Evaluation of T and B lymphocytes in liver infiltrates of patients with chronic active hepatitis.

The proportions of T and B lymphocytes in the liver infiltrates of 23 patients with chronic active hepatitis have been determined. The results were compared with the values obtained from peripheral blood and with the presence of HB virus markers and alpha-fetoprotein in liver tissue. A group of patients with chronic liver disease other than chronic active hepatitis were studied as controls. In chronic active hepatitis the percentage of hepatic T cells was 49 +/- 8 SD (control patients 61 +/- 8) (P less than 0.01), whereas the percentage of B cells was 40 +/- 10 (control patients 18 +/- 8) (P less than 0.01). No correlation was observed between hepatic T and B cells and the presence of HB virus. The numbers of T cells in liver tissue was significantly higher, the numbers of B cells lower, in patients whose biopsies were positive for alpha-fetoprotein than in those whose biopsies were negative. In peripheral blood, only the patients with chronic active hepatitis and established cirrhosis presented lower absolute values of T cells, whereas surface immunoglobulin-positive lymphocytes were within the normal range.     

Spontaneous and interferon-induced activity of natural killer cells in patients with liver cirrhosis and chronic active hepatitis

We investigated spontaneous and gamma-interferon stimulated natural killer (NK) cell activity and antibody-dependent cellular cytotoxicity (ADCC) in patients with chronic liver disease: A significantly reduced NK activity was found in alcoholic liver cirrhosis (LZ) and in chronic active hepatitis (CAH), as compared to healthy control individuals (p less than 0.001 and p less than 0.005, respectively). Patients with primary biliary cirrhosis (PBC) or with alcoholic non-cirrhotic liver disease had normal NK activities. Furthermore, ADCC was found to be significantly reduced in patients with CAH (p less than 0.05). The addition of gamma-interferon to peripheral blood mononuclear cells in vitro induced a similar increase in NK cell activity in all studied groups of patients. The reduced NK cell activity found in patients with LZ and CAH could constitute an important mechanism in the pathogenesis and contribute to the high incidence of virus-associated hepatic malignancies in these disorders.     

Cytotoxic activity of lymphocytes from patients with Hodgkin's disease.

Blood lymphocytes from twenty-three untreated patients with Hodgkin's disease and twelve healthy controls were studied for their ability to lyse tissue culture cells (Chang cells) labelled with [51Cr]chromate. Lysis induced by patients' lymphocytes in the presence of PHA or rabbit IgG antibodies to Chang cells (ACS) was impaired in some, and higher than normal in others. ACS-induced lysis showed some corlation with the content of lymphocytes carrying receptors for human complement (CRL) in the effectory population. No correlation with immunoglobulin-bearing cells was noted. PHA-induced cytoxicity did not correlate with lymphocyte subpopulations. The observations are consistent with the assumption that effector cells of antibody-induced lymphocyte-mediated cytotoxicity (K cells) may be present among CRL. The K-cell activity and PHA cytotoxicity by lymphocytes tended to decrease at high age, but impaired cytotoxity was also noted in young patients. A preliminary follow-up of patients 1-2 years after the beginning of treatment revealed almost abolished K-cell activity in four patients who died 6--13 months after testing. Patients in incomplete remission or with relapse after treatment had lower mean K-cell activity than those in complete remission. A similar, but less pronounced tendency was found for PHA-induced cytotoxicity. A prognostic role of impaired K-cell activity in Hodgkin's disease is suggested from these data, but requires confirmation in a larger clinical follow-up.     

Cytotoxic activity of antibodies to a collagen-like synthetic polytripeptide on cells in tissue culture

Antibodies to the ordered collagen-like synthetic polymer (Pro-Gly-Pro)_n, as well as antiserum to rat tail collagen, have a similar specific cytotoxic activity on primary cultures of rat muscle cells and on rat embryo fibroblasts. This cytotoxic effect is complement-dependent, and could be observed both in monolayers and in cell suspensions.     

Selective sensitization of peripheral blood T lymphocytes to hepatitis B core antigen in patients with chronic active hepatitis type B.

