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Biomedical subjects

J Andersson

Publications and source records attributed to J Andersson.

At least 307 records · Page 17Linked to original sources

T-cell-dependent B-cell stimulation is H-2 restricted and antigen dependent only at the resting B-cell level.

Cloned lines of helper thymus-derived (T) cells produce help for bone marrow-derived (B) cell growth and Ig secretion in the presence of histocompatible adherent cells and of specific antigen. This help stimulates histocompatible as well as histoincompatible B-cell blasts polyclonally. Thus, neither antigen nor histocompatibility, but antigen-unspecific factor(s) for growth and Ig secretion are required to stimulate a B-cell blast through the next round of division. On the other hand, only histocompatible, resting, small B cells, and only those binding their specific antigen, can be stimulated by antigen-activated T-cell help to initiate growth and Ig secretion. The preference of the resting B cells for such collaboration with T-cell help is mapped to the K end of the H-2 histocompatibility locus, and probably constitutes the antigen expressed on B cells by the immune response (I) region. It appears that a resting B cell is excited by the binding of specific antigen to surface Ig and by the interaction of its surface Ia antigen with helper T cells. After this dual recognition the excited B cell can be stimulated by the antigen-unspecific factor(s) generated by the interaction of helper T-cells, adherent cells, and antigen to initiate replication.

Animals↗

The purified protein derivative of turberculin, a B-cell mitogen that distinguishes in its action resting, small B cells from activated B-cell blasts.

The purified protein derivative of tuberculin (PPD tuberculin) stimulates approximately one of two lipopolysaccharide (LPS)-activated B-cell blasts of C57BL/6J nu/nu spleen cells to continued clonal growth and maturation to IgM and IgG secretion. It alwo stimulates background, in vivo-activated large cells of normal C57BL/6J nu/nu spleen to growth and Ig secretion, at a frequency of approximately 1 of 100 large spleen cells. PPD tuberculin, therefore, is a polyclonal B-cell activator for B-cell blasts. Many single murine splenic B cells (approximately 50%) appear to have reactivities, and therefore probably receptors, for LPS and PPD tuberculin. PPD tuberculin does not stimulate small, resting B cells to growth as measured by the number of cells in culture and by thymidine uptake. However, it stimulates approximately one-fourth of all spleen cells to blast transformation. The large-size blast cells secrete IgM and, therefore, form plaques in the protein A plaque assay. IgG-secreting, plaque-forming cells develop at later stages of stimulation, indicating that the switch from IgM to IgG may occur without division in single, stimulated B cells. Stimulation of resting B cells to maturation by PPD tuberculin is polyclonal. Thus, approximately 1 in 10(2) IgM-secreting plaque-forming cells form plaques with trinitrophenyl-substituted sheep erythrocytes, 1 in 450 do so with horse erythrocytes, and 1 in 10(3) with sheep erythrocytes. Furthermore, the number of Ig-secreting cells developing from small, resting cells without growth in cultures with or without filler thymus cells suggests polyclonal activation by PPD tuberculin to maturation only of at least one out of four small, splenic B cells.

Animals↗

Mitogen-activated B-cell blasts reactive to more than one mitogen.

One of three splenic B cells of 2- to 3-mo-old C57BL/6J or C3H/Tif mice are activated either by lipopolysaccharide (LPS) or by lipoprotein (LP) to grow and to mature to IgM-secreting cells. When mitogen-activated blast cells, after a 2-d activation period, are separated from nonactivated, small cells by velocity sedimentation, and no mitogen is readded, the blasts do not continue to grow but do continue to secrete IgM for several days. When the homologous mitogen is added for restimulation, the blast cells continue for several days to divide every 18 h and to develop IgM-secreting, plaque-forming cells. Frequency analyses at limiting dilutions of reactive B-blasts show that one cell in approximately equal to 1.2--1.5 blasts continue to grow and mature in the presence of the homologous mitogen, either LPS or LP. When the B-cell blasts obtained in a first activation period with either LPS or LP are restimulated with a heterologous mitogen, LPS, LP, Nocardia mitogen or mitogens contained in fetal calf serum a high proportion of the blasts continue to grow and mature to IgM-secreting cells. Frequency analyses show for LPS- or LP-blasts, restimulated in a heterologous fashion with either LP or LPS, that one cell in 1.35--1.5 blasts continue to divide and to mature to IgM-secreting cells. C3H/HeJ-splenic B-cells, which are LPS nonresponders, can only be activated to blast cells by LP. These LP-activated blasts can be restimulated by the homologous LP and by Nocardia mitogen and mitogens of fetal calf serum, but not by LPS. The results indicate that the majority of splenic B cells of 2- to 3-mo-old C57BL/6J or C3H/Tif mice are reactive to more than one B-cell mitogen. B cells, therefore, can possess in their surface membrane multireactive mitogen-receptor complexes which regulate growth and maturation.

