out under aspetic conditions. Sterility of the mixture was tested by in­oculation on blood. agar. Fresh antigen was prepared for each batch of injections to guard against denaturation and alteration of antigenicity during storage.

Freunds adjuvant* (complete) was used throughout the study. The antigen adjuvant mixture was prepared by mixing the two in the ratio 7:3 (Aronson et al 1963 a). The mixture was prepared each time just before injecting.

Sensitization was carried out through intra-peritoneal and sub­cutaneous routes by four weekly injections of the angiten. Intrao­ocular, intramuscular or intracar­diac challenge was given 2 weeks after the last immunization injec­tion.

The study was divided into the following groups :­

GROUP I

Intraocular injections:-
to serve as data to determine the type of ocular response to primary injection of the antigen, as con­trols for comparison with experi­ments in group II & III.

a. 0.2 cc of uveal tissue antigen - 6 animals.

b. 0.2 cc of Freund's adjuvant (complete) mixed with saline in the ratio of 3:7 - 6 ani­mals.

c. 0.2 cc. of adjuvant-antigen mixture in the ratio of 3:7 as in b. - 6 animals.

GROUP II

Intraperitoneal sensitization :­


by 3 injections of the antigen given intraperitoneally at weekly intervals.

a. 1 cc of the uveal tissue anti­gen - 9 animals.

b. 1 cc of adjuvant-saline (3:7) mixture - 3 animals.

c. 1 cc of adjuvant-antigen (3:7) 3 animals.

GROUP III

... Subcutaneous sensitisation :-
9 animals, by three injections of 1 cc of uveal tissue antigen given sub­cutaneously at weekly intervals.

The animals in each group were challenged with the uveal antigen two weeks after the last sensitiz­ing injection in the following ways :­

i. intraocular challenge 0.02 cc -3 animals.

ii. intracardiac challenge 1.00 cc - 3 animals.

iii. intramuscular challenge 1.00 cc - 3 animals.

All intraocular injections were given in the anterior chamber of the right eye.

The animals were observed for a period of 22 days following the challenge injections.

Histopathological studies of the eyes were carried out on the days of sacrificing the animals i.e. 5th, 14th and 28th days. The eyes were embedded in paraffin and the sec­tions stained with haematoxylin and eosin.

Immunological studies of the blood and aqueous were done on the days of challenge and of sacri­ficing the animals. Aqueous studies on the day of challenge were limi­ted only to those animals getting an intraocular injection. This was to avoid unnecessary trauma to the eye which might interfere with the picture of ocular response. Double diffusion in agar by the technique of Agarwal, Gupta and Madan Mohan (1963) and immuno­electrophoresis by the technique of Schiedegger (1955) were the methods employed for the study of antibody response. The antigen for immune diffusion studies was cent­rifuged to remove the particulate matter. It was concentrated so that protein concentration was at least 10 mg./cc.


  Observations Top


In all the groups, animals were observed clinically for a period of 28 days. Observations were record­ed under two heads - I. Clinical and histopathological, II. Immuno­logical.

I. Clinical and Histopathological Group I (a)

1st to 5th
day : All injected eyes showed inflammation, characteriz­ed by ciliary congestion, miosis and corneal oedema with mild flare but no K. P.'s. Fundus was normal. Inflammation reached its peak on the 3rd day, and started regressing by the 5th day.

Histopathology of the eye enuc­leated on the 5 th day showed polymorphonuclear reaction in the cornea, iris and ciliary body with pigmented particles sticking on the back of cornea. The choroid show­ed areas of pigment proliferation.

6th-28th day : The mild signs of inflammation which were present si.arted disappearing progressively. On the 7th day there was no sign of inflammation. The eye remained normal till the last day of observation.

Histopathology of eyes enucleat­ed on 14th day and 28th day did not reveal any sign of inflamma­tion. The choroid showed areas of pigment proliferation.

Group I b

1st to 5th
day : All the injected eyes showed signs of inflammation in the form of ciliary congestion, miosis and corneal oedema. As the anterior chamber contained the injected material it was, therefore, not possible to comment on the presence of K.P's and aqueous flare. Fundus could not be seen. Inflammation progressively increa­sed. There was marked increase in ciliary congestion which formed a girdle around the limbus. Corneal haziness resembled a ground glass. The anterior chamber contained in addition to the injected material tats of exudates and in one animal it had formed hypopyon. The vitre­ous and fundus could not be visualized. Clinical picture did not show any abatement.

Histopathology on the 5th day showed diffuse polymorphonuclear cellular infiltration of the cornea, iris and ciliary body with exuda­tion of cells into the anterior chamber. Stromal vessels of the iris and ciliary body showed dila­tation. The choroid showed areas of pigment proliferation.

