Hepatitis B virus markers and lymphocyte populations in the asymptomatic multiple-drug addict

The authors have studied in particular hepatitis B virus markers and the ratio of OKT4+/OKT8+ cells in two subgroups of 52 deprived asymptomatic drug addicts. In the group 1 (deprived for 19 days on an average), 52% of the subjects showed a ratio of OKT4+/OKT8+ less than 1, whereas in the group 2 (deprived for 30 months on an average), this percentage is only of 25%. HBV markers were present in 90% of the subjects in each group. We would like to point out the high frequency of the anti-HBc positivity without other markers in these two groups, respectively 20% in the group 1 and 32% in the group 2. These results emphasize the interest in screening systematically this marker in all blood donors.     

Peripheral T-cell subsets in chronic type B hepatitis: correlation with biochemical and histological activities and hepatitis B e antigen/antibody status

Peripheral T-cell subsets in 77 patients with hepatitis B surface antigen (HBsAg)-positive chronic liver diseases were studied by indirect immunofluorescence using murine monoclonal antibodies against all peripheral T cells (OKT3), T-helper/inducer cells (OKT4), and T-cytoxic/suppressor cells (OKT8). OKT4/OKT8 ratios were significantly reduced in patients with hepatitis B e antigen (HBeAg)-positive chronic liver diseases, including 28 patients with chronic active hepatitis (CAH) (P less than 0.001) and 15 with chronic persistent hepatitis (CPH) (P less than 0.001). OKT4/OKT8 ratios were significantly lower in 21 HBeAg-negative patients with CAH (P less than 0.05), as compared to those of 17 normal controls, while T-cell subsets in 13 patients with HBeAg-negative CPH were essentially normal. Low OKT4/OKT8 ratios significantly correlated with HBeAg positivity (P less than 0.001) and CAH (P less than 0.05), as assessed with multiple regression. There was a significant negative correlation between OKT4/OKT8 ratios and serum glutamic-pyruvic transaminase (SGPT) levels (r = -0.37; P less than 0.01). It was concluded that in chronic hepatitis B virus infection, low OKT4/OKT8 ratios are closely related to active viral replication and more severe histological and biochemical activity.     

Peripheral T-cell subsets in asymptomatic hepatitis B-virus carriers

To ascertain whether the abnormalities of circulating T-cell subsets in patients with hepatitis B virus (HBV)-related chronic liver diseases represent the primary immunological process or are secondary to liver disease process, peripheral T-cell subsets were analyzed by indirect immunofluorescence using monoclonal antibodies against total T cells (OKT3), T helper/inducer cells (OKT4), and T suppressor/cytotoxic cells (OKT8), in 30 asymptomatic HBV carriers without biochemical or histological evidence of liver disease, and the results were compared to 15 HBV-induced chronic active liver diseases. The results revealed that OKT4/OKT8 ratios were significantly reduced in 15 hepatitis B e antigen (HBeAg)-positive asymptomatic carriers as compared with controls, with decreased OKT4-positive cells and increased OKT8-positive cells, while T-cell subsets and ratios were normal in 15 hepatitis B e antibody (anti-HBe)-positive asymptomatic carriers. The changes of circulating T-cell subsets in 15 HBe-Ag-positive asymptomatic carriers showed no significant difference from those of 15 HBeAg-positive patients with chronic active liver diseases. These findings suggest that the deranged T-cell subsets in chronic HBV infection are not secondary to liver cell damage, but might represent the underlying immunological abnormalities which are closely related to HBeAg/anti-HBe status, and that the pathogenetic mechanism of liver cell damage in chronic HBV infection may not be simply related to circulating T-cell subsets.     

Secretions of anti-myelin-associated glycoprotein antibodies by B cells from patients with neuropathy and nonmalignant monoclonal gammopathy

Four patients with peripheral neuropathy and nonmalignant monoclonal gammopathy with anti-myelin-associated glycoprotein (MAG) antibodies were studied to determine whether secretion of anti-MAG IgM antibodies by B cells was autonomous, or whether the monoclonal B cells were responsive to T cells. Secretion of anti-MAG IgM by isolated B cells was stimulated by the addition of increasing numbers of pokeweed mitogen (PWM)-activated autologous OKT4+ helper T cells in all four patients. Secretion of anti-MAG IgM by peripheral blood lymphocytes was dependent on the ratio of OKT4+ T helper cells to OKT8+ T suppressor/cytotoxic cells. In three patients with an OKT4+ to OKT8+ T-cell ratio of 2:1, PWM activation stimulated secretion of anti-MAG IgM; in one patient with an OKT4+ to OKT8+ ratio of 1:2, activation by PWM suppressed anti-MAG IgM secretion. These studies suggest that the monoclonal B cells that secrete anti-MAG IgM are responsive to regulatory T cells.     

