Identification of the Lymphokines, Inferferon-Gamma and Lnterleukin-2, in Inflammatory Eye Diseases

Identification of the Lymphokines, Inferferon-gamma and lnterleukin-2, in Inflammatory Eye Diseases

John J Hooks,* C. C. Chon,f and Barbara Derrickf

The exact pathogenic mechanisms involved in autoimmune and inflammatory eye diseases are not known. However, studies during the past few years indicate that a T cell infiltrate, T cell sensitization to retinal antigens and expression of major histocompatibility complex (MHC) class II antigens are associated with this process. In this report we show that the lymphokines, IL-2 and IFN-gamma, are present in the human eye during inflammatory and autoimmune diseases. The presence of these lymphokines is associated with a lymphocyte infiltrate, predominantly of T cell origin, and with the expression of MHC class II antigens on both the infiltrating cells and ocular resident cells, that is, retinal pigment epithelial (rpe) cells and retinal vascular endothelial cells. Furthermore, in vitro studies demonstrate that IFN-gamma can enchance the expression of the HLA-DR determinant on both of these cell types. These observations suggest that lymphokine induced class II antigen expression may serve as a local amplification system in autoimmune and inflammatory eye diseases. A better understanding of the role of lymphokines in the mechanisms involved in the development of autoimmunity and inflammation may be beneficial in the treatment of these diseases. Invest Ophthalmol Vis Sci 29:1444-1451, 1988

Lymphokines, such as IFN-gamma and IL-2, are cess. 16 The nature of the infiltrating cells, some of believed to play a major role in facilitating immune the ocular antigens involved in the autoimmune pro- responses.1'2 For example, IL-2, which is produced by cess and the activated expression of MHC class II T-lymphocytes, can activate T cells and NK cells in antigens have all been described. cytotoxic responses. IFN-gamma, a major immuno- Studies evaluating systemic immunologically re- regulatory protein, induces or augments MHC class lated disorders suggest that lymphokines may be am- II antigen expression on a variety of cells.3"5 These plifying immune responses and thus in this way influ- MHC class II antigens enable cells to present antigens ence the pathogenesis of those diseases.17"9 In order to T-lymphocytes resulting in the initiation and per- to investigate the immunoregulatory processes in au- petuation of immune responses.6"9 Through these toimmune and inflammatory eye diseases, we exam- processes, it appears that lymphokines may help to ined ocular tissue for the presence of lymphokines. In mediate certain immunopathologic processes and this report we show by immunocytochemical staining may be required to augment the host defense against that the lymphokines, IFN-gamma and IL-2, are in- certain infections and malignancies.12 Moreover, it is deed present in the eye during inflammation. More- becoming increasingly clear that a better understand- over, their presence correlates with the expression of ing of the mechanisms involved in the development class II antigens on infiltrating cells and on the retinal of autoimmunity and inflammation may be benefi- pigment epithelial and the retinal vascular endothe- cial in the treatment of these diseases. lial cells. The exact pathogenic mechanisms involved in idiopathic inflammatory eye diseases are not known. Materials and Methods However, studies during the past few years have Patient Population begun to shed light on various aspects of this pro- Fresh enucleated eyes were obtained from four patients with inflammatory eye diseases and from three From the *Immunology and Virology Section and the "flmmu- individuals without eye disease. The patients with in- noregulation Section, Laboratory of Immunology, National Eye flammatory eye diseases consisted of one patient with Institute, National Institutes of Health, Bethesda, Maryland. idiopathic uveitis and three patients with sympathetic Submitted for publication: October 21, 1987; accepted April 7, ophthalmia. The case histories and histopathologic 1988. findings of these patients have been previously de- Reprint requests: John J Hooks, PhD, Chief, Immunology Virol- 1416 ogy Section, National Eye Institute, National Institutes of Health, scribed. Informed human consent was obtained Bldg. 10, Room 6N228, Bethesda, MD 20892. prior to undertaking the study.

