Evaluation of HBV and HCV antigens expression in correlation with HBV DNA and HCV RNA occurrence in liver biopsy specimens of patients with chronic hepatitis type B and chronic hepatitis type C, respectively.

MATERIAL AND METHODS: The material studied consisted of single liver biopsy specimens and serum samples from 97 patients diagnosed by clinical and serological criteria as having chronic hepatitis of either type B (38 patients seropositive for HBsAg) or type C (59 patients seropositive for anti-HCV). Diagnosis of chronic hepatitis, including grading and staging of the process, was established by histopathological examination of routinely stained serial paraffin sections. The expression of HBV and HCV antigens was detected in frozen sections by amplified immunoperoxidase method (EnVision, DAKO) with the use of monoclonal anti-HBs antibodies (DAKO), polyclonal anti-HBc antibodies (Biogenex) and FITC-labeled human antibodies to HCV antigens. HBV DNA and HCV RNA were searched for in tissue homogenates with PCR (In Gen; Terpol, Sieradz) and RT PCR (Cobas Amplicor HCV RNA, Roche), respectively. RESULTS: HBsAg was detected in all and HBcAg in 26 of the 38 biopsy specimens with chronic hepatitis type B from patients who were seropositive for HBsAg and anti-HBc, and for either HBeAg (25 cases) or anti-HBe (13 cases). HBV DNA was identified in 32 out of 38 liver biopsy specimens. Out of the 32 HBV DNA positive specimens, 6 that had been obtained from patients seropositive for anti-HBe, lacked HBcAg expression. HCV antigens were found in 27 (50%) out of 54 specimens which showed the presence of HCV RNA. CONCLUSIONS: These results are indicative for a strong positive correlation between the expression of HBV antigens and that of HBV DNA in liver biopsy specimens. The detection of HCV antigens has proved to positively correlate with that of HCV RNA in 50% of the specimens examined. It appears, therefore, that testing of liver biopsy specimens for HBV and HCV antigens may, in a significant proportion of cases, extend the histopathological diagnosis as to include identification of the etiologic factor.

[Evaluation of cyclosporin A efficacy in the treatment of steroid-resistant nephrotic syndrome in the course of membranoproliferative glomerulonephritis--case report]. / Ocena skutecznosci cyklosporyny A w leczeniu steroidoopornego zespolu nerczycowego w przebiegu bloniastoproliferacyjnego klebuszkowego zapalenia nerek--opis przypadku.

On the case of 35 old patient with steroid-resistant nephrotic syndrome in the course of membranoproliferative glomerulonephritis complicated with renal failure, we paid the attention to the therapeutic efficacy of CsA in this case with remission of nephrotic syndrome and significant improvement of renal function, which enabled us to withdraw the patient from the hemodialysis programme.

AIDS pathology: infections and neoplasms in 55 fatal AIDS cases. A postmortem study.

The study evaluated the incidence of infections and neoplasms in 55 out of 104 patients with AIDS who died in Poland from January 1986 to April 1994 (the estimated autopsy rate-52.8%). Histopathological examination revealed 103 infections and 11 neoplasms. In 40 persons (73%) either multiple infections or a neoplasm and an infection were diagnosed. Cytomegalovirus infection was most common. (65.5% of cases) followed by Pneumocystis carinii (24% of cases). These infections were the leading cause of death in 20% and 16% of cases, respectively. The results of this study showed a significantly lower incidence of Pneumocystis carinii, Kaposi's sarcoma and non-Hodgkin's lymphoma in comparison with the results of similar studies in countries with a large number of AIDS cases.

[Etiology of chronic hepatitis as evaluated by liver biopsy and serum samples submitted to the Department of Immunopathology of the National Institute of Hygiene in 1993-1995]. / Etiologia przewleklych zapalen watroby na podstawie badan biopsji watroby i surowic nadeslanych do Zakladu Immunopatologii Panstwowego Zakladu Higieny w Latach 1993-1995.

