Biomedical subjects

B Axelsson

Publications and source records attributed to B Axelsson.

At least 55 records · Page 3Linked to original sources

PubMed1989

Persistent superphosphorylation of leukosialin (CD43) in activated T cells and in tumour cell lines.

CD43 (leukosialin) is a highly sialylated, single-chain molecule expressed on most human leucocytes. Regulatory signals appear to be transduced through the molecule as suggested by the ability of anti-CD43 antibodies to induce aggregation and proliferation of T cells and to enhance B-cell proliferation and natural killer cell activity. Activation of protein kinases is an essential event in signal transduction. We were therefore interested to study whether CD43 may function as a substrate for protein kinases during mitogenic activation of lymphocytes. We show that CD43 was rapidly superphosphorylated (within minutes) on serine residues following addition of phorbol ester (PMA) to peripheral blood lymphocytes. PMA treatment of the cells was not followed by rapid down-regulation of CD43. Activation of the lymphocytes by concanavalin A or anti-CD3 antibodies (OKT-3) also resulted in superphosphorylation of CD43. However, the phosphorylation was delayed as compared to that induced by PMA and was detected 3-4 h after the addition of the reagents. A plateau was reached after 24-48 h of stimulation. Interestingly, the high level of phosphorylation of CD43 was maintained in long-term cultures of T cells activated by various means. Furthermore, CD43 was found to be constitutively superphosphorylated (on serine and tyrosine) in continuously growing cell lines of T, B, and non-lymphoid origin. Taken together, the results suggest that CD43 has an important role during both early and late phases of T-cell activation and that modulation of its biochemical properties by protein kinases may be associated with progression through the cell cycle and with cellular growth.

Antigens, CD↗

PubMed1988

Induction of aggregation and enhancement of proliferation and IL-2 secretion in human T cells by antibodies to CD43.

CD43 (large sialoglycoprotein) is a heavily glycosylated protein expressed on virtually all thymus-derived lymphocytes, on a subpopulation of B cells and on granulocytes. Recently, an anti-CD43 mAb (L10) was shown to induce proliferation in T cells comparable to that induced by anti-CD3. The L10 antibody was reported to react with both sialylated and desialylated CD43. In order to further elucidate the role of CD43 in various T cell functions we have studied the biologic properties of two other mAb (B1B6 and E11B, IgG1) directed against sialic acid-dependent epitopes on CD43. Addition of low amounts of antibody (5 to 10 ng/ml) to freshly isolated T cells or to T cell lines resulted in a rapid clustering of the cells. Fab fragments were also active albeit at a 10-fold higher concentration. Aggregation was dependent on active cell metabolism (inhibited by azide and at low temperatures), on the presence of divalent cations (Mg2+) and was inhibited by antibodies to CD18 but not by antibodies to CD11a (leukocyte function-associated Ag-1 alpha). B1B6 and E11B were poorly mitogenic when added alone in soluble form to PBL or to T cells. However, supernatants from cultures of PBL treated with B1B6 for 2 days contained IL-2 activity. No increase in the number of CD25+ cells was seen during the same period. Exogenously added IL-2 did not synergize with B1B6 or E11B in activation of PBL, whereas proliferation was significantly increased by the addition of the antibodies to activation systems with low endogenous production of IL-2 (PMA or soluble anti-CD3). The anti-CD43 antibodies amplified T cell proliferative responses induced by Con A or leukoagglutinin from Phaseolus vulgaris. F(ab')2 fragments enhanced proliferation significantly better than Fab fragments suggesting that cross-linking of CD43 molecules was an essential features of the amplifying signal. Compared with cultures activated by Con A alone, an increased number of CD25+ cells and of blast cells as well as an increased IL-2 production was observed in cultures activated by B1B6-Con A. The results indicate that regulatory signals, which may function to modify homo- or heterotypic T cell adhesion as well as autocrine production of IL-2, can be transduced through CD43.

Antibodies, Monoclonal↗

PubMed1988

Organ distribution of Adriamycin after intravesical instillation with or without Tween 80 in the rat.