1444

Downloaded from iovs.arvojournals.org on 09/30/2021 No. 9 INTERFERON AND IL-2 IN INFLAMMATORY EYE DISEASES / Hooks er ol 1445

Case 1: A 45-year-old woman underwent retinal inducer subset of T cells.19 Anti Leu 14 binds to the reattachment surgery followed by vitrectomy and CD22 molecule present on B-lymphocytes. Anti- lensectomy in her right eye. A year later, she was OKM 1 reacts with the CR3 complement/receptor treated with laser photocoagulation for a retinal tear and Leu M3 reacts with the CD1 lb molecule on pe- in her left eye. Following laser treatment, she devel- ripheral monocytes.20'21 Anti-Leu 7 binds to the oped bilateral panuveitis. A diagnosis of sympathetic CD 16 molecule present on NK cells. Anti-TAC ophthalmia was made and her right eye was then monoclonal antibody identifies with the IL-2 recep- enucleated. The complete case history and histo- tor described on activated T cells. pathologic evaluation are reported as case 4 in reference 14. Immunoperoxidase Procedure Case 2: A 38-year-old man suffered blunt trauma Freshly enucleated eyes were immediately embed- leading to a ruptured right eye. Surgical repair was ded in OCT at -80°C. Four micron serial frozen performed immediately. However, 6 months later the sections were cut and placed on gelatinized slides. patient presented with symptoms and clinical fea- Immunocytochemical evaluation of these sections tures consistent with sympathetic ophthalmia. His were performed by the avidin-biotin-peroxidase right eye was enucleated. The complete case history (ABC) complex technique.22 Slides were fixed for 5 and histopathologic findings are reported as case 5 in min in acetone, washed in tris-buffered saline (pH reference 14. 7.6) and immersed for an additional 5 min in 10% Case 3: A 60-year-old man underwent four unsuc- normal horse serum. Slides were overlaid with ap- cessful retinal reattachment operations in his left eye. propriate mouse monoclonal antibodies or with Two months prior to his last surgery he developed mouse ascites fluid, or supernatant fluid which served typical bilateral sympathetic ophthalmia. His left eye as the negative control. After 1 hr incubation in a was enucleated 6 years later. A complete case history moist chamber, the slides were washed in tris-buf- and histopathologic evaluation are reported as case 4 fered saline and then overlaid with biotin-conjugated in reference 16. horse anti-mouse IgG (1:200) (Vector Lab, Burlin- Case 4: This is a 40-year-old man with a 14-year game, CA). This was followed by another 1 hr incu- history of bilateral progressive chronic uveitis. Diag- bation in a moist chamber, and slides were then nostic vitrectomy revealed nonspecific chronic in- washed in tris-buffered saline and overlaid 45 min flammation. His right eye became phthisical and with a 1:100 dilution of avidin-biotin-peroxidase painful and was enucleated. complexes. The slides were washed again in tris-buf- The nature of the cell type infiltrating the ocular fered saline and developed in diaminobenzide-8% tissue from these patients was evaluated using mono- nickel sulfate-3% hydrogen peroxidase solution. The clonal antibodies directed against T-lymphocytes, B- slides were then counterstained with methyl green lymphocytes, monocytes and NK cells. The majority (1% in methanol), dehydrated, cleared and mounted. of infiltrating cells in each case demonstrated cell surface markers for T-lymphocytes.1416 Preparation of Cells Blood was drawn from healthy human volunteers Antibodies into heparinized syringes and the mononuclear cells Monoclonal anti-HLA-DR antibodies, directed were isolated by Ficoll-Hypaque centrifugation. The against the common region of the HLA-DR mole- mononuclear cells were suspended at a concentration cule, were obtained from Ortho Pharmaceutical Co. of 2.5 X 106 cells/ml in RPMI 1640 medium supple- (OKla+) (Raritan, NJ) and Becton Dickinson mented with 10% fetal calf serum. Cells were incu- (L-243) (Mountainview, CA). The specificity of these bated with medium or conconavalin A (Con A) (10 antibodies has been previously described.17 Mono- ng) for 48 hr. The cell suspensions were cytocentri- clonal anti-IL-2 antibody was obtained from Genen- fuged onto slides and analyzed by immunoperoxidase zyme (Boston, MA), and monoclonal anti-IFN staining. Supernatant fluids were assayed for IFN ac- gamma immunoglobulin was kindly provided by Dr. tivity. Monocytes were also obtained from Ficoll-Hy- 18 Jan Vilcek (New York University, NY). paque-treated blood and used as the control cell type The following monoclonal antibodies prepared in the complement mediated cytotoxicity assay. against human lymphocytes were used in this study. Anti-Leu 4 monoclonal antibody reacts with the CD3 Interferon Assay molecule present on pan T-lymphocytes. Anti-Leu 2 Antiviral activity was determined by the reduction binds to the CD8 molecule present on the suppres- in vesicular stomatitis virus plaque formation on sor/cytotoxic subset of T cells and anti-Leu 3 reacts human amnion (WISH) cells (ATCC; CCL25) grown with the CD4 molecule on the surface of the helper/ in microtiter plates.2324 The antiviral activity, ex-

