Assessment of the immune status of chronic bronchitis patients in the remission phase during prophylactic treatment with levamisole and staphylococcal vaccine

The work presents the results of the study of the immune status in 18 patients with chronic bronchitis at the stage of remission, subjected to prolonged prophylactic treatment with the inhalations of levamisole solution, and in 16 patients receiving the inhalations of staphylococcal vaccine. These results indicate that the inhalations of levamisole have proved to be more effective than the inhalations of staphylococcal vaccine. In patients treated with levamisole an essential decrease in the ratio of theophylline-resistant to theophylline-sensitive E-rosette-forming lymphocytes has been noted. The determination of the sensitivity of lymphocytes and neutrophils to incubation with levamisole by means of the rosette-formation test makes it possible to prognosticate, taking into account the initial level of spontaneous E-rosette formation of these cells, the effectiveness of the prophylactic treatment of patients with chronic bronchitis. The determination of the sensitivity of the cells to incubation with staphylococcal toxoid is ineffective for the prognostication of the result of the treatment of such patients with staphylococcal vaccine.     

Immunologic studies in asymptomatic hemophiliac patients

Various immunological parameters were studied in 20 asymptomatic patients with hemophilia A, 3 patients with hemophilia B and 1 patient with von Willebrand disease. Patients were treated with cryoprecipitate or fresh frozen plasma. Significantly decreased mean percentage and absolute count (p less than 0.01) of peripheral blood E-rosette-forming cells compared to controls was found. There were normal mean percentages and absolute counts of lymphocytes, T-helper inducer, T-suppressor cytotoxic and natural killer cells. The proportion and absolute number of B cells was slightly increased. Significantly decreased natural killer cell activity (p = 0.02) of peripheral blood lymphocytes was observed. Our results indicate that asymptomatic patients with hemophilia may have early evidence of immunodeficiency.     

Generation of killer lymphocytes in vitro against human autologous leukemia cells with leukemic blasts and BCG extract

Summary Acute lymphoblastic leukemia patients under 18 years of age were studied to determine the ability of their remission lymphocytes to kill autologous leukemic blasts (ALB) following in vitro sensitization with their leukemic cells and/or soluble extract of BCG (BCG-SE). Remission lymphocytes, when cultured together with the mitomycin-treated ALB, became significantly lytic for ALB but not for autologous remission lymphocytes. The ALB were usually immunogenic at low concentrations and no cytotoxic lymphocytes were generated at a ratio of 1:1 of responding lymphocytes to stimulating leukemic cells. T-leukemic cells appeared to immunize more effectively than null-cell leukemic cells. In some cases, when ALB alone could not generate killer lymphocytes (KL) the combination of ALB and BCG-SE induced more intense cytotoxicity than was induced by BCG-SE alone. In a few other cases, the addition of BCG-SE to mixed lymphocyte leukemic cell cultures potentiated the immunization of lymphocytes by leukemic cells. Inhibition of cytotoxicity induction was noted in one case when remission lymphocytes were cultured together with ALB and BCG-SE. Leukemic cellssensitized lymphocytes from some cancer patients and normal persons were cytotoxic to several but not all patients' leukemic cells tested. Nylon wool-nonadherent, non-E-rosette-forming, and E-rosette-forming cells became cytotoxic following in vitro stimulation with autologous leukemic cells.     

Lack of effect of levamisole on the immune function in melanoma patients

Summary The immune function of 24 patients with local and regional melanoma receiving levamisole as adjuvant treatment was followed during 12–36 months. There were only minimal changes in the number of the peripheral blood lymphocytes and of E-rosette-forming cells. There was no essential improvement in the responses of peripheral blood lymphocytes to phytohemagglutinin (PHA), concanavalin A (Con A), and purified protein derivative of tuberculin (PPD) during levamisole treatment. Only eight of 24 patients were without recurrence during the observation time of 12–36 months. Our results indicate that adjuvant levamisole therapy in melanoma patients does not exert significant beneficial effects.     