The absorption of Adriamycin (ADM) into the systemic circulation and into different organs including the urinary bladder was investigated in the rat after intravesical instillation of ADM with or without a surface active detergent, Tween 80. The postinstillation plasma concentration of ADM increased significantly with increasing dose of the drug. Even though the leakage of ADM into the systemic circulation and into extravesical organs in general was slight a significant increase was observed after addition of Tween 80. The uptake of ADM into the urinary bladder wall was also significantly enhanced by Tween 80 thus the role of this agent in conjunction with intravesical chemotherapy should be further investigated.

Absorption↗

PubMed1988

Induction of CD43 expression during activation and terminal differentiation of human B cells.

Only a small population (25-30%) of human peripheral blood B lymphocytes expresses large sialoglycoprotein (LSGP) (CD43). However, in the presence of autologous T cells and pokeweed mitogen (PWM) a majority (50-90%) of the immunoglobulin-producing cells (cIg+ cells) that develop from these B cells express CD43 is detected with anti-CD43 monoclonal antibodies (MoAb) B1B6, and the proportion of CD43+cIg+ cells increases with time of culture. Furthermore, a relatively larger proportion (60-80%) of the IgG-producing cIg+ cells are CD43+ compared with IgM-containing cIg+ cells (30-50%). In human tonsils, significantly more CD43+ cells (35%) are found in the in vivo-activated fraction of B cells than in the fraction of resting B cells (5%). A majority of the cIg+ cells that develop from the resting or the in vivo-activated tonsillar B cells in a PWM-induced B-cell differentiation system are CD43+ (80-100%). Furthermore, tonsillar B cells depleted of CD43+ cells give rise to cIg+ cells, of which the majority are CD43+, and the proportion of such cells increases with time of culture (60-90%). Taken together, these results indicate that LSGP belongs to a group of B-cell membrane molecules that are induced and upregulated upon activation and differentiation.

Antigens, CD↗

PubMed1988

Characterization of human CD4+ T-cell clones that secrete helper factor(s) for B-cell proliferation and maturation.

Human peripheral blood lymphocytes (PBL) were activated with K46M, a m mitogenic monoclonal antibody against La-reactive T lymphocyte surface structures. The cultures were expanded in the presence of interleukin 2 (IL-2). After 1 month of culture, the activated T cells were cloned by limiting dilution at 0.5 cells/well. Five clones with the CD3+CD4+ phenotype and one clone with the CD3+CD8+ phenotype were obtained. The CD3+CD8+ clone (K99) displayed a strong major histocompatibility complex (MHC)-unrestricted cytolytic activity against MOLT-4 and a weaker reactivity against the bladder tumour cell lines T24 and RT4. The natural killer (NK)-susceptible K562 cells were not lysed. Two of the CD3+CD4+ clones (K91 and K914) showed a helper activity in pokeweed mitogen (PWM)-induced IgG production by B cells. These cells differed in the expression of CD45R and CDw29 antigens, as defined by the monoclonal antibodies 2H4 or D10D11 and 4B4. When stimulated with PWM for 48 or 72 h, clone K91 and an additional CD4-positive clone (K913) secreted a factor into the supernatants which helped B cells to produce IgG. The K913 supernatant also induced some IgM production. The supernatant obtained after similar stimulation of K914 cells was inactive. None of these supernatants induced B cells to proliferate when tested together with phorbol myristate acetate (PMA). However, when K91 and K914 cells were activated with phytohaemagglutinin (PHA) for 48 or 72 h, the supernatant from K91 was strongly helpful in B-cell proliferation, whereas the supernatant from K914 cultures was only moderately active. In conclusion, we have established human T helper clones that release different factors supporting either B-cell proliferation or maturation when stimulated with PWM or PHA.

Antigens, CD↗

PubMed1988

Studies on the role of lymphocyte function-associated antigen 1 (LFA-1) in T cell activation.