Downloaded from iovs.arvojournals.org on 09/30/2021 1446 INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE / September 1988 Vol. 29

:*>

.-. v Fig. 1. Immunocyto- chcmical identification of lymphokines and MHC class II antigens on peripheral mononuclear cells. Pe- ripheral mononuclear cells were incubated with Con A IL-2, for 48 hr. Cells were then analyzed by immunocytochemical analysis with mouse ascitic fluid (C)> or with monoclonal antibodies prepared against IL-2 (IL-2), IFN-gamma (IFN) or MHC class II antigens (HLA-DR).

,».•»* IFN

pressed in interferon units, was calculated as the re- onstrated in peripheral mononuclear cells by immu- ciprocal of the highest dilution of the sample that nocytochemical assays. Mononuclear cells, obtained reduced the number of viral plaques by 50%. Human as described in Materials and Methods, were incu- reference interferon-gamma (20,000 units; obtained bated with media alone or with Con A (10 ng/ml) for from the Research Reagent Branch, National Insti- 48 hr. Cells were then analyzed by the ABC immuno- tute of Allergy and Infectious Disease, NIH, Be- peroxidase assay with monoclonal antibodies pre- thesda, MD) contained 10,000 units when tested in pared against T cells, B cells, macrophages, NK cells, our assay system. and the lymphokines, IFN-gamma and IL-2. When cells incubated with media were analyzed, IFN- Results gamma could not be detected in any of the preparations. However, cells from these same preparations IFN-gamma and IL-2 in Peripheral Mononuclear occasionally stained for IL-2. In contrast, all the prep- Cells arations containing cells treated with Con A showed Studies were initially designed to determine if the reactivity with IFN-gamma and IL-2 (Fig. 1). Al- lymphokines, IFN-gamma and IL-2, could be dem- though nearly all of these cells reacted with IFN-

Downloaded from iovs.arvojournals.org on 09/30/2021 No. 9 INTERFERON AND IL-2 IN INFLAMMATORY EYE DISEASES / Hooks er ol 1447

V* v

Fig. 2. Immunoperoxidase staining of a frozen section of normal human eye. (A) A negative reaction with mouse ascitic fluid(control) . (B) A negative reaction with monoclonal anti-IL2 antibody. (C) A negative reaction with monoclonal anti-IFN-gamma antibody.

