Comparison of the biological properties of purified natural and recombinant human interleukin-2

Naruo, Ken‐ichi; Hinuma, Shuji; Kato, Koichi; Koyama, Masaru; Tada, Hayato; Shiho, Osamu; Tsukamoto, Kikuo

doi:10.1016/0006-291x(85)91672-9

Cited by 49 publications

(12 citation statements)

References 27 publications

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“…No differences in the biological activities of the glycosylated (natural) and nonglycosylated (recombinant) forms of IL-2 have been observed in vitro (26,30), but no reports of in vivo differences have been published. Now that we have obtained several high-producer cell lines of IL-2, it will be possible to purify glycosylated as well as non-glycosylated IL-2, and to compare their physiological properties and therapeutic values.…”

Section: Resultsmentioning

confidence: 99%

The establishment of IL-2 producing cells by genetic engineering.

Sasada

Onda

Igarashi

1987

Cell Struct. Funct.

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ABSTRACT. Expression plasmids containing human interleukin-2(IL-2) cDNA under the control of viral promoters (SV40 early region, MuLV LTR, HTLV-I LTR, and ASV(Y73) LTR) were introduced into TK-mouse L cells and human FL cells to establish IL-2 producing cells. The highest levels of IL-2 producing clones were obtained in TK+ mouse L cells transformed with a recombinant plasmid having MuLV LTR as a promoter, whereas transformed cells of human FL cells (G418r) were revealed to produce IL-2 at the highest level when the cells were transfected with a plasmid containing HTLV LTR as a promoter. These results suggest that these promoter/enhancer regions possess different cell specificities in gene expression. To obtain higher levels of IL-2 production using gene amplification, the hybrid plasmids containing the hamster DHFR and human IL-2 genes were constructed and transfected into DHFR-CHO cells. DHFR+ colonies produced IL-2 at about the same level as that produced by TK+ L cells transformed with the recombinants containing MuLV LTR. Selection of methotrexate-resistant cells resulted in a 5-to 30-fold increase of IL-2 production. These cells produced IL-2 stably for at least 3 months, even in the absence of methotrexate.Studies on and the use of the physiologically active proteins are, in many cases, hampered by the limited amounts of the purified substance. Recombinant DNA technology is now widely used to produce such proteins, for instance, interferons, lymphokines and growth factors, in prokaryotes in high amounts. However, many of these secretary proteins from natural sources receive post-transcriptional modifications, such as glycosylation, which are not performed by prokaryotes. Several systems (7, 38) have been developed to express cloned DNA in mammalian cells, in which the resulting proteins are believed to be glycosylated and secreted. To obtain established cell lines that express the introduced foreign genes at high levels, the most suitable promoter-vector system has to be used for each host cell type. Furthermore, the gene amplification procedures have been known to be rather effective for obtaining a high level of gene expression (12,16,25,31).Here, we report the comparative studies on the suitable promoters for the expression of cDNA of human IL-2, a lymphokine which plays an essential role in immunoregulatory networks (3, 28) in mouse or human cells, and the establishment of cell lines which constitutively synthesize and secrete human IL-2. We also describe the expression of this gene in CHO cells by using two kinds of amplifiable vectors.

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Section: Resultsmentioning

confidence: 99%

The establishment of IL-2 producing cells by genetic engineering.

Sasada

Onda

Igarashi

1987

Cell Struct. Funct.

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“…Some investigators have reported that NK cells express IL 2 receptors and proliferated by exogeneous IL 2 (10,17,26 (25). A number of investigators have reported that IL 2 augmented the induction of not only antigen-specific but also nonspecific killer cells (5,18). Recent studies have revealed that IL 2 induced nonspecific killer cells, so-called lymphokineactivated killer (LAK) cells, which were cytotoxic against not only NK-sensitive but also NK-resistant tumor cells (15,23).…”

Section: Discussionmentioning

confidence: 99%

“…macrophages (4,9,18,26,28). Through these actions, IL 2 influences a variety of immune responses, including the induction of antigen-specific and -nonspecific killer cells, antibody production, and other lymphokine productions (4,18).…”

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confidence: 99%

“…Through these actions, IL 2 influences a variety of immune responses, including the induction of antigen-specific and -nonspecific killer cells, antibody production, and other lymphokine productions (4,18). In addition, early studies indicated that IL 2 receptors were expressed on T cells after stimulation with antigens or lectins in vitro (5,16), but recent studies indicate that IL 2 receptor-positive cells exist in the absence of any prior stimulation and are directly responsive to exogenous IL 2 (6,10,15,17,22,26).…”

