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Notes: An IgE-binding factor(s) (IgE-BF(s)) was partially purified from the supernatant of human HTLV-II carrying T-cell line MO, This IgE-BF(s) was shown to increase the IgE synthesis in the human myeloma cell line U-266, but did not affect its viability or growth. The of effect of the IgE-BF(s) was dose-dependent and selective for IgE protein synthesis as β2-microglobulin synthesis in the U-266 and the immunoglobulin production in the U-1958 IgG-secreting human myeloma cell line were unaffected. The IgE-BF(s) increased the production of the c heavy chain but not the X light chain production. The IgE-BF(s) was distinct from IL-1β. IL-3, IL-4, IL-5, lL-6, TNF-α, IFN-α -β, -γ, M-CSF, and fragments of CD23.

Notes: Rats were immunized with ovalbumin, either subculaneously or by aerosol inhalation. The lymphocyte distribution in lymph nodes, peripheral blood, and spleen was investigated by flow cytometry after labelling with T pan (0X19 and W3/13), T helper lymphocytes (W3/25), T cytotoxic/suppressor lymphocytes (OX8), k light chain (MAR 18-5), or MHC class II (0X6) monoclonal antibodies. The influence of the neurotoxic agent capsaicin on the lymphocyte distribution was also analysed. Subcutaneous immunization resulted in an increased number of OX8+ cells in mesenteric lymph nodes, spleen, and peripheral blood but not in the draining lymph nodes, axillary, brachial, and mediastinal lymph nodes. The number of positive cells for the other cell markers used were not affected by immunization. The neuromodulatory effect of capsaicin had no effect on the lymphocyte distribution. The results showed that the type of immunization used, low amounts of antigen without adjuvant given during a prolonged period, selectively induced OX8+ cells. The patterns were unaffected by neuromodulation using capsaicin.

Notes: Baltic salmon Salmo salar females displaying wiggling behaviour had significantly lower (P〈0.05) hepatic and ovarian thiamine (vitamin B1) concentrations than the normal females, confirming that they suffered from a thiamine deficiency. A significantly (P〈0.05) increased monoaminergic activity was found in the telencephalon and the hypothalamus of the wiggling individuals as indicated by [5-hydroxyindoleacetic acid (5-HIAA)]: [5-hydroxytryptamine (5-HT)] and [3,4-dihydroxyphenylacetic acid (DOPAC)]: [dopamine (DA)] ratios. The 5-HIAA concentrations of wiggling individuals were significantly (P〈0.05) higher in the telencephalon and the hypothalamus compared to normal fish. Wiggling fish showed significantly (P〈0.05) higher concentrations of the DA metabolite DOPAC in the hypothalamus and the brain stem compared to normal fish. Furthermore, the brain stem in wiggling fish contained significantly (P〈0.05) less 5-HT than in normal individuals, which was also reflected in a significant (P〈0.05) increase in the (5-HIAA): (5-HT) ratio. These results demonstrate an increased serotonergic and dopaminergic activity in wiggling compared to normal fish. The altered monoaminergic activity may be directly related to altered brain thiamine metabolism, but a general stress caused by thiamine deficiency and an inability to regulate swim bladder inflation may contribute. Furthermore, a changed brain monoaminergic activity may contribute to the behaviour characterizing wiggling fish.

Notes: We have studied the influence of substance P (SP) on the proliferative response of concanavalin A (ConA)-activated peripheral blood lymphocytes from 16 birch pollen-allergic patients, sampled before and during the pollen season, and from 15 normal individuals. The median response to ConA 3μg/ml in the presence of SP 10-11 -10-6 M, was in most instances within ±10% of the control value for cells from both healthy and atopic individuals. However, the individual differences were considerable. Analysis of the proliferative responses to ConA of the cells from the allergic patients sampled before and during season, revealed higher responses in the presence of 10-6 than of 10-2 M SP. This was in contrast to the findings in the normal individuals: only half of their cells showed such increased responses. This difference in response frequency was statistically significant between allergic patients before season and normal individuals (P 〈 0.05) and between allergic patients during season and normal individuals (P 〈 0.01). The difference in proliferation rate in the SP concentration interval, 10-6 to 10-7 M, for the cells from allergic patients, sampled both before and during season, was significantly different from the cells from healthy individuals (P 〈 0.03 and P 〈 0.001 respectively). The cells sampled from four allergic subjects during the birch pollen season showed a more profoundly decreased response to ConA in the presence of SP 10-8 M, compared with their cells sampled before season. Such responses were never seen with cells sampled before season and with cells from normal individuals. The results suggest an involvement of SP in the immunoregulation, particularly in patients exhibiting allergic reactions to birch pollen.

