Assessment of skin test with varicella-zoster virus antigen for predicting the risk of herpes zoster.

The Shozu Herpes Zoster (SHEZ) Study was designed to clarify the incidence of and predictive and immunological factors for herpes zoster in a defined community-based Japanese population. As part of this series, a total of 5683 residents aged ≥50 years received a varicella-zoster virus (VZV) skin test with VZV antigen, and 48 h later, the erythema and oedema were assessed by measuring the longest diameter. The diameters of both the erythema and oedema decreased with the increasing age of the subject. Sixty-three subjects contracted herpes zoster within a year after receiving the VZV skin test. Analysis of the herpes zoster incidence rate vs. the skin test reaction revealed that the shorter the diameter of erythema or oedema, the greater the likelihood of herpes zoster. These results demonstrated that the VZV skin test is an excellent surrogate marker for predicting the risk of herpes zoster.

Studies on intrahepatic cholestatic: intrahepatic cholestasis induced in rats by the culture supernatant of activated lymphocytes.

When lymph node lymphocytes from sensitized guinea pigs were stimulated with a specific antigen in vitro, and their culture supernatants injected into the mesenteric vein of rats, a marked reduction of bile flow and bile secretion was seen. Gel filtration of this active principle revealed that the active material has a molecular size similar to that of the macrophage migration inhibitory factor (MIF). Histologically, dilated bile canaliculi with decreased microvilli were observed by electron microscopy in rat liver after injection of culture supernatant. No such change was observed in rats after injection of the supernatant of lymphocyte cultures prepared from non-sensitized guinea pigs. These results strongly suggest that the sensitized lymphocytes produce a factor (or factors) which causes intrahepatic cholestasis when stimulated with a specific antigen.

Immunological studies on chronic active hepatitis: possible involvement of macrophage-mediated cytotoxicity in its immunopathogenesis.

The possible involvement of cell-mediated immune responses to liver-specific protein in the pathogenesis of liver injury was investigated. The subjects consisted of seven patients with acute hepatitis, 12 cases with chronic active hepatitis, four cases with chronic inactive hepatitis, and three cases with liver cirrhosis. When peripheral blood lymphocytes from these patients were cultured in the presence of liver specific protein, and lymphocyte transformation was determined by measuring the uptake of [3H]thymidine into acid-insoluble materials, positive blastogenesis was seen in two cases with acute hepatitis and in six cases with chronic active hepatitis. The macrophage activating factor (MAF), a kind of lymphokine, was also detectable in the culture medium of activated lymphocytes from six patients who showed positive blastogenesis by estimating [3H]glucosamine incorporation into macrophages. Furthermore, the MAF-activated macrophages wer shown to be cytotoxic for the isolated liver cells causing marked inhibition of albumin synthesis. This macrophage-mediated cytotoxicity was detected in eight cases that showed positive lymphocyte transformation. These observations suggest that macrophage-mediated cytotoxicity plays a role in the pathogenesis of chronic active hepatitis.

Possible involvement of macrophage-mediated hepatocytotoxicity in chronic active hepatitis: the induction of liver cell injury by culture supernatant of MAF-activated macrophages.

When peripheral blood lymphocytes from patients with various types of hepatitis were stimulated in vitro with liver specific protein, lymphocyte transformation and MIF production were demonstrated in many cases, especially in chronic active hepatitis. The culture supernatant of these activated lymphocytes was also shown to contain MAF, a kind of lymphokine, which activated the peritoneal macrophages of guinea pigs. When the culture fluid of MAF-activated macrophages was added to isolated liver cells, a significant inhibition of their albumin biosynthesis was detected. The active principle which caused the impairment of liver function was fractionated by gel filtration using a Sephadex G-75 column, and it was found that the active material was a protein-like substance with a molecular weight of about 10,000-40,000. The results suggest the possibility that the soluble substance released from the activated macrophage may be involved at least partially in the immunological pathogenesis of chronic active hepatitis among many other immunological processes.

Studies on intrahepatic cholestasis in drug-induced allergic hepatitis: intrahepatic cholestasis induced in the rat by the culture supernatant of activated lymphocytes.