T-lymphocyte subset interactions in the cell-mediated immune response to Epstein-Barr virus

The T-lymphocyte subset interactions in the cell-mediated response to Epstein-Barr virus (EBV) were studied in an in vitro system in which the ability of T lymphocytes to inhibit outgrowth of autologous EBV-transformed B lymphocytes is assessed. Inhibition could be demonstrated within lymphocytes of both the T4 and T8 surface phenotypes. Outgrowth inhibition was observed more frequently when the effector T-cell population contained cells of both surface phenotypes. In blocking experiments the OKT3 antibody completely prevents development of outgrowth inhibitory activity; the OKT4 and OKT8 antibodies were less effective in interfering with outgrowth inhibitory function. Maximal blocking activity occurred when antibody addition occurred in the early phase of generation of suppressor function. Pharmacologically achievable concentrations of interferon-alpha restored outgrowth inhibitory activity after blocking with monoclonal antibody. EBV reactivation was easily demonstrable in a group of 10 renal allograft recipients who received OKT3 antibody for treatment of acute rejection. These studies suggest that the immunoregulatory control of proliferation of EBV-transformed B lymphocytes is complex, and involves a collaborative interaction of lymphocytes of both the T4 and T8 surface phenotypes.     

Functional analysis of monkey lymphocyte subsets defined by OKT4 and OKT8 monoclonal antibodies

The percentages of rhesus monkey blood lymphocytes (PBL) reactive with OKT4 and OKT8 antibodies and the $\text{OKT}\text{4}\text{OKT}\text{8}$ ratio showed significant correlations with the log of the immunoglobulin plaque-forming cell (PFC) response after stimulation with pokeweed mitogen (PWM). These correlations suggested that monkey OKT4⁺ cells function as “helper” cells and OKT8⁺ cells function as “suppressor” cells for the PFC response. This was confirmed by separation and study of enriched T- and B-cell subpopulations. OKT8-depleted (OKT4⁺) and OKT4-depleted (OKT8⁺) cells were obtained by treatment of purified T cells with antibody and complement. OKT4⁺ cells augmented the PWM-induced B-cell differentiation into PFC but OKT8⁺ cells did not. OKT8⁺ cells suppressed the PFC response by mixtures of B cells and OKT4⁺ cells. OKT8 antibodies also detected a suppressive cell subset in African green monkeys since the percentage of OKT8⁺ cells showed a negative correlation with the log PFC response. OKT4 antibodies failed to bind to African green monkey PBL.     

Characterization of human cytotoxic lymphocytes directed against cells infected with typhus group rickettsiae: evidence for lymphokine activation of effectors

An in vitro culture and assay system was used to determine whether cytotoxic lymphocytes are generated in humans after rickettsial infection. Peripheral blood mononuclear cells (PBMC) were obtained from six individuals with serologic evidence of prior infection with typhus group rickettsiae and from six nonimmune individuals. After PBMC from immune individuals were stimulated in vitro for 7 days with rickettsial antigen, they were capable of lysing typhus group rickettsia-infected, autologous phytohemagglutinin (PHA)-induced blasts, but not uninfected PHA-blasts. No cytotoxic effector cells were generated when either PBMC from immune individuals were placed in culture for 7 days without antigenic stimulation, or when PBMC from nonimmune individuals were stimulated in vitro with antigen for 7 days. Freshly isolated PBMC from immune donors were also unable to lyse typhus group rickettsia-infected autologous PHA-blasts or an autologous rickettsia-infected lymphoblastoid cell line (LCL). Neither supernatants from antigen-stimulated cultures of PBMC from immune donors nor recombinant human interferon-gamma were capable of significantly lysing typhus group rickettsia-infected PHA blasts by this assay. Populations of cytotoxic effector cells depleted of OKT3, OKT4, or OKT8-positive cells by treatment with the respective monoclonal antibodies and complement were assayed for their cytotoxic capacity. The results suggest that the cytotoxic effector cell population is predominantly OKT3 and OKT8-positive, but OKT4-negative. Positive selection with the use of a fluorescence-activated cell sorter also suggested that most of the cytotoxic effector cells are OKT8-positive. PBMC from immune donors after in vitro stimulation with rickettsial antigen were capable of significantly lysing infected autologous LCL or infected HLA-mismatched LCL as compared with the respective uninfected controls. In addition, PBMC from either immune donors or nonimmune donors after stimulation in vitro for 7 days with media containing purified lymphokines were capable of significantly lysing autologous infected LCL as compared with the uninfected autologous control. We conclude that lysis of cells infected with typhus group rickettsiae is mediated by a lymphokine-activated killer.     

