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Visceral leishmaniasis (VL) or kala-azar, is a disease caused by the parasite Leishmania donovani, which is transmitted by the bite of sandflies. The clinical disease is characterized by fever, anemia, leukopenia (reduced white cell count), and hepatosplenomegaly (enlargement of the liver and spleen), and in full blown cases, by marked immunologic dysfunction. Cachectin, or tumor necrosis factor-alpha (TNF-alpha), is a cytokine, a nonantibody protein produced in response to specific antigenic stimuli. It acts as an intercellular mediator, and is believed to play a role in the mediation of normal cellular function, inflammation, and in the body's response to bacterial or parasitic infection. The levels of TNF-alpha in patients with VL were measured to assess the possibility of using these levels as indicators of the progress of the disease. The study group consisted of 58 patients with VL; 28 patients with acute disease, and 30 with cryptic disease. Cryptic disease patients had no symptoms of VL, but had a positive skin test, indicating an immune response to the infection. Fifteen healthy volunteers comprised the control group. Blood levels of TNF-alpha were elevated in the patients with acute VL prior to treatment and fell afterward. In cases refractory (resistant) to treatment, TNF-alpha levels remained high. Normal levels were observed in the asymptomatic and control groups. Levels of TNF-alpha appear to be a suitable marker for measuring the progress of VL and patient response to therapy. (Consumer Summary produced by Reliance Medical Information, Inc.)

Interferons (IFNs) initiate their effects by interacting with specific high-affinity cell surface receptors, but little is known about the physiology of IFN receptor interaction in vivo. Treatment of patients suffering from hairy cell leukemia (HCL) with human recombinant alpha IFN results in significant tumor regression, with clinical improvement in a high percentage of cases. To investigate a possible relevance of binding parameters as response markers, IFN receptor interaction on tumor cells responsive to IFN in vivo was studied. Binding of human alpha 2 IFN to circulating hairy cells was analyzed before and during IFN therapy in ten patients selected on the basis of high numbers of peripheral hairy cells. Binding experiments were carried out on Ficoll- Paque fractionated peripheral cell samples containing a majority of hairy cells. All patients reacted to recombinant alpha IFN treatment with a striking decrease in binding capacity within 12 hours after the first injection. As demonstrated by using a monoclonal antibody able to recognize alpha 2 IFN bound to its receptor, this decreased binding capacity was not due to blocking by circulating IFN but rather to a decrease in receptor number. This receptor “down-regulation” was partially reversible after the first IFN injection. However, upon prolonged IFN therapy, all patients displayed a stable state of decreased receptor expression. Down-regulation of IFN receptors can be regarded as a response marker to IFN treatment. This response marker, however, was not correlated with the clinical response within the first months of IFN therapy.

Seventeen patients with hairy cell leukemia (HCL) were treated with low doses of recombinant alpha interferon (IFN) for over 4 months. Marked improvement was observed in peripheral blood and bone marrow in 15 of 17 patients. Comparison of pretreatment values and hemograms obtained after 4 months of treatment showed a marked decrease in circulating hairy cells (P less than .01), a decrease in the number of lymphocytes (P less than .01), a rise in the number of platelets (P less than .05), granulocytes (P less than .05), and monocytes (P less than .01), and a rise in the hemoglobin level (P less than .01). Transient reduction in the number of granulocytes was noted during the first month. Correction of thrombocytopenia often appeared within 2 months and usually preceded improvement of anemia, monocytopenia, and neutropenia. Bone marrow biopsy specimens were taken before treatment and 2, 4, and 7 months after its initiation. The volumes occupied by hairy cells, cells of the myeloid lines, and adipocytes were studied by stereological analysis of semithin sections. Decrease in the volume occupied by hairy cells was seen after 4 months of treatment (P less than .01), and the volume continued to decrease at the seventh month (P less than .05). Hairy cells were no longer detected on bone marrow biopsies of 4 of 17 patients by the fourth month and in 3 of 8 additional patients by the seventh month. A rise in the volume occupied by normal myeloid cells was visible by the second month of treatment (P less than .01). Nevertheless, the volume occupied by granulocytes remained lower than in the normal controls (P less than .01). After an initial increase during the first 2 months of treatment (P less than .01), the overall cellularity remained unchanged at 4 months and decreased significantly (P less than .05) at 7 months. Except for biopsies at 2 months, mean cellularity was below that of control biopsies (P less than .01).

Hairy cells are stimulated to DNA synthesis by low molecular weight B cell growth factor (LMW-BCGF) and this proliferative response is suppressed by interferon (IFN)-alpha, both in vitro and in vivo. The suggestion that the CD23 molecule (Fc epsilon II receptor) might be involved in the signalling pathway of LMW-BCGF prompted us to study the expression of this molecule on hairy cells and its modulation by IFN- alpha. By flow cytometry and direct binding experiments with anti CD23 monoclonal antibodies, the presence of the CD23 antigen was detected in 7 of 12 cases tested, on variable percentages of cells, ranging from low to medium expression. In vitro incubation of hairy cells with IFN- alpha, which elicits a suppression of the proliferative response of these cells to LMW-BCGF, induced a parallel significant reduction of CD23 expression in only three cases. Similarly, a transient in vivo decrease of CD23 expression, concommitant with an inhibition of the LMW- BCGF response, could be detected in only one of three patients injected with IFN-alpha. Soluble sCD23/IgE-binding factor (BF) was quantitated in the serum from six other patients with hyperleukocytic hairy cell leukemia (HCL) undergoing a clinical trial of IFN-alpha therapy. Before treatment, these patients presented higher concentrations of the cleaved soluble form of the CD23 molecule than normal controls. Within a few weeks of IFN-alpha administration, these levels markedly decreased, paralleling a diminution of blood leukemic cells. Of interest, no such diminution was noticed for another patient resistant to IFN-alpha therapy. These results show that the proliferative response of hairy cells to LMW-BCGF is not linked to the expression of the CD23 marker. Besides, when the latter molecule was present, its decrease following IFN-alpha treatment, which could be detected in some cases, was not necessarily required for the suppression of the LMW-BCGF response and is thus not mandatory for the therapeutic efficacy of IFN- alpha. Our results point out that quantitation of serum sCD23/IgE-BF, whether related to a process of autocrine proliferation or not, is a parameter of potential importance for therapy monitoring.