gamma and IL-2 antibodies, the specific staining pat- The most intense staining for the lymphokines was terns for the two lymphokines were very distinct. As observed in those cases characterized by a diffuse in- is seen in Figure 1, the IFN-gamma staining pattern flammation (cases 1 and 2), whereas weak staining for was diffuse, whereas the IL-2 staining pattern con- both IFN-gamma and IL-2 was detected in the cases sisted of discrete, pinpoint areas of dark stain in the characterized by mild or focal inflammation (cases 3 cytoplasm. When the cells were evaluated by immu- and 4). As is seen in Figure 3 and Figure 4, both IL-2 noperoxidase staining it was noted that the majority and IFN-gamma were detected in the lymphocytes of the cells were T cells, while only a small percentage infiltrating the choroid. The staining pattern ob- of the cells were B cells and monocytes. Over 90% of served in these cells was similar to the staining noted these cells marked with anti-HLA-DR antibody. in the mononuclear cells which were activated in We next attempted to correlate the immunocyto- vitro. Both lymphokines were detected in the cyto- chemical localization of IFN-gamma in the lympho- plasm. Once again the IL-2 displayed a pinpoint pat- cytes with the generation of antiviral activity in the tern (Figs. 3b, 4b) while IFN-gamma displayed a dif- supernatant fluids. Supernatant fluids from the fuse pattern (Figs. 3 c, 4c). media treated lymphocytes did not display antiviral activity, whereas the Con A-treated lymphocytes Relationship Between IFN-gamma Production and contained 160 IFN units. This antiviral activity was MHC Class II Antigen Expression During shown to be due to IFN-gamma antibody since the Inflammatory Eye Diseases antiviral activity was neutralized by monoclonal Since IFN-gamma and IL-2 are products of acti- anti-IFN-gamma antibody and not by anti-IFN- vated T cells and IFN-gamma modulates MHC class alpha antibody. These studies demonstrate that the II antigen expression, we next correlated the presence immunocytochemical identification of IFN-gamma of T cells and class II antigen expression within the in activated T cells corresponds to the development inflamed eye. As can be seen in Figure 4d, the major- of biologically active IFN-gamma. ity of the infiltrating cells were shown to be T cells and to express class II antigens. In addition, the pres- Detection of Lymphokines in the Eye During ence of both lymphokines was correlated with the Inflammatory Eye Diseases expression of class II antigens on two types of ocular We next evaluated both patient and control tissue resident cells, the retinal vascular endothelial cell and for the presence and tissue distribution of IFN- the rpe cell (Fig. 4). These studies clearly show that T gamma and IL-2. Using the immunocytochemical cells and two T cell-induced lymphokines are present staining procedure we were not able to detect IFN- within the inflamed eye. gamma or IL-2 in three eyes from normal individuals (Fig. 2). In contrast, IFN-gamma and IL-2 were de- Discussion tected in all the frozen eye sections from patients with In this study we show that the lymphokines, IFN- inflammatory eye diseases (Figs. 3, 4). gamma and IL-2, are present in the eye during the

Downloaded from iovs.arvojournals.org on 09/30/2021 1448 INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE / September 1988 Vol. 29

