Jerzy A. Georgiades

Purified human immune interferon has more potent anticellular activity than fibroblast or leukocyte interferon

Abstract Human immune interferon preparations have anticellular activity on human cell lines (WISH and HEp-2). This anticellular activity copurified with the human immune interferon and appears to be a function of the immune interferon molecule. On the basis of a unit of antiviral activity, purified human immune interferon had about 20 and 100 times more anticellular activity than purified fibroblast or leukocyte interferon, respectively. The possible implications of this finding in the treatment of human neoplasia are discussed.

Classification of interferons with antibody to immune interferon

Abstract Mouse immune Interferon, induced by the T-cell mitogen staphylococcal enterotoxin A (SEA), was partially purified and used to immunize rabbits. The resulting antiserum neutralized all immune interferon preparations tested, including interferon induced in vitro by SEA, concanavalin A, phytohemagglutinin P, and pokeweed mitogen, and in mixed lymphocyte cultures. Interferon produced in vivo with specific antigen was also neutralized. The antiserum was equally potent against all these interferon preparations. The serum did not neutralize any virus-type interferon preparation tested, but immune interferon induced by SEA in athymic nude mouse spleen cells was neutralized. The neutralizing activity was precipitable by 33% ammonium sulfate, and was not removed by absorption of the serum with mouse cells. The data suggest that immune interferons produced under diverse conditions are antigenically the same or closely related.

Modulation of 4HNE-mediated signaling by proline-rich peptides from ovine colostrum

In previous studies we showed that colostrinin (CLN), a complex of proline-rich polypeptides derived from ovine colostrum, induces mitogenic stimulation, as well as a variety of cytokines in human peripheral blood leukocytes, and possesses antioxidant activity in pheochromocytoma (PC12) cells. In this study we investigated the effects of CLN on 4-hydroxynonenal (4HNE)-mediated adduct formation, generation of reactive oxygen species (ROS), glutathione (GSH) metabolism, and the modification of signal transduction cascade that leads to activation of c-Jun N-terminal kinase (JNK) in PC12 cells. Here we demonstrate that CLN (1) reduced the abundance of 4HNE-protein adducts, as shown by fluorescent microscopy and Western blot analysis; (2) reduced intracellular levels of ROS, as shown by a decrease in 2′,7′-dichlorodihydro-fluorescein-mediated fluorescence; (3) inhibited 4HNE-mediated GSH depletion, as determined fluorimetrically; and (4) inhibited 4HNE-induced activation of JNKs. Together, these findings suggest that CLN appears to down-regulate 4HNE-mediated lipid peroxidation and its product-induced signaling that otherwise may lead to pathological changes at the cellular and organ level. These findings also suggest further that CLN could be useful in the treatment of diseases such as Alzheimer’s, as well as those in which ROS are implicated in pathogenesis.

Nonsensitized lymphocytes produce leukocyte interferon when cultured with foreign cells.

Abstract Nonsensitized human lymphocytes produce interferon when cultured with either transformed or normal heterologous cells. De novo RNA and protein synthesis was not required in a foreign cell for it to act as the inducer. The interferon produced has been characterized as being leukocyte type rather than immune or fibroblast type.

[21] Large-scale induction and production of human and mouse immune interferons

Publisher Summary Production of large quantities of immune interferon in cultures is greatly facilitated when a suitable inducer is available. Using the T cell mitogen staphylococcal enterotoxin A, a system for production of sufficient human and mouse immune interferon for partial purification, antibody production, and initiation of clinical trials in humans has been developed. There are two general methods for in vitro induction of immune interferon in lymphocyte cultures. One method involves the use of specific antigen usually added to cultures containing lymphocytes previously sensitized to the antigen. This method is generally impractical with poor yields of immune interferon. The second method involves the use of T cell mitogens. Three such mitogens, phytohemagglutinin P (PHA-P), concanavalin A (Con A), and staphylococcal enterotoxin A (SEA) are compared. An alternative way to produce high concentrations of human immune interferon is to treat lymphocyte cultures with the enzyme galactose oxidase. This method has not yet been adapted to large-scale production, but it shows strong potentiality for such use because of the high yields and the fast kinetics of production.

