Lymphocyte Blast Transformation Research Articles

Effect of adamantane amino derivatives on the immune response

In the previous paper [ 1 ] we reported that remantadine and deutiphorin exhibit moderate immunosuppressive activity and are capable of inhibiting the cytotoxicity of natural killer cells and the reaction of lymphocyte blast-transformation in the mononuclear fraction of peripheral blood l~om healthy donors. Earlier, Clark et al. [2] reported similar properties for l-aminoadamantane. In our work [1], it was established that the immunosuppressive activity of the main amino derivatives of adamantane is due to their ability to inhibit activation of Ca2+-phospholipid dependent protein kinase (protein kinase C), and it was suggested that this property may also account for the antiviral activity of these compounds. The ability ofadamantane amino derivatives, as anti-influenza preparations, to inhibit the activation of protein kinase C (thus suppressing the lymphocyte blast-transformation and the functional activity of natural killer cells) suggest that these derivatives may also be capable of affecting the humoral immune response and the post-infection response related to the formation of immune memory cells. Both these processes include the interaction of various T and B cell subclasses, whereby the activated T-lymphocytes belonging to different subclasses may activate or suppress the functional activity of the B cells [3, 4]. As is known, the activation of Tand B-lymphocytes is related to the activation of protein kinase C [5, 6]. In this connection, we have studied the effect of remantadine, 1-aminoadamantane, and deutiphorin on the formation of humoral immunity to bacterial antigen, a living antituberculous BCG vaccine, in CBA line mice. At the same time, we have studied the effect of these compounds on the delayed hypersensitivity that develops simultaneously with the formation of antibodyproducing cells and reflects the T-cell immunity reaction.

Long-term enhancement of Na,K-ATPase pump during blasttransformation of human lymphocytes is controlled first by translational, then by transcriptional mechanisms

The transition of phytohemagglutinin-activated human lymphocytes from resting state to proliferation is accompanied by a long-term increase in ouabain-sensitive Rb(K) influx which is closely related to a cyclosporin A-sensitive step of G0/G1/S progression. At least two distinct phases of the up-regulation of cation pump has been revealed: the initial stage (5–20 h) which is cycloheximide-inhibitable and actinomycin D (α-amanitin)-unaffected, and the later stage (after 20 h) which is cycloheximide- and actinomycin D (α-amanitin)-inhibitable. Thus, the enhanced Na,K-ATPase pump during the cell progression from quiescence to proliferation is controlled both at translational and transcriptional levels.

Cellular immunity to the P6 outer membrane protein of nontypeable Haemophilus influenzae

Cellular immunity to nontypeable Haemophilus influenzae in a population of 10 healthy, immune adults was determined by measuring lymphocyte blast transformation and antibody secretion in response to the P6 outer membrane protein. P6 (200 microliters/ml) induced lymphocyte blast transformation that peaked on day 10 of incubation. The peak induction of antibody-secreting cells occurred on day 8 of incubation. In comparison with the response to tetanus toxoid stimulation, the peak lymphocyte blast transformation response to P6 was reduced (mean counts per minute +/- standard error of the mean [SEM], 3,457 +/- 503 versus 9,414 +/- 1,464; P = 0.0051) and delayed (mean days +/- SEM, 10.3 +/- 0.4 versus 8.4 +/- 0.5; P = 0.0169); however, P6 was a better stimulus of antibody secretion from lymphocytes, particularly antibody of the immunoglobulin M (IgM) class (mean peak numbers of antibody-secreting cells per 10(5) peripheral blood mononuclear cells +/- SEM: IgG, 85 +/- 29 versus 42 +/- 16 [P = 0.0469]; IgM, 81 +/- 20 versus 25 +/- 7 [P = 0.0125]; IgA, 24 +/- 8 versus 16 +/- 6 [P = 0.0526]). Thus, lymphocytes from immune individuals recognize P6 of nontypeable H. influenzae as an immunogen. These data provide a basis for future studies with otitis-prone children who fail to develop a normal antibody response to P6 antigen (N. Yamanaka and H. Faden, J. Pediatr. 122:212-218, 1993).

Immunological consequences of burn injury in children.

