Definition of a novel cellular constituent of the bone marrow that regulates the response of immature B cells to B cell antigen receptor engagement.

Previously we defined a Thy1(dull) bone marrow-derived cell population that regulated fate decisions by immature B cells after Ag receptor signaling. The microenvironmental signals provided by this cell population were shown to redirect the B cell Ag receptor -induced apoptotic response of immature B cells toward continued recombination-activating gene (RAG) expression and secondary light chain recombination (receptor editing). Neither the identity of the cell responsible for this activity nor its role in immature B cell development in vivo were addressed by these previous studies. Here we show that this protective microenvironmental niche is defined by the presence of a novel Thy1(dull), DX5(pos) cell that can be found in close association with immature B cells in vivo. Depletion of this cell eliminates the anti-apoptotic effect of bone marrow in vitro and leads to a significant decrease in the number and frequency of bone marrow immature B cells in vivo. We propose that, just as the bone marrow environment is essential for the survival and progression of pro-B and pre-B cells through their respective developmental checkpoints, this cellular niche regulates the progression of immature stage B cells through negative selection.

Negative selection of immature B cells by receptor editing or deletion is determined by site of antigen encounter.

Immature B cells that encounter self-antigen are eliminated from the immune repertoire by negative selection. Negative selection has been proposed to take place by two distinct mechanisms: deletion by apoptosis or alteration of the antigen receptor specificity by receptor editing. While convincing evidence exists for each, the two models are inherently contradictory. In this paper, we propose a resolution to this contradiction by demonstrating that the site of first antigen encounter dictates which mechanism of negative selection is utilized. We demonstrate that the bone marrow microenvironment provides signals that block antigen-induced deletion and promote RAG reinduction. In the periphery, the absence of these signals allows the immature B cell to default to apoptosis as a result of BCR engagement.

B cell receptor-induced apoptosis in primary transitional murine B cells: signaling requirements and modulation by T cell help.

Self-reactive immature B cells may be eliminated in the bone marrow (BM) after B cell receptor (BCR) engagement in a process known as negative selection. Immature B cells emigrating from the BM, the so-called transitional cells, remain sensitive to negative selection and are likely to be important targets of tolerance towards peripheral antigens. Transitional cells are deleted through apoptosis after BCR cross-linking in vitro. Using anti-Ig as a surrogate antigen, we determined the signaling requirements for the induction of apoptosis in transitional cells. Treatment with anti-Ig for only 20 min causes most cells to be apoptotic 16 h later. Furthermore, apoptosis of transitional cells is induced with low doses of anti-Ig while mature cell proliferation requires extended culture at 30-fold higher concentrations. For both populations of B cells, total surface Ig expression is equivalent, therefore indicating that the threshold of BCR signaling required to elicit these responses is different. T cell help can modulate B cell tolerance. However, specific help may not be available when apoptosis is triggered by a peripheral antigen. The opportunity to reverse apoptosis of transitional cells is surprisingly long. Even 8 h after anti-Ig treatment, IL-4 or anti-CD40 antibody can block apoptosis. The upper time limit of protection is concurrent with irreversibility of apoptosis as measured by DNA fragmentation. These findings indicate that B cell negative selection is more easily triggered than activation, and that the induction of apoptosis and its reversal by T cell help can be events that occur in distinct microenvironments.

Cluster-oriented discriminant analysis; taxonomic classification of the thyroid function.

A computer assisted procedure for the diagnosis of thyroid diseases, based on seven clinical chemical parameters, is proposed. The population studied consisted of 592 consecutive outpatients with a tentative diagnosis of thyroid disease. Thyroxine, triiodothyronine, T3 uptake test, TSH before and after TRH application, its difference and thyroxine binding globulin have been determined. The patients were clinically examined and in each case a Tc-scintigram was obtained. As a first step, 20 biochemical patterns were defined by cluster analysis (pattern cognition). T check how good in grouping process was, linear discriminant analysis was applied after which the reclassification rates were very satisfactory. The clusters found corresponded well to the pathophysiological situations with some exceptions. As a second step, patients were assigned to these patterns by use of the derived discriminant functions (pattern recognition). The proposed method seems to have some advantages over other diagnostic models published hitherto.