Animals↗

Studies on T lymphocyte activation. I. Requirements for the mitogen-dependent production of T cell growth factors.

Conditioned media (CM) obtained from concanavalin A (Con A)-stimulated spleen cell cultures maintain mitogen-derived T cell blasts in exponential growth for indefinite periods of time. Such purified T cell blasts do not respond to Con A, and the growth-promoting activity in CM is independent of the mitogenic lectin used for its production. However, the appearance of activity in CM is Con A dose-dependent. Furthermore, the production of these T cell growth factors is independent of B cells, while it requires both T cells and nontheta-bearing, probably Ia-positive cells present in normal and nude spleens. On the other hand, the T cell blasts which are reactive to the growth factors, neither produce CM by themselves nor can they cooperate with nude spleen cells for its production, in the presence of a mitogenic lectin. Since the concentration of growth factors in CM determines the extent of T cell growth, we propose that the growth of some T cells is necessarily dependent upon the activity in CM described here and, consequently, that the basis for T cell activation is the induction of such growth factors.

Animals↗

Studies on T lymphocyte activation II. The target cells for concanavalin A-induced growth factors.

Growth factors contained in Con A(concanavalin A)-conditioned media (CM) maintain exponential growth in T cell blasts derived from mitogen stimulation of spleen cells with Con A, phytohemagglutinin, lentil lectin and pokeweed mitogen (PWM), as well as from mixed lymphocyte reactions to H-2D or I-C-encoded determinants and from non-H-2 Mls locus-controlled reactions. Such growth factors are strictly T cell blast-specific, inasmuch as they do not stimulate resting, small T or B lymphocytes nor B cell blasts generated by lipopolysaccharide or lipoprotein activation. The responsiveness of T cell blasts to CM appears to be the result of the availability on the cell surface of an acceptor site for growth factors which is not expressed in resting cells, because T cell blasts, but not small spleen cells, absorb out the growth-promoting activity contained in CM. Furthermore, lectins such as Con A and PWM interfere with the blast surface membrane in such a way that they inhibit the response to growth factors. Finally, there is no detectable allotypic or isotypic restriction in the activity of Con A-CM on a variety of target T cell blasts.

Animals↗

Metabolic changes in blood and skeletal muscle in reconstructive aortic surgery.

The declamping shock has been attributed to reactive hyperemia with pooling of blood in the legs, metabolic acidosis following lactate accumulation in the ischemic leg muscles and hyperkalemia. We have studied eight patients undergoing arterial reconstruction with temporary aortic occlusion, in six cases because of obstructive arteriosclerotic disease and in two cases for aortic aneurysm. Muscle biopsies from the lateral vastus muscle as well as blood samples from the iliac vein, the superior caval vein and the radial artery were taken before clamping of the aorta, just before declamping, and 20 minutes after restitution of leg blood flow. ATP, ADP, AMP, phosphocreatine (PCr), creatine (Cr), glycogen, glucose, lactate, pyruvate and NH4+ from the immediately frozen muscle biopsies as well as lactate and pyruvate in the iliac vein and central venous blood were determined with an enzymatic, fluorometric technique. Only one patient reacted with a temporary hypotension at declamping. Arterial pH was unaffected during the operation, and no hyperkalemia was noted. During clamping of the aorta an increased lactate/pyruvate ratio in muscle indicated tissue hypoxia. Energy charge (EC) was, however, unchanged and the adenylate pool was maintained possibly due to a decrease in PCr. In spite of the restitution of blood flow after declamping of the aorta, no normalization of the metabolic state was seen, not even 20 minutes after the release of the clamp. EC was still unchanged but a significant decrease of both the adenylate and creatine pools was seen. These findings might indicate a damage of the mitochondria and the cellular membranes in skeletal muscle after temporary arterial occlusion.