The effect of polyclonal (anti alpha and beta chain) and monoclonal (anti alpha-chain) antibodies against lymphocyte function-associated antigen 1 (LFA-1) on T cell activation was studied. When added at the beginning of activation but not after 24 h or later the antibodies as well as the F(ab')2 or Fab fragments of polyclonal antibodies inhibited concanavalin A (Con A)-induced proliferation, interleukin 2 (IL-2) production, and the expression of receptors for IL-2 and transferrin. The inhibitory effect reached a maximum at the same time as optimal proliferation (72 h). Inhibition of proliferation lasted for 5 days or longer, although IL-2 production was only inhibited during the first 48 h of culture. Receptors for IL-2 and transferrin were re-expressed to the original level after 3 days of activation. Addition of external IL-2 at the beginning of the anti-LFA-1 containing culture prevented the inhibition of IL-2 receptor expression, while inhibition of transferrin receptor expression was unaffected, supporting the conclusion that the expression of these two receptors is regulated by partially independent signals. The polyclonal and monoclonal anti-LFA-1 antibodies also inhibited phorbol ester (PMA)-dependent OKT3 activation of highly purified T cells. The results suggest that the LFA-1 antibodies block an early step in the reactions necessary for IL-2 production, and that the LFA-1 molecule participates not only in T cell-accessory cell interaction but also in T-T interaction during the early phases of the activation process.

Antibodies, Monoclonal↗

PubMed1988

Enhancement of human spontaneous cell-mediated cytotoxicity by a monoclonal antibody against the large sialoglycoprotein (CD 43) on peripheral blood lymphocytes.

A murine monoclonal antibody, MoAb B1B6 (IgG1 chi), which recognizes the large sialoglycoprotein (LSGP) on human peripheral blood lymphocytes (PBL) effectively enhanced the spontaneous cytotoxicity of these cells against the natural killer (NK)-sensitive target cells K562 and Molt-4. Whereas preincubation of the lymphocytes with MoAb B1B6 resulted in increased cytotoxicity, preincubation of the target cells had no effect, indicating that the MoAb amplified cytotoxicity at the effector cell level. Kinetic analysis of the data revealed no differences between the control and the MoAb-treated lymphocytes with regard to Vmax, usually considered to reflect the overall lytic potential of the cells. The slopes of the saturation curves, however, differed significantly for the two cell populations, indicating a substantial increment in the activity of the MoAb-treated cells. When studied at the single cell level and with K562 as targets, treatment of PBL with the MoAb resulted in the recruitment of new effector lymphocytes from the pool of non-binding cells. In contrast, when Molt-4 cells were employed as targets, no additional effector cells were recruited. These results indicate that the enhanced cytotoxicity induced by MoAb B1B6 is the result of either recruitment of new effector lymphocytes or of an increased recycling capacity of preexisting effector cells. Together with previous observations, these findings support the conclusion that LSGP belongs to the set of surface molecules which regulate human lymphocyte activation.

Antibodies, Monoclonal↗

PubMed1987

Absorption of Adriamycin into the systemic circulation after intravesical instillation with or without Tween 80 in the rat.

The absorption of Adriamycin (ADM) into the systemic circulation after intravesical administration, using an experimental model in the rat, was closely related to the volume of the instilled solution and the concentration of the drug. The post instillation plasma concentration of ADM was increased after addition of Tween 80 to the instilled solution but the magnitude of the absorption was still discreet and, from a clinical point of view, negligible.

Absorption↗

PubMed1987

Functional characterization of human B cells carrying the lymphocyte large sialoglycoprotein gp150.

Human B cell-enriched populations were prepared from buffy coats of healthy donors. By means of affinity chromatography, the B cells were separated into two fractions, one enriched in and the other depleted of cells expressing gp150, the large sialoglycoprotein of lymphocytes. In the presence of autologous T cells, monocytes and pokeweed mitogen B cell populations enriched for gp150+ cells gave rise to significantly more plasma cells (cIg+ cells) and secreted significantly more IgG than gp150-depleted populations. In contrast, more or an equal amount of IgM was secreted in cultures containing gp150-depleted cells. The differences between the fractions could not be ascribed to uneven distribution of T3+ cells, OKM1+ cells or B1+ (CD20) cells. However, the gp150-enriched population contained significantly more B2+ (CD21) cells than the gp150-depleted population. These results suggest that the gp150+ B cells differ from gp150- B cells, not only in their responsiveness to T cell differentiation signals but also in their commitment to Ig heavy chain isotype secretion.