The authors investigated immunological changes after burn injury in children. Relevant findings were observed in the following immunological factors: phagocytose activity, T-lymphocytes, active T-lymphocytes, test of blastic transformation of lymphocytes and immunoglobulins. The findings confirm from an immunological point of view the requirement of early necrectomy (at the latest on day 4 or 5 after injury). Later necrectomy cannot prevent immunosuppression after burn injury but itself causes deterioration of the immunological status of patient. Adequate therapy can improve the immunological status of burned patients.

Immunosuppressive and Antiproliferative Effects of Somatostatin Analog SMS 201–995

The effects of long acting somatostatin analog SMS 201-995 were examined in vivo on: 1) lymphoid morphostasis and functional reactivity of cells obtained from SMS treated donors, 2) on humoral, and 3) cellular type of immunity; and in vitro on: 1) blastic transformation of lymphocytes stimulated by activators of different transmembrane pathways (CD2 by PHA and CD3/TCR by anti-CD3 monoclonal antibody and by allogeneic cells) and 2) on growth and secretory activity of several hybridoma cell lines. The data have shown that SMS in vivo decreases the proportion of CD4+, CD5+ and Ig+ cells in spleen. The reactivity of these cells to Con A was suppressed, but their spontaneous blastic transformation was increased. SMS suppressed also the plaque forming cells generation and proliferation of cells in popliteal lymph nodes during the local host versus graft reaction. The former immunosuppression was abrogated with the use of growth hormone, while in the latter, the time dependent changes in spleen composition were also noticed. The data obtained in vitro revealed that SMS may inhibit only the CD2-induced blastogenesis (in early and late interval after the use of PHA). SMS inhibited also the spontaneous growth and/or secretion of antibodies in some hybridoma cell lines.

薬剤性リンパ球幼若化試験における各種薬剤希釈濃度の比較

薬剤性リンパ球幼若化試験における各種薬剤希釈濃度の比較

Blast transformation of lymphocytes and the activity of natural killer cells in the presence of γ-globulinin vitro

The cytotoxic activity of natural killer cells against3H-uridine-labeled target cells (human erythromyeloleukosis cells K-562) and the intensity of spontaneous blast transformation are studiedin vitro in the presence of human serum γ-globulin. It is shown that spontaneous blast transformation is 49–51% due to the presence of aggregated γ-globulin, while the aggregate-free γ-globulin fraction does not induce this reaction. The cytotoxic activity of natural killer cellsin vitro declines in the presence of native γ-globulin, which is related to the influence of aggregated γ-globulin, the intensity of whose formation may increase upon a manyfold decrease in the γ-globulin content of the preparation.

Effect of autologous sera on mitogenic stimulation of peripheral blood lymphocytes of mini-pigs chronically intoxicated with alcohol

There are no statistically significant differences in the stimulation index for purified lymphocytes of control mini-pigs and mini-pigs with chronic alcohol intoxication. Autologous sera of control and experimental animals strongly suppress mitogen-induced blast transformation of lymphocytes without death of these cells. There are no statistically significant intergroup differences in the absolute number (per mm3) of T lymphocytes in peripheral blood.

A Phase I Study of Interferon-α2b in Combination with Interleukin-2 in Patients with Human Immunodeficiency Virus Infection

Interferon-alpha (IFN-alpha) can inhibit human immunodeficiency virus (HIV-1) replication and is effective in treating Kaposi's sarcoma; interleukin-2 (IL-2) can increase circulating lymphocytes in HIV-1-infected patients. The safety of combination treatment with recombinant (r)IFN-alpha 2b and IL-2 was evaluated in HIV-1-infected patients with > 200 CD4+ T cells/mm3. A maximal tolerated dose of rIFN-alpha 2b was determined for 17 patients; then they received in combination 3, 6, or 12 x 10(6) IU/day rIL-2, given intravenously over 21 days. Twelve patients ultimately received the combination, 9 for the full 21 days. Significant toxicities included flu-like symptoms, anemia, transaminemia, and depression. Transient increases in CD4+ T cell percentages and spontaneous lymphocyte blast transformation were observed. Quantitative microcultures demonstrate a decline in HIV titers in patients receiving rIFN-alpha 2b (5/9) with a further decline on addition of rIL-2 (7/9). In summary, continuous rIL-2 at 6 x 10(6) IU/day in combination with rIFN-alpha 2b was reasonably tolerated and provided preliminary evidence of immunomodulatory and antiviral activity.