Adult↗

The switch from IgM to IgG secretion in single mitogen-stimulated B-cell clones.

The frequency of mitogen-reactive B cells yielding an IgG plaque-forming cell (PFC) response has been determined in vitro by limiting dilution analysis under culture conditions which allow every growth-induced B cell to grow and mature into a clone of Ig-secreting cells. The frequencies of lipopolysaccharide (LPS)-and lipoprotein-reactive precursors for IgG-secreting cells in the spleen of 6--8 wk old C3H/Tif and of C57BL/67 mice were found to be between 1 in 30 and 1 in 40 B cells and, therefore, only one tenth of the frequencies of mitogen-reactive precursors of clones secreting IgM. All IgG-secreting cells developed by switching in clones which previously contained IgM-secreting cells. This was shown in two experiments where the total number of mitogen-reactive precursor yielding IgM-secreting cell clones was limited such that 82 or 90% of all responding cultures originated from one precursor. Thus, of 480 cultures in the first and 720 cultures in the second experiment, 86 and 98 cultures were found positive, yielding IgM-secreting cells at day 5. When the same cultures were assayed at day 7 for IgG-secreting cells 9 and 10 cultures were found positive. All 19 cultures with IgG-secreting cells previously had contained IgM-secreting cells. The probability that IgG-secreting cells and IgM-secreting cells would have arisen from independent precursors can be calculated using Fisher's exact test of independence. For the two experiments those probabilities are 3.4 X 10(-7) and 4.0 X 10(-9). Since we have previously shown that each cell in a mitogen-stimulated, growing B-cell clone divides, and that each dividing cell secretes Ig, we conclude from these experiments that the large majority--in our experiments all--of the IgG-secreting cells in mitogen-stimulated B-cell clones develop by switch from IgM-secreting cells. IgG-secreting cells develop either early or late during growth of a single IgM-secreting cell clone. The switch to IgG secretion, therefore, is not fixed in the time of clonal growth after mitogenic stimulation.

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Frequencies of mitogen-reactive B cells in the mouse. I. Distribution in different lymphoid organs from different inbred strains of mice at different ages.

Frequencies of mitogen-reactive B cells have been determined in vitro under culture conditions which allow every growth-inducible B cell to grow and mature into a clone of Ig-secreting PFC. The frequencies of LPS-reactive B cells in the spleen of 6- to 8-wk old mice were between 1 in 3 and 1 in 10 splenic B cells from the following inbred strains of mice: C3H/Tif; BALB/c; BALB/c nu/nu; C57BL/6J; DBA/2J; C57BL/6J x DBA/(2J)F(1); and CBA and A/J. Very similar frequencies are found for lipoprotein-reactive B cells in BALB/c, BALB/c nu/nu, C3H/Tif, and C3H/HeJ mice. No LPS-reactive cells but normal frequencies of lipoprotein-reactive cells were found in C3H/HeJ mice, genetically nonreactive to LPS. SJL mice had significantly lower frequencies of LPS- and of lipoprotein-reactive B cells (1 in approximately 30 B cells). The number of LPS- and of lipoprotein-reactive B cells in spleen was dependent upon the age of the mouse. Newborn spleen contained approximately 10 percent of the number of reactive cells found at 6- to 8-wk of age. From there the frequencies declined again to drop below 5 percent of the maximal number at ages beyond 11 mo. LPS-reactive B cells yielding IgM- and IgG-PFC responses could be found in mesenteric lymph nodes, bone marrow, thymus, thoracic duct, and peripheral blood of 6- to 8-wk old mice. Their frequencies were one in three to five lymph node cells, 1 in 50 to 100 bone marrow cells, one in 10(5) thymus cells, and 1 in 20 to 40 thoracic duct or peripheral blood cells.

Aging↗