[Radioimmunoassay of beta2-microglobulin in monoclonal gammopathies (author's transl)]. / Radioimmunologische Bestimmung von beta2-Mikroglobulin bei monoklonalen Gammopathien.

Serum concentration of beta2m were measured by radioimmunoassay in 78 healthy subjects and 80 patients with monoclonal gammopathies. The beta2m levels of normal sera were normally distributed with a mean concentration of 1610 microgram/l. 77% of values were between 1200 and 2000 microgram/l. The serum beta2m levels of patients with monoclonal gammopathies were significantly higher than those of the healthy subjects, but there was no significant difference when comparing the Ig class or the light-chain type. Urinary beta2m excretion of patients with monoclonal gammopathies (3--5400 microgram/l) were slightly higher than normal (15--113 microgram/l). Serum beta2m levels did not correlate with the serum levels of monoclonal immunoglobulins. This supports the hypothesis of the mutual independence of beta2m and Ig production.

Performance of various mathematical methods for computer-aided processing of radioimmunoassay results.

Interpolation and regression methods are available for computer aided determination of radioimmunological end results. We compared the performance of 6 algorithms (weighted and unweighted linear logit log regression; quadratic logit log regression, smoothing spline interpolation with a large and small smoothing factor, respectively, and polygonal interpolation and the manual curve fitting on the basis of three radioimmunoassays with different reference curve characteristics (digoxin, estriol, human chorionic somatomammotrophin (HCS)). Great store was set by the accuracy of the approximation at the intermediate points on the curve, i.e. those points that lie midway between two standard concentrations. These concentrations were obtained by weighing and inserted as unknown samples. In the case of digoxin and estriol the polygonal interpolation provided the best results, while the weighted logit log regression proved superior in the case of HCS.

[Studies of activity kinetic of isoenzyme CK-MB in serum after myocardial infarction (author's transl)]. / Untersuchungen zur Aktivitätskinetik des Isoenzymes CK-MB im Serum nach Myokardinfarkt.

We investigated the activity kinetics of CK-total and CK-MB in 83 patients with proven myocardial infarctions. Serial serum samples were taken at intervals of 2--6 h. The activity of isoenzym CK-MB was determined by means of the immunological inhibition method. CK-MB activity was determined in all patients. The mean peak activity of CK-MB was 65 U/l (range: 9-241 U/l). At the time of peak CK-MB activity the mean percentage CK-MB activity was 13.2% (range: 3.4--21.7%). The CK-MB activity reached its peak at 17.4 h (range: 3.0--32.5 h) after the onset of retrosternal pain. This is 1.4 h after peak CK-total activity. The mean disappearance rate constant for CK-MB (n = 31) was found to be 9.3 X 10(-4) U/min with a large individual variation. This value corresponds to a half life of 12.5 h (CK-total: 15.5 h). The determination of CK-MB activity is therefore only of diagnostic significance within 48 h of possible myocardial occurrence. Moreover, isoenzyme CK-MB is not found exclusively in myocardium. For this reason it is better to use the percentage CK-MB activity in the differential diagnosis of myocardial infarction. With 80% of the patients this value is greater than 6% within 36 h of proven myocardial infarction.

[Effect of total fasting on serum protein concentrations (author's transl)]. / Einfluss totaler Nahrungskarenz auf Serumproteinkonzentrationen.

During a period of total fasting restricted to three weeks, haptoglobin, transferrin and beta1A-globin showed a marked, continuous decline. The complement was finally distinctly below the reference range. Macroglobulins and coeruloplasmin rised until the tenth day and decreased thereafter. IgG globulin remained essentially unchanged while immunoglobulins gammaA and gammaM showed an increase. On account of the considerable fluctuations observed in the protein fractions fasting in this form at least beyond a period of 21 days does not seem justified.