Immune changes during long-duration missions

Immune changes during space flights in excess of two weeks have been almost exclusively studied by Russian investigators. Most of these studies have compared postflight values with those obtained before flight. In a very few cases, analyses have also been attempted during flight or with samples collected during flight. Studies of cosmonauts during spaceflight have shown that IgG levels were unchanged, whereas IgA and IgM levels were sometimes increased. Additionally, inflight delayed type hypersensitivity testing demonstrated a decrease below the warning level in 1/3 of the cosmonauts tested. Pre- vs. postflight analyses have often revealed a postflight decrease in: PHA-triggered lymphocyte blast transformation; the proliferation index of T-lymphocytes in the xenogeneic graft versus host reaction; the mitogen-induced production of interleukin-2; the presence of certain leukocyte sub sets; and cytotoxic activity of natural killer cells. Other factors that either did not change, or changed in an apparently random manner after flight included: production of alpha and gamma interferon; autoimmune tests; and globulin classes.

Cell-mediated immunity to Chlamydia pneumoniae measured as lymphocyte blast transformation in vitro

The purpose of the present study was to analyze Chlamydia pneumoniae-induced, antigen-specific, cell-mediated immunity. Peripheral blood mononuclear cells of four persons infected with C. pneumoniae Kajaani 6 and 17 healthy volunteers were stimulated with antigen composed of whole elementary bodies of C. pneumoniae Kajaani 6 (CP-Ag). Definitive antigen-specific lymphoproliferation (LP) responses were developed after recent infection. The LP responses of healthy people to CP-Ag varied considerably. There was no clear correlation between LP responses to CP-Ag and those to an antigen prepared from Chlamydia trachomatis serotype L2 (r > or = 0.50, P < 0.1). A larger study is required to demonstrate whether the LP responses to CP-Ag can be used for the diagnosis of C. pneumoniae infection.

Immunomodulating Effects of Various Phenothiazines and Related Tricyclic Compounds in Vitro and in Vivo

A biotechnological approach to the immunomodulating ability of various phenothiazines and related tricyclic compounds in vitro and in vivo was made. The effect of these compounds was studied on various mononuclear leukocyte functions such as antibody-dependent cell-mediated cytotoxicity, blast transformation of lymphocytes by three mitogens, phytohemagglutinin, concanavalin A and pokeweed mitogen (PWM), rosette formation with erythrocytes and with antibody coated erythrocytes (EA), as well as on polymorphonuclear leukocyte functions including phagocytosis and chemiluminescence emission. Moreover, the ability of these compounds to modulate the bioluminescence emission capacity of Escherichia coli cloned with lux genes was measured. Also in vivo studies were made by measuring the number of haemolytic plaque forming cells in the spleen of mice after drug administration. The electrochemical structure of these tricyclic compounds was shown to be in good correlation with their immunomodulating activity on all the functions except EA rosette formation and on blast transformation by PWM showing the role of charge transfer complex formation in the drug-target cell interaction. These results suggest that the biological effectiveness of phenothiazines and related tricyclic compounds can be presumed from their electrochemical properties and a tricyclic compound with a given biological effect can be theoretically designed.

Influence of ganglioside GM3 and its breakdown products on lymphoblastic transformation and T‐suppressor activity

The influence of ganglioside GM3 and some of its breakdown products on phytohemagglutinin-induced blast transformation of human lymphocytes and concanavalin-A-induced T-suppressor activity was studied. The structures of two major hydrolysis products of GM3 were established by negative-ion fast-atom-bombardment mass spectrometry as neuraminyllactosylsphingosine (NeuLacSph) and neuraminyllactosylceramide (NeuLacCer). Both substances were shown to be potent inhibitors of mitogen-induced lymphoblastic transformation whereas their acetylation products NeuAcLacSphAc and GM3 did not affect the proliferative response of lymphocytes to phytohemagglutinin. On the other hand, only GM3 and NeuLacSph were able to enhance concanavalin-A-induced T-suppressor activity. On the basis of these data, it is suggested that the effects of GM3 and its breakdown products on lymphoblastic transformation and T-suppressor activity must rest on different mechanisms and that N-deacylation of GM3 appears to be an essential step in conversion of the ganglioside into an inhibitor of lymphocyte blast transformation.

