Int. |. Exp. Path. (1990), 7I, 8I5-821
Interleukin-ii and -2 in sera of patients with chronic hepatitis (type B) Tsuneo Ozeki*, Kenichi Imanishi,t Hideaki Unoki*, Keisuke Funakoshit and
Yoshiyasu Kiya§
*Third Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health, Kitakyushu; tDepartment of Pathology, Kyushu Dental College; tDepartment of Bacteriology, Tokyo Women's Medical College, and §Clinical Research Center, Dokukyo University School of Medicine, Japan Received for publication I2 June I990 Accepted for publication i8 July 1990
Summary. IL-if? and 1L-2 values in serum from 28 patients with chronic hepatitis B diagnosed by liver biopsy and 23 healthy controls were measured by the radioimmunoassay (RIA) method. Though both values in patients seropositive for anti-HBe were slightly higher than those in patients seropositive for HBeAg, the differences between the two groups were not statistically significant. In contrast, the relationship between IL-ifA and 1L-2 in patients with chronic hepatitis was statistically significant (P < o.0oo I). In general, high values of serum ILifP and I1-2 were seen in patients with chronic persistent hepatitis and low values of serum ELif? and IL-2 were seen in patients with chronic active hepatitis (severe) and in some of these with chronic persistent hepatitis. Serum IL-i# and 11-2 values of all patients with chronic hepatitis were higher than in healthy controls (P< o.ooi). Serum IL-if? and IL-2 in chronic hepatitis (B) are important indicators of the grade of the inflammatory state of the liver. Keywords: interleukin-i and -2, chronic hepatitis B, liver. It is thought that hepatitis B results from an immunological reaction between HBV associated antigen and T cells (Thomas et al. I98I; Eddleston et al. I974). Interleukin-i (IL-i) is related to the first step in the immunological reaction and is secreted by stimulated macrophages (Bayne et al. I986). Interleukin-2 (IL-2) is then secreted by T cells which are stimulated by IL-i (Gillis et al. I983; Farrar et al. I980). IL-2 has the important function of inducing the proliferation and differentiation of Tc cells (Gillis & Smith 1977). IL-i and IL-2 in serum have been measured by biological assay (Law-
rence et al. I979; Yoshioka et al. I984). However, the value obtained by biological assay is not absolute because it is influenced by the biological activity of the target cell used. Furthermore, for biological assay, a large volume of blood is needed to collect a sufficient fraction of lymphocyte or mononuclear cells. An alternative method, the RIA assay which uses monoclonal antibodies, has no such problem. We have used RIA to investigate the relationship between serum IL-i and EL-2, seroconversion in HBeAg and anti-HBe, and the grade of chronic hepatitis (B). Evidence is reported
Correspondence: Dr Tsuneo Ozeki, i-i Maruomachi, Yahata Nishi-ku, Orio Post Office, PO Box I2, Kitakyushu City, Japan. 8I5
Tsuneo Ozeki et al. which suggests that IL-i and IL-2 are impor- incubation tube and the kit procedure subsetant indicators of the grade of the inflamma- quently followed. The method employed by tory state of chronic hepatitis (B). these kits is based upon the principle of 8I6
Materials and methods Liver biopsy specimens from 28 Japanese hospitalized patients with chronic hepatitis (B) were studied. The patients were inpatients in the college hospital (the 3rd Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health). Serum HBsAg was measured by reversed passive haemagglutination (R-PHA); antibody to HBsAg (antiHBs) was measured by passive haemagglutination (PHA) or RIA; antibody to HBcAg (anti-HBc) was measured by immune adherence haemagglutination (IAHA) or RIA; and serum HBeAg and anti-HBe were measured by RIA. There were 2 5 men and three women with ages ranging between 2I and 53 years (average 39 years).
