Lymphocyte subpopulations in patients with systemic lupus erythematosus

A discrepancy between E-rosette-forming cells and total T cells identified by monoclonal antibodies (Leu-1+) was observed in patients with systemic lupus erythematosus (SLE). E-Rosette-forming cell percentages of peripheral blood mononuclear cells (MNC) were significantly lower than those observed in normal individuals, in contrast to the percentage of Leu-1+ cells in SLE patients which were not different from those of normal values. The cells which did not form E rosettes, but stained positively with the monoclonal anti-Leu-1 antibody (E-, Leu-1+) had certain functional capability evidenced by their ability to amount a proliferative response to T-cell mitogens, phytohemagglutinin and concanavalin A, but failed to provide T-helper-cell activity to support the differentiation of normal B lymphocytes into immunoglobulin-secreting cells following stimulation with pokeweed mitogen. Inhibition of E-rosette formation, but not of the staining for Leu-1, was demonstrated by incubating sera from SLE patients with normal MNC. These studies suggest that the T-cell markers, E+ and Leu-1+, do not necessarily characterize identical subpopulations of T cells. When referring to E- cells in studies with MNC from SLE patients, it should be realized that this population includes, in addition to B cells and macrophages, cells staining positively for Leu-1 antigen and reactive to T-cell mitogens.     

Delta infection in patients with chronic viral hepatitis B

The results of examination for anti-delta antibodies in the blood of 86 patients (33 children and 53 adults) with chronic persisting hepatitis (CPH) and chronic active hepatitis (CAH) verified by morphological examinations of liver biopsy specimens are presented. Delta infection was found in 43 (31.5%) of patients with CAH, only anti-HBe being demonstrable in the blood. In this group of patients, marked enzymatic disorders and more intensive changes in the morphological picture of the liver were demonstrated.     

Characterization of viral kinetics in patients with hepatitis B e antigen-positive chronic hepatitis B

A study was conducted during a 1 year follow-up to characterize the viral kinetics in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B and to develop a model of predicting the probability of spontaneous HBeAg seroconversion. Fifty-seven patients with HBeAg-positive chronic hepatitis B were enrolled with monthly follow-ups from three Phase III clinical trial placebo groups. According to serial viral loads, 30 patients (52.6%) with the stationary pattern maintained stable HBV DNA levels with fluctuations of less than 1.5 log copies/ml. Twenty patients (35.1%) with the declining pattern exhibited a spontaneous decline of more than 1.5 log copies/ml without a following rebound of at least 1.5 log copies/ml. The remaining seven patients (12.3%) had the wavering pattern. Both declining and wavering patterns, when compared with the stationary pattern, had significantly higher hepatic necroinflammation in terms of ALT and Knodell scores at the baseline and peak ALT levels during the follow-up period. The declining pattern had a significantly better clinical outcome in terms of the lowest final HBV DNA and a reduction in the necroinflammatory score after 1 year. Furthermore, the declining pattern had a favorable HBeAg seroconversion rate (40%) compared with the wavering (14.3%) and stationary patterns (0%). A regression equation, incorporating simultaneous serum bilirubin, ALT, and HBV DNA levels, predicted the probability of HBeAg seroconversion with a sensitivity of 76.8% and a specificity of 74.7%. In conclusion, different viral kinetic patterns in patients with chronic hepatitis B implicate distinct clinical significance and immunologic perspective.     

Lymphocyte subpopulations in patients with hydroxocobalamin responsive megaloblastic anaemia.

Lymphocyte subpopulations and intrinsic factor and gastric parietal cell antibodies have been measured in 23 patients with megaloblastic anaemia who responded to treatment with hydroxocobalamin. The ratio of helper (OKT4) to suppressor (OKT8) lymphocytes was significantly increased in patients with intrinsic factor antibody compared with those who lacked the antibody. No such correlation was found for gastric parietal cell antibody. Alterations in the lymphocyte helper to suppressor (OKT4:OKT8) ratio may be associated with pernicious anaemia.     

Lymphocyte subpopulations and reactivity to mitogens in patients with scleroderma.

