Biomedical subjects

T Logtenberg

Publications and source records attributed to T Logtenberg.

78 records · Page 5Linked to original sources

PubMed1986

Antigen-specific activation of autoreactive B cells in normal human individuals.

The purpose of our study was to determine if normal human B cells can be activated to autoantibody production using an antigen-specific system. For this purpose we investigated the in vitro antibody response to the autoantigen thyroglobulin (Tg) employing soluble or insolubilized Tg (i-Tg) and a B cell growth and differentiation factor (BDGF) to stimulate lymphocytes from healthy individuals. Similar experiments were carried out with the xenoantigen ovalbumin (OVA). The presence of Tg and OVA-reactive B cells was demonstrable by stimulating lymphocytes from tonsil, spleen and blood with a combination of pokeweed mitogen and formalinized Staphylococcus aureus: mitogen stimulation resulted in the generation of IgM anti-Tg and IgM anti-OVA antibody-forming cells (AFC) as detected in a spot enzyme-linked immunosorbent assay. Soluble antigen failed to induce autoantibody production. However, i-Tg or i-OVA did activate normal tonsil and spleen B cells. Differentiation of these activated B cells to IgM AFC required the presence of BGDF, derived from a human T hybrid clone. Preincubation experiments with the particulate autoantigen show that a specific activation signal is provided by the antigen which subsequently renders the B cells responsive to BGDF. i-Tg-dependent stimulation of B lymphocytes could be inhibited by adding free Tg to the cultures; the same applied to i-OVA stimulation. We conclude that the normal human B cell repertoire contains B cells that can be activated to autoantibody production by the autoantigen Tg if the necessary T cell signals are provided. Thus, these B cells are not in an inherently anergic state. Similar mechanisms seem to play a role in the activation of B cells responding to i-OVA and i-Tg.

Antibody-Producing Cells↗

PubMed1985

Enumeration of (auto)antibody producing cells in human using the "spot-ELISA".

An enzyme-linked immunosorbent assay (spot-ELISA) for individual immunoglobulin secreting cells, which became recently available, was applied to the enumeration of human B lymphocytes secreting specific antibodies of thyroglobulin. Polyclonally activated B cells from patients with auto-immune thyroid disease are incubated in thyroglobulin coated plates. After removal of the cells specific antibodies are visualized by means of an immunoenzyme technique employing agarose to localize converted substrate. Individual specific antibody secreting cells are counted as blue spots using an inverted microscope. Numbers and isotype of spots correlate well with the amount and isotype of secreted antibody as detected with a conventional ELISA. This easy to perform, complement-independent technique offers a useful alternative to conventional plaque forming cell assays.

Antibody-Producing Cells↗

PubMed1985

Human peripheral blood lymphocytes from recently vaccinated individuals produce both type-specific and intertypic cross-reacting neutralizing antibody on in vitro stimulation with one type of poliovirus.

An in vitro system of poliovirus-specific antibody production by peripheral blood B cells on stimulation by the virus has been developed. Virus-neutralizing antibodies in culture supernatant fluids, or virus-specific antibody-secreting cells (ASC) were detected by microneutralization assay and ELISA-SPOT test, respectively. After booster immunization with polio vaccine, anti-poliovirus-neutralizing ASC were present in circulation. This response was measurable between 5 and 12 days after booster vaccination. At between 12 and 90 days, another subset of B cells was found in peripheral blood that only produced poliovirus-specific neutralizing antibody after in vitro antigenic stimulation. The in vitro virus-induced response required B cells, monocytes, and T4+ (T helper) cells, and was shown to result from de novo protein synthesis. The anti-poliovirus-neutralizing response in vitro could be dissected in a type-specific and intertypic cross-reactive response by using various antigen concentrations for in vitro stimulation. Evidence was obtained by absorption studies for the existence of intertypic cross-reactive neutralization-inducing epitopes.