Inhibition of rosette formation by antithymocyte globulin

Summary The peripheral blood leukocytes from 29 patients with adenocarcinoma of the colon were studied sequentially for the T-cell level, the rosette-inhibition titer of antithymocyte globulin and the blastogenic response to PHA and Con A to evaluate the T-cell immunocompetence. The level of E-rosette-forming cells and the blastogenic response did not reflect the immunocompetence of the T-cell population. Fluctuations in the rosette-inhibition titer of antithymocyte globulin (ATG) were observed; an increased ATG titer indicated an unfavorable clinical course, while a decreased ATG titer was observed with patients who had a favorable clinical course. The rosette-inhibition titer with antithymocyte globulin was observed to be an important indicator of competent T cells in patients with colorectal cancer.Supported by the Physicians' Medical Education and Research Foundation and General Research Supports FundsAbbreviations used that are not spelled out in the text are: AET 2-aminoethylisothiouronium bromide; ATG Antithymocyte globulin; ALG Antilymphocyte globulin; E-RFC Erythrocyte rosette-forming cells; PHA Phytohemagglutinin; Con A Concanavalin A; E Sheep erythrocyte     

The mixed lymphocyte reaction (MLR) stimulatory capacity of human lymphocyte subpopulations.

Using various cell separation techniques and combinations of these, suspensions were obtained highly enriched or depleted with respect to their content of E-rosette-forming T cells, Ig-bearing B cells, Fc-receptor-bearing cells, or monocytes. These purified populations were tested for their capacity to stimulate allogeneic cells in a mixed lymphocyte reaction (MLR). It could be demonstrated that the Ig-bearing B cells provide the strongest stimulus in the MLR.     

Cellular immunity factors of tick-borne typhus patients in northern Asia

In patients with North Asian tick-borne typhus fever the deficiency of the T-system of immunity, most pronounced in the acute period of the disease, is observed, which is manifested by low values of lymphocyte blast-transformation and rosette-formation. A decrease in the number of active E-rosette-forming cells depends on the severity of the disease. The shifts in the T-system of immunity, appearing in patients with North Asian tick-borne typhus fever, are compensated for comparatively quickly, the normalization of the functional activity of T-lymphocytes occurring sooner than the restoration of their number. The specific dynamics of the characteristics of cell-mediated immunity in patients with North Asian tick-borne typhus fever is accompanied by a comparatively mild course of the disease and its favorable outcome.     

A unique antigen (sigma) on sézary cells

Summary Lymphapheresis was performed on a patient with Sézary syndrome. The Sézary cells were purified by removing E-rosette-forming and Fc receptor-bearing cells. Antiserum against these purified Sézary cells was raised in rabbits. This antiserum had cytotoxicity against Sézary cells as well as against normal peripheral blood lymphocytes. Absorption was carried out with chronic lymphocytic leukemia (CLL) and normal lymphocytes. The absorbed antiserum maintained cytotoxicity against Sézary cells but lost cytotoxicity against CLL and normal peripheral blood lymphocytes.Indirect immunofluorescence assay showed that the antiserum reacted against purified Sézary cells and a high percentage (66%) of peripheral blood mononuclear cells from five patients with Sézary syndrome. It also reacted against 5.7% of normal lymphocytes, 8% of CLL cells, 5% of the lymphocytes from a patient who had undergone splenectomy, 2% of lymphocytes from a patient with multiple myeloma, 5% of lymphocytes from a hairy cell leukemia patient, and 1% of acute lymphocytic leukemia cells (T cell). The antiserum did not react against thymocytes but reacted against 34.6% of the bone marrow lymphocytes. This unique marker was designated as sigma () antigen. It was suggested that Sézary syndrome may represent proliferation or malignant transformation of normally present antigen-positive lymphocytes.     