T lymphocyte subpopulations were studied in 40 patients with scleroderma (PSS), 26 of whom were studied simultaneously for lymphoproliferative responses to phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM). PSS patients exhibited a reduction relative to 42 age- and sex-matched controls in the absolute number and percentage of early E rosettes, late E rosettes and E rosettes formed with aminoethylisothiouronium bromide (AET) treated sheep red blood cells. There was no difference between patients and controls in the proportions of B lymphocytes. PSS patients exhibited normal lymphocyte transformation responses to PHA and ConA and an augmented response to PWM. The mitogen responses did not correlate with the absolute number or percentage of lymphocytes or T and B lymphocyte subpopulations. No correlation was observed between any immunological variable studied and the extent of skin or organ involvement, disease duration or therapy.     

Lymphocyte subpopulations in peripheral blood of patients with non-Hodgkin lymphoma

Various lymphocyte subpopulations of peripheral blood of 65 patients with aleukaemic non-Hodgkin lymphoma were studied. The clinically active state and remission in both low and high grade malignancy groups were compared. An elevated lymphocyte count was found in the blood of aleukaemic patients, too. Lymphocytosis in low grade malignancies in the clinically active states was found to be higher than in the corresponding group of the high grade cases. The data showed that the most important factor in the increase of total blood lymphocyte count is the increase in the amount of B-lymphocytes. The TA/TT ratio was significantly reduced in all subgroups. The OKT4/OKT8 ratio was normal in all cases.     

Lymphocyte subpopulations in the skin of patients with chronic urticaria

In order to characterize the nature of the mononuclear cells in the perivascular infiltrates in the skin of 11 patients with CU, skin biopsy specimens were analyzed in situ by an avidin-biotin immunoperoxidase technique. Serial frozen sections were stained for total T cells, helper-inducer T cells, suppressor-cytotoxic T cells, B cells, monocytes/macrophages, and HLA-DR antigen. The infiltrates were found to consist mainly of T cells, whereas B cells and macrophages were rarely seen. Most of the T cells possessed the T4+ helper phenotypes, whereas smaller numbers of infiltrating cells were defined as suppressor-cytotoxic cells. Most of the helper-inducer T cells coexpressed the Ia (HLA-DR) antigen. Several potential pathogenic mechanisms could be implicated in CU based on these observations.     

Detection of hepatitis B viral DNA by polymerase chain reaction in patients with hepatitis B surface antigen

Hepatitis B virus (HBV) DNA was assayed using the polymerase chain reaction in serum samples of 116 hepatitis B surface antigen (HBsAg) carriers, including 30 positive for hepatitis B e antigen (HBeAg) and 86 negative for HBeAg. In the HBeAg-positive group, all were positive for HBV DNA. In the HBeAg-negative group, 80.2% were positive for HBV DNA (80.0% in the healthy carrier group, 90.0% in the chronic active liver disease group, and 69.2% in patients with cirrhosis). This study indicated that every HBeAg-positive carrier as well as the majority of HBeAg-negative carriers were infectious and, in the latter group, that viral replication is most active in patients with chronic active liver disease.     

Lymphocyte subpopulations in sarcoidosis

Highly purified blood lymphocytes from sarcoidosis patients were characterized by rosette technique for the presence of receptors for complement and by indirect immunofluorescence for the presence of membrane-bound immunoglobulin (B cells). T cells were estimated by spontaneous rosette formation with sheep red blood cells. The proportion of B cells was significantly higher and the proportion of T cells significantly lower than in the control group. Marked lymphocytopenia was found in the patient group. As a consequence the total number of circulating B cells was normal whereas the total number of circulating T cells was significantly depressed. Large atypical mononuclear cells were found in most patients. One-third of these cells had receptors for human complement.     

Markers of viral replication in patients with chronic hepatitis B virus infection

Serum samples from 56 patients with biopsy-proven chronic B viral hepatitis without superimposed delta hepatitis were analyzed for the various markers of viral replication, including serum hepatitis B e Ag (HBeAg), hepatitis B virus deoxyribonucleic acid (HBV-DNA), and hepatitis B core antigen (HBcAg) in the liver tissues. Twenty-seven patients had persistent viral hepatitis (PH) and 29 patients had chronic active hepatitis (CAH) with or without cirrhosis. HBV-DNA was identified in the sera of 81% of patients with PH and 60% of patients with CAH. Significantly higher levels of HBV-DNA were found in patients with PH than in those with CAH. Both HBeAg in serum and HBcAg in liver correlated positively with serum HBV-DNA. Nine patients had serum HBV-DNA in the absence of HBeAg (four had anti-HBe), and seven of these nine patients had stainable HBcAg in the liver (two did not have staining). None of these patients had hepatic HBcAg in the absence of serum HBV-DNA. When these patients were stratified according to their epidemiologic background, serum HBV-DNA was present in a significantly higher number of male homosexuals than in any other groups. This was unrelated to their status of human immunodeficiency viral serology.     