A defect in the suppressor circuits among OKT4+ cell populations in patients with systemic lupus erythematosus occurs independently of a defect in the OKT8+ suppressor T cell function

The autologous mixed lymphocyte reaction (MLR) is thought to be part of a regulatory role of T cells on B cell function. OKT4+, but not OKT8+, cells can proliferate in response to autologous non-T cells. Moreover, the OKT4+ cell population activated early in the course of autologous MLR functioned as inducer cells for the differentiation of B cells, whereas later in the response, the activated OKT4+ cells were particularly enriched in suppressor cells. A part of the autologous MLR appears to be an important pathway for the activation of feedback suppression mechanisms among cells contained within the OKT4+ populations. Patients with systemic lupus erythematosus (SLE) were studied with regard to the following OKT4+ cell functions in vitro after activation in the autologous MLR: a) proliferative response, and b) helper and suppressor activities for differentiation of B cells. A marked reduction in the proliferative response of OKT4+ cells was observed in SLE patients. SLE OKT4+ cells activated in the autologous MLR could function as helper cells but could not exert any suppressor activity. This OKT4+ cell abnormality was present regardless of the disease activity, and occurred in the absence of autoantibodies including anti-T cell antibodies. Instead, SLE anti-T cell antibodies could preferentially eliminate cells bearing the OKT8+ phenotype characteristic of suppressor cells in populations of normal T cells. These results suggest that the defect in the suppressor circuits among OKT4+ cell populations is intrinsic to SLE lymphocytes and that the OKT8+ suppressor T cell defect is caused by antibodies produced by the B cells of SLE patients.     

T lymphocyte subsets in patients with malignant melanoma

Summary T lymphocyte subset profiles were determined by monoclonal antibodies on cryopreserved peripheral blood lymphocytes from 57 patients with malignant melanoma and 19 healthy controls. Quantitation of percentages of total T cells (OKT3.PAN), helper (OKT4.IND) or suppressor (OKT8.SUP) cells, and the ratio of helper/suppressor subsets revealed no correlation of these markers with stage of disease or clinical outcome. A sequential study of these markers on peripheral blood lymphocytes from three stage I melanoma patients with subsequent recurrent disease showed no fluctuations that could be correlated to tumor progression. This study indicates that there is no systemic imbalance in T cell subsets in malignant melanoma and that quantitation of these subsets cannot predict the clinical course of this disease.     

Characterization of the acquired immune deficiency syndrome at the cellular and molecular level

Summary The acquired immunodeficiency syndrome (AIDS) is a new disease characterized by severe dysfunction of both the T cell and B cell systems, occurring in previously healthy individuals. Affected individuals may have recurrent and chronic opportunistic infections and/ or Kaposi's sarcoma or other malignancy. Analysis of the cellular and subcellular components of immunity demonstrates a profound depression in the number and function of helper/ inducer T cells bearing the OKT4 (Leu 3) differentiation antigen and a concomitant defect in the synthesis of the immuno-enhancing soluble growth factor, interleukin 2 (IL-2). Hypotheses to explain the etiology of the immunological dysfunction and implications for future therapy of AIDS are discussed.     