This study underscores the prominent role these soluble mediators may demonstrate in the pathophysio- logical chain of events leading to inflammation in general and ocular inflammation in particular. Although gaps in our knowledge of the pathogenic mechanisms involved in human ocular inflammatory diseases exist, exploration into the morphology and immunopathology of these conditions has yielded valuable information. Using a variety of monoclonal antibodies, investigators identified the lymphocyte population associated with the inflamed eye. In most cases the T-lymphocyte was found to be the predomi- nant cell type.10"16'25 Likewise, the development of animal models has helped to facilitate the identification of ocular antigens, such as S-antigen and interphotoreceptor retionid-binding protein (IRBP). Both of these proteins have been shown to be instru- mental in invoking experimental autoimmune uveitis (EAU).26'27 EAU is a T cell-mediated disease which is presently considered by many to be the rodent counterpart of the human disease, sympathetic ophthalmia. Re- \ cently, the role of class II antigen expression has been implicated in this disease. Two cell types in the retina, rpe cells and retinal vascular endothelial cells, were demonstrated to atypically express la antigens during the course of EAU.28'29 In fact, the activation of la antigens on rpe cells was noted 2-4 days prior to < clinical and histopathological evidence of disease and appeared to persist long after the disease had sub- sided. In vitro data from our earlier studies demon- \ strate that treatment of rpe cells and vascular endothelial cells with IFN-gamma can enhance their class II antigen expression. This information suggests that these retinal resident cells may function as an antigen-presenting cell in the ocular microenvironment.5 These findingsar e not unlike those recently described in other systems, such as experimental autoimmune Fig. 3. Immunoperoxidase staining of a frozen section of an eye encephalomyelitis and multiple sclerosis, where class from a patient with sympathetic ophthalmia (case 2). (A) A nega- II positive resident cells have been suggested to play a tive reaction with the mouse ascitic fluid (control) (R = retina, C 30 31 = choroid). (B) A positive reaction with the monoclonal anti-IL-2 role in inflammatory CNS diseases. ' Finally, in a antibody (dark blackish stain), on infiltrating cells (arrow) in the very recent study, we demonstrated that EAU could choroid (C) A positive reaction with the monoclonal anti-lFN- be significantly modified with antibodies to the la gamma on the infiltrating cells (arrows) in the choroid. molecule (Wetzig et al, unpublished data), further substantiating the role for class II positive ocular cells course of two ocular inflammatory disorders, in three in the regulation of immune reactivity in the eye. cases of sympathetic ophthalmia and in a case of Clearly, the elaboration of cytokines represents a idiopathic uveitis. In contrast, these lymphokines are necessary element in the evolution of an immune not detected in normal eye tissue. Furthermore, the response. Both IFN-gamma and IL-2, which are pro- presence of these inflammatory mediators is asso- duced as part of this cascade of events, are known to ciated with a mononuclear infiltrate consisting pre- have a variety of divergent functions. IFN-gamma is dominantly of T-lymphocytes. Concomitant with one of the major agents responsible for inducing the this T cell infiltrate is the expression of class II anti- expression and biosynthesis of the class II antigens on both infiltrating cells and selected ocular resi- gens.13"5'32'33 In addition, it can enhance T cell cyto- dent cells (ie. rpe cells and vascular endothelial cells). toxicity, modify B cell immunoglobulin production

Downloaded from iovs.arvojournals.org on 09/30/2021 No. 9 INTERFERON AND IL-2 IN INFLAMMATORY EYE DISEASES / Hooks er ol 1449

Fig. 4. Immunoperoxidase staining of a frozen section of an eye from a patient with sympathetic ophthalmia (case I). (A) A negative reaction with mouse ascitic fluid (C = choroid). (B) A positive reaction with monoclonal anti-IL2 antibody on the infiltrating cells in the choroid (arrows) and retina. (C) A positive reaction with monoclonal anti-IFN-gamma antibody in the infiltrating cells in the choroid (arrows) and retina. (D) A positive reaction with monoclonal anti-HLA-DR antibody on the infiltrating cells (arrows) and the rpe cells (open arrows).

Downloaded from iovs.arvojournals.org on 09/30/2021 1450 INVESTIGATIVE OPHTHALMOLOGY 6 VISUAL SCIENCE / September 1988 Vol. 29

