TISSUE, TEAR, AND SERUM IgE CONCENTRATIONS IN VERNAL CONJUNCTIVITIS M A T H E A R. ALLANSMITH,

M.D.

Boston, Massachusetts GARY S. H A H N ,

B.S.

Stanford, California AND M E R E D I T H A. S I M O N ,

M.S.

Boston, Massachusetts

We postulated that perhaps vernal con­ junctivitis had as part of its mechanism a localized hyperplasia of IgE plasma cells in the cobblestone excrescences; and also that perhaps locally produced IgE raised the tear IgE level. To evaluate these pos­ tulations, tarsal conjunctiva from patients with vernal conjunctivitis was stained for all immunoglobulins, and IgE was meas­ ured in tears and serum of patients and controls. M A T E R I A L AND M E T H O D S

Subjects—Eleven patients with vernal conjunctivitis were selected for the tearserum portion of the study on the basis of availability. All had severe itching, tear­ ing, disease for more than two seasons, typical cobblestone excrescences over the entire upper tarsal conjunctiva (Fig. 1), and many eosinophils in scrapings of the conjunctiva. Ten patients (seven males and three females) were aged 6 to 33 years From the Department of Cornea Research, Eye Research Institute of Retina Foundation (Dr. Allansmith and Ms. Simon), and the Department of Oph­ thalmology, Harvard Medical School (Dr. Allansmith), Boston, Massachusetts arid the Division of Ophthalmology, Stanford University School of Medicine, Stanford, California (Mr. Hahn). This study was supported in part by training grant EY000208 and Research grant EY-00043 from the Na­ tional Eye Institute, by National Institutes of Health grant EY-00445 and Research Career Developmen­ tal Award EY-11682 (Dr. Allansmith), and by the Massachusetts Lions Eye Research Fund, Inc. Reprint requests to M. R. Allansmith, M.D., 20 Staniford St., Boston, MA 02114.

with an average age of 14 years. The last patient was a 75-year-old man with clini­ cally typical disease including many eo­ sinophils in the conjunctival scraping. Four patients had tissue removed from the upper tarsal conjunctiva. One of these, a 23-year-old man, was also in the tear-serum group. Two other males and one female, aged 8 to 33, comprised the group. All had at least moderately severe disease. Ten normal subjects, six females and four males, served as controls for the tear-serum group and were aged 15 to 53 years. None had ocular disease. Three had mild hay fever with no ocular symptoms. Upper tarsal tissue from two cadavers served as controls for the tissue group. Both subjects had died of cancers not involving the eyes or orbits at 31 and 53 years of age.

Fig. 1 (Allansmith, Hahn, and Simon). Clinical picture of vernal conjunctivitis. The upper palpebral conjunctiva has many giant papillary excrescences resembling cobblestones (arrow).

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VERNAL CONJUNCTIVITIS

Tissue — Cobblestone excrescences were shaved from the upper tarsal plates under topical anesthesia. Tissues were fixed for at least four hours in a freshly prepared mixture of 19 parts absolute ethanol and 1 part glacial acetic acid. Fixed specimens were embedded in de­ gassed paraffin, cut at 4 μ, and stained by the immunofluorescence method as previ­ ously described for IgG, IgA, IgM, IgD, and IgE with appropriate controls. 1 Tears and serum—Paired samples were obtained. Blood was taken from an arm vein, allowed to clot, and serum was removed. Tears were collected by posi­ tioning a chip of Week cell sponge about 1 x 1 x 3 mm between the upper and lower lacrimal puncta. The sponge ex­ panded with accumulated tears. The tears were extracted by recompressing the satu­ rated sponges into the bottom of a small self-capping microcentrifuge tube with a metal probe. Approximately six soaked sponges yielded several hundred microliters of tears that were stored at —5°C until analyzed. Serum was collected from venous blood. Serum and tear IgE concentrations were determined using the radioimmunosorbent technique (Phadebas IgE test). The mean error of the technique, express­ ed as the standard deviation of 44 dupli­ cated assays, was 15%. The value used for the amount of IgE corresponding to one unit was taken to be 2.39 ng/ml. This value was derived from the numerical average of the value 2.35 ng/unit, ob­ tained from 20 analyses of World Health Organization IgE reference standard 68-341 containing 10,000 units/ml as measured by Yunginger and Gleich 2 and the value 2.42 ng/unit measured by Bazaral and Hamburger. 3 All mean values, correlation coefficients, and statistical comparisons were calculated with the amounts transformed to their common logarithm. This type of transformation, which has precedence in biological sys­

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tems, 4 resulted in a more normal distribu­ tion and a relatively linear relationship between serum and tear IgE concentra­ tions. RESULTS

Plasma cells were seen in all tissues stained with hematoxylin and eosin. Plas­ ma cells of three types were seen by immunofluorescence stain in tissues from two of the four patients. There was no predominance of IgE staining cells. Rath­ er, cells staining for IgA, IgD, and IgE were found (Figs. 2 and 3) with plasma IgA cells predominating over others in a ratio of approximately 4:1:2 for IgA: IgD:IgE. No staining plasma cells were seen in the upper tarsal conjuhctival tissues from the two normal subjects. Control staining was negative. 1 In the control-stained slides, the many eosinophils were easily visible (Fig. 4). Abundant extracellular immunoglobulin was present in both groups (Fig. 5). This was thought to be extravascular serum protein as is present in the normal conjunctiva. 1 More IgE seemed to be in the extracellular stroma of the tissues

Fig. 2 (Allansmith, Hahn, and Simon). Upper tarsal cobblestone tissue stained for IgE. Arrow indiates plasma cell containing IgE (x250).

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AMERICAN JOURNAL OF OPHTHALMOLOGY

APRIL, 1976

Fig. 3 (Allansmith, Hahn, and Simon). Tissue stained for IgA. Arrow in­ dicates plasma cells con­ taining IgA (x250).

from the patients than of the normal sub­ jects. The mean serum IgE concentration of the patient group was 1,031 ng/ml with a 95% confidence interval of 18 to 59,346

Fig. 4 (Allansmith, Hahn, and Simon). Tissue from patient with control stain shows no specific fluorescence. Many autofluorescent eosinophils are seen (arrow) (x250).

ng/ml (Table 1). The control group had a mean serum IgE concentration of 201 ng/ml with a 95% confidence interval of 52 to 781 ng/ml (Table 2). Student's i-test revealed a significant difference between the means of the two groups (P

Tissue, tear, and serum IgE concentrations in vernal conjunctivitis.

Abnormal upper tarsal tissue from four patients with vernal conjunctivitis stained immunofluorescently contained abundant IgA-, IgD-, and IgE-forming ...
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