Search

Inflammatory exudate (SE) cells were collected from subcutaneous coverslips in mice and transferred into lethally irradiated (1,000 r) recipients. Eight days after transplantation 59Fe incorporation in the spleen and bone marrow was significantly greater than in controls treated with the suspending medium only. One hundred percent of mitoses were of the T6T6 karyotype in the marrow and spleen when SE cells were obtained from CBA/T6T6 donors. The repopulating potential of SE cells, however, lagged significantly behind that of bone marrow cells and the failure to observe consistently macroscopic spleen colonies calls into question whether the observed regeneration was due to pluripotent stem cells. Radioautographic studies with 3H-TdR showed that the majority of SE cells had recently been generated, but long-lived, noncycling cells of lymphoid and monocytoid morphology were also present in the exudate.

The proliferative kinetics of the intraepithelial lymphocytes (IL) of the mouse intestine have been evaluated. By inducing mitotic arrest it was found that large IL - constituting about 50% of the IL - showed a mitotic rate of 2.3. Autoradiographic results obtained after two different schedules of 3H-thymidine injections showed that 30% of the large IL were in DNA synthesis, and that the large IL were renewed at a rate comparable to that of blast cells from Peyer's patches, mesenteric lymph nodes and thoracic duct lymph. The small IL were renewed very rapidly compared to small lymphocytes of peripheral lymphoid tissues, although small lymphocytes with lifespans of several weeks were also present in the epithelial sheet. By the use of intestinal perfusion, in vivo, it was estimated that the loss of lymphocytes from intestinal villi into the lumen of the gut was negligible, and it is concluded that the most probable kinetic model for the majority of IL is: B and T lymphoblasts invade the epithelium and undergo mitosis. B lymphoblasts give rise predominantly to plasma cells, and T lymphoblasts give rise to small lymphocytes - probably long-lived - which reenter the circulation.

Lymphocytes in thymic cortex and germinal centers of lymphoid tissues are labeled intensely with generally labeled tritiated deoxycytidine [G-3H]dCyd whereas they are weakly labeled with methyl tritiated deosythymidine [methyl3H]dThd of the same specific activity, not only by single injection but also by an intensive injection schedule. [G-3H]dCyd can be used to label short-lived lymphocytes strongly, although not specifically. The distribution patterns of labeled lymphocytes were different depending on the injection schedules of [G-3H]dCyd. [G-3H]dCyd can be used as a precursor molecule for cytosine and also thymine found in DNA. The ratios of radioactive thymine to crytosine measured biochemically on DNA extracted from radioactive lymphocytes labeled by the various schedules indicate strongly that short- and long-lived lymphocyte populations have different abilities to utilize pyrimidine nucleosides for DNA synthesis.

Autoradiography and scintilation counting have been used after various schedules of 3H-thymidine injections to evaluate lymphocyte kinetics in normal and thymus-deprived BALB/C mice. The thymus was found to be an active production site of small lymphocytes, the majority of these cells having thymic residence times of three to five days. Peripheral lymphoid tissues--including spleen, lymph nodes and Peyer's patches--were dominated by long-lived lymphocytes and produced very few small lymphocytes. Nearly identical percentages of long-lived lymphocytes with comparable grain counts were found in the peripheral tissues. In adult thymectomized animals, and in mice thymectomized and reconstituted with bone marrow cells following total body irradiation, percentages and lifespan of long-lived lymphocytes were found to be of the same order as in normal or sham-operated controls. It is concluded that T cells in the peripheral long-lived pool can be formed outside the thymus and that the bone marrow probably produces long-lived B lymphocytes.

The kinetics of lymphoid cells within the epithelium of the small gut has been studied in various thymus-deprived mice and in antigen-deprived mice by the use of 3H-thymidine injections and radioautography. In thymus-deprived mice--including adult thymectomized, thymectomized and irradiated, neonatally thymectomized, and nude mice - and in germ-free mice decreased numbers of intraepithelial lymphocytes (IL) were found. On the other hand, the radioautographic results indicated that the remaining IL populations included both newly formed and long-lived lymphoid cells in the same percentages as found in sham-operated controls and normal mice. It is concluded that although the presence of the thymus and the antigen content of the gut is of importance to the maintenance of the numbers of cells in the lymphoid populations of the intestinal wall, the basic kinetics of these cell populations are preserved in deprived mice.

Healing skin wounds were studied in a series of parabiotic rats. The femurs of one parabiont of each pair were shielded whilst both animals were given 800 r from a Co(60) source. The animals were wounded 3 days after irradiation. Each animal with partially shielded marrow was then given tritiated thymidine intraperitoneally daily while the cross-circulation was arrested by clamping. After the thymidine-(3)H had cleared the blood, the clamp was released. Animals were sacrificed, and wounds were prepared for radioautography 1, 2, and 6 days after wounding. In the wounds of the shielded animals thymidine-(3)H was observed in epidermis, endothelium, leukocytes, fibroblasts, and mast cells. Only neutrophilic leukocytes, monocytes, and lymphocytes were labeled, as determined by light and electron microscope radioautography, in the wounds of each nonshielded parabiont. None of the many fibroblasts present were found to contain label in the wounds of the nonshielded parabionts through the 6 day period. These observations provide further evidence that wound fibroblasts do not arise from hematogenous precursors and, therefore, must arise from adjacent connective tissue cells.

When Lewis rats were exposed to antilymphocytic serum, produced in rabbits, to Lewis lymphoid cells, the transformation of their thoracic duct lymphocytes in response to foreign lymphoid cells was markedly reduced in comparison with that in response to similar cells from donors treated with saline or normal rabbit serum. It is suggested that the mixed lymphocyte reaction may be used as an in vitro test for the in vivo activity of antilymphocytic serum.