Blood transfusion and acquired immunodeficiency syndrome (AIDS)

This report was presented at the June 1983 meeting of the Blood Transfusion National Consultative Commission at the French Ministry of Health. Clinical and epidemiological data on AIDS as well as problems raised by the lack of specific tests for screening of blood donors were briefly summarized. Out of 49 AIDS patients recorded in France up to April 1983, only one had a history of previous blood transfusion given in Haiti, 4 years before the clinical onset of the disease. Blood donors, all Haitians, had no sign and symptom of AIDS. Retrospective review of 2 300 hemophiliacs followed up in France until April 1983 disclosed no AIDS. However, in 6 patients, the following features, more or less associated, were found to be present: thrombocytopenia, neutropenia, micropolyadenopathy, splenomegaly, hypergammaglobulinemia and low OKT4/OKT8 ratio. No clear correlation could be found between these abnormalities and the origin, commercial or national, of the coagulation factor concentrates used for the treatment. Three main recommendations were proposed: -- information of blood donors and experimental evaluation of some non specific screening tests, in the at risk donor population. -- more cautious use of coagulation factor concentrates -- reduction of importations aiming at complete national self sufficiency concerning factor VIII concentrates.     

Characteristics of human NK clones: target specificity and phenotype

Clones derived from purified human large granular lymphocytes (LGL) of three different donors were expanded in culture medium supplemented with interleukin 2 (IL 2). Their cytotoxic activity was tested in a 51Cr-release cytotoxicity assay against a panel of three to five NK-susceptible tumor cell lines. Of 196 LGL clones tested, only 44 (22.4%) displayed significant cytotoxic activity. A heterogeneous pattern of reactivity was seen; 26 clones (59%) killed all the targets within the panel tested, whereas 18 clones (41%) had a more restricted specificity. Among these 18 clones, 12 lysed only one target (K562, six clones; ADCC, three clones; Daudi, two clones; MOLT-4, one clone), whereas the other six killed two different targets (ADCC and A1ab, one clone; K562 and MOLT-4, five clones). Clones derived from LGL preselected for positive reactivity with the monoclonal antibodies (MoAb) alpha OKM1, alpha OKT10 and alpha B73.1 also demonstrated either broad or restricted patterns of cytotoxicity. The LGL reactive with MoAb alpha B73.1 gave a high percentage of cytotoxic clones. Phenotype analysis showed that clones could express both antigens associated with T cells (i.e., OKT3, OKT4, and OKT8) and antigens shared by LGL (i.e., OKM1, OKT10, and B73.1). The pattern of surface markers varied considerably among the clones; however, no clear correlation between the pattern of antigenic phenotype and cytotoxic activity was seen. These data show that clones derives from purified preparations of LGL present different functional and antigenic characteristics, and support the hypothesis that the heterogeneity of the entire NK population is attributable, at least in part, to a mixture of clones that vary substantially in their target specificities and phenotypes.     

IL-2 secretion by soluble antigen-reactive human T-cell clones

Human tetanus toxoid specific T-cell lines and clones capable of producing IL-2 were established. IL-2 production occurred only when the antigen-specific T cells were cultured with both tetanus toxoid antigen and an autologous, irradiated adherent cell population. The T-cell lines and clones remained strictly dependent on exogenous IL-2 for proliferation at all other times. Phenotypic characterization with monoclonal antibodies recognizing T-cell subsets revealed that the antigen-specific lines and clones bore predominantly OKT3 and OKT4 markers with essentially no OKT8 positive cells present. T-cell clones which were demonstrated to secrete IL-2 activity could also partially deplete media of IL-2 if cultured in the absence of soluble antigen and irradiated adherent cells.     

T-lymphocyte subpopulations in the peripheral blood and bone marrow of patients with aplastic anemia

A decrease in the absolute number of total lymphocytes, OKT3+ and OKT4+ lymphocytes, and a normal number of OKT8+ lymphocytes were found in the peripheral blood of patients with aplastic anemia. The OKT4:OKT8 ratio was decreased in patients due to a reduction in the percentage of OKT4+ cells and 3 out of 18 patients had a ratio less than 1. The values of the OKT4:OKT8 ratio were not associated either with the severity of the disease or with treatment with androgens. There was no correlation between the OKT4:OKT8 ratio and the number of transfusions received by patients. On the other hand, studies performed with bone marrow lymphocytes showed that the OKT4:OKT8 ratio for both patients and controls was lower than that of the peripheral blood. Since the ratio of OKT4:OKT8 cells in aplastic and control bone marrow was similar no direct pathogenic role can be assigned to the marrow for the imbalance detected in the peripheral blood.     

