Short Papers
Effect of Antiepileptic Drugs on Serum and Salivary IgA J A. AARLI Si O. TONDER Departments of Neurology and Microbiology and The Broegelmann Research Laboratory tor Microbiology, School of Medicine, University of Bergen, Bergen, Norway
Aarli, J, A. & Tonder, O. Effect of Antiepileptic Drugs on Senun and Salivary IgA. Scand. } . Immunol. 4, 391-396, 1975. IgA, IgG, and IgM concentrations were determined in sera from 100 patients under treatment for epilepsy for at least 6 months. Low IgA was found in 25% of adult sera and in an even larger percentage of sera from children. IgM concentrations were also low, whereas there was no significant difference in the IgG concentrations compared with normal sera. IgA values were normal in untreated patients but fell during treatment with phenytoin, in one patient from 1.5 to 0.2 mg/ml within 2 months. Low or undetectable salivary IgA was 3 common finding, especially in gingival hyperplasia. /. A. Aarli, Department of Neurology, S0I6 HaukeUnd sykehus, Betgen, Norway
Phenytoin and phenobarbital are the most widely used antiepileptic drugs. Among complications attributed to these compounds are reactions from the lymphoid tissue and bloodforming organs. These include benign lymphadenopathy and eosinopliilia as well as throtnbocytopenia, agranulocytosis, allergic reactions, and even malignant lytnphomas. Disturbances of semni immunoglobulins have also been reported, but with conflicting data (7, 12, 14, 15), The present report therefore concerns serum and salivary immunoglobulins in patients with epilepsy and the effect of antiepileptic drugs on immunoglobulin levels.
MATERIALS AND METHODS Sera. Sera were taken from 100 patients with epilepsy treated for at least 6 months at the outpatient clinic. Of these, 73 were adults and 27 children (between 9 and 16 years of age).
In addition, sera from 12 epilepsy patients, all children or young adults who had not taken anticonvulsant drugs, were included. Patients suffering from acute infections of intercurrent nature were excluded from the study. Sera from healthy individuals within the same age groups as the patietits served as controls (normal sera), A standard serum from Behringwerke AG, Marburg/Lahn, West Germany, served as reference serum for all immunoglobulin (Ig) determinations. Rabbit antisera to IgA, IgG, IgM, and to free secretory component (SC) were purchased from Behringwerke. Saliva. Whole saliva was collected at the same time as blood was drawn. It was cleared by centrifugation at 1000 g for 20 min and stored at -20°C until use. The protein concentration was determined by a modified Folin-Ciocalteau method ( I I ) . Ig determinations. Immunodiffusion plates were purchased from Behringwerke. IgA, IgG,
392
/. A. Aarli & O. Tonder
NUMBS! OF RMIENTS 20 T
Fig. 1. Distribution of number of epilepsy patients and healthy controls according to concentration of IgA in their sera. The bases for the various columns are inttn'als of < 0.29 mg/ml. 0.31^-0,59. 0.6lM).89, and so forth.
IgA
• Patients n Controls
15
10
0.3 06
09
15
18
21
33
and TgM were (]uantitated using Tri-Partigen plates. All sera with IgA concentrations below 0.4 mg/ml were also tested using S-Partigen plates. Ig concentrations in saliva were determined using LC-partigen plates and various dilutions of standard serum as reference. The lower level of quantitation wa.s 1 mg/100 ml. Secretory component was demonstrated by double diffusion in agar atid immunoelectrophocesis. All determinations were performed under identical conditions. The values were read with sera or salivas coded. Patient aiid control sera/ salivas were always included on each plate together with standard serum. When more than one sample from one patient was examined, all samples were tested on the same immunodiffusion plate.
n.
36 39 mg/ml
For statistical evaluation of the results, the difference between groups was tested by Wiicoxon's two-sample test (two-tailed). Preparative ultracentrifuf^atiort. Density gradient centrifugation was performed, using a 10%-40'}h sucrose gradient (9). Fourteen successive 0.35-ml fractions numbered from top to bottom were collected.