Identification, Characterization, and Quantitation of Soluble HLA Antigens in the Circulation and Peritoneal Dialysate of Renal Patients

Human lymphocyte antigen (HLA) class I and class II antigens and beta 2 microglobulin (B2M) were identified in peritoneal dialysate (PD) and serum from patients with end-stage renal disease (ESRD) using monoclonal antibodies in an enzyme-linked immunoassay. The HLA class I and class II antigens each exhibited approximate molecular weights of 50,000 to 60,000 daltons by chromatography on Sepharose CL 6B. Class I antigens in serum and PD fluid were associated with B2M. Free B2M (Mr 11,500) also was detected in both sera and PD fluids. Unlike class I antigens, class II antigens were not found to have attached B2M. Class I and class II antigens eluted from 2-diethylaminoethanol ion exchange gradient columns at 0.07 mol/L (molar) phosphate buffer pH 7.2 and migrated with alpha 2-beta 1 mobility in agarose electrophoresis. Class I antigens were purified from ESRD patients' PD fluid by solid-phase immunoaffinity chromatography. Enzyme-linked immunoassay demonstrated that this purified protein was composed of a class I heavy chain and B2M. Class I allospecificity was confirmed by neutralization on known HLA typing antisera in a microcytotoxicity assay. Soluble HLA class I antigen preparations specifically inhibited blast transformation of responder lymphocytes in mixed lymphocyte culture reactions. Inhibition was dose dependent and ranged from 0% to 95%. The presence of soluble HLA antigens in body fluids may play an important part in the immunologic tolerance to self. This study demonstrates a ready source of large quantities of soluble HLA for detailed analysis.

Exsudative Reaktion nach Implantation einer künstlichen Linse

Results of clinical observation over post-operative period and degree of exsudative reaction in 240 patients (262 eyes) with implantation of intraocular lenses are analysed. Three types of postoperative course are distinguished: a reactive, postoperative with early exsudative reaction and postoperative with late exsudative reaction. Immunologic investigations (determination of percentage and absolute amount of T-lymphocytes in peripheral blood, reaction of blast-transformation of lymphocytes on phytohemagglutinin and on antigens of the lens, the choroid and the retina) performed in 184 patients with artiphakia allowed to make a conclusion that postoperative course with late exsudative reaction is dependent on preliminary sensibilization of the body to proteins of eye tissues and has immune nature. The treatment with indometacin proved to be effective and prevented development of late exsudative reaction in 40 patients with implantation of intraocular lenses.

Development of Francisella tularensis antigen responses measured as T-lymphocyte proliferation and cytokine production (tumor necrosis factor alpha, gamma interferon, and interleukin-2 and -4) during human tularemia

The lymphocyte immune reactivity of 12 tularemia patients to Francisella tularensis antigens prepared from the bacterial cell envelope was examined during a 14-week follow-up study. Lymphocyte blast transformation responses of peripheral blood mononuclear cells (PBMC) to different protein antigens appeared simultaneously 2 weeks after the first symptoms of tularemia, indicating that none of these antigens had any special role at the early phase of immunization. While the lymphocyte blast transformation responses of total lymphocytes to all bacterial antigens were negative in the week 1 samples, continuously growing F. tularensis-specific T-lymphocyte lines were obtained from PBMC at the same time, indicating that an immune response had already occurred. Later, the T-lymphocyte lines and lymphocyte blast transformation responses were similar. Lymphocyte activation among the PBMC was reflected in an increased number of HLA-DR antigen-expressing, CD4-positive T lymphocytes (CD4+ DR+). The mean secretion of soluble CD8 from F. tularemia antigen-stimulated PBMC increased 2 weeks after tularemia onset, but the mean number of CD8+ DR+ T lymphocytes did not vary during the study period and no correlation could be found between the soluble CD8 and number of CD8+ DR+ T lymphocytes. F. tularemia antigen-induced cytokine production was measured from the PBMC supernatants. High levels of tumor necrosis factor alpha were detected from the first week onwards. The highest levels of interleukin-2 and gamma interferon were recorded during the second and third weeks, respectively, after tularemia onset. Interleukin-4 could not be demonstrated in the lymphocyte supernatants.