Syntheses of two deacetyl-thymosin alpha(1) analogues and their effects on low E-rosette-forming lymphocytes of uraemic patients

Two deacetyl-thymosin alpha(1) analogues containing Phe (4Br) or D-Phe (4Br) residue-[D-Phe(4Br)(21)]deacetyl-thymosin alpha(1) and [Phe(4Br)(21)]deacetyl-thymosin alpha(1), respectively-were synthesized by the manual solid-phase method and their immunological effects on the low E-rosette-forming lymphocytes of uraemic patients were examined. One of the synthetic analogues, [Phe(4Br)(21)deacetyl-thymosin alpha(1), demonstrated a restorative effect on the low E-rosette-forming lymphocytes of uraemic patients, which was stronger than that of deacetyl-thymosin alpha(1), but the other analogue, [D-Phe(4Br)(21)]deacetyl-thymosin alpha(1), showed no restorative effect under the same conditions.     

Atypical T cells in rheumatoid synovial membranes

Atypical mononuclear cells (AMC) with Sézary-cell-like morphology were demonstrated to be present in rheumatoid synovial membranes. These cells have exclusively T cell membrane characteristics, like identical cells previously found in rheumatoid synovival fluids. AMC were predominantly present in transitional areas between nodular lymphocytic aggregates and plasma cell rich areas. In ultrathin sections AMC accounted for approximately 30% of all lymphocyte cells; in studies of cells isolated from rheumatoid synovial membranes AMC comprised 30--40% of the E-rosette-forming cells. The hypothesis that these cells represent reactive T cells is discussed.     

Evaluation of three E-rosette assays in melanoma patients

Summary The percentage of E-rosette-forming cells (E-RFC) was determined repeatedly in the peripheral blood of 43 patients with malignant melanoma. Three methods of E-rosette assaying were used: total E-rosette test, active E-rosette test, and 29° C E-rosette assay.Depressed E-RFC values were found in fewer assays than normal values. The mean values of E-RFC and the proportion of depressed E-RFC in patients in stage I did not differ from the values in healthy control persons whatever method of assay was used.The frequency of depressed E-RFC values was higher in advanced stages. Significant differences were demonstrated between stage I and stages II and III combined in the values for total and for active E-rosettes (P<0.01).The active E-rosette test and the 29° C E-rosette assay gave no more positive results (i.e., depressed E-RFC values) than the total E-rosette test in melanoma patients.     

Comparative analysis of the heterogeneity of mononuclear cells present in adult and cord blood by simultaneous examination of multiple phenotypic characteristics

In an attempt to better relate specific membrane characteristics of human adult and cord blood lymphocytes to specific functional activities, the phenotypic differences that exist in these two populations have been examined. Cord blood cells have considerably more spontaneous suppressor cell activity than adult cells. A technique that allows cells to be examined simultaneously for their ability to ingest latex beads, react with specific monoclonal antisera, bind sheep erythrocytes, or react with the Fc portion of IgG was used. As well as assessing fresh populations, phenotypic changes that occur when such cells are held in culture or stimulated with phytohemagglutinin for 3 days were sought. Many differences were found when comparing these mononuclear populations. These included the observations that 12% of adult and 9% of cord blood E-rosette-forming cells ingest latex beads and that 9% of OKT3 reactive cells in both populations did not form E rosettes. In cord blood 58% of T cells that bind OKT8 do not form E rosettes. A similar percentage of cord blood T8-positive cells express a receptor for Fc gamma, such cells being very uncommon in adult blood. Four "monocyte" subpopulations were identified in both samples. One such population (an OKM1- and Fc gamma-positive, nonphagocytic cell) was three times more common in cord blood. In cord blood some OKM1-positive cells also appeared to be simultaneously OKT8 positive. These phenotypic variations forward populations that may be candidates responsible for the functional differences noted in vitro.     

Human lymphocyte cytotoxicity against mumps virus-infected target cells. Requirement for non-T cells.