Adolescent↗

PubMed1985

Synergistic action of A23187 and phorbol ester on human B cell activation.

We have investigated the existence of a synergy occurring between the calcium ionophore A23187 and phorbol myristic acetate (PMA) with respect to human B cell proliferation and differentiation. The combination of A23187 (250 to 500 nM) with nonmitogenic concentrations of PMA (1 to 3 ng/ml) resulted in a strong proliferative response in human tonsillar, spleen, and peripheral blood B cells. This proliferation could not be blocked by anti-Tac antibody at concentrations that effectively inhibited T cell proliferation under similar culture conditions, suggesting that IL 2 and its receptor are not involved in B cell proliferation in this system. During a 3-day culture period, A23187 (500 nM) did not activate B cells in terms of changes in cell size or in the expression of transferrin receptor, HLA-DR, and Tac antigen. PMA at a nonmitogenic concentration (3 ng/ml) enhanced the expression of the first two markers. Combination of the ionophore with PMA induced the occurrence of Tac and further increased the expression of transferrin receptor and HLA-DR. A23187 similarly enhanced the PMA-mediated increase in cell size. PMA and A23187 did not induce differentiation to lg production. However, when cells were prestimulated with a combination of the two agents and were recultured in the presence of a preparation containing B cell differentiation factor, a strong increase in IgM, IgG, and IgA production was found. We conclude that PMA and A23187 synergistically trigger intracellular events in human B cells, leading to proliferation and to responsiveness to differentiation factors.

B-Lymphocytes↗

PubMed1984

Cloning of nitrate reductase genes from the cyanobacterium Anacystis nidulans.

Anacystis nidulans, a non-nitrogen-fixing cyanobacterium, can fulfill its nitrogen requirement by the assimilation of nitrate. The first step in the pathway, the reduction of nitrate to nitrite, is catalyzed by the molybdo-protein nitrate reductase. In this study, newly developed techniques for gene cloning in A. nidulans R2 were used for the isolation of two genes involved in nitrate reduction. One gene was cloned by complementation of the corresponding mutant; the other gene was picked up from a cosmid gene library by using a restriction fragment containing the transposon-inactivated gene as a probe. Both genes were unlinked single-copy chromosomal genes. Transformation studies provided evidence for the existence of a third locus involved in nitrate reduction.

Cloning, Molecular↗

PubMed1982

Peanut agglutinin (PNA) binding as a marker for immature human B lymphocytes. Is bone marrow not the complete bursa-equivalent?

To investigate the nature of peanut agglutinin (PNA) binding cells in various human lymphoid tissues a double marker assay was performed using fluorescent PNA and rosetting with anti-mu or anti-delta coated ox red blood cells for detection of B cells or rosetting with sheep red blood cells for detection of T cells. In human bone marrow 60.5 +/- 8.6% of the surface mu+ve (smu+ve) B cells did bind PNA whereas only a small minority of the surface delta +ve (delta +ve) B cells (5.0 +/- 4.2%) and none of the T cells were PNA+ve. In peripheral blood most of the PNA+ve cells appeared to be monocytes. Only a small proportion of the smu+ve B cells (9.7 +/- 2.7%) and none of the s delta +ve B cells or T cells did bind PNA. Contrarily in tonsils a relatively high proportion of smu+ve B cells (33.2%) of s delta +ve B cells (26.3%) and of T cells (17.2%) were PNA+ve. These results indicate that PNA binding is also a marker for immature B cells. Moreover in human bone marrow at least two populations of B cells may be distinguished, an immature population of smu+ve, s delta-ve, PNA+ve B cells and a mature populations of smu+ve, s delta +ve, PNA-ve B cells, the latter probably representing recirculating B cells. We hypothesize that the first population comprises immature B cells, that leave the bone marrow in an early stage and complete the maturation to immunocompetent B cells in peripheral lymphoid organs like tonsils.

Animals↗