The proliferative response of peripheral blood T cells to hepatitis B surface antigen (HBsAg), pre-S antigen and hepatitis B core antigen (HBcAg) has been studied in 20 patients with hepatitis B virus induced chronic active hepatitis (CAH) and in 12 control subjects. Eleven of the 20 CAH patients showed a significant T lymphocyte proliferative response to HBcAg, whereas no proliferation was detectable in response to envelope antigens (HBs and pre-S) in any patient. T cell subset fractionation revealed that HBcAg specific proliferation was limited to the CD4+ (helper/inducer) population. CD8+ (suppressor/cytotoxic) T cells were unresponsive to HBcAg and did not suppress the proliferative response of autologous CD4+ cells. The HBcAg specific T cell proliferative response did not correlate with serum anti-core antibody titres or with biochemical evidence of liver disease. The present results show the selective presence of HBcAg-sensitized T cells in the peripheral blood of patients with HBsAg positive CAH and suggest that the peripheral lymphoid compartment may not reflect immunopathogenetically important cellular events operative at the site of tissue injury.     

Telomerase activity of peripheral blood lymphocytes in patients with chronic hepatitis B

Chronic hepatitis B is the immunocompromising condition. The decrease of lymphocyte telomerase is linked to immunosenescence in hosts. To know whether telomerase activity of lymphocytes is involved in immunopathogenesis in patients with chronic hepatitis B, telomerase activity of peripheral lymphocytes was determined in such patients. The results showed that telomerase activity in resting peripheral lymphocytes of healthy subjects was detectable at low level, and obviously increased (P<0.001) after stimulation in vitro with phytohaemagglutinin (PHA). Telomerase activity of lymphocytes decreased with age in both groups with or without PHA stimulation. Telomerase activity of resting lymphocytes in patients with chronic hepatitis B was also observed at detectable level and markedly upregulated after PHA stimulation. The decreased telomerase activity of resting lymphocytes was found in patients with chronic hepatitis B (n=14, 0.32+/-0.27) compared to that in healthy subjects (n=17, 0. 52+/-0.28; P<0.05). However, there was no difference present between these two groups in telomerase activity of activated lymphocytes with PHA. In addition, no effect of recombinant human interleukin-12 (rhIL-12) on telomerase expression was observed in either the patient group or the healthy group. We concluded that the decreased telomerase activity of lymphocytes in chronic hepatitis B patients is present, which may be partly responsible for immunosuppressive condition in such patients.     

T and B lymphocytes in patients with chronic active hepatitis (CAH).

Absolute numbers of T and B lymphocytes as well as active E rosette-forming cells were measured in twenty-seven patients with chronic active hepatitis (CAH), and in thirty control patients. In patients with CAH without cirrhosis, active E rosette-forming cells (a subpopulation of T lymphocytes considered to be actively involved in cell-mediated immune reactions) as well as lymphocytes with surface markers for IgA, IgM and IgG were increased. In patients with CAH and cirrhosis, total T lymphocytes were decreased. These results emphasize the significance of lymphocytes in CAH, and suggest the importance of monitoring T- and B-cell populations in patients with this disease.     

Studies of lymphocyte subpopulations in the liver tissue and blood of patients with chronic active hepatitis (CAH)

Monoclonal antibodies to antigens on the surfaces of mononuclear cells (MNC) were used to characterize lymphocyte subpopulations infiltrating portal areas and parenchyma of livers in 31 patients with chronic active hepatitis (CAH). The distribution and numbers of infiltrating lymphocytes were determined in serial sections immunostained by the avidin-biotin-peroxidase complex method. T lymphocytes were the major component of inflammatory cells in the portal tracts. In the peripheral blood and portal areas, T helper-inducer (T4+) cells were the more numerous subpopulation. However, the hepatic lobules and areas of "piecemeal" necrosis always contained more T suppressor-cytotoxic (T8+) cells. The latter were demonstrated in contact with HBsAg-containing hepatocytes in tissues of patients with HBsAg-positive CAH. The mean numbers of T lymphocytes infiltrating the portal and periportal areas of livers from patients with HBsAg-negative and HBsAg-positive CAH were not different. Large numbers of B cells forming distinct follicles were seen in tissues from patients with HBsAg-positive CAH. The presence of increased numbers of portal T and B lymphocytes correlated with progressive liver damage as observed in two patients studied at yearly intervals.     

Detection of lymphocytes with increased DNA-Synthesis in the peripheral blood of patients with active chronic hepatitis

Journal of Molecular Medicine - The first observation of “in vivo activated lymphocytes” [IVAL] in cases of active chronic hepatitis is reported. Lymphocytes with an elevated metabolism...     

Disordered immunoregulatory functions in patients with chronic active hepatitis.