Immune mechanisms in beryllium lung disease

The role of the immune system in the pathogenesis of beryllium lung disease has been suspected for years. The observation of cutaneous hypersensitivity to beryllium led to the development of the lymphocyte blast transformation test; the test clearly distinguishes between healthy subjects, who show little or no blast transformation response, and patients with beryllium lung disease, who demonstrate significant responses. The degree of blast transformation also correlates with the severity of the clinical disease. Animal studies have demonstrated the importance of histocompatibility antigens in development of the disease, and support the participation of cellular immune mechanisms.

Correlation of serum antigen and antibody concentration with clinical features in HIV infection.

Serum antigen and antibody values were studied in 164 infants and children infected perinatally with HIV. HIV antigens p17, p24, gp41, and gp120 were determined in sera by immunoblot and antigen capture assays. Lymphocyte blast transformation, serum immunoglobulins, and circulating immune complexes were also evaluated. Altogether 50 patients had HIV antigens measured: 31 (62%) patients had p17 antigen in the serum and 29 (58%) had p24 antigen present. In 19 (38%) and nine (18%) patients, respectively, gp120 and gp41 were detected. All four HIV antigens were detected in seven (14%) patients. There was a positive correlation between the concentration of each HIV sequential specimens were outcome. When sequential specimens were analysed, 120 (73%) patients had p24 antigen present. Patients with stage P2B and P2D (Centers for Disease Control classification) had the highest concentrations of p24 antigen with a mean of approximately 200 pg/ml. Altogether 70% of patients with a p24 antigen concentration of greater than 30 pg/ml eventually died or had severe clinical disease within six to 24 months. Infants under 15 months of age with a p24 antigen concentration as low as 5 pg/ml also did poorly. Increased immunoglobulins and decreases in mitogenic responses and absolute CD4+ lymphocyte counts were more prevalent in patients with raised p24 antigen. Raised concentrations of circulating immune complexes were seen in the symptomatic phase of the disease whereas in the terminal stage of the disease raised serum antigen and a decrease in circulating immune complexes and absolute CD4+ lymphocyte count were evident. Loss of p24 and/or p17 antibody as well as a decreasing ELISA optical density for HIV antibody also signalled progression of the disease.

Inhibition of Cluster Formation and Lymphocyte Proliferation by Anti-Fibronectin Antiserum

Dendritic cells (DC) confer concanavalin A responsiveness to accessory cell-depleted canine lymphocytes. DC-lymphocyte cluster formation is followed by blast transformation and proliferation of lymphocytes. We investigated whether fibronectin, which is known to be involved in cell:cell interactions, might also be involved in the interaction between canine DC and lymphocytes. The addition of rabbit anti-dog fibronectin antiserum to concanavalin A-stimulated peripheral blood mononuclear cells decreased cluster formation and reduced 3H-thymidine incorporation by 57-70%. Similarly, when cell fractions, enriched for DC, were pretreated with anti-fibronectin antiserum before being added back to accessory cell-depleted lymphocytes, cluster formation was reduced and lymphocyte proliferation after concanavalin A stimulation decreased by 18-46% as measured by 3H-thymidine uptake. Ferritin-conjugated anti-fibronectin antibody was bound to the surface of DC, primarily at the dendritic processes. We conclude that fibronectin, located on the surface of canine DC, participates in the accessory cell function of these cells.

Effect of ergot-alkaloids dihydroergotoxine and dihydroergosine on lymphocyte blastic transformation

Effect of ergot-alkaloids dihydroergotoxine and dihydroergosine on lymphocyte blastic transformation