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K Hasløv

Publications and source records attributed to K Hasløv.

33 records · Page 2Linked to original sources
PubMed1990

Inactivation of suppressor T cell activity by the nontoxic lipopolysaccharide of Rhodopseudomonas sphaeroides.

Antibody responses of mice immunized with type III pneumococcal polysaccharide were examined with and without treatment with nontoxic lipopolysaccharide from Rhodopseudomonas sphaeroides (Rs-LPS). The results obtained were similar to those described previously for mice treated with monophosphoryl lipid A (MPL) except that lower amounts of Rs-LPS were needed. Both were without effect when given at the time of immunization with type III pneumococcal polysaccharide but elicited significant enhancement when given 2 to 3 days later. Such enhancement was T cell dependent and not due to polyclonal activation of immunoglobulin M synthesis by B cells. Treatment with either Rs-LPS or MPL abolished the expression but not induction of low-dose paralysis, a form of immunological unresponsiveness known to be mediated by suppressor T cells (Ts). The in vitro treatment of cell suspensions containing Ts with extremely small amounts of Rs-LPS or MPI completely eliminated the capacity of such cells to transfer suppression to other mice. These findings indicate that the immunomodulatory effects of both MPL and Rs-LPS are mainly the result of eliminating the inhibitors effects of Ts; this permits the positive effects of amplifier T cells to be more fully expressed, thereby resulting in an increased antibody response. The significance of these and other findings to the use of Rs-LPS as a pharmacotherapeutic agent for gram-negative bacterial sepsis is discussed.

Animals↗
PubMed1990

Reactivity of lymphoid cells isolated from the tracheobroncheal lymph nodes of two guinea-pig strains with high or low susceptibility to respiratory anaphylaxis.

The mechanisms underlying the different sensitivity to respiratory anaphylaxis were studied in two inbred strains of guinea pigs with high (IMM/S) or low (IMM/R) sensitivity to ovalbumin (OA) aerosol-induced respiratory anaphylaxis. Guinea pigs were immunized with OA in Freund's complete adjuvant and lymphocyte stimulation tests were done with cells from the tracheobroncheal lymph nodes (TBL) draining the airways, from peripheral blood lymphocytes and from the axillary and inguinal lymph nodes. The TBL of the IMM/R strain had a significantly lower response than those of the IMM/S strain. This selectively low response, which was not antigen specific, could be increased by cyclophosphamide pretreatment. The high TBL response in the IMM/S strain was decreased by OA inhalations to the level of the IMM/R strain. OA inhalations decreased the IgE response in the IMM/S but not the IMM/R strain. Different interpretations of these findings are discussed, with emphasis on the possibility that an inherent suppressor cell population in the TBL are involved.

Anaphylaxis↗
PubMed1989

Estimation of guinea pig antigen-specific and non-specific suppressor cell activity.

A functional assay for the quantitative estimation of suppressor cell (SC) activity in guinea pigs has been developed. Cultures of antigen-stimulated peripheral blood lymphocytes (PBL) from sensitized guinea pigs develop SC activity. The suppression of proliferation can be demonstrated in antigen-stimulated autologous co-cultures of precultured and freshly isolated PBL. The extent of suppression is dependent on the preculture antigen concentration but not the preculture period and it consists, as demonstrated with PBL from doubly sensitized guinea pigs, of an antigen-specific and a non-specific component. The observed SC activities were not due to an alteration of the kinetics of the co-cultures. The estimates of suppression are highly dependent on corrections for the values of the control cultures. The present method may prove useful in immunological studies of mycobacterial infections in guinea pigs.

Animals↗
PubMed1989

The generation of guinea pig T-cell lines reactive to antigens from Mycobacterium tuberculosis. Selected lines induce erythematous skin reactions.

We describe methods for the development and partial characterization of antigen-specific T-cell lines from the guinea pig, which is the classical experimental animal in tuberculosis research. T cells were obtained from strain 2 guinea pigs immunized with BCG vaccine or with killed Mycobacterium tuberculosis in oil. T-cell lines were obtained from limited dilution cloning of antigen-stimulated, blast-enriched lymphocyte cultures. The lines were grown with weekly reseedings in the alternating absence and presence of mycobacterial antigen. Antigen reactivity of the cell lines was studied with lymphocyte stimulation tests. With these methods we have consistently obtained antigen-reactive cell lines. When injected in small numbers intradermally in the presence of antigen in syngeneic guinea pigs, some of these cell lines gave rise to antigen-specific erythematous tuberculin-like skin reactions. The skin reactions, which were usually without induration, were most pronounced after 24 h. Histological examinations of skin undergoing such reactions showed that the erythemas were not accompanied by mononuclear infiltrations. We expect that transfer experiments with guinea pig T-cell lines will prove useful tools in the analysis of the contribution of defined mycobacterial antigen preparations to tuberculosis immunity.