The purpose of this study was to assess the relative incidence of chronic hepatitis in a population of patients with chronic liver disease and to determine the etiological spectrum of this syndrome with special reference to its defined histopathological forms. Histopathology aided by immunohistochemistry, and serology aided by the PCR method were employed in studies of liver biopsy specimens and serum samples, respectively. Out of 1150 patients with chronic liver disease examined, chronic hepatitis was diagnosed in 685 (60% of all cases examined). In this group, there were 308 males aged 18-74 yrs (mean 32 yrs), 153 females aged 18-71 yrs (mean 43 yrs), and 213 children aged 1-17 yrs (mean 8 yrs). Viral infections documented in these patients included HBV (50.4%), HCV (36.2%), HBV/HCV (7.2%) and HBV/HDV (0.7%); cryptogenic and autoimmune hepatitis (AIH) accounted for 2.9% and 2.6% all cases, respectively. In the group of minimal hepatitis (16.1%), HBV infection was documented in 66.4% of cases, HCV-in 29.1%, HBV/HCV-in 3.6% (one case of AIH was included into this group). In the group of mild hepatitis (44.2%), HBV infection accounted for 47.3% of cases, HCV-for 41.9%, HBV/HCV-for 9.9%, and 0.9% was diagnosed as cryptogenic. In the group of moderate hepatitis (19.6%), HBV infection accounted for 50% of cases, HCV-for 37.3%, and HBV/HCV-for 4.5%; cases of cryptogenic and AIH accounted for 3.7% and 4.5%, respectively. In the group of severe hepatitis (20.1%), HBV etiology was found in 44.9% of cases, HCV-in 28.3%, HBV/HCV-in 6.5% and HBV/HDV-in 3.6%; cryptogenic and AIH accounted for 6.5% and 8.0% of cases, respectively. There was a high incidence of low-titer autoantibodies (SMA, ANA and LKM) ranging from 75% in cryptogenic hepatitis and 51% in each HBV and HBV/HCV hepatitis to 46.3% in HCV hepatitis.

HIV lymphadenopathy--a histopathological and immunomorphological study of 65 cases.

Lymph nodes from 65 HIV-infected persons were studied by morphological and immunomorphological methods. Early stages of HIV lymphadenopathy (follicular hyperplasia without fragmentation or follicular hyperplasia with severe fragmentation) were revealed in 92% of patients with PGL and the most advanced stadium of HIV-related lymphadenopathy (diffuse pattern) in 92% of AIDS patients. Striking B-cell activation and polyclonal proliferation, found in 19 lymph nodes (35% of advanced lymphadenopathy cases--follicular atrophy or diffuse pattern) justified separation of these cases as a particular subgroup of HIV lymphadenopathy.

Expression of multivalent pre-S1 antigen of hepatitis B virus in Escherichia coli (synthetic oligodeoxynucleotides, surface antigens, recombinant DNA, fusion protein, beta-galactosidase).

The nucleotide sequence encoding 30 amino acids (aa) of the pre-S1 envelope region of the human hepatitis B virus has been constructed from twenty chemically synthesized oligodeoxynucleotides by simultaneous ligation. The DNA fragment containing four repeated sequences encoding the pre-S1 region (aa 20-49) has been inserted into the lacZ gene of the plasmid pWR450.1, yielding the recombinant pWX4 plasmid. The Escherichia coli DH5 strain transformed with pWX4 produces a beta-galactosidase-[-pre-S1(20-49) x 4] fusion protein. The hybrid protein containing 127 aa of repeated pre-S1 region has been isolated from Escherichia coli as inclusion bodies and purified by anion exchange chromatography. The antigenic properties of this fusion protein were confirmed by immunoblotting with pre-S1-specific monoclonal antibodies.

[Hepatitis due to HDV infections]. / Zapalenie watroby wywolane zakazeniem HDV.

From May 1985 to the end of February 1988 the Department of Immunopathology carried out serological studies of 518 samples of the blood serum and made an immunomorphological analysis of 518 hepatic specimens taken by needle biopsy from chronic HBsAg carries. In four cases we found the presence of antigen delta (HDV Ag) in the nuclei of the hepatocytes. These are the first fully documented by immunomorphological and serological methods cases of HDV infection in Poland. We also discussed the epidemiology and biology of viral infection in delta hepatitis.

[Incidence of various types of viral hepatitis in Poland 1988]. / Czestosc wystepowania typów wirusowego zapalenia watroby w Polsce w 1988 roku.

109 sera from patients in the early phase of acute hepatitis non-B were tested by immunoenzymatic methods for presence of serologic markers of infections with HBV, HAV and CMV. The analysis of the results showed that 14 patients were infected with HBV, 20 with HAV and 2 with CMV. The remaining 73 cases could apparently be attributed to the infection with HCV. It has been estimated that in 1988, 69.5% of hepatitis cases in Poland could have resulted from the infection with HBV, 24%--from the infection with NANB viruses and only, and only 6.5%--from the infection with HAV. These data even if representing a slight overestimate of the incidence of hepatitis due to the parenteral spread of infections agents support the postulate that measures to prevent these infections should be strengthened.