Subpopulations of human lymphocytes were tested for their capacity to kill mumps virus-infected target cells in a 51-chromium release asaay. Using two different cell fractionation techniques, lymphocytes were fractionated into T cell-enriched (primarily T cells) and T cell-depleted (primarily B cells) subpopulations. Filtration of lymphocytes through columns coated with human immunoglobulin and rabbit anti-human-immunoglobulin (Ig-anti-Ig) rendered the resulting T-cell preparation inactive as effector cells against target cells carrying mumps virus antigens. In the second technique, lymphocytes were fractionated by centrifugation into two fractions according to their ability to form spontaneous rosettes with sheep erythrocytes (E). The E-rosette-forming population (primarily T cells) was shown to lack cytotoxic activity against mumps virus-infected target cells. This activity was present in the nonrosetting population. The results suggest that the effector cells involved in this cytotoxic system are of a non-T variety.     

Immune depression--a possible cause of the unfavorable course of pneumonia in chronic alcoholics

In 27 patients, suffering with chronic alcoholism and hospitalized for pulmonary diseases in the Clinic of Pulmonology and Phthisiology, the following immunological characteristics were checked up: the functional activity of polymorphonuclear leukocytes and in 12 patients also that of alveolar macrophages were evaluated on the basis of the study of the phagocytic index and the phagocytic number, myeloperoxidase and the nitro blue tetrazolium test; the levels of serum IgG, IgA and IgM, the titer of the complement, E-rosette-forming cells (active and total) were also evaluated; the deficiency of cell-mediated immune response was determined by means of intradermal tests with the use of P.P.D., phytohemagglutinin, candidin, trichophytin. In all these investigations the depression of the functional activity of polymorphonuclear leukocytes and alveolar macrophages, dysimmunoglobulinemia, the increased level of circulating immune complexes and the suppression of cell-mediated immunity characteristics were revealed in the patients. Frequent infections and the severe course of bacterial and viral infections observed in such patients can be probably attributed to deficient cell-mediated immune response and to disturbances in phagocytosis.     

Early and total E-rosette-forming lymphocytes in furunculosis patients

In 228 furunculosis patients a decrease in the level of total and early E-rosette-forming lymphocytes (E-RFL) was observed. Early E-RFL underwent greater changes, as these lymphocytes, being more closely related to the clinical picture of furunculosis, better reflected the severity of the clinical course of this infection.     

Inhibitory activity of interleukin 2-activated lymphocytes on human granulocyte precursors (CFU-g)

Interleukin 2-activated lymphocytes (lymphokine-activated killer [LAK] cells) cultured from 2 to 14 days were added to the cultures of granulocyte precursors (CFU-g). The LAK cells inhibited colony formation of granulocyte precursors; LAK cells cultured for five days showed the strongest inhibitory activity on colony formation. The presence of cell-to-cell interaction between LAK cells and bone marrow mononuclear cells (BMNC) in CFU-g assays emphasized the LAK cell-derived colony inhibitory activity (LAK-CIA), but cell-to-cell interaction was not always a requirement for LAK-CIA, since LAK cells were also found to inhibit colony formation without such interaction. This report shows that LAK cells can inhibit in vitro colony formation of granulocyte precursors. We therefore concluded that the observed CIA is caused by soluble factor(s) derived from LAK cells, and that E-rosette-forming cells are manifesting LAK-CIA.     

The suppression of mitogen responses associated with resistance to experimental autoimmune encephalomyelitis requires adherent and T cells

Resistance to experimental autoimmune encephalomyelitis (EAE) in Hartley guinea pigs has previously been reported to be associated with disease-specific antigen-induced suppression of mitogen responses in vitro. The present studies were initiated to investigate the requirement for different cell populations in this suppression. Intact and adherent-cell-depleted cultures of spleen cells from experimental and control animals were incubated with myelin basic protein (MBP), the major antigen of EAE, with the T-cell mitogen concanavalin A (Con A) alone or with Con A in the presence of MBP. In agreement with previous studies, MBP-induced suppression of the Con A response was observed only in cultures derived from resistant animals. In addition, it was observed that this suppression was abrogated by depletion of adherent cells. When cells from resistant and susceptible animals were mixed, suppression occurred only in the presence of nonadherent cells from resistant guinea pigs. Adherent cells from either resistant or susceptible animals functioned equally well. Cultures of purified E-rosette-forming cells (E+) from resistant animals (i.e., T cells) showed no suppression. Similarly, cells from these same animals which were depleted of E+ cells (i.e., non-T cells) did not demonstrate suppression in vitro. Upon reconstitution of spleen cell populations from resistant guinea pigs by mixing E+ and E- cells, suppression was restored. These experiments show that this model of suppression in vitro requires adherent cells as well as T cells and suggests that antigen-induced suppression of mitogen responses is dependent upon a cell-mediated immunologic mechanism.     