Animals↗
PubMed1987

Production, assay and partial characterization of guinea pig interleukin 2.

Optimum conditions for the production and assay of guinea pig interleukin-2 (IL-2) have been established. The mitogenic activities of serial dilutions of guinea pig IL-2 preparations were compared in cultures of guinea pig peripheral blood lymphocytes prestimulated for 7 days with phytohemagglutinin (PHA) used at 1 microgram/ml. Parallel log dose-log response curves were used for quantitative comparisons. Optimum IL-2 yields were obtained from cultures of lymph node lymphocytes stimulated for 20 h with Concanavalin A (ConA) at 5 micrograms/ml. Guinea pig T cell lines reactive to mycobacterial antigens were propagated for several months using our IL-2 preparations. The molecular weight of guinea pig IL-2 was estimated to be 30,000 using S-200 gel filtration. The species specificities of guinea pig, human, mouse and rat IL-2s were examined. It was shown that guinea pig T lymphocyte blasts were stimulated only weakly with human IL-2 and not at all with mouse and rat IL-2.

Animals↗
PubMed1987

Biological activity in sensitized guinea pigs of MPB 70, a protein specific for some strains of Mycobacterium bovis BCG.

MPB 70 is a protein found in large quantities in the culture filtrate (CF) of the Tokyo and some other strains of Mycobacterium bovis BCG, and it has a remarkable degree of specificity for these strains. We estimated the molecular weight of MPB 70 to 22,000 by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE and immunoblotting showed that MPB 70 was present in high quantities in CF from BCG Tokyo, that it could be also demonstrated in BCG Copenhagen, and that it was absent in CF from M. tuberculosis H37Rv. When the purified MPB 70 preparation used in the present study was run in SDS-PAGE, blotted and stained with a polyclonal rabbit or a monoclonal mouse anti-MPB 70 antibody, several bands in addition to the main 22 kDa band were seen, indicating a tendency of the MPB 70 molecules and/or fragments thereof to form very stable aggregates with themselves. The biological activity of MPB 70 was studied in groups of guinea pigs sensitized with live BCG of the Tokyo and Copenhagen strains. Guinea pigs from both groups developed reactivity to tuberculin PPD as assessed by skin tests and lymphocyte stimulation tests with peripheral blood or lymph node lymphocytes. In addition, a strong and persistent reactivity to MPB 70 was demonstrated in the BCG Tokyo group with both methods. Guinea pigs sensitized with the Copenhagen strain were only weakly reactive to MPB 70. Skin reactions in guinea pigs that had been repeatedly tested with MPB 70 and tuberculin were compared with reactions in animals tested only once. Reactions to MPB 70 in BCG Tokyo sensitized guinea pigs were suppressed by repeated tests, whereas tuberculin reactions were boosted by the interim tests. The levels of specific anti-MPB 70 antibodies were higher in BCG Tokyo- than in BCG Copenhagen-sensitized guinea pigs. MPB 70 has a high degree of specificity and is a strongly immunogenic protein, which may prove useful in studies of mycobacterial immunology.

Animals↗
PubMed1987

Allergenic and blastogenic reactivity of three antigens from Mycobacterium tuberculosis in sensitized guinea pigs.

Three antigens from a culture filtrate of Mycobacterium tuberculosis H37Rv were purified by affinity chromatography, using monoclonal antibodies. The molecular weights of the purified antigens are 17,000 to 19,000, 32,000 to 33,000, and 39,000, respectively, and by their migration patterns in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions, they all appeared to be single-chain polypeptides. Western blot and enzyme-linked immunosorbent assay analyses indicated that the antigens are non-cross-reactive. All antigens generated an intermediate to strong skin reaction when tested in guinea pigs previously immunized with a live M. bovis BCG vaccine or with an oil emulsion preparation of phenol-or heat-killed M. tuberculosis. Lymphocytes isolated from peripheral blood or lymph nodes of similarly immunized guinea pigs could be stimulated by purified protein derivative and the purified antigens. Qualitative differences in stimulatory capacity between the preparations were demonstrated. The antigens may prove useful in further studies of the immunology and pathogenesis of tuberculosis.

Animals↗
PubMed1986

Comparison in sensitized and unsensitized guinea-pigs of tuberculin PPDs RT 23 and PPD-M by skin tests and lymphocyte stimulation tests. Effect of immunization time.