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confidence: 99%

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Characteristics of Mouse Lymphoid Cells Proliferating in vitro by Recombinant Human Interleukin 2 (TGP‐3): Comparison between Normal and Nude Mice

Hinuma

Naruo

Shiho

et al. 1989

Microbiology and Immunology

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Various lymphoid cells obtained from BALB/c and BALB/c nu/nu mice were cultured in vitro with recombinant human interleukin 2 (rIL 2), and the characteristics of responder cells to rIL 2 were analyzed. Spleen cells, lymph node cells, and thymocytes except for bone marrow cells obtained from BALB/c mice remarkably proliferated in response to rIL 2. On the other hand, among lymphoid cells obtained from BALB/c nu/nu mice, only lymph node cells showed significant proliferation by rIL 2. Flow cytometric analyses revealed that mainly two types of lymphoid cells were proliferating in response to rIL 2 in BALI* mice, i.e., Thy 1+, Lyt 1-, Lyt 2-and Thy 1+, Lyt 1-, Lyt 2+ cells. On the other hand, most of the proliferating cells were Thy 1+, Lyt 1-, Lyt 2-cells in BALB/c nu/nu mice. Treatment with various antibodies plus complement revealed that the majority of IL 2-responsive cells in BALB/c mice were Thy 1+, Lyt 1+, and Lyt 2+, although a minor part of them were Thy 1-, Lyt 1-, and Lyt 2-. On the other hand, a predominant type of the IL 2-responsive cells in BALB/c nu/nu mice were Thy 1-, Lyt 1-, and Lyt 2-, though some were Thy 1+. Nonspecific killer activity against tumor cells increased to variable extents in all of the lymphoid cells of both strains after culture with rIL 2. Our results indicate that mouse responder cells to rIL 2 have the following characteristics. First, the responder cells exist abundantly among spleen, lymph nodes, and thymus in normal mice, though their cell lineages are heterogeneous; one is of T cell lineage and the other of natural killer (NK) cell lineage. Second, nude mice are defective in the responder cells of T cell lineage but not of NK cell lineage. Moreover, the responder cells in nude mice predominantly accumulate in the lymph nodes but not other lymphoid organs.Since Morgan et al first reported a lymphokine as a growth factor to T cells (16), numerous findings on interleukin 2 (IL 2) have appeared (11,27). Nowadays, the importance of IL 2 in the immune system is widely accepted, although the nature of responder cells to IL 2 remains to be fully characterized. In early studies, IL 2 was thought to support only the growth of T cells stimulated with lectins or antigens (5, 16), but recent studies have demonstrated that IL 2 regulates proliferation and differentiation in not only T cells but also other cells such as B cells, NK cells, and 761

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“…The biological and biochemical properties o f the rlL-2 have been described in detail elsewhere [2][3][4], One unit o f Takeda 11^2 is equivalent ot 383 U o f the Japan Reference Unit (JRU) and Jurkat Unit (NIH, USA). In the present study, JRU is expressed in all the results.…”

Section: Ril-2mentioning

confidence: 99%

Treatment of Advanced Primary Lung Cancer Associated with Malignant Pleural Effusion by the Combination of Immunotherapy and Chemotherapy

Miyao¹,

Chou²,

Ito³

et al. 1994

Oncology

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Twelve patients with advanced primary lung cancers associated with malignant pleural effusion were treated with intrathoracic instillation of recombinant interleukin-2 with or without in-vitro-sensitized cells. Two cases achieved complete response, and 7 partial response. The adverse effects seen in the protocol were marginal, and the protocol was well-tolerated and feasible. Furthermore, 4 cases were treated with the combination of systemic chemotherapy and adoptive immunotherapy. Of these, 3 cases responded well to the therapy and have shown a complete response for more than 20 months, indicating that adoptive immunotherapy together with chemotherapy might be a beneficial treatment for advanced lung cancer patients.

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Comparison of the biological properties of purified natural and recombinant human interleukin-2

Cited by 49 publications

References 27 publications

The establishment of IL-2 producing cells by genetic engineering.

The establishment of IL-2 producing cells by genetic engineering.

Characteristics of Mouse Lymphoid Cells Proliferating in vitro by Recombinant Human Interleukin 2 (TGP‐3): Comparison between Normal and Nude Mice

Treatment of Advanced Primary Lung Cancer Associated with Malignant Pleural Effusion by the Combination of Immunotherapy and Chemotherapy

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