Determinants of Protection by Human Immune Globulin against Experimental Herpes Neonatorum

Abstract There is no established prophylaxis of Herpes neonatorum. In experimental, newborn animals protection can be achieved by sufficient passive antibody given shortly after infection. Animal model data was sought to estimate a realistic and practical dose of human immune globulin for prophylaxis of newborn humans at risk. We used a neonatal mouse model and type II herpes simplex virus (HSV) to evaluate factors which govern the efficiency of antibody prophylaxis. Antibodies prepared in mice, rabbits, or man were equally protective. Timing experiments showed that these antibodies gradually lose their protective effect when administered more than 12 hr after infection. The first dose of multiple doses of antibody is the most crucial for protection. Intraperitoneal or subcutaneous injections of human immune globulin were equally effective. About 65 units of human anti-HSV antibody in 1-g newborn mice was the minimum dose which resulted in maximum protection. Comparison of this minimum protective dose with levels of neutralizing antibody in commercial immune globulin indicated that approximately 100 ml would be needed to achieve similar protective serum antibody levels in man. Six percent of potential blood donors were determined to have sufficient HSV antibody titers (≤1000 units) to prepare hyperimmune globulin at least 10 times more potent than commercially available immune globulin. Only 10 ml or less of such hyperimmune globulin is estimated to be protective. This reduced volume of hyperimmune globulin is easily injected into the loose subcutaneous tissue in the backs of newborn children.

[75] Partial purification of human immune interferon

Publisher Summary This chapter describes the purification of human interferon produced under large-scale culture conditions. The procedure results in both purification and potentiation of interferon activity. Matrex Blue does not always significantly increase the specific activity of the interferon, but this step appears to be necessary for the potentiation of activity observed in interferon eluted from the Ultrogel gel filtration column. The specific activity of the starting material is 10 2.6 units of interferon per milligram of protein, whereas that of the Ultrogel pool was10 5.53 units per milligram of protein, with the peak fraction containing 10 6.3 units per milligram of protein. Thus, a 13,800-fold increase in specific activity is obtained. This is the highest specific activity obtained for human immune interferon, and purified interferon should be useful for biological and functional studies.

Separation of Immune Interferon and MIF

Summary A preparation of mitogen-induced immune interferon and MIF was subjected to sequential salt precipitation with first 55% and then 80% saturated ammonium sulfate. At least 95% of the interferon activity was found in the reconstituted 55-80% precipitate, while MIF activity was found in both precipitates, but most active in the reconstituted 55% salt precipitate. The immune interferon and MIF activities of the 55-80% reconstituted salt precipitate were shown to be different by BSA-Affi-Gel 10 affinity chromatography. Both lymphokines were also shown to be heterogeneous by the above techniques.

[12] Large-scale production, concentration, and partial purification of human immune interferon

Publisher Summary This chapter discusses the large-scale production, concentration, and partial purification of human immune interferon. Induction of the synthesis of gamma interferon by lymphocytes is connected with the activation of a variety of peripheral white blood cells (PWBC) with nonspecific mitogens. Normally PWBC from a large number of donors are employed for large-scale production. In such situations, a mixed lymphocyte reaction occurs in addition to mitogen activation. As a result of the combined activation, different types of PWBC release into the media gamma interferon plus a variety of enzymes (proteases, phosphatases, oxygenases, phosphorylases, and lipases) and other lymphokines (interleukins 1, 2, and 3, inhibitors, other molecules, and growth factors). Isolation of gamma interferon from the products of activated PWBC and the proteins and mitogens introduced in the incubation medium creates formidable purification problems. To achieve success in purification, it is necessary to achieve adequate induction of gamma interferon by the PWBC. Gamma interferon batches gave an initial titer below 300 units/m. The above purification method is simple, effective, and relatively fast (purification time does not exceed 48 hr). This method can be applied to large quantities and can be totally automated without need for intermediate steps that would otherwise require adjustment of the material between steps of purification.

[77] Partial purification and characterization of mouse immune interferon

Publisher Summary This chapter discusses the methodology for the purification of mouse immune interferon that separates it from some of the other well-characterized lymphokines. Data on the use of the partially purified interferon for antibody production are presented for the purpose of characterizing mouse immune interferons that are produced under various conditions. The immune (type II) interferons are induced under a variety of conditions, in primed lymphocytes with specific antigens or in unprimed lymphocytes with T cell mitogens. Newcastle disease virus (NDV)-induced L cell interferon, interferon induced in mouse peripheral leukocytes stimulated with NDV, and interferon induced in mice by lipopolysaccharide (LPS) are all neutralized by anti-L cell interferon. The anti-immune interferon serum did not affect these interferons. In an effort to determine whether immune interferons induced in various ways are neutralized equally well by anti-immune interferon, relatively constant amounts of various interferons were mixed with different dilutions of the antiserum.