Peripheral T lymphocytes from patients with chronic active hepatitis (CAH) showed a significantly decreased suppressor cell (or increased helper cell) effect on differentiation of allogenic B cells to Ig-producing cells (Ig-PC). Spontaneous helper cell activity as measured after irradiation of T cells appeared normal, while Concanavalin A (Con A)-induced suppressor cell activity was significantly reduced. Some patients of chronic persistent hepatitis (CPH) also showed mild depression of Con A-induced suppressor cell activity. Poor suppressor cell activity in CAH was much more often seen in HBsAg negative, autoantibody positive patients than in HBsAg positive autoantibody negative ones. Autologous mixed lymphocyte reaction (AMLR) was significantly decreased in patients with CAH. Also, a serum factor(s) that decreased Con A-induced suppressor cell function of healthy subjects could be demonstrated in some patients with CAH and CPH. Our results suggest that altered immune responses observed in CAH may be due to defective suppressor cell function, partly attributable to serum factor(s).     

Assessment of mitochondrial function in cells grown in tissue culture.

To assess mitochondrial function (pyruvate dehydrogenase [PDH] activity), cells were grown in the appropriate media to confluence, rinsed and incubated in glucose free media containing 25 microM L-lactate and [1-14C]-D,L-lactate. Lactate oxidation was measured as the amount of lactate oxidized in nmol of 14CO2 generated per mg of protein per minute. Basal activity varied with cell number and the cell type studied: fibroblast 2.26 +/- 0.01; Chinese hamster ovary (CHO) 42 +/- 0.4; BC3H-1 52 +/- 2.1 nmol per mg per minute. The CHO cells screened for PDH activity decreased their dependence on lactate as a substrate in the presence of 5mM glucose by 60 percent. Increasing the cold lactate concentration diluted the labelled lactate available for pyruvate oxidation in a dose dependent manner. The mitochondrial inhibitor rotenone (25 microM) decreased assay activity by > 75 percent in CHO and BC3H-1 cells. The lactate oxidation assay was shown to be sensitive enough to measure insulin stimulation of PDH in a dose dependent manner with maximum activity occurring at concentrations between 1 microU per ml and 100 microU per ml.     

Immunological aspects in patients with chronic active hepatitis--cellular immune responses

We studied host immune parameters which might be related to the activity and the pathogenetic mechanism of chronic active hepatitis. The subjects consisted of 45 cases with hepatitis B virus surface antigen (HBsAg)-positive chronic active hepatitis (CAH), 44 HBsAg-negative CAH, 22 with inactive chronic hepatitis, and 45 cases of normal persons, hepatitis B virus (HBV) carriers, or the patients with acute myocardial infarction. The in vitro assay for the in vivo activated lymphocytes was performed by measuring spontaneous thymidine uptake (SLT) of lymphocytes isolated from peripheral blood. SLT was significantly (p < 0.001) elevated in cases with HBsAg-positive (1227 +/- 806 cpm) and-negative CAH (1017 +/- 559 cpm) compared to the patients with inactive chronic hepatitis (347 +/- 79 cpm) and to the control group (320 +/- 106 cpm). SLT values observed in 7 cases with active disease (group I and II), in which remission and relapsing phase could be assessable, were elevated from 648 +/- 121 cpm in remission phase to 1548 +/- 606 cpm one to two weeks before the appearance of biochemical evidence (SGPT) of relapse. This pattern of SLT elevation, however, was not observed in patients with inactive hepatitis. Neither the abnormal distribution of T-cell subsets nor the presence of conventional HBV markers were related to the elevated SLT value. Our findings may therefore indicate that SLT might be useful in assessment of the disease activity in patients with CAH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Studies on experimental chronic active hepatitis in the rabbit. II. Immunological findings

The role played by humoral and cellular immune response to liver antigens in the pathogenesis of experimental chronic active hepatitis (CAH) was studied in rabbits which had been immunized with the two human liver-specific proteins LSP and LP2, with LSP alone, or with an extract of human skeletal muscle. Rabbits immunized with LSP, alone or with LP2, developed skin test reactivity and circulating antibody to homologous LSP; liver biopsy revealed immunoglobulin bound to the hepatocyte cell surface. It has been suggested that cellular immunity to homologous LSP or alternatively, antibody to an antigenic determinant shared by human and rabbit LSP, may play a role in the pathogenesis of experimental CAH, but both abnormalities were present in two rabbits which did not develop CAH despite observation for at least 18 months. Five normal rabbits given an intravenous injection of serum pooled from rabbits with CAH did not develop significant hepatic lesions. Immunity to homologous LSP or other hepatocyte cell surface antigens could not be detected in any rabbit which had been immunized with skeletal muscle, and hepatocytes from these rabbits did not have immunoglobulin on their cell surface. The pathogenesis of CAH in these animals is obscure. These findings suggest that mechanisms other than an auto-immune response to LSP play an important role in the pathogenesis of experimental chronic active hepatitis.     