Antigens, Surface↗

PubMed1987

Radioactive iodine and cesium in travellers to different parts of Europe after the Chernobyl accident.

Thyroid uptake of 131I was measured in 130 volunteers following the nuclear power plant accident at Chernobyl in April 1986. Ninety of these volunteers had been travelling in different parts of eastern Europe at the time of or immediately after the accident while 40 persons were permanently in Sweden. Also, 28 additional healthy volunteers, living in Sweden, were chosen for a long-term follow-up of the time-course of 134Cs and 137Cs whole body uptake. The highest levels of 131I were found in persons having visited Poland (mean value 3.27 kBq +/- 3.68 SD, extrapolated to April 27) while persons that had stayed in other parts of eastern or northern Europe showed significantly lower levels (p less than 0.01). The whole body burdens of cesium radionuclides were barely detectable immediately after the accident but increased gradually throughout the observed period. After five months nine farmers from a high fallout area in central Sweden had reached mean values of 4.20 kBq (+/- 3.34 SD) of 134Cs and 137Cs while six nonfarmers from the Stockholm area showed significantly lower levels, 0.64 kBq (+/- 0.24 SD, p less than 0.05). The radiation doses from the observed amounts of iodine and cesium isotopes reported in this study reflect only a marginal addition to the already existing dose from the natural environmental background radiation.

Accidents↗

PubMed1987

Generalized scatter correction method in SPECT using point scatter distribution functions.

A new two-dimensional (2-D) scatter correction technique in single photon emission computed tomography (SPECT) based on convolution or frequency filtering with a 2-D scatter distribution function is described. A scatter distribution function of the form A exp(-Br), has been derived from measurements of a point source in a water phantom. Both the amplitude A and the slope B of this function, were approximately invariant with source position except near phantom surface. The accuracy of the 2-D correction technique was compared with that of the previous one-dimensional (1-D) scatter correction technique. As could be expected the latter technique was shown to be less accurate due to its dependence on axial distribution of radioactivity. Phantom SPECT studies showed a clear superiority of the 2-D over the 1-D scatter correction in quantitative imaging. Images derived from clinical studies of regional bloodflow with 99mTc-HM-PAO and liver uptake showed significant contrast improvement by both techniques.

Scattering, Radiation↗

PubMed1986

Separation of human B lymphocytes on Helix pomatia A haemagglutinin into two major fractions differing in responsiveness to T-dependent mitogen (pokeweed mitogen) or antigen (tetanus toxoid).

A B-cell fraction consisting of 70% of cells carrying the B-cell-associated B1 antigen, 15-20% of M1+ non-B cells, and less than 3% of T cells was prepared from the peripheral blood of healthy human donors, previously vaccinated with tetanus toxoid (TT). As assessed by immunofluorescence after treatment with neuraminidase, approximately 40-50% of the B cells had surface structures binding to Helix pomatia A haemagglutinin (HP). The cells were separated into three fractions by affinity chromatography on HP conjugated to Sepharose (P, non-retained cells; EI, cells eluted with 0.1 mg/ml N-acetyl-D-galactosamine (D-GalNac); EII, cells eluted with 1 mg D-GalNac/ml). The majority of B cells in fraction EII were HP+ and were rich in cells expressing the B2 differentiation antigen. Sixty per cent of the B cells in this fraction also expressed the major HP-binding glycoprotein, gp 150. In the presence of autologous T cells, these B cells were strongly responsive to activation by either pokeweed mitogen (PWM) or antigen (TT), as reflected by differentiation into plasma cells, secretion of polyclonal IgG and IgM, or IgG anti-TT antibodies. In contrast, fraction P, which contained more than 90% HP-B cells, and which was partially depleted of B2+ cells, responded poorly or not at all to both PWM and TT. Fraction EI was a mixed fraction that responded in an intermediate fashion. When the preparations were depleted of contaminating non-B cells carrying the monocyte or large granular lymphocyte associated M1 antigen, their response to the two stimulating agents did not alter. The results suggest that HP+ B cells differ from HP-B cells in their responsiveness to T-cell signals. Fractionation on unsolubilized HP offers a simple and efficient way of separating B cells into at least two subsets differing in their responsiveness to T-cell-derived differentiation and maturation signals.