and enhance the proliferation and differentiation of presence of these two mediators can result in the NK cells. With regard to monocytes and macro- coincident generation of other cytokines at the in- phages, IFN-gamma stimulates phagocytosis, inflammatory site. Once set into motion, this cyclic creases tumoricidal activity and inhibits monocyte pattern of lymphokine production and cell activation migration in a fashion analogous to that of MIF. Sim- leads to destruction of ocular tissue. ilarity, IL-2, first described as a T cell growth factor, is In summary, we have used in situ immunocyto- also a product of activated T cells. It has been shown chemical studies of the intraocular inflammatory re- to increase the function of cytotoxic T cells, enhance sponse to delineate those cells and cellular products the production of IFN-gamma and augment NK ac- responsible for ocular destruction. We have corre- tivity. U2M lated this finding with in vitro studies on rpe cells and To date, the tissue localization of IFN-gamma or vascular endothelial cells, to provide information other lymphokines to the eye has not been described. about the mechanisms involved.5 The data presented Until recently, the presence of these mediators has indicate a strong correlation among the development been supported mainly by in vitro data.1 Using im- of ocular inflammation, the acquisition of HLA-DR munohistochemical analysis, some lymphokines antigen and the presence of IFN-gamma and IL-2. have been identified in selected inflamed tissues. These observations suggest that lymphokine-induced Husby and Williams identified IFN-gamma and IL-2 HLA-DR antigen expression may serve as a local in association with HLA-DR expression and T cell amplification system in autoimmune and inflamma- infiltration in synovial tissue from rheumatoid arthri- tory eye diseases. tis patients, but not in renal lesions of systemic lupus erythematosis patients.35'36 Likewise, Isenberg and Key words: interferon-gamma, interleukin-2, lymphokines, sympathetic ophthalmia, uvetitis, inflammation associates detected these same components in muscle biopsy specimens from patients with polymyositis,37 while others reported that IL-2 was localized to a Acknowledgments small number of lymphocytes infiltrating the salivary The authors thank Ms. C. Percopo and Dr. Y. Dalavanga glands of patients with Sjogren's syndrome.38 In these for assistance in the immunoperoxidase assays. We also reports the investigators postulate that the presence of thank Dr. Jan Vilcek (New York University) for generously providing the monoclonal anti-IFN-gamma immunoglob- these lymphokines may be responsible for perpetuat- ulin and Dr. R. Nussenblatt for his helpful discussions and ing the inflammation associated with these diseases. review of the manuscript. We have now extended some of these studies to the eye. References Based on these and other recent reports, we pro- 1. Hooks JJ and Detrick B: Immunoregulatory functions of in- pose the following hypothesis for the role of IL-2 and 10 16 terferon. In Biological Response Modifiers (BRM'S), Torrence IFN-gamma in inflammatory eye diseases. " Al- P, editor. New York, Academic Press, 1985, pp. 57-75. though our studies were not designed to evaluate the 2. Rosenberg SA, Lotze MT, Muul LM, Lehman S, Chang AE, earliest events in the pathogenic process, we suggest Ettinghausen E, Matory YL, Skibber J, Shiloni E, Vetto JJ, that an initiating event, such as trauma or a virus Seipp CA, Simpson C, and Reichert CM: Observations on the systemic administration of autologous lymphokine-activated infection, may sufficiently traumatize the posterior killer cells and recombinant interleukin-2 to patients with met- pole of the eye, leading to the release of mediators astatic cancer. N Eng J Med 313:1485, 1985. that increase vascular permeability. This breach in 3. Basham TY and Merigan TC: Recombinant interferon in- the endothelial cell barrier of the microvasculature creases HLA-DR synthesis and expression. J Immunol and the ability of infiltrating cells to enter the sur- 130:1492, 1983. 4. Pober JS, Gimbrone MA Jr, Cotron RS, Reiss CS, Barakoff SJ, rounding tissue is of prime importance in the devel- Fiers W, and Auits KA: la expression by vascular endothelium opment of an inflammatory reaction. Once mononu- is inducible by activated T cells and by human gamma inter- clear cells invade the retina, the release of lympho- feron. J Exp Med 157:1339, 1983. kines can help to sustain the inflammatory reaction. 5. Detrick B, Newsome D, Percopo CM, and Hooks JJ: Class II For example, sensitized T cells can interact with reti- antigen expression and gamma interferon modulation of monocytes and retinal pigment epithelial cells from patients nal antigens in the context of MHC class II antigens. with retinitis pigmentosa. Clin Immunol Immunopathol As a consequence of this interaction, IFN-gamma 36:201, 1985. and IL-2 are released. IL-2 can activate T cell prolif- 6. Unanue ER: The regulatory role of macrophages in antigenic eration and IFN-gamma can induce the synthesis and stimulation: II. Symbiotic relationship between lymphocytes expression of class II antigens on monocytes, endo- and macrophages. Adv Immunol 31:1, 1981. 7. Bottazzo GF, Pujol-Borrel R, Hanafusa T, and Feldman M: thelial cells and rpe cells. Moreover, IFN-gamma can Hypothesis: Role of aberrant HLA-DR expression and antigen enhance the cytotoxicity of cytotoxic T cells and en- presentation in induction of endocrine autoimmunity. Lancet hance antibody formation by B cells. Finally, the ii: 1111, 1983.