A marked reduction in bile flow and bile acid excretion was whenever peripheral lymphocytes from patients with drug-induced allergic intrahepatic cholestasis were stimulated with a specific drug in vitro in the presence of a soluble liver-specific antigen fraction, and their culture supernatant injected into the mesenteric vein of rats. A gel filtration study of the active fraction of the supernatant that caused a reduction in bile flow, suggested that the molecular size of this active principle is similar to that of the macrophage migration inhibitory factor (MIF). Histologically, dilated bile canaliculi with decreased microvilli were observed via electron microscopy in rat liver after injection of culture supernatant. No such changes were observed in rats after injection of the supernatant of a lymphocyte culture similarly prepared from normal individuals. These results strongly suggested that sensitized lymphocytes obtained from patients with drug-induced intrahepatic cholestasis produce a factor (of factors) causing cholestasis when stimulated with a specific drug in the presence of liver-specific antigen fractions.

Immunological studies on the drug-induced allergic hepatitis.

The possible involvement of cell-mediated immunity in the pathogenesis of drug-induced allergic hepatitis was investigated in 21 patients; 6 patients with cholestasis, two cases with the hepatitis resembling viral hepatitis and 13 cholestatic hepatitis. The peripheral blood lymphocytes from all these patients showed the positive lymphocyte transformation and MIF production when stimulated by the offending drug in the presence of liver specific lipoprotein. By injection of the culture medium prepared from activated lymphocytes into mesenteric vein of rat, a marked reduction of bile flow and bile acid secretion was observed in 12 cases among 17 patients tested. Active material which caused the reduction of bile flow was fractionated by a gel filtration and was identified to have similar molecular size to MIF. Morphologically, a dilated bile canaliculus with diminution of microvilli and vesicles around the dilated bile canaliculus were observed by an electron microscopy after injection of culture supernatant or their fractionated material into mesenteric vein of rat. No such changes could be seen in rats by administering the supernatant of lymphocytes from normal individuals prepared as above. Macrophage activating factor (MAF), a kind of lymphokines, was also detected in the culture medium of activated lymphocytes from seven patients among eight cases tested. The MAF-activated macrophages were shown to exhibit a cytotoxic effect on the separated liver cells by judging from the inhibition of albumin biosynthesis. Moreover, the antibody-dependent cell-mediated cytotoxic reaction as well as lymphocyte-mediated cytotoxicity were also demonstrated in three cases among nine patients tested. These observations suggest that diverse immune reactions were possible correlated to the pathogenesis of the drug-induced allergic hepatitis although their exact participation or relative significances are remained to be elucidated.

Partial purification of the cholestatic factor derived from the lymphocytes of tuberculin-sensitized guinea pigs.

When lymph node lymphocytes from the tuberculin-sensitized guinea pigs were stimulated in vitro with PPD (purified protein derivatives) and the culture fluid was injected into the mesenteric vein of rats, a marked reduction of bile flow was observed. These culture supernatants contained cholestatic activity were fractionated by gel filtration using a Sephadex G-75 column followed by DEAE-cellulose column chromatography. Both the cholestatic activity and the macrophage migration inhibitory factor (MIF) activity were detected predominantly in the fourth fraction of gel filtration. By further fractionation using a DEAE-cellulose column chromatography, the cholestatic activity was separated into two fractions; one of them was shown to have both cholestatic activity and MIF activity, but the other did not have any detectable MIF activity. These results suggest that the cholestatic factor may be different at least partially from the MIF.

Immunological studies on drug-induced allergic hepatitis--hepatocellular injury by macrophage-mediated cytotoxicity.

When the peripheral blood lymphocytes from patients with drug-induced allergic hepatitis were stimulated with a specific drug in vitro in the presence of a liver cytosol fraction containing liver specific antigen, lymphocyte transformation was seen in eight out of 11 patients. The macrophage activating factor (MAF), a kind of lymphokines, was also detectable in the culture medium of activated lymphocytes from seven out of eight patients who showed positive blastogenesis evaluated the uptake of 3H-glucosamine into macrophages. MAF-activated macrophages exhibited a cytotoxic effect on separated liver cells resulting in a marked inhibition of albumin synthesis. This macrophage-mediated cytotoxicity was also observed in eight out of 11 patients who showed positive lymphocyte transformation. These observations suggest that macrophage-mediated cytotoxicity may play some role in the pathogenesis of drug-induced allergic hepatitis.