Animals↗

PubMed1985

The large sialoglycoprotein of human lymphocytes. I. Distribution on T and B lineage cells as revealed by a monospecific chicken antibody.

Chickens were immunized with highly purified large sialoglycoprotein of human lymphocytes (L-LSGP; gp 150) which was isolated from neuraminidase-treated normal peripheral blood lymphocytes by affinity chromatography to HP-Sepharose and further purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antibodies isolated from plasma and egg yolk were highly specific for desialylated L-LSGP (apparent molecular mass approximately 150 kDa). The antigenic sites recognized by the antibodies are probably located in the peptide moiety of the molecule since antibody binding to lymphocytes was not inhibited by a variety of different sugars or abrogated by absorption on various erythrocytes. In immunofluorescence experiments, greater than or equal to 75% of neuraminidase-treated thymocytes and peripheral blood lymphocytes, virtually all E+ cells and T4+ or T8+ T chronic lymphocytic leukemia (CLL) cells were stained by anti-gp 150. A small fraction (approximately 10%) of thymocytes and a larger fraction (greater than or equal to 30%) of T CLL cells in some patients were stained before neuraminidase treatment. Thymocytes appear to contain considerably lower amounts of a less sialylated form of L-LSGP than peripheral blood lymphocytes. In contrast to blast cells of 5-day concanavalin A or leucoagglutinin cultures (greater than or equal to 90% anti-gp150+) only about 50% of the blast cells generated in 5-day mixed leukocyte cultures were anti-gp150+. The large majority (greater than or equal to 75%) of both the anti-gp150+ and anti-gp150- cells were T3+ and T11+.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1985

The large sialoglycoprotein of human lymphocytes. II. Biochemical features.

Large sialoglycoprotein of human lymphocytes (L-LSGP) from thymocytes and from peripheral blood lymphocytes (PBL) of normal donor and of B chronic lymphocytic leukemia (CLL) patients was purified by affinity chromatography to Maclura pomifera agglutinin (MPA)-Sepharose followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). L-LSGP from the three different sources was very similar in amino acid composition. It contained a high proportion of acidic and hydroxy amino acids and also significant amounts of cysteine. No reduction in mobility in SDS-PAGE was noted for unreduced L-LSGP. The molecule may already in its native form have an extended conformation containing either free sulfhydryl groups or small S-S loops not affecting mobility in SDS-PAGE. L-LSGP was found to be highly glycosylated, the thymocyte glycoprotein containing somewhat less carbohydrate by weight (44%) than that of PBL (normal PBL 53% and B CLL 52%). This was due primarily to a lower content of sialic acid. The molecules contained mannose, galactose, N-acetyl galactosamine, N-acetylglucosamine and sialic acid in molar ratios 1.0:3.0:1.1:1.2:1.3 (thymocyte L-LSGP), 1.0:3.8:1.2:1.0:1.7 (PBL L-LSGP) and 1.0:3.5:2.2:1.3:2.8 (B CLL L-LSGP). The weak interaction of L-LSGP with lentil lectin, concanavalin A (Con A) and leucoagglutinin (La), its unchanged mobility in SDS-PAGE after tunicamycin treatment and its high amount of hydroxy amino acids suggest that most carbohydrate chains are O-glycosidically linked to the peptide chain. Native as well as Nase-treated L-LSGP show size microheterogeneity. This is probably due to small chemical differences in the L-LSGP molecules from different lymphocyte subsets.