Histological diagnosis and findings Histological diagnosis of chronic hepatitis by HE and Azan staining were made according to the International Group Classification (International Group 1977). The following histological findings were recorded; (i) piecemeal necrosis (PN), (ii) spotty necrosis (SN), (iii) portal hepatitis (PH), (iv) P-P connection (P-P), (v) hepatocyte swelling (HS). Measurement of IL-i and IL-2 Intravenous blood samples were taken from patients and healthy controls and the serum was separated and stored at - 200C until analysed. Serum IL-i/ from 22 patients and 23 healthy controls was measured with the IL-if RIA Kit (Cistron Ltd, USA) and serum IL-i of six patients was measured by IL-i RIA Kit (Medgenix Ltd, Belgium). Serum IL-2 of all patients and healthy controls was measured with the IL-2 RIA Kit (Medgenix Ltd, USA). O.I ml of each collected sample was transferred to a 12 x 75 mm polystyrene
competition between a labelled antigen and an unlabelled antigen for specific monoclonal antibodies.
Results Tables i and 2 show results from patients with chronic hepatitis (positive for HBeAg and positive for anti-HBe, respectively). Patients with very high values of IL-ifl had only a low grade ofchronic hepatitis (chronic persistent hepatitis). By contrast, those with very low values of IL-i,f and IL-2 had high grades of chronic hepatitis (chronic active hepatitis (CAH) severe) and chronic persistent hepatitis (CPH). Severe piecemeal necrosis was also seen in patients with low values of IL-i/p, whereas spotty necrosis (chronic lobular hepatitis) was found in patients with relatively low IL-2 values. The other histological findings of portal hepatitis, P-P connection, and hepatocyte swelling did not appear to relate closely to IL-i or IL-2 values. Table 3 shows IL-if and IL-2 values in patients with chronic hepatitis (seropositive for HBeAg, and for anti-HBeAg, respectively). There were somewhat higher values of the IL-i,B and IL-2 value in patients seropositive for anti-HBe than in patients positive for anti-HBeAg. However, the differences were not statistically significant. The difference between both groups (seropositive for HBeAg (+) and anti-HBe (+)) and healthy controls was statistically significant (P
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Interleukin-ii and -2 in sera of patients with chronic hepatitis (type B) Tsuneo Ozeki*, Kenichi Imanishi,t Hideaki Unoki*, Keisuke Funakoshit and
Yoshiyasu Kiya§
*Third Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health, Kitakyushu; tDepartment of Pathology, Kyushu Dental College; tDepartment of Bacteriology, Tokyo Women's Medical College, and §Clinical Research Center, Dokukyo University School of Medicine, Japan Received for publication I2 June I990 Accepted for publication i8 July 1990
Summary. IL-if? and 1L-2 values in serum from 28 patients with chronic hepatitis B diagnosed by liver biopsy and 23 healthy controls were measured by the radioimmunoassay (RIA) method. Though both values in patients seropositive for anti-HBe were slightly higher than those in patients seropositive for HBeAg, the differences between the two groups were not statistically significant. In contrast, the relationship between IL-ifA and 1L-2 in patients with chronic hepatitis was statistically significant (P < o.0oo I). In general, high values of serum ILifP and I1-2 were seen in patients with chronic persistent hepatitis and low values of serum ELif? and IL-2 were seen in patients with chronic active hepatitis (severe) and in some of these with chronic persistent hepatitis. Serum IL-i# and 11-2 values of all patients with chronic hepatitis were higher than in healthy controls (P< o.ooi). Serum IL-if? and IL-2 in chronic hepatitis (B) are important indicators of the grade of the inflammatory state of the liver. Keywords: interleukin-i and -2, chronic hepatitis B, liver. It is thought that hepatitis B results from an immunological reaction between HBV associated antigen and T cells (Thomas et al. I98I; Eddleston et al. I974). Interleukin-i (IL-i) is related to the first step in the immunological reaction and is secreted by stimulated macrophages (Bayne et al. I986). Interleukin-2 (IL-2) is then secreted by T cells which are stimulated by IL-i (Gillis et al. I983; Farrar et al. I980). IL-2 has the important function of inducing the proliferation and differentiation of Tc cells (Gillis & Smith 1977). IL-i and IL-2 in serum have been measured by biological assay (Law-