6th to 15th day : The clinical picture did not show signs of decline. The cornea showed increa­sed opacificaton, a few limbal blood vessels were seen invading super­ficial layers of corneal stroma. Ex­udation showed signs of organiza­tion forming synechea between iris and back of cornea.

Histopathological picture was essentially the same as on the 5th day except that the cornea showed sub-epithelial vascularization in addition to infiltration. The choroid, iris and ciliary body showed pig­ment proliferation.

16th to 28th day : The Clinical picture remained unchanged. His­topathology showed increased irre­gularity of corneal lamellae, with subepithelial and stromal vascula­rization. In additon to diffuse poly-morphonuclear cellular reac­tion, some mononuclear and plasma cells could be seen in the region of the iris and ciliary body. Substance of the iris showed hya­line degeneration with pigment proliferation.

Group I c : ....

Clinical and histological studies were similar to those in group I b

Group II: (Intraperitoneal injec­tion group).

Group II a (1)

Clinical, histopathological and immunological pictures were the same as in group Ia.

Group II a (ii & iii)

There was no clinical or histopa­thological evidence of uveitis in any of the animals in these groups.

Group 11 b (i)

The observations were the same as in group 1 a (i).

Group II b (ii & iii)

No clinical or histopathological evidence of uveitis was seen.

Group II c (i, ii & iii) -

Observations were comparable to those in II a (i, ii & iii) respective­ly.

Group III : (Subcutaneous Sensiti­zation)

Group III a (i) - Clinical, his­topathological and immunological pictures were similar to that in group I a (i).

Group III a (ii & iii) The eyes remained normal clinically as well as histopathologically during the period of observation.

IMMUNOLOGICAL STUDIES

No antiuveal antibodies could be demonstrated in either blood or aqueous of all the groups examined.


  Discussion Top


Contradictory reports in litera­ture are available on the immune nature of heterologous, homolog­ous and autologus uvea. Elschnig (1910 a & b ) Kummel (1912 & 1913) and Woods (1916 and 1917) supported the allergic theory of sympathetic ophthalmic by show­ing that the homologous uvea is capable of an immune response in experimental animals. Von Szily (1916) and Naquin (1955) have, however, failed to demonstrate such a reaction.

Collins (1949) Aronson, Hogan and Zweigart (1963, a, b & c) have succeeded in producing immune uveitis but have either failed to or have not demonstrated the pre­sence of antiuveal antibies in the serum of the animals.

Immunological studies in our series have failed to confirm the earlier works of ELschning (1910 b). Kummel (1912) and Woods (1916) who have shown that uveal tissue pigment is antigenic. We sensitis­ed the animals with whole uveal tissue antigen, but have failed to find demonstratable antiuveal anti­bodies in either serum or aqueous. We believe that the failure to show demonstratable antibody titres is either due to the fact that homologous uveal tissue is not an­tigenic at all or is so weakly anti­genic that it has failed to produce demonstrable antibody titres. It is, further surmised that the anti­bodies may not have been in sufficient concentration to produce an immune response which could be observed clinically or histo­logically.

Intraocular injection of uveal tissue antigen alone or in combina­tion with Freund's adjuvant in the anterior chamber could not produce any clinical, histopatho­logical or immunological evidence of ocular or general sensitization. This finding is however, contradi­ctory to the earlier work `(Silverstein and Zimmerman (1959). We feel that difference could be due to two factors. The latter observers used bovine serum albumin which is a strong antigen as compared to uveal tissue and the injection was given into the vitreous which has the capacity to store and then slowly release the antigen.

Histological picture in which antigen and adjuvant or adjuvant alone was injected showed reaction in the early phase to which later mononuclear lymphocytes and plasma cells were added but no epithelioid cell. We attribute this granulomatous reaction to the pre­sence of tubercle bacilli (dead) and the oils in Freund's adjuvant.

Our clinical and histological studies do not support the earlier works of Vannas et al (1935), Collins (1949) and Aronson and Hogan (1963 a, b, c.)

In our present study, the clini­cal and histopathological picture on intraocular challenge with uveal tissue antigen alone in animals sensitized either with uveal tissue antigen or with Freund's adjuvant was similar and it compared favou­rably with that in group I a. (without sensitization). This pro­bably indicates that the reactions noticed have probably the same aetiological basis. This is further confirmed by the absence of im­munolocial response in all these animals. The lack of antibody pro­duction in intraperitoneal sensiti­zation group is also further supported by the absence of reaction following even intracar­diac challenge and the absence of antiuveal antibodies in blood and aqueous in all the groups. The animals sensitized with a combi­nation of uveal tissue antigen and adjuvant showed similar reaction which compares well with group Ia.