RESULTS l^ in serum The median of the IgA concentrations wa.s 1.18 mg/ml for the adult epilepsy group and 1.71 mg/ml in the adult controls. The difference is statistically significant (P < 0.005)- Whereas the range of results in the normal group was
Table I. IgA concentrations in sera from 27 children with epilepsy (P) and 21 healthy children (C) Age groups (years) 9-11 P C 12-15 P C
No. of patients with various IgA concentrations (mg/ml)
Effect of Antiepileptic Drugs on Serum and Salivary IgA J A. AARLI Si O. TONDER Departments of Neurology and Microbiology and The Broegelmann Research Laboratory tor Microbiology, School of Medicine, University of Bergen, Bergen, Norway
Aarli, J, A. & Tonder, O. Effect of Antiepileptic Drugs on Senun and Salivary IgA. Scand. } . Immunol. 4, 391-396, 1975. IgA, IgG, and IgM concentrations were determined in sera from 100 patients under treatment for epilepsy for at least 6 months. Low IgA was found in 25% of adult sera and in an even larger percentage of sera from children. IgM concentrations were also low, whereas there was no significant difference in the IgG concentrations compared with normal sera. IgA values were normal in untreated patients but fell during treatment with phenytoin, in one patient from 1.5 to 0.2 mg/ml within 2 months. Low or undetectable salivary IgA was 3 common finding, especially in gingival hyperplasia. /. A. Aarli, Department of Neurology, S0I6 HaukeUnd sykehus, Betgen, Norway
Phenytoin and phenobarbital are the most widely used antiepileptic drugs. Among complications attributed to these compounds are reactions from the lymphoid tissue and bloodforming organs. These include benign lymphadenopathy and eosinopliilia as well as throtnbocytopenia, agranulocytosis, allergic reactions, and even malignant lytnphomas. Disturbances of semni immunoglobulins have also been reported, but with conflicting data (7, 12, 14, 15), The present report therefore concerns serum and salivary immunoglobulins in patients with epilepsy and the effect of antiepileptic drugs on immunoglobulin levels.
MATERIALS AND METHODS Sera. Sera were taken from 100 patients with epilepsy treated for at least 6 months at the outpatient clinic. Of these, 73 were adults and 27 children (between 9 and 16 years of age).
In addition, sera from 12 epilepsy patients, all children or young adults who had not taken anticonvulsant drugs, were included. Patients suffering from acute infections of intercurrent nature were excluded from the study. Sera from healthy individuals within the same age groups as the patietits served as controls (normal sera), A standard serum from Behringwerke AG, Marburg/Lahn, West Germany, served as reference serum for all immunoglobulin (Ig) determinations. Rabbit antisera to IgA, IgG, IgM, and to free secretory component (SC) were purchased from Behringwerke. Saliva. Whole saliva was collected at the same time as blood was drawn. It was cleared by centrifugation at 1000 g for 20 min and stored at -20°C until use. The protein concentration was determined by a modified Folin-Ciocalteau method ( I I ) . Ig determinations. Immunodiffusion plates were purchased from Behringwerke. IgA, IgG,
392
/. A. Aarli & O. Tonder
NUMBS! OF RMIENTS 20 T
Fig. 1. Distribution of number of epilepsy patients and healthy controls according to concentration of IgA in their sera. The bases for the various columns are inttn'als of < 0.29 mg/ml. 0.31^-0,59. 0.6lM).89, and so forth.
IgA
• Patients n Controls
15
10
0.3 06
09
15
18
21
33
and TgM were (]uantitated using Tri-Partigen plates. All sera with IgA concentrations below 0.4 mg/ml were also tested using S-Partigen plates. Ig concentrations in saliva were determined using LC-partigen plates and various dilutions of standard serum as reference. The lower level of quantitation wa.s 1 mg/100 ml. Secretory component was demonstrated by double diffusion in agar atid immunoelectrophocesis. All determinations were performed under identical conditions. The values were read with sera or salivas coded. Patient aiid control sera/ salivas were always included on each plate together with standard serum. When more than one sample from one patient was examined, all samples were tested on the same immunodiffusion plate.
n.
36 39 mg/ml
For statistical evaluation of the results, the difference between groups was tested by Wiicoxon's two-sample test (two-tailed). Preparative ultracentrifuf^atiort. Density gradient centrifugation was performed, using a 10%-40'}h sucrose gradient (9). Fourteen successive 0.35-ml fractions numbered from top to bottom were collected.
RESULTS l^ in serum The median of the IgA concentrations wa.s 1.18 mg/ml for the adult epilepsy group and 1.71 mg/ml in the adult controls. The difference is statistically significant (P < 0.005)- Whereas the range of results in the normal group was
Table I. IgA concentrations in sera from 27 children with epilepsy (P) and 21 healthy children (C) Age groups (years) 9-11 P C 12-15 P C
No. of patients with various IgA concentrations (mg/ml)