The biological activities of tuberculin PPD RT 23 and the International Standard for Purified Protein Derivative of Mammalian Tuberculin (PPD-M) were compared in sensitized and unsensitized guinea-pigs by skin tests and lymphocyte stimulation (LS) tests. Estimates of relative potency (RP) from skin test results were dependent on the dose level, on the immunogen used, and, in guinea-pigs immunized with killed tubercle bacilli in oil, also on the immunization time. Relative potency estimates from LS results were dependent on the source of the lymphocytes and were different from estimates obtained from skin tests. Lymphocyte stimulation dose-response curves for the tuberculins were qualitatively different. In contrast to RT 23, PPD-M gave rise to non-specific skin reaction in unsensitized guinea-pigs. Both tuberculins were mitogenic to lymph node lymphocytes isolated from unsensitized guinea-pigs, PPD-M being the more mitogenic of the two tuberculins. The present results confirm that qualitatively different tuberculins cannot be unambiguously calibrated in identical terms and thus emphasize that the uncritical use of (international) standards should be avoided in tuberculin calibration.

Animals↗
PubMed1986

Interspecies reactivity of five monoclonal antibodies to Mycobacterium tuberculosis as examined by immunoblotting and enzyme-linked immunosorbent assay.

Five different murine monoclonal antibodies (MAbs) to Mycobacterium tuberculosis were examined for degree of cross-reactivity with other mycobacterial species by enzyme-linked immunosorbent assay and immunoblotting. One MAb reacted solely with M. tuberculosis and M. bovis BCG. Two of the MAbs reacted with all mycobacterial species examined, whereas two MAbs demonstrated a limited reactivity pattern. The epitopes are located on molecules susceptible to protease treatment, and two of these molecules possess concanavalin A-binding moieties. Two of the antigens defined by these five MAbs are present in tuberculin purified protein derivative.

Animals↗
PubMed1984

In vivo and in vitro boosting effects of tuberculin skin tests in guinea-pigs immunized with living BCG or with killed Mycobacterium tuberculosis.

Eight weeks after the onset of immunization with living BCG vaccine or with heat-killed, dried Mycobacterium tuberculosis in paraffin oil (TB), guinea-pigs were skin tested with small doses of tuberculin PPD. Three weeks later the effect of these tests on skin reactions and on lymphocyte transformation (LT) responses of lymph node lymphocytes (LNL) or peripheral blood lymphocytes (PBL) was estimated by a comparison with non-tested groups. For both immunogens, the skin test after 8 weeks significantly enhanced skin reactions, particularly to low tuberculin doses. In the BCG-vaccinated guinea-pigs the LT responses of both cell types were significantly enhanced by the skin test after 8 weeks, whereas the LT responses from the TB-immunized guinea-pigs were not affected. Therefore, in the planning and interpretation of in vitro tests of cellular immunity, possible effects from previously applied skin tests should be taken into consideration.

Animals↗
PubMed1984

Studies on the development of tuberculin sensitivity in immunized guinea pigs with demonstration of a close relationship between results of skin tests and the lymphocyte transformation technique.

The development of tuberculin sensitivity in groups of guinea pigs immunized with living BCG vaccine or with heat-killed Mycobacterium tuberculosis suspended in oil (TB) was measured by skin tests and lymphocyte transformation (LT) tests with tuberculin PPD. LT tests on lymphocytes from peripheral blood (PBL) and lymph nodes (LNL) and skin tests were carried out on different groups of guinea pigs for 1 year following immunization. Three ways of expressing the LT results were considered, i.e. as stimulation index, delta cpm and uncorrected cpm values. Since neither of the first two methods were equally applicable to all of the data, we decided to express the data as geometric means of cpm values for stimulated and control cultures, respectively. For both immunogens the sensitivity measured by PBL LT and skin tests showed a closely parallel development; LNL reactivity appeared earlier and remained at a higher level than PBL reactivity. Antibody levels in the guinea pigs were measured using a radioimmunoassay technique, were found to be dependent on the immunization period and not correlated to the cellular immunity.

Animals↗
PubMed1983

Effect of 2-mercaptoethanol and mycostatin on guinea pig lymphocyte transformation: interpretation of results depends on the method of calculation.

The stimulating effects of 2-mercaptoethanol (2-ME) reported in murine cell cultures were confirmed for guinea pig lymphocyte transformation (LT). 2-ME was mitogenic for peripheral blood lymphocytes (PBL) and lymph node lymphocytes (LNL) and enhanced PPD-induced LT. The effect on LNL was dependent on the magnitude of the response of non-2-ME treated cultures. The 2-ME effect on PPD-stimulated LNL was reversed when stimulation index (SI) replaced the delta cpm estimate. Mycostatin (MYC) inhibited the blastogenic response of PBL control cultures, whereas its effect on LNL control cultures varied. The interpretation of the effect of MYC on PPD-stimulated cultures was dependent on the use of the delta cpm or SI estimates. The interpretation of LT experiments is therefore highly dependent on whether delta cpm or SI is used for expression of results. Analysis of effects on control and stimulated cultures should precede the addition of 2-ME or MYC to cultures for LT.

Animals↗