Interleukin-2 production

As was the case for most other cytokines, medical appraisal/use of IL-2 was initially impractical due to the minute quantities in which it is normally produced. Some transformed cell lines, most notably the Jurkat leukaemia cell line, produces IL-2 in increased quantities, and much of the IL-2 used for initial characterization studies was obtained from this source. Large-scale IL-2 production was made possible by recombinant DNA technologies. Although the IL-2 gene/cDNA has now been expressed in a wide variety of host systems, it was initially expressed in E. coli, and most products being clinically evaluated are obtained from that source. As mentioned previously, the absence of glycosylation on the recombinant product does not alter its biological activity. 

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Anti-hepatitis B virus activity in vitro and in vivo was also found in wogonin and baicalein (Fig. 4), the major active constituents of the traditional Chinese medicine Scutellaria radix.More recently, Blach-Olszewska et al investigated the effect of baicalein and wogonin on two important mechanisms of innate immunity The secretion of cytokines, and the natural resistance of human leukocytes to viral infection. The results obtained indicate that these fiavonoids modulate cytokine production, that is they inhibit interferons-a and -y, and stimulate tumor necrosis factor-a and interleukin production. They also augment the resistance of peripheral blood leukocytes to the vesicular stomatitis virus. 

Warner, R.L., Brooks, K., Pestka, J.J. (1994). In vitro effects of vomitoxin (deoxynivalenol) on T-cell interleukin production and IgA secretion. Food Chem. Toxicol. 32 617-25. 

A knockout mutant of the same L. acidophilus NCFM strain, lacking the S-layer A protein (SlpA) was significantly impaired in its binding to dendritic cells as L. acidophilus NCFM attaches to dendritic cells and induces changes in interleukin production. This mutant incurred a chromosomal rearrangement, which caused the expression of a second S-layer protein, SlpB. In the SlpB-expressing strain, the impact on the immune system was the opposite of the wild type, as proinflammatory cytokines were produced by the action of this strain with dendritic cells, with an opposite action detected in the wild type. 

Experts from academia, industry and government agencies reviewed past practices and experiences in design and evaluation of preclinical studies, including a discussion of relevant human experiences, for a variety of interferon and interleukin products. Acute phase responses common for many of these types of product include malaise, headache, myalgia, fever and anaemia. These responses have been generally more important in patients than in experimental animal studies. 

Table 13.1 Studies recommended for the safety evaluation of interferon and interleukin products...

Jujo K, Renz H, Abe J, Gelfand EW, Leung DYM. Decreased interferon gamma and increased interleukin production in atopic dermatitis promotes IgE synthesis. J Allergy Clin Immunol 1992 90(3 Pt 1) 323-331. 

Therapeutics. Therapeutic materials represent a class of polypeptides that are a low volume, high value product. The production system need not be very efficient but the quaHty of the recombinant protein has to be extremely pure (33,34). Thus high cost mammalian production systems can be tolerated. However, some of the therapeutic proteins such as insulin, human growth hormone, interleukins, interferon, and streptokinase are produced microbially. 

One component of the age-ielated decline in immune function is decreased production of the lymphokine that promotes the growth of T-ceUs, interleukin 2 (IL-2). Administration of recombinant-derived IL-2, both in vitro and in vivo, appears to restore certain immune functions in aged mice. Recovery of T-regulatory effects on B-ceU differentiation has been reported in human cells from elderly patients treated with IL-1 and/or IL-2 (42). Similar effects have been observed in the presence of the pentapeptide thymopentin [69558-55-0] (Arg Lys Asp Val Tyr), a weU-known IL-2 inducer. Recombinant IL-2 adrninistered to aged mice for three weeks has been shown to correct the T-ceU functional deficiency associated with antigen-specific immunoglobulin production by certain lymphoid tissue (43). 

Reconstitution of T-ceU deficiencies with thymic hormones has not been successhil even though the various hormone preparations induce prothymocyte differentiation and functions of mature T-ceUs. They do not regulate the maturation of thymocytes in the thymus. In contrast, IL-2, endotoxin, thymic epithehal cell products, but not interleukin 1, were found to promote functional maturation of immature thymocytes. Two classes of dmgs show thymomimetic actions (Table 2). Levamisole [14769-73-4], sodium salt of diethyl dithiocarbamate (imuthiol) and certain... 

This drug also is reported to activate macrophages, to iaduce polyclonal B-ceU activation as well as enhance specific antibody production m vivo, and to iaduce the synthesis of iaterferon and interleukin 1 (52). The iaduction of these important cytokiaes (and others) largely accounts for the profile of biological activity displayed by the pyrimidinones. Bropirimine is currentiy ia clinical evaluation for cancer, arthritis, and immunorestoration ia AIDS patients. 

DiaZepin Nucleosides. Four naturally occurring dia2epin nucleosides, coformycin (58), 2 -deoxycoformycin (59), adechlorin or 2 -chloro-2 -deoxycoformycin (60), and adecypenol (61), have been isolated (1—4,174,175). The biosynthesis of (59) and (60) have been reported to proceed from adenosine and C-1 of D-ribose (30,176,177). They are strong inhibitors of adenosine deaminase and AMP deaminase (178). Compound (58) protects adenosine and formycin (12) from deamination by adenosine deaminase. Advanced hairy cell leukemia has shown rapid response to (59) with or without a-or P-interferon treatment (179—187). In addition, (59) affects interleukin-2 production, receptor expression on human T-ceUs, DNA repair synthesis, immunosuppression, natural killer cell activity, and cytokine production (188—194). 

Inhibition of inflammatory cytokines (Fig. 2) Humanized monoclonal anti-TNF antibodies (Infliximab (Remicade ), Adalimumab (Humira )) bind with high selectivity to human TNF-a and neutralize its activity. Thereby, infliximab decreases the effects of enhanced TNF levels during inflammatory disease such as production of proteases, chemokines, adhesion molecules, cyclooxygenase products (prostaglandins), and proinflammatory molecules such as interleukin-1 and -6. The antibodies may also recognize membrane-bound TNF-a on lymphocytes and other immune cells. These cells may subsequently become apoptotic or are eliminated via Fc-receptor-mediated phagocytosis. 

Thrombopoietic factors (no recombinant TPO product in clinical use at this time IL-11 [recombinant product oprelvekin] has marketing approval) stimulate the production of megakaryocyte precursors, megakaryocytes, and platelets [8]. Interleukin-11 has many effects on multiple tissues, and can interact with IL-3, TPO, and SCF. AMG 531, a recombinant peptibody in that binds to the thrombopoetin receptor Mpl and stimulates the production of platelets, is in phase 1 and 2 studies and has been shown to safely increase platelet counts in patients with immune thrombocytopenic purpura [9]. 

Fig. 14 Effect of the molecular weight of tamarind seed xyloglucan depolymerized by ( ) 7-irradiation, ( ) ultrasonication, and ( ) endo-glucanase treatment on the production of various cytokines (Tumor necrosis factor a, TNE-a Interleukin 8, IL-8 Interleukin 10, IL-10 and Interleukin 12, IL-12) in HaCaT cells (Immortalized keratinocytes line) [301]...

Marshall JS. Leal-Berumen I. Nielsen L. Glibetic M. Jordana M Interleukin (IL)-IO inhibits long-term IL-6 production but not preformed mediator release from rat peritoneal mast cells. J Clin Invest 1996 97 1122-1128. 

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