Downloaded from iovs.arvojournals.org on 09/30/2021 No. 9 INTERFERON AND IL-2 IN INFLAMMATORY EYE DISEASES / Hooks er ol 1451

8. Hall BM, Duggin GG, Philips J, Bishop AG, Horvath JS, and Notkins AL: Immune interferon in circulation of patients with Tiller DS: Increased expression of HLA-DR antigens on renal autoimmune diseases. N Engl J Med 301:5, 1979. tubular cells in renal transplants: Relevance to the rejection 25. Nussenblatt RB, Palestine AG, and Chan CC: Proposed im- response. Lancet ii:247, 1984. mune mechanisms in uveitis and their modulation. In Interna- 9. Aichinger G, Fill H, and Wick G: In situ immune complexes, tional Ophthalmology Clinics: Immunological Ocular Dis- lymphocyte subpopulations and HLA-DR positive epithelial eases. Vol. 25. Friedlaender MH and Tabbano KF, editors. cells in Hashimoto thyroiditis. Lab Invest 52:132, 1985. Boston, Little, Brown, 1985, pp. 81-93. 10. Jacobiec FA, Marboe CC, Knowles DM II, Iwamoto, T, Herri- 26. Wacker WB, Donoso LA, Kalsow CM, Yankeelov JA, and son W, Chang S, and Coleman J: Human sympathetic oph- Organisciak DT: Experimental allergic uveitis: Isolation, char- thalmia, an analysis of the inflammatory infiltrate by hybrid- acterization and localization of a soluble uveito-pathogenic oma-monoclonal antibodies, immunochemistry, and correla- antigen from bovine retina. J Immunol 119:1949, 1977. tive electron microscopy. Ophthalmology 90:76, 1983. 27. Gery I, Wiggert B, Redmond TM, Kuwabara T, Crawford MA, 11. Muller-Hermelink HK, Kraus-Machiw E, and Daus W: Early Vistica BP, and Chader GJ: Uveoretinitis and pinealitis in- stage of human sympathetic ophthalmia, histologic and immu- duced by immunization with interphotoreceptor retinoid- nopathologic findings.Arc h Ophthalmol 102:1353, 1984. binding protein. Invest Ophthalmol Vis Sci 27:1296, 1986. 12. Kaplan HJ and Waldrep JC: Immunologic insights into uveitis 28. Chan CC, Hooks JJ, Nussenblatt RB, and Detrick B: Expres- and retinitis. Ophthalmology 91:655, 1984. sion of la antigen on retinal pigment epithelium in experimen- 13. Kaplan HJ, Waldrep JC, Chan WC, Nicholoson JKA, and tal autoimmune uveoretinitis. Curr Eye Res 5:325, 1986. Wright JD: Human sympathetic ophthalmia: Immunologic 29. Fujikawa LS, Chan CC, McAllister C, Gery I, Hooks JJ, De- analysis of the vitreous and uvea. Arch Ophthalmol 104:240, trick B, and Nussenblatt RB: Retinal vascular endothelium 1986. expresses fibronectin and class II histocompatibility complex 14. Chan CC, Nussenblatt RB, Fujikawa LS, Palestine AG, Ste- antigens in experimental autoimmune uveitis. Cell Immunol vens G, Parver LM, Luckenback MW, and Kuwabara T: Sym- 98:139, 1987. pathetic ophthalmia: Immunopathologic findings. Ophthal- 30. McCarron RM, Kempsks O, Spatz M, and McFarlin DE: Pre- mology 93:690, 1986. sentation of myelin-basic proteins by murine cerebral vascular 15. Detrick B, Rodrigues MM, Chan CC, Palestine AG, Nussen- endothelial cells. J Immunol 134:3100, 1985. blatt RB, and Hooks JJ: Expression of HLA-DR antigen on 31. Traugott N, Scheinberg LC, and Reine CS: On the