Presence of areas of pigment aggregation with lymphocytes which were considered diagnostic by Collins (1949) were, however, found in the animals in the control group and were considered to be of normal occurrence. Such nonspeci­fic aggregations have already been reported by Naquin (1955). It was therefore. considered that an intra­peritoneal injection of uveal tissue antigen alone or in combination with Freund's adjuvant (complete) can neither evoke an immune res­ponse nor can produce a clinical and histological picture of allergic uveitis.

Encouraged by the results of subcutaneous immunization with corneal antigen (Agarwal, Gupta and Mohan, 1963) we tried this route for immunization in group III. The final results were in no way different from those in group II.

We, therefore, feel that homolo­gous uveal tissue antigen alone or in combination with Freund's adjuvant (complete) is neither capable of producing a detectable immune response nor can it pro­duce a clinical picture of uveitis resembling sympathetic ophthal­mia.


  Summary and Conclusions Top


Response of ocular tissue to re­peated injections of homologous uvea has been studied in guinea pigs. Animals were immunized to uveal tissue antigen alone and in combination with Freund's adjuv­ant (complete) through intraperi­toneal and subcutaneous routes. They were observed clinically, histopathologically and immuno­logically. We draw the following conclusions :­

(i) Intraocular injection into the anterior chamber of uveal tissue antigen alone produces a transitory reac­tion which completely sub­sides in 7 days. No ocular or general sensitization could be demonstrated.

(ii) Intraocular injection of Freund's adjuvant alone or in combination with uveal tissue antigen produces a very severe inflammatory reaction which does not show any sign of abatement during the period of obser­vation. No ocular or gene­ral sensitization could be demonstrated.

(iii) Repeated intraperitoneal or subcutaneous injection of uveal tissue alone or in com­bination fails to produce any general or ocular sensitivity.

(iv) Picture resembling sym. pathetic ophthalmia could not be produced by repeated immunization with uveal tissue

(v) Pigment proliferation in choroid is of normal occur­rence in guinea pig uveal tissue and, therefore, can­not be regarded as pathog­nomonic of sympathetic ophthalmia in these ani­mals[20].

 
  References Top

1.
Agarwal, L.P.; Gupta, A.K. and Mohan M. (1963) Orient. A. Ophth.; 1. 217.  Back to cited text no. 1
    
2.
Aronson S.D., Hogan M.J. and Zweigart, P. (1963 a) A.M.A. Arch. of Ophth. 69, 105.  Back to cited text no. 2
    
3.
Aronson S.D., Hogan M.J. and Zweigart, P. (1963 b) ibid 69, 203.  Back to cited text no. 3
    
4.
Aronson S.D.. Hogan M.J. and Zweigart, P. (1963 c) ibid 69, 208.  Back to cited text no. 4
    
5.
Cellins, R.C. (1949) Am. J. Ophth. 32. 1687.  Back to cited text no. 5
    
6.
Eischnig, A. (1910a) Arch. Fur. Ophth. 75. 459.  Back to cited text no. 6
    
7.
Elschnig, A. (1910b) Arch. Fur. Ophth. 76, 499.  Back to cited text no. 7
    
8.
Fuchs, A. and Meller, J. (1914) Arch. F. Ophth. 88, 280.  Back to cited text no. 8
    
9.
Kummel, R. (1912) Arch. F. Ophth, 81. 486.  Back to cited text no. 9
    
10.
Kummel, R. (1913) Arch. F. Ophth. 84, 440.  Back to cited text no. 10
    
11.
Naquin G.A. (1955) Amer. J. Ophth. 39. 196.  Back to cited text no. 11
    
12.
Rados, A. (1913) Z. f. Immun Und. Exp. Therap. 19, 579.  Back to cited text no. 12
    
13.
Silver-Stein, A.M. and Zimmer-man L.E. (1959) Am. J. Ophth. 48, 447. (Nov. I & II)  Back to cited text no. 13
    
14.
Szily, A. Von (1916) Klin. Monatsbl. F. Augenh, 56, 79.  Back to cited text no. 14
    
15.
Schiedegger, J.J. (1955) Arch. Allergy. App. Immunol. 7, 103.  Back to cited text no. 15
    
16.
Vannas, S; Wedman, E; and Tier, N. (1935) Acta. Ophth. Kbh. 36, 618.  Back to cited text no. 16
    
17.
Woods, A.C. (1916) Arch. of Ophth. 45. 557.  Back to cited text no. 17
    
18.
Woods, A.C. (1917) Arch. of Ophthal. 46, 8.  Back to cited text no. 18
    
19.
Woods A.C. (1921) Tr. Sect. Ophthal. A M.A. 105.  Back to cited text no. 19
    
20.
Wissman R. (1911) Arch. of Ophth. 80, 399.  Back to cited text no. 20
    




 

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