Lepromin-induced lymphoproliferative response of experimental leprosy monkeys: regulatory role of monocyte and lymphocyte subsets

We investigated the immunological status of seven normal, control Mangabey monkeys and 23 Mangabey monkeys experimentally inoculated with Mangabey-origin Mycobacterium leprae. Clinically, these monkeys were divided into three broad groups: a recently inoculated group, a resistant group, and a susceptible group. The resistant group included 11 monkeys, seven of which showed no clinical sign of disease to date and four of which had shown local disease that partially regressed spontaneously. The susceptible group included eight monkeys, five of which have disseminated disease and three with local but stable disease. When peripheral blood mononuclear cells of these monkeys were cultured with Dharmendra-type human lepromin, one of seven normal monkeys, four of four of the recently inoculated group, seven of 10 resistant monkeys, and three of eight susceptible monkeys showed significant responses. In this experimental monkey model, we studied possible regulatory mechanisms by using OKT4- and OKT8-enriched lymphocytes, and Fc receptor-positive (FcR+) and FcR- monocyte (M phi) subsets. The OKT4+ subset was the main lepromin-responsive cell type. High percentages of OKT8+ cells showed a good negative correlation with the lymphoproliferative responses of T-enriched cells supplemented with unfractionated M phi. But the depletion of OKT8+ cells could not increase the response of nonresponding monkeys' lymphocytes. The resistant group and susceptible group did not differ in their percentages of OKT8+ cells. Because OKT8+ cells negatively regulate the response of lymphocytes and OKT4+ cells are the main responding cells, OKT8+ cells are phenotypically and functionally suppressor cells and OKT4+ cells are the helper/inducer cell population in this system. The FcR- M phi population mainly includes antigen-presenting activity, but high percentages of FcR- M phi showed a significant negative correlation with lymphoproliferative responses in the resistant group. A weak but significant lymphocyte response to Dharmendra lepromin was obtained by depleting FcR+ M phi from cultures of some susceptible monkeys, whereas lymphocytes of other susceptible monkeys remained unresponsive to lepromin. By these criteria, we could find an array of immunological defects in monkeys with experimental leprosy. The data suggest that some immunological defects may exist in the OKT4+ lymphocytes or FcR- M phi of leprosy monkeys.     

Immune deficiency in the X-linked lymphoproliferative syndrome. II. Immunoregulatory T cell defects

Surface phenotypic markers and the function of lymphocytes in patients affected with the X-linked lymphoproliferative syndrome (XLP) were studied. This syndrome is characterized by a defective response to infection with Epstein Barr virus (EBV). Normal numbers of B and T cells were detected with anti-Ig and monoclonal OKT3 antisera, respectively. T cell subset values, however, were persistently altered: cells reacting with OKT8 were significantly elevated in five of nine patients, accompanied by a slight decrease in the percentage of OKT4-positive cells, leading to abnormally low OKT4 to OKT8 ratios. One patient had a high OKT4 to OKT8 ratio due to low number of OKT8-positive cells. Lymphocytes from patients showed normal proliferation after stimulation with T and B cell mitogens. In contrast, Ig synthesis by lymphocytes after stimulation with B cell mitogens was markedly deficient: low or undetectable levels of one or all classes of Ig were detected, whereas cell lines established from EBV-infected B lymphocytes from patients produced normal quantities of Ig. These studies imply immune regulatory impairments in the patient with XLP.     

Changes in the ratio of T-lymphocyte subpopulations in various cytological samples of Plummer's thyroid adenoma

In order to correlate possible alterations of cell-mediated immune response with the evolutive phases of Plummer Adenoma (P. A.), T lymphocytes subpopulations in FNA samples and in peripheral blood lymphocytes (PBL) have been studied in 5 patients with autonomous nodules. The lymphocyte component in FNA and peripheral blood has been isolated by Lymphoprep gradient centrifugation; the analysis of T helper and T suppressor subpopulations was made by indirect immunofluorescence with OKT8 and OKT4 monoclonal antibodies. Our results show a reduction in OKT4/OKT8 ratio in cytological samples compared with PBL in patients with P. A., while in control subjects there was not statistically significant difference. In the patients with P. A., the relative increase of OKT8 lymphocytes in FNA compared with PBL is correlated with the functional state, that is toxic adenomas have a lower OKT4/OKT8 ratio compared with nodules in pre-toxic phase. In conclusion: T lymphocyte subpopulations typing in FNA demonstrate that, even in this type of hyperthyroidism, immune response disorders are present and consist of relative increase of suppressor/cytotoxic T cells, compared to T helper cells.     