Amino Acids↗

PubMed1985

Importance of particulate antigen for the induction of dual bronchial reaction in guinea-pigs.

There has been a lack of small animal models for the secondary allergic response (SAR) seen after bronchial challenge in many asthmatic patients. We have found that challenge with particulate instead of soluble antigen will provoke an SAR-like bronchial obstruction in the guinea-pig. The particulate form was obtained by coupling the antigen covalently to Sepharose beads (approximately 100 microns). Different experiments suggest that SAR is obtained only when the challenge is induced via IgE-mediated mechanisms and when the antigen is sufficiently large to provoke frustrated phagocytosis by the invading inflammatory cells. As judged in lungs sections SAR was related to bronchiolitis.

Airway Resistance↗

PubMed1985

Characterization of chronic lymphocytic leukemia (CLL) cells with low density of surface membrane bound immunoglobulins (smIg).

The study consists of 6 CLL patients with leukemic blood lymphocytes lacking T-cell characteristics and with smIg on only a very small fraction of the cells detected by microscopy or FACS analysis after direct immunofluorescence (IFL) staining. Using B-cell specific monoclonal antibodies all leukemias were found to be of the B-cell type. SmIg, mainly of IgD-class and the monotypic light chain, was detected on a large number of cells in all cases when using direct IFL. No lymphocyte clone was judged to be of the pre-B-cell type or represented fully differentiated terminal B-cells. CLL cells from the individual cases probably represent intermediate B-cell maturation steps. The results lend no support to the suggestion that this subtype of CLL necessarily belongs to an early stage of differentiation just beyond the pre-B-cell level.

Antibodies, Monoclonal↗

PubMed1985

A method for combined quantitative pertechnetate and bone scintigraphy of the sacro-iliac joints.

A technique for combined quantitative bone and pertechnetate (perfusion) scintigraphy of the sacro-iliac joints is described. The method is based on localization of the joints in the image obtained after the injection of a small dose of bone-seeking technetiated methyldiphosphonate. This image is later subtracted from the image obtained after the injection of pertechnetate. This combination of methods is believed to constitute a more accurate way of localizing the sacro-iliac joints than the previously used method, based on fluoroscopy. A minor comparison between these two ways of localising the joints is reported too. It is possible that the combination of quantitative bone and pertechnetate scintigraphy will improve the overall sensitivity of the scintigraphic method.

Adult↗

PubMed1984

Inhibition of proliferative and cytotoxic activities of human T lymphocytes with rabbit antibodies directed against leucoagglutinin-reactive T cell surface components.

Three rabbit antisera (870, 872 and 873) were raised against leucoagglutinin-reactive components from the surface of human T cells. The antibodies reacted with two major glycoproteins of 175 kDa and 105 kDa. None of the antibodies triggered peripheral blood lymphocytes or T cells to proliferation when tested under different culture conditions. All antibodies inhibited the proliferative response to concanavalin A or to allogeneic lymphocytes in mixed lymphocyte culture when whole IgG fractions were used. Complete inhibition of cytotoxic activity was obtained in cell-mediated lympholysis and in natural killer cell cytotoxicity (NK) when fresh peripheral blood lymphocytes were used as effector cells. Weak inhibition was also obtained in NK when mixed lymphocyte culture-activated effector cells were used. The inhibition was stronger, when NK activity was determined against MOLT4 target cells as compared to K562. Whereas F(ab')2 fragments of 873 IgG inhibited cytotoxic T lymphocyte activity completely, Fab fragments of 873 IgG neither inhibited proliferation nor cytotoxic T lymphocyte activity, but gave some inhibition of NK against MOLT4 targets. The results indicate that antibodies against these leucoagglutinin-reactive structures reacted with polypeptides similar to or identical with the human "leukocyte function-associated antigen-1" (LFA-1) considered to be an important mediator of cell-cell interactions and nonspecific adherence.

Agglutinins↗