非甲-非戊型慢性病毒性肝炎患者隐匿性HBV感染的研究

目的 观察非甲-非戊型慢性病毒性肝炎患者隐匿性HBV感染的状况,探讨荧光定量聚合酶链反应(FQ-PCR)技术对隐匿性HBV感染的诊断价值.方法 应用FQ-PCR技术对57例非甲-非戊型慢性病毒性肝炎患者进行了血清、肝组织HBV-DNA定量检测,并将肝组织HBV DNA定量水平与肝脏炎症活动度的关系进行了分析.结果 血清、肝组织HBV DrqA定量阳性分别为13例(22.81%)、22例(38.60%).13例血清HBV DNA定量阳性患者其肝组织定量亦均阳性,但9例肝组织HBV DrqA定量阳性患者其血清定量为阴性,差异有统计学意义(P<0.01);同时13例血清与肝组织定量均阳性患者比较.显示肝组织HBV DNA定量水平显著高于血清定量水平[(6.62±1.21)拷贝,gvs.(4.03±1.06)拷贝/ml,(P<0.01)].肝组织HBV DNA水平与肝脏炎症活动度并无相关性,10例G2,7例G3,5例G4患者HB'q DNA定量分别为(6.13±1.65)拷贝/g、(5.92±1.81)拷贝,g、(5.83±1.89)拷贝/g,(P0.05),但HBV DNA定量阳性患者均为活动性肝脏病变.结论 HBV隐匿性感染是部分非甲-非戊型慢性病毒性肝炎患者的病因.单纯检测血清免疫学标志物对HBV感染诊断存在漏诊,对非甲-非戊型慢性病毒性肝炎患者应用FQ-PCR技术开展血清定量尤其是肝组织中HBV DNA定量检测可提高HBV感染的诊断.对隐匿性HBV感染的慢性病毒性肝炎亦应给予有效的抗病毒治疗.     

Association of hepatitis B viral precore mutations with fulminant hepatitis B in Japan

We studied the precore DNA sequences of hepatitis B viral genomes in five patients with fulminant hepatitis B and in five with acute self-limited hepatitis B from Japan. Using the polymerase chain reaction, three to four independent HBV DNA clones from each patient were obtained and analyzed. We demonstrated that patients with fulminant hepatitis B carried HBV genomes with a G to A mutation at nucleotide positions 1898 (five of five patients; 18 of 18 clones, 100%) and 1901 (five of five patients; 12 of 18 clones, 66%) in the precore region. The first mutation results in an in-phase stop codon (TAG) in the precore open reading frame and the absence of HBeAg production. In contrast, a G to A mutation was found in 6 of 16 clones (37%) in position 1898 and in 0 of 16 clones (0%) in position 1901 from patients with acute self-limited hepatitis. We concluded that both of the precore mutations are commonly associated with fulminant hepatitis B and may contribute to the pathogenesis of fulminant hepatitis. A hypothetical model for the biological significance of these two mutations is proposed.     

Lymphocyte stimulation in hepatitis B infections.

Cell-mediated responses in patients with acute and chronic forms of Type B hepatitis were tested by lymphocyte transformation to purified hepatitis surface antigen (HBS Ag) and to phytohemagglutinin, by dinitrochlorobenzene sensitization and by response to skin-test antigens. Although absent during acute Type B hepatitis, lymphocyte transformation by HBS Ag appeared during early convalescence and remained detectable for six years after recovery. Patients with Type B chronic active hepatitis with increased serum transaminase levels showed a partial lymphocyte transformation with HBS Ag, whereas those with normal transaminase values demonstrated no transformation. Patients with Type B chronic persistent hepatitis and carriers of HBS Ag did not have lymphocyte transformation. Normal responses to dinitrochlorobenzene, intradermal skin tests and phytohemmaglutinin were found in all patients with acute, chronic active or chronic persistent hepatitis and carriers. These results indicate that recovery from Type B hepatitis is associated with the ability to elicit a specific lymphocyte response to HBS Ag.     