Phytohemagglutinin-induced interferon-gamma (IFN-gamma) was reported to act on mouse leukemic L1210 R cells resistant to IFN-alpha and -beta. Results reported here show that these cells are also sensitive to various preparations of murine IFN-gamma derived from different sources and purified to different degrees and that lymphokines present in the preparations are not involved in the antiviral effect of IFN. In addition, IFN-gamma preparation increases concanavalin A binding to L1210 S and L1210 R cells indicating that the sensitivity of L1210 R cells to IFN-gamma is not limited to its antiviral effect.

Three fractions of Nocardia, Nocardia water-soluble mitogen (NWSM), Nocardia water-soluble mitogen pellet (NWSMP), and the cell wall peptidoglycan, which are mitogenic for B lymphocytes, were able to induce circulating interferon in mice, NWSMP and NWSM being the most active. The peak of interferon appeared about 2 h after injection. The interferon induced by NSWMP and NWSM was acid stable and antigenically related to viral interferon, as shown by neutralization with antibodies directed against Newcastle disease virus-induced interferon.

Different interferons can be obtained from the same animal species depending on the cells and (or) the inducers used. Interferons of type I and type II differ not only antigenically but also in molecular weight and stability at low pH. We have investigated whether mouse type I and type II interferons also differ in properties relating to their biological action. We present evidence which suggests that the molecular mechanism leading to antiviral and antigrowth effects induced by both types of interferon in susceptible cells must differ in at least one important step. Antiviral and antigrowth activities of type I but not of type II interferon are both inhibited when gangliosides are added to cell cultures together with the interferon. Whereas type I interferon strongly binds to ganglioside affinity columns and can be eluted with solutions of N-acetylneuraminyllactose, type II interferon passes through such columns unretarded. L-1210 mouse leukemia cells (L-1210S) respond equally well to antiviral and antigrowth activities of type I and type II interferons. Type I interferon-resistant L-1210 cells (L-1210R), derived from L-1210S cells after continuous culture in the presence of mouse fibroblast interferon, lack antiviral and antigrowth response to mouse type I interferon [Gresser, I., Bandu, H.T. & Brouty-Boyé, D. (1974) J. Natl. Cancer Inst. 52, 553-559]. However, these cells display the same sensitivity toward type II interferon as do the parent L-1210S cells from which they were derived and respond equally well to its antiviral and antigrowth activities.

Phytohemagglutinin is able to trigger interferon synthesis in spleen cell cultures from nude (nu/nu) mice as effectively as in splenic cell cultures from haired, control (nu/+), thymus-bearing mice. A minor theta-bearing cell population present in the spleen of nude mice appears essential to phytohemagglutinin interferon production, although cooperating cells are also required. The properties of nude mouse phytohemagglutinin interferon are indistinguishable from those displayed by the interferon induced in thymus-bearing mouse spleen cell cultures. Both interferons are unstable at pH 2 and cannot be neutralized by an antiviral interferon serum; hence, their characteristics correspond to those described for type T interferon. As in the case of viral interferon, pretreatment of L cells with nude phytohemagglutinin interferon induced specific enhanced phosphorylation of a 67,000-molecular-weight protein in vitro when cell extracts were incubated with double-stranded RNA and gamma-[32P]ATP.

Mengo virus double-stranded ribonucleic acid (dsRNA) was obtained on a semi-industrial scale from infected cultures of BHK-21 cells grown in suspension. Yield of the extraction and purification operations was small (about 22 mg from 1011cells in a 100-liter culture). Physicochemical characterization of this dsRNA gave an estimated molecular weight close to 4 × 106, a density of 1.59 (similar to that of the poliovirus dsRNA), and a thermal transition midpoint of 94 C. This product was a little more toxic for the mouse, by the intravenous route, than polyriboinosinic · polyribocytidylic acid (poly I:C) and strictly comparable in this respect to poliovirus dsRNA. The interferon-inducing capacity in the mouse and the antiviral activities in the mouse (infected with encephalomyocarditis, Semliki Forest, influenza, foot-and-mouth disease, and murine hepatitis viruses) and in the rabbit (Shope fibroma virus) of the ultraviolet light-inactivated product were practically identical, on a quantitative basis, with those of poly I:C. In vitro and in vivo experiments showed the dsRNA from Mengo virus to be slightly but significantly more resistant than poly I:C to the inactivating effect of human serum.

When noninfected L-cell suspension cultures are treated with interferon (specific activities superior to 106reference units per mg of protein), the cell-free cytoplasmic extracts obtained are inactive for the translation of exogenous natural mRNAs. The dose-response curve shows that comparable amounts of interferon are required to produce a 50% reduction of Mengo virus multiplication in vivo and Mengo RNA translation in vitro. With higher doses of interferon, Mengo RNA translation is completely abolished, while poly U translation and endogenous protein synthesis are only slightly affected. The inactivation of Mengo RNA translation is reversible; after removal of interferon, normal translation activity is regained together with the ability to support Mengo virus multiplication. Fractionation of the cell-free extracts shows that the effect is localized in the fraction which can be washed off the ribosomes by high salt. These results establish that interferon induces a block in genetic translation in noninfected L cells.