Functional analysis of human T cell subsets defined by monoclonal antibodies. III. Regulation of helper factor production by T cell subsets

In the present report we extended our previous studies demonstrating that obligatory T-T interactions are important in regulating human immune responses in vitro. Functionally distinct human T cell subsets were isolated by complement-mediated lysis using the monoclonal antibodies OKT4 and OKT8. Evidence was obtained that during allogeneic interactions, OKT4+, but not OKT8+, responder T cells are required to generate helper factor(s) capable of polyclonally activating human B cells independent of additional T cell help. Importantly, the alloantigen-induced helper factor(s) production and/or release was found to be suppressed by addition of graded numbers of radiosensitive OKT8+ cells. On the other hand, no evidence was obtained that supernatant derived from alloactivated OKT8+ cells could counterbalance the helper activity generated in the presence of supernatant from alloactivated OKT4+ cells. Furthermore, OKT8+ cells, known to suppress PWM-driven B cell differentiation in the presence of OKT4+ cells, do not suppress B cell differentiation induced by preformed helper factor even in the presence of OKT4+ cells. These data further underscore the importance of functional T-T interactions in immunoregulation in vitro and support the idea that the target of suppression of B cell differentiation, induced either by alloantigen-triggered helper factor or PWM, are OKT4+ cells and not B cells themselves.     

Use of monoclonal antibodies to human lymphocytes to identify lymphocyte subsets in lymph nodes of the rhesus monkey and the dog

Using commercially available monoclonal antibodies that bind to human lymphocyte subsets, we examined lymph nodes from the rhesus monkey and the dog for their binding ability to these monoclonal antibodies. The avidin biotin-peroxidase immunostaining procedure was used, and the following antibodies were reactive in the rhesus monkey: Leu 4, Leu 3a, OKT4, Leu 2a, OKT8, T8, Leu 5b, T11, Leu 14, B1, and Leu 12. No immunoreactivity was observed in the dog lymph node except for moderate reactivity of OKT8. The following antibodies failed to react in both the rhesus monkey and the dog: OKT3, T1, T2, T1B, T4, T8A, Pan B, and T29/33. Kappa and lambda immunoglobulin light chains were positive in both the dog and monkey.     

接触猪职业人群和对照人群戊型肝炎感染率比较研究

近年来戊型肝炎（Hepatitis E,HE）被怀疑是一种人兽共患病,猪是其重要的宿主.通过在HE流行区比较接触猪人群和对照人群HE感染率,评价接触猪对人群的HE感染的风险.在浙江省北部某县采用限制研究对象的群组匹配的横断面调查,对同地区同年龄段的接触猪人群和对照人群进行问卷调查,采集血液并检测抗HE抗体.研究发现,接触猪人群和对照人群的年龄大致相当,但前者男性比例较高.接触猪人群抗HE抗体感染率（254/340,74.71%）明显高于对照人群（319/512,62.30%）.经过分层分析和多因素非条件Logistic回归分析,平衡性别和年龄因素后,接触猪仍能增加约50%人群的HE感染风险.随着接触年数的增加,其风险也增加,接触猪5～14年和15年以上人群分别比对照人群增加65%和157%的HE感染风险.接触猪人群男性、女性和合计的标化感染率分别为72.30%、65.67%和70.92%,均高于对照人群（69.87%、51.82%和60.80%）,尤其以女性更为明显.在HE流行区,接触猪可能增加人群HE感染风险,对女性的影响可能较大,随着接触猪年数的增加HE感染的风险也增加.由于本研究采用病因和结果同时出现的横断面研究,确切的接触猪和HE感染的关系需要建立合适的接触猪人群队列并结合详细的接触猪暴露调查得到阐明.