Crystalline aggregates of hepatitis B core particles in cytoplasm of hepatocytes.

Large crystalline accumulations of hepatitis B core antigen (HBcAg) particles were detected in the cytoplasm of hepatocytes from a patient with mild reactive hepatitis. Within these crystals, the core particles showed a symmetrical hexagonal arrangement with the center-to-center particle distance estimated at 24 nm. The possible mechanism of aggregation and the significance of HBcAg crystalline lattice in the preservation of hepatitis B virus are discussed.

Pathogenetic aspects of hepatitis A virus infection in enterally inoculated marmosets.

Experimental hepatitis A virus (HAV) infection was studied in marmosets after enteral (intragastric) inoculation with special reference to the primary sites of HAV replication and immunopathology of the disease. The experiment was carried out using 28 Saguinus mystax negative for antibody to HAV (anti-HAV) and with statistically uniform baseline values of serum isocitrate dehydrogenase (SICD) activity. Each animal was infected with 1 ml of a 15% w/v stool suspension that was derived from marmosets infected with the third or fourth passage of the MS-1 strain of HAV. The incubation period measured by the first significant SICD elevation was 32 days in 11 of 13 marmosets. The animals were sacrificed 2, 4, 7, 11, 14, 18, 23, 28, and 32 days after inoculation and 1, 4, 8, 14, 21, 28, and 35 days after SICD elevation. HAV antigen, immunoglobulins, complement, and fibrin were identified in the liver, eight segments of the gastrointestinal tract, lymphoid system, and kidneys. HAV antigen was found only in the cytoplasm of hepatocytes and in gallbladder bile. These findings indicated that the liver was the sole and primary site of virus replication. Combined immunomorphologic and histopathologic observations also revealed that HAV antigen localization was associated with the sites of hepatocellular damage. There was no immunomorphologic evidence for humoral immune clearance of HAV antigen in the liver, lymphoid system, or kidneys.

Hepatitis B surface antigen and albumin in human hepatocytes. An immunofluorescent and immunoelectron microscopic study.

A close correlation between the presence of hepatitis B surface antigen and albumin in the cytoplasm of hepatocytes infected with hepatitis B virus was established by immunofluorescence and immunoelectron microscopy in 52 liver biopsy specimens of various forms of hepatitis and liver cirrhosis. Albumin deposits usually accompanied cytoplasmic content of hepatitis B surface antigen, but were less frequently observed together with hepatitis B antigen localized in or on the membranes. Ultrastructural observations demonstrated albumin on the tubular and spherical forms of hepatitis B surface antigen in the endoplasmic reticulum. The hepatocytes with the content of hepatitis B surface antigen and albumin showed the ability of binding with the fluorescein-labeled preparation of polymerized human serum albumin. The affinity of polymerized albumin to hepatitis B surface antigen was considerably increased after preincubuation of liver sections with 2-mercaptoethanol that removed most of the originally present albumin. This may be indicative for the role of disulfide bonds in the formation of hepatitis B surface antigen-albumin complexes. These results justify the hypothesis that albumin may be incorporated into the viral coat protein during its synthesis in the cytoplasm of infected hepatocytes.

Membranous glomerulopathy associated with hepatitis B core antigen immune complexes in children.

Direct immunofluorescence, immunoelectron microscopy, and special immunohistochemical procedures including guinea pig complement fixation, differential elution, and in situ antigen binding were employed in an immunomorphologic analysis of kidney biopsy specimens from 98 children with clinically diagnosed nephrotic syndrome and/or glomerulonephritis (GN). Glomerular deposits of hepatitis B virus (HBV) antigens, immunoglobulins, and complement were detected in specimens from 24 children, all seropositive for hepatitis B surface antigen (HBsAg) and antibody to hepatitis B core antigen (HBcAg). Of these, 21 cases were diagnosed as membranous glomerulopathy (MGN), 1 as membranoproliferative GN, and 2 as diffuse mesangial proliferative GN. HBaAg was identified as the only HBV antigen in about a third of the cases of MGN, whereas in another third it was accompanied by HBsAg. HBsAg was the only HBV antigenic component detected in the glomerular deposits in the remaining third of the cases of this GN form. The results of this study indicate that apart from, or in addition to, HBsAg immune complexes, HBcAg immune complexes may also participate in the pathogenesis of a significant number of MGN cases in children subclinically infected with HBV. A possibility that these complexes include nonparticulate, presumably low-molecular-weight HBaAg components and that they are found in an environment of antibody-excess is discussed.