rence et al. I979; Yoshioka et al. I984). However, the value obtained by biological assay is not absolute because it is influenced by the biological activity of the target cell used. Furthermore, for biological assay, a large volume of blood is needed to collect a sufficient fraction of lymphocyte or mononuclear cells. An alternative method, the RIA assay which uses monoclonal antibodies, has no such problem. We have used RIA to investigate the relationship between serum IL-i and EL-2, seroconversion in HBeAg and anti-HBe, and the grade of chronic hepatitis (B). Evidence is reported
Correspondence: Dr Tsuneo Ozeki, i-i Maruomachi, Yahata Nishi-ku, Orio Post Office, PO Box I2, Kitakyushu City, Japan. 8I5
Tsuneo Ozeki et al. which suggests that IL-i and IL-2 are impor- incubation tube and the kit procedure subsetant indicators of the grade of the inflamma- quently followed. The method employed by tory state of chronic hepatitis (B). these kits is based upon the principle of 8I6
Materials and methods Liver biopsy specimens from 28 Japanese hospitalized patients with chronic hepatitis (B) were studied. The patients were inpatients in the college hospital (the 3rd Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health). Serum HBsAg was measured by reversed passive haemagglutination (R-PHA); antibody to HBsAg (antiHBs) was measured by passive haemagglutination (PHA) or RIA; antibody to HBcAg (anti-HBc) was measured by immune adherence haemagglutination (IAHA) or RIA; and serum HBeAg and anti-HBe were measured by RIA. There were 2 5 men and three women with ages ranging between 2I and 53 years (average 39 years).
Histological diagnosis and findings Histological diagnosis of chronic hepatitis by HE and Azan staining were made according to the International Group Classification (International Group 1977). The following histological findings were recorded; (i) piecemeal necrosis (PN), (ii) spotty necrosis (SN), (iii) portal hepatitis (PH), (iv) P-P connection (P-P), (v) hepatocyte swelling (HS). Measurement of IL-i and IL-2 Intravenous blood samples were taken from patients and healthy controls and the serum was separated and stored at - 200C until analysed. Serum IL-i/ from 22 patients and 23 healthy controls was measured with the IL-if RIA Kit (Cistron Ltd, USA) and serum IL-i of six patients was measured by IL-i RIA Kit (Medgenix Ltd, Belgium). Serum IL-2 of all patients and healthy controls was measured with the IL-2 RIA Kit (Medgenix Ltd, USA). O.I ml of each collected sample was transferred to a 12 x 75 mm polystyrene
competition between a labelled antigen and an unlabelled antigen for specific monoclonal antibodies.
Results Tables i and 2 show results from patients with chronic hepatitis (positive for HBeAg and positive for anti-HBe, respectively). Patients with very high values of IL-ifl had only a low grade ofchronic hepatitis (chronic persistent hepatitis). By contrast, those with very low values of IL-i,f and IL-2 had high grades of chronic hepatitis (chronic active hepatitis (CAH) severe) and chronic persistent hepatitis (CPH). Severe piecemeal necrosis was also seen in patients with low values of IL-i/p, whereas spotty necrosis (chronic lobular hepatitis) was found in patients with relatively low IL-2 values. The other histological findings of portal hepatitis, P-P connection, and hepatocyte swelling did not appear to relate closely to IL-i or IL-2 values. Table 3 shows IL-if and IL-2 values in patients with chronic hepatitis (seropositive for HBeAg, and for anti-HBeAg, respectively). There were somewhat higher values of the IL-i,B and IL-2 value in patients seropositive for anti-HBe than in patients positive for anti-HBeAg. However, the differences were not statistically significant. The difference between both groups (seropositive for HBeAg (+) and anti-HBe (+)) and healthy controls was statistically significant (P
++++++
+ ++ ++ + ++~~~
+U co
O&
O
SH
v+ >
>
o
Z
-H 0
H
++
tNI)H0
*-
LA rAei rA i 4 -i rl ~ci16 l + ++ ++ ++ ++ ++ + ++
CZ
0~~~~~~~~~~~~~~ ¢ q 00 N ar) e t 0- 00me&x8laFTYjV$ stX;.yr,2i*a>-
*R}(t,2-|;A*X>.^M.12)§kf~|e;i3\:1zib,fj-Sb..~J+ r.l } i ' ~ ~ ~ ~ ~ ~ .1 Tsuneo Ozeki et a]. M .
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