Leukocyte-adherence inhibition: a specific assay of cell-mediated immunity dependent on lymphokine-mediated collaboration between T lymphocytes.

The leukocyte-adherence inhibition (LAI) assay was studied to determine its immunologic relevance and identify the cell populations on which it depends. Two systems were employed: peripheral blood leukocytes from humans immunized with KLH, and lymph node cells from rats immunized with DNP-BCG. In both cases, LAI responses appeared about 3 to 4 days after immunization, reached a peak about 3 to 4 weeks later, and diminished thereafter. Reimmunization resulted in a booster-like response. LAI analysis in both systems showed dose-response dependency. Responses could be elicited only with the immunizing antigen. Virtual depletion of phagocytic cells had no effect on the response. E-rosette-forming cells gave an excellent response to KLH and also produced an active supernatant (lymphokine). Cells not forming spontaneous E-rosettes were inactive and could not produce active supernatants. Only those nonimmune cells that formed E-rosettes could respond to active supernatants. Thus, the LAI response is a specific indicator of cell-mediated immunity. T lymphocytes probably are required both at the antigen-reactive stage and at the stage of responding to the T cell-dependent lymphokine.     

Lymphocyte subpopulations in sensitization and specific immunotherapy in an experiment. Lymphocyte subpopulations in sensitization to staphylococci, Artemisia pollen and their combinations

The distribution of T- and B-lymphocytes in the body of guinea pigs was studied in different groups of the animals. As shown in this study, in delayed hypersensitivity to staphylococci the number of PE- and E-rosette-forming cells increased in the blood, the spleen, and the lymph nodes and decreased in the thymus; the number of EA- and EAC-rosette-forming cells decreased in the bone marrow and the spleen, the number of T gamma-suppressors decreased in the bone marrow and the distant lymph node. Immediate hypersensitivity to tarragon pollen induced the general increase of the content of T- and B-lymphocytes; the number of T gamma-cells decreased in the thymus, the bone marrow, and the lymph nodes and increased in the spleen. The characteristic features of combined microbial-pollen sensitization were the high content of B-cells in all lymphoid organs (except the thymus), a low level of T-lymphocytes in the blood and the peripheral lymphoid organs, the decreased number of T gamma-cells in most of the immunogenetic organs.     

Human natural cytotoxic lymphocytes: definition by a monoclonal antibody of a subset which kills an anchorage-dependent target cell line but not the K-562 cell line

A monoclonal mouse antibody (HNC-1A3) which defines a subset of human lymphocytes with natural cytotoxic activity was produced and studied. HNC-1A3+ cells represent 12 +/- 3% of peripheral blood mononuclear leukocytes. When sorted out using a fluorescence-activated cell sorter, they consist of 60% small lymphocytes, 35% large (predominantly agranular) lymphocytes, and 5% monocytes. They contain 30 +/- 6% E-rosette-forming cells, 6 +/- 1% OKT4+ cells, 17 +/- 6% OKT8+ cells and less than 2% OKT10+ or Leu-7 (HNK-1)+ cells. They are responsible for most of the natural cytotoxic activity against the MA-160 prostatic adenoma cell line but mediate an insignificant amount of cytotoxicity against the NK prototype target K562 cell line. Conversely, Leu-7+ cells which mediate most NK activity against K562 are weakly active against MA-160. Our data suggest a heterogeneity among leukocytes mediating natural cytotoxicity, with restricted specificities for the recognition sites on target cells.