Notes: The cell line HMC-1, derived from a patient with mast cell leukaemia, is the only established cell line exhibiting a phenotype similar to that of human mast cells. This paper reports on a detailed characterization of the expression of a panel of markers for various types of immature and mature haematopoietic cells in the HMC-1. We also studied the potential of HMC-1 to differentiate upon treatment with conditioned media from the human T-cell line Mo, retinoic acid or DMSO.HMC-1 was found to express several mast cell-related markers. A high expression of Kit, the receptor for stem-cell factor, was detected. The majority of the cells were stained with a MoAb against the mast cell-specific serine protease tryptase. Of particular interest was the finding that β-tryptase mRNA, but not a-tryptase mRNA, was expressed in HMC-1. Using enzyme-histochemistry we were able to show that the β-tryptase was enzymatically active, indicating that tryptase can form active homotetramers. Both heparin and chondroitin sulfate were found to be present in approximately equal amounts. HMC-1 lacked surface expression of the high-affinity IgE receptor, which was confirmed by the absence of mRNA of the α- and β-chains of the IgE-receptor complex. However, a strong expression of the 7-chain of the IgE-receptor complex was detected. A positive staining of the monocyte/macrophage marker CD68 was obtained, as well as a strong hybridization signal for the eosinophilic/basophilic-related differentiation marker the Charcot-Leyden crystal. Treatment of HMC-1 with conditioned media from the human T-cell line Mo, retinoic acid or DMSO induced only moderate changes in the surface or intracellular expression of the studied markers. The agents tested neither induced any of the monocyte/ granulocyte markers examined, nor expression of the FceRIα-chain.

Notes: Expression of a mast cell tryptase mRNA was detected in two human monocytic cell lines, the U-937 and the Mono Mac 6, and in normal human peripheral blood(PB) monocytes. In the U-937 cell line but not in normal PB monocytes, the tryptase expression was upregulated 3–50 fold following phorbol ester (PMA)-induced differentiation, but no such induction was seen after retinoic acid, interferon-γ or vitamin D3 exposure.The tryptases expressed in PMA-induced and non-induced U-937 and in Mono Mac 6 were characterized by PCR amplification and nucleotide sequence analysis. The U-937 cell line was found to express a tryptase identical to one of the previously cloned mast-cell β tryptases (Tryptase I), and the tryptase expressed in Mono Mac 6 was found to be nearly identical to the previously cloned α tryptase. By northern blot analysis with oligonucleotide probes specific for the α and β tryptases both cell lines were found to express only one type of tryptase. Densitometric quantifications of tryptase mRN A levels, in the two cell lines, showed approximately 80 times higher mRNA levels in Mono Mac 6 compared to non-induced U-937. Immunohistochemical staining for tryptase showed a marked heterogeneity in the Mono Mac 6 cell line. Only one out of 10 cells were positive for the protein but the levels in these cells were very high, equivalent, or even higher than the levels seen in the human mast cell line HMC- 1. This shows that the expression of a single tryptase, in this case the a tryptase, is sufficient for the production of a stable protein and probably also a stable proteolytically active tetramer.The family of human mast-cell tryptases has been considered to represent a class of proteases specifically expressed in mast cells and basophilic leucocytes. The expression of tryptases in two monocytic cell lines and in normal PB monocytes indicate that in humans, the lineage specificity of these serine proteases is less restricted than earlier expected.The cloning of a full length cDNA for the murine counterpart to the human mast cell tryptases, the MMCP-6, is presented. No expression of the MMCP-6 was detected in a panel of mouse monocyte or macrophage cell lines indicating a species difference in the lineage specificity of the ‘mast cell tryptases’.

Notes: The human leukaemia cell line KU812 has previously been used to study basophil differentiation. In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts. KU812 cells were treated with tetradecanoyl-phorbol-13-acetate (TPA), conditioned medium from the human T-cell line Mo (Mo-CM), or cultured under serum free conditions. After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity. Mo-CM and serum starvation increased the expression while TPA treatment down-regulated the expression of FcεRI-α chain. An increase in tryptase content in cell extracts was detected after 4 days of culture in serum-free medium or in the presence of Mo-CM. KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA. Mast cell-derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells. Furthermore, affinity-purified lung tryptase did not show any mitogenic activity. Platelet-derived growth factor was also excluded. Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.

Notes: LAMA-84, a human leucocytic cell line, which upon establishment was described as having megakaryocytic, erythroid and granulocytic characteristics, was analysed for expression of various differentiation markers. In addition to some of the previously described phenotypic characteristics, this cell line was found to express mRNA for several proteins characteristic for basophilic leucocytes and mast cells. The authors show that LAMA-84 cells express mRNA for the mast cell tryptase, the proteoglycan core protein, carboxypeptidase A and the α and β chains of the high affinity IgE receptor (FcεRI). The authors examined the potential of LAMA-84 to differentiate in serum-free medium or after DMSO or PMA treatment. Depending on the inducing factor, surface expression of the FcεRI α-chain was increased from 20% to 35–50% of the cells and mRNA levels for tryptase were increased in serum-free medium and after DMSO treatment. LAMA-84 was found to express CD13, CDw17, CD29, CD33, CD40, CD45 and CD117. Furthermore, mRNA for the eosinophil/basophil markers Charcot–Leyden crystal (CLC) protein and the major basic protein (MBP), as well as the erythrocyte differentiation marker α-globin, was detected. However, the authors observed only trace amounts of mRNA for another erythroid differentiation marker (glycophorin), trace amounts of the megakaryocytic marker GPIIIa, and no detectable level of GPIbα. By comparing the expression pattern of a panel of differentiation markers in LAMA-84, and a second human cell line (KU812) expressing a basophil phenotype, it is evident that these cell lines, which presently are the only two cell lines identified with basophilic characteristics, share a large number of phenotypic characteristics.