Lymphocyte blast transformation to HBsAg and HBcAg in patients with various clinical forms of hepatitis B

Sensitization of the peripheral blood lymphocytes to HBsAg and HBcAg in 51 patients with acute hepatitis B (HB) in the time course of infection, in 13 with chronic active hepatitis B (CAH) and 8 HBsAg carriers was studied by lymphocyte blastogenesis assay. In patients with mild or moderately severe forms of acute HB at the peak of the disease lymphocyte blastogenic response to HBcAg was observed; sensitization to HBsAg was lacking and could be detected only in the stage of convalescence when the specific response to HBcAg was already undetectable in most patients. No lymphocyte sensitization to HBsAg either at the peak of the disease or in convalescence was observed in patients with a lingering form of acute HB as well as in those with CAH and HBsAg carriers. At the peak of severe HB blastogenic response was demonstrated to both antigens under study but was more marked to HBcAg. Most patients with CAH showed lymphocyte blastogenic response to HBcAg. It is concluded that specific lymphocyte response to HBV antigens varies in relation to the severity and course of the infection.     

Unusual electron microscopy findings in liver biopsies from patients with viral hepatitis B

Electron microscopy of 57 needle biopsies from livers of 55 patients with chronic hepatitis, mostly of type B, revealed, in addition to typical hepatitis B virus nucleocapsids in 34 cases, also atypical intranuclear inclusions in 9 biopsies. These inclusions consisted of electron-dense particles similar in size to hepatitis B virus nucleocapsids. The possibility of an involvement of a hepatitis non-A non-B virus is discussed.     

Hepatitis B virus mutations associated with in situ expression of hepatitis B core antigen, viral load and prognosis in chronic hepatitis B patients

In this retrospective study, we investigated the prevalence and significance of mutations in part of the hepatitis B virus (HBV) x gene, and tried to clarify their relationship with clinicopathological or histopathological characteristics and prognosis in patients with chronic hepatitis B (CHB). A total of 83 consecutive CHB patients (1986-1994) were chosen for the present study. Sequence analysis was performed using polymerase chain reaction (PCR) and the direct sequencing method. The histological activity index was described using Scheuer scores. Two-step immunohistochemical staining showed the expression of viral antigens in situ. Tissue HBV DNA levels were determined by fluorescence quantitative real-time PCR. For the prognostic study, all the patients were followed up using clinical and laboratory data. Mutation at nt1726-1730 correlated significantly with decreased expression of HBcAg in situ (P = 0.006) and with lower HBV DNA levels in the liver (P = 0.004). In particular, the CTGAC mutation showed the strongest decrease of the viral load (P = 0.007). By contrast, nt1762/1764 mutation correlated with increased HBcAg (P = 0.005) and higher HBV DNA levels (P = 0.006). The mutants with the wild-type of nt1726-1730 or nt1762/1764 mutation were more prevalent in hepatocellular carcinoma (HCC) patients than in CHB patients. Although the mutations did not correlate with cirrhosis, the frequency of nt1762/1764 mutation in patients with hepatocarcinogenesis was significantly higher than in those without hepatocarcinogenesis (P = 0.011). Mutations at nt1726-1730 and nt1762/1764 are associated with in situ expression of HBcAg and viral load. Higher HBV DNA levels in the liver may be associated with hepatocarcinogenesis. Mutation at nt1762/1764 remarkably increases the risk of hepatocarcinogenesis.     

Lymphocyte subpopulation composition in hepatic tissue and autoimmune manifestations in viral hepatitis

This work had the aim to study the role of disbalance of inflammatory procytokines, lymphocyte subpopulation composition in peripheral blood, and cell populations of hepatic tissue in pathogenesis of viral hepatitis. A total of 159 patients with chronic hepatitis B and C were examined. They proved to exhibit signs of autoimmune processes (ASMA, ANA, AMA, anti-LKM-1, RF, CG) and cytokine (IL-1b, TNFa) disbalance. The number of lymphocytes expressing HLADR+ was shown to increase and CD+ lymphocyte to decrease in proportion to the severity of periportal necrosis, portal inflammation, intralobular degeneration and fibrosis compared with healthy donors. Hepatic biopsies contained T-regulatory cell complex (CD4+ CD25+). These data give evidence of the morphofunctional basis for autoimmune manifestations in patients with persistent viral hepatitis infection.