presence of retinal pigment epithelial cells in retinitis pigmentosa. Am J la positive endothelial cells and astrocytes in multiple sclerosis Ophthalmol 101:584, 1986. lesions and its relevance to antigen presentation. J Neuroim- 16. Chan CC, Detrick B, Nussenblatt RB, Palestine AG, Fujikawa munol 8:1, 1985. L, and Hooks JJ: Expression of HLA-DR antigens on RPE 32. Schneeberger EE, DeFarrari M, Skoskiewicz MJ, Russell PS, cells from patients with uveitis. Arch Ophthalmol 104:725, and Colvin RB: Induction of MHC-determined antigens in the 1986. lung by interferon-gamma. Lab Invest 55:138, 1986. 17. Lampson LA and Levy R: Two populations of la-like mole- 33. Skoskiewicz MJ, Colvin RB, Schneeberger EE, and Russell PS: cules on a human B cell line. J Immunol 125:293, 1980. Widespread and selective induction of major histocompatibil- 18. Le J, Barrowclough BS, and Vilcek J: Monoclonal antibodies ity complex-determined antigens in vivo by interferon- to human immune interferon and their application for affinity gamma. J Exp Med 162:1645, 1985. chromatography. J Immunol Methods 69:61, 1984. 34. Kasahara T, Hooks JJ, Dougherty SF, and Oppenheim J: In- 19. Engleman EG, Benike CJ, Grumet FC, and Evans RL: Acti- terleukin 2 mediated immune interferon (IFN-7) production vation of human T lymphocyte subsets: Helper and suppers- by human T cells and T cell subsets. J Immunol 130:1784, sor/cytotoxic T cells recognize and respond to distinct histo- 1983. compatibility antigens. J Immunol 127:2124, 1981. 35. Husby G and Williams RC: Immunohistochemical studies of 20. Breard J, Reinherz EL, Kung PC, Goldstein G, and Schloss- interleukin-2 and interferon-gamma in rheumatoid arthritis. man SF: A monoclonal antibody reactive with human periph- Arthritis Rheum 28:17, 1985. eral blood monocytes. J Immunol 124:1943, 1980. 36. Husby G, Williams RC, Ramirez F, and Tung KSK: Absence of interferons alpha and gamma in renal lesions of systemic 21. Shaw S: Characterization of human leukocyte differentiation lupus erythematosus and membranous glomerulonephritis. antigens. Immunol Today 8:1, 1987. Clin Immunol Immunopathol 39:68, 1986. 22. Hsu SM, Raine L, and Fangeor H: The use of avidin-biotin- 37. Isenberg DA, Rowe D, Shearer M, Novick D, and Beverley peroxidase complex (ABC) in immunoperoxidase technique: PCL: Localization of interferons and interleukin-2 in poly- A comparison between ABC and unlabeled antibody (PAP) myositis and muscular dystrophy. Clin Exp Immunol 63:450, procedure. J Histochem Cytochem 29:577, 1982. 1986. 23. Detrick-Hooks B, Borsos T, and Rapp HJ: Quantitative com- 38. Fox RI, Theofilopoulos AN, and Altman A: Production of parison of techniques used to measure complement-mediated interleukin-2 by salivary gland lymphocytes in Sjogren's syn- cytotoxicity of nucleated cells. J Immunol 114:287, 1975. drome: Detection of reactive cells by using antibody directed to 24. Hooks JJ, Moutsopoulos HM, Geis S, Stahl N, Decker J, and synthetic peptides of IL-2. J Immunol 135:3109, 1985.

Downloaded from iovs.arvojournals.org on 09/30/2021