Immunological studies in children with acute viral hepatitis.

Sera from 116 consecutive unselected cases of sporadic acute viral hepatitis in children were examined for hepatitis B antigen (HBAg), smooth-muscle autoantibodies (SMA), other autoantibodies and immunoglobulins, and skin tests were performed with dinitrochlorobenzene (DNCB). HBAg was detected in twenty-one and SMA in ninety-eight out of 116 sera that had been obtained during the 1st or 2nd week from the onset of jaundice. Hepatitis B antigen was present in seventeen out of the eighteen SMA negative patients (94-4%) and in only four out of the ninety-eight SMA-positive patients (4-1%). The presence of SMA was not related to the sex and age of the patients or to the serum bilirubin and transaminase levels. SMA did not persist for more than 6 weeks from the onset of jaundice in most of the cases. In twenty-eight out of forty-one sera which were tested the IgM level was found to be elevated during the acute phase of illness and within normal limits during the recovery stage. A negative correlation between the presence of SMA and the elevated serum IgM level and the presence of HB Ag in the same patients was observed. The DNCB skin test was found to be positive in all fifty-two patients who did not have HBAg in their serum and in twenty out of the twenty-one patients who had circulating HbAg. From these findings there appears to be no gross impairment of cell-mediated immunity in acute viral hepatitis, and hepatitis A is associated with SMA production and an increase in serum IgM levels, when compared to hepatitis associated with HBAg.     

Studies on experimental chronic active hepatitis in the rabbit. II. Immunological findings.

The role played by humoral and cellular immune response to liver antigens in the pathogenesis of experimental chronic active hepatitis (CAH) was studied in rabbits which had been immunized with the two human liver-specific proteins LSP and LP2, with LSP alone, or with an extract of human skeletal muscle. Rabbits immunized with LSP, alone or with LP2, developed skin test reactivity and circulating antibody to homologous LSP; liver biopsy revealed immunoglobulin bound to the hepatocyte cell surface. It has been suggested that cellular immunity to homologous LSP or alternatively, antibody to an antigenic determinant shared by human and rabbit LSP, may play a role in the pathogenesis of experimental CAH, but both abnormalities were present in two rabbits which did not develop CAH despite observation for at least 18 months. Five normal rabbits given an intravenous injection of serum pooled from rabbits with CAH did not develop significant hepatic lesions. Immunity to homologous LSP or other hepatocyte cell surface antigens could not be detected in any rabbit which had been immunized with skeletal muscle, and hepatocytes from these rabbits did not have immunoglobulin on their cell surface. The pathogenesis of CAH in these animals is obscure. These findings suggest that mechanisms other than an auto-immune response to LSP play an important role in the pathogenesis of experimental chronic active hepatitis.     

血清和肝组织TGF-β1与慢性乙型肝炎肝纤维化的关系

目的 探讨血清和肝组织TGF-β1水平与慢性乙肝肝纤维化程度的关系,为肝纤维化诊断提供依据.方法 以肝活检病理诊断区分131例慢性HBV感染者纤维化程度(S0～S4),用ELISA法检测血清TGF-β1水平,免疫组化法检测肝组织TGF-β1表达并半定量.分析血清TGF-β1和肝组织TGF-β1表达与肝纤维化程度的关系.结果 血清和肝组织TGF-β1均与肝纤维化程度具有非常显著性正相关(r分别是0.74和0.89,P＜0.01).血清TGF-β1各组间比较差异有统计学意义(P＜0.01).组问分割比较,S0和S1分别与S4比较差异均有统计学意义(P＜0.005);各组与S0组比较差异均有统计学意义(P＜0.005);S1组和S3组之间比较有统计学意义(P＜0.005).肝组织TGF-β1表达在S3和S4组之间比较差异无统计学意义(P＞0.05),其余组间比较差异均有统计学意义(P＜0.01).血清TGF-β1和肝组织TGF-β1表达具有非常显著性意义相关(r=0.61,P＜0.01).结论 血清TGF-β1和肝组织TGF-β1水平与慢乙肝肝纤维化程度相关,血清TGF-β1有希望成为临床判断轻度或重度肝纤维化的无创伤性诊断指标.