Geneviève Lespinats

Histamine and serotonin suppression of lymphocyte response to phytohemagglutinin and allogeneic cells

Histamine added to murine spleen cells suppressed in vitro proliferation of lymphocytes induced by PHA or allogeneic spleen cells. Another vasoactive amine, serotonin (5-hydroxytryptamine), exerted a similar inhibitory activity on PHA- or allogeneic cell-induced lymphocyte proliferation. Anti-H2 histamine antagonists, cimetidine, metiamide, and ranitidine, blocked the histamine and serotonin suppressive effect. Cyproheptadine, an anti-H1 histamine and anti-serotonin antagonist, and methysergide, an anti-serotonin antagonist, also blocked histamine and serotonin inhibitory activities. These data suggest the presence, on lymphocytes, of receptors for serotonin which might be related to histamine receptors.

B-cell mitogenicity of carragheenan in mouse.

Abstract Carragheenan, a reported toxic agent for macrophages, has been studied for its properties of stimulation of mouse lymphocytes, by measuring the amount of incorporation of tritiated thymidine in cultures of lymphocytes in vitro . Unequivocal evidence has been obtained, concluding that carragheenan selectively activates mouse “B” cells from each strain tested, with the exception of C3H/He mice. Carragheenan probably acts on a subpopulation of B cells different from the populations stimulated by dextran sulfate and LPS.

Evidence for serotonin (5HT) binding sites on murine lymphocytes

The binding of 3H-labeled serotonin (or 5-hydroxytryptamine: 5HT) to mouse lymphocytes was investigated. It was shown to be highly specific, time-dependent, saturable and partly reversible. Saturation analysis demonstrated a Kd of 198 nM and B max of 3.53 nM. We studied receptor specificity by using different types of serotonin antagonists, and numerous other substances. Serotonin was found to be the most effective drug among those tested in inhibiting the binding of 3H-5HT, having an IC50 of 194 nM. The fact that 5HTP, a 5HT precursor, had no inhibitory capacity indicated the high specificity of these 5HT binding sites. Dopamine was somewhat able to competitively inhibit 5HT fixation (IC50 = 27,000 nM), whereas norepinephrine and histamine had no effect. Lastly, we investigated the cellular specificity of this binding, and observed that nonmacrophage peritoneal cells extensively bound serotonin under the same conditions as spleen cells. This is the first direct demonstration of 5-hydroxytryptamine receptors on mouse lymphocytes. The presence of these binding sites can contribute to the understanding of the suppressive effect of 5HT on mouse immunoreactivity.

Selective increased tissue histamine levels in tumour-bearing rodents

Histamine levels increased in the fundus of mice bearing a primary 3-methylcholanthrene-induced fibrosarcoma, and in the ventral skin, skeletal muscle and rumen of rats bearing a D.M.B.A.-induced mammary adenocarcinoma; they did not increase in the tissues of mice bearing a McC3-1 fibrosarcoma (38th passage) or a Lewis lung carcinoma before the appearance of metastasis, but an increase in histamine levels was observed in dorsal skin, ventral skin and fundus, after the appearance of metastasis.

Tissue histamine levels and mast cell numbers in tumour-bearing mice

In C57BL/6 mice bearing the 3LL carcinoma and in C3H mice bearing the McC3-l fibrosarcoma (18th passage), the increase in skin histamine levels was correlated with the increase in mast cell number. The number of cells able to incorporate tritiated thymidine was proportional to the mast cell number. These results strengthen the notion that, in tumour-bearing rodents, the increase in tissue histamine is an active phenomenon.

In vivo andin vitro anti-tumour activity of dimaprit

Dimaprit, a histamine H2-receptor agonist, injected daily i.p. to fibrosarcoma-bearing mice, induced a decrease in tumour growth and an increase in survival. Dimaprit, added to tumour cell cultures (10−4M), inhibited the incorporation of3H-thymidine while embryonic cell cultures were unaffected. This particular anti-tumour activity is probably H2-independent as histamine and impromidine have no effect on tumour cell cultures.

Cytotoxic activity of lymphoid cells from mice immunized with semiallogeneic hybrid cells: Requirement of in vitro lymphoid cells culture for expression of cytotoxicity against a syngeneic chemically induced tumor

Abstract Semiallogeneic somatic hybrid cells (AB2) derived from fusion of a C57B1/6 chemically induced fibrosarcoma (MCB6-1) and a fibroblastic cell (A9) of C3H origin were used to immunize C57B1/6 mice against the parental MCB6-1 tumor cells. In vitro immune lymphocytes were directly cytotoxic against AB2 hybrid cells and A9 allogeneic parental cells, but could not lyse the syngeneic MCB6-1 parental tumor cells. Nevertheless, after a 4-day culture of these immune lymphocytes, a cytotoxic activity against the syngeneic MCB6-1 tumor cells appeared; expression of such a cytotoxic activity did not require the presence of stimulator cells (mitomycin-treated MCB6-1 tumor cells) during the culture. This cytotoxicity is mediated by T cells, as it was completely abrogated by treatment with anti-Thy 1–2 antiserum and complement. These results suggest that a maturation or a differentiation of immune T lymphocytes occurs during in vitro culture, and is necessary for the expression of antitumor cytotoxicity.