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Oncol Rep. 1999 Mar-Apr;6(2):427-32.
Constitutive secretion of hematopoietic cytokines by human carcinoma cell lines and its up-regulation by interleukin-1 and phorbol ester.

Steube KG, Meyer C, Drexler HG.

DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen, GmbH, D-38124 Braunschweig, Germany.

We investigated secretion of hematopoietic growth factors in ten human tumor cell lines, derived from 5 different tissue types. The cell lines were stimulated transiently with interleukin-1 (IL-1 ), IL-6, interferon-c (IFN-c), lipopolysaccharide (LPS) and tetradecanoyl phorbol 13-acetate (TPA), respectively. Conditioned media (CM) were screened in a bioassay using an indicator cell line which is growth factor-dependent and needs the supply of at least one hematopoietic cytokine. The CM of almost all tumor cell lines analyzed induced proliferation of this indicator cell line as documented by incorporation of [3H]-thymidine. The majority of constitutively cytokine-producing tumor cell lines could be induced to further increase their amounts of secreted proliferation-inducing activity. The strongest inducers were TPA and IL-1 ; weakly or not effective at all were IFN-c, LPS and IL-6. Enzyme-linked immunosorbent assay (ELISA) identified several cytokines and macrophage-colony-stimulating factor (M-CSF) and IL-6 were secreted at the highest concentrations. Especially in kidney cell lines, the levels of granulocyte-macrophage-CSF (GM-CSF), M-CSF and IL-6 were further strongly increased by the TPA and IL-1 pretreatment. The by far highest amounts of granulocyte-CSF (G-CSF) were elaborated by the bladder cell line T-24 and could be further more than doubled by TPA. Other cell lines, derived from esophagus, lung and pancreas, responded less strongly to the pretreatment with the biomodulators. Our results demonstrating secretion of functionally active cytokines by human solid tumor cell lines suggest that these, to date so-called, hematopoietic cytokines are not entirely hematopoietic specific and might play a fundamentally important role in tumor cell biology.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10023015&dopt=Abstract

Hum Gene Ther. 1999 Jan 1;10(1):77-84.
In vivo tumor suppression by adenovirus-mediated interferon alpha2b gene delivery.

Ahmed CM, Sugarman BJ, Johnson DE, Bookstein RE, Saha DP, Nagabhushan TL, Wills KN.

Canji, Inc., San Diego, CA 92121, USA.

A replication-deficient adenovirus encoding human interferon alpha2b, driven by the human cytomegalovirus (CMV) promoter, was constructed and characterized. This construct was used to infect human cells derived from different types of cancer. The production of protein and its secretion into the culture medium were tested by Western blotting and immunoassay. Inhibition of cell proliferation and antiviral activity, two of the most important biological activities of interferon, were observed with this construct. PC-3 cells, derived from human prostatic cancer, or Hep3B cells, derived from human hepatocellular carcinoma, were injected subcutaneously to generate and establish in vivo tumors in athymic nude mice. Intratumoral injection with the recombinant adenovirus expressing interferon alpha2b resulted in complete regression of tumor growth. Our results demonstrate that interferon gene delivery using recombinant adenoviral vectors may be a useful approach to treat a variety of cancers.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10022532&dopt=Abstract

J Biol Chem. 1999 Mar 5;274(10):6698-703.
Interleukin-12 induces expression of interferon regulatory factor-1 via signal transducer and activator of transcription-4 in human T helper type 1 cells.

Coccia EM, Passini N, Battistini A, Pini C, Sinigaglia F, Rogge L.

Laboratory of Immunology, Istituto Superiore di Sanita, I-00161 Rome, Italy.

IRF-1-deficient mice show a striking defect in the development of T helper 1 (Th1) cells. In the present report, we investigate the expression of IRF-1 during differentiation of human T helper cells. No significant differences of IRF-1 mRNA expression were found in established Th1 and Th2 cells; however, interleukin 12 (IL-12) induced a strong up-regulation of IRF-1 transcripts in Th1 but not in Th2 cells. We demonstrate that IL-12-induced up-regulation of IRF-1 is mediated by signal transducer and activator of transcription-4, which binds to the interferon (IFN)-gamma-activated sequence present in the promoter of the IRF-1 gene. Strong IL-12-dependent activation of a reporter gene construct containing the IRF-1 IFN-gamma-activated sequence element provides further evidence for the key role of signal transducer and activator of transcription-4 in the IL-12-induced up-regulation of IRF-1 transcripts in T cells. IRF-1 expression was strongly induced after stimulation of naive CD4(+) T cells via the T cell receptor, irrespective of the cytokines present at priming, indicating that this transcription factor does not play a major role in initiating a Th1-specific transcriptional cascade in differentiating helper T cells. However, our finding that IRF-1 is a target gene of IL-12 suggests that some of the IL-12-induced effector functions of Th1 cells may be mediated by IRF-1.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10037767&dopt=Abstract

Antimicrob Agents Chemother. 1999 Mar;43(3):514-9.
Effects of the Chinese traditional medicine mao-bushi-saishin-to on therapeutic efficacy of a new benzoxazinorifamycin, KRM-1648, against Mycobacterium avium infection in mice.

Shimizu T, Tomioka H, Sato K, Sano C, Akaki T, Dekio S, Yamada Y, Kamei T, Shibata H, Higashi N.

Department of Microbiology and Immunology, Shimane Medical University, Japan.

The Chinese traditional medicine mao-bushi-saishin-to (MBST), which has anti-inflammatory effects and has been used to treat the common cold and nasal allergy in Japan, was examined for its effects on the therapeutic activity of a new benzoxazinorifamycin, KRM-1648 (KRM), against Mycobacterium avium complex (MAC) infection in mice. In addition, we examined the effects of MBST on the anti-MAC activity of murine peritoneal macrophages (M phi s). First, MBST significantly increased the anti-MAC therapeutic activity of KRM when given to mice in combination with KRM, although MBST alone did not exhibit such effects. Second, MBST treatment of M phi s significantly enhanced the KRM-mediated killing of MAC bacteria residing in M phi s, although MBST alone did not potentiate the M phi anti-MAC activity. MBST-treated M phi s showed decreased levels of reactive nitrogen intermediate (RNI) release, suggesting that RNIs are not decisive in the expression of the anti-MAC activity of such M phi populations. MBST partially blocked the interleukin-10 (IL-10) production of MAC-infected M phi s without affecting their transforming growth factor beta (TGF-beta)-producing activity. Reverse transcription-PCR analysis of the lung tissues of MAC-infected mice at weeks 4 and 8 after infection revealed a marked increase in the levels of tumor necrosis factor alpha, gamma interferon (IFN-gamma), IL-10, and TGF-beta mRNAs. KRM treatment of infected mice tended to decrease the levels of the test cytokine mRNAs, except that it increased TGF-beta mRNA expression at week 4. MBST treatment did not affect the levels of any cytokine mRNAs at week 8, while it down-regulated cytokine mRNA expression at week 4. At week 8, treatment of mice with a combination of KRM and MBST caused a marked decrease in the levels of the test cytokines mRNAs, especially IL-10 and IFN-gamma mRNAs, although such effects were obscure at week 4. These findings suggest that down-regulation of the expression of IL-10 and TGF-beta is related to the combined therapeutic effects of KRM and MBST against MAC infection.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10049260&dopt=Abstract

Biochem Biophys Res Commun. 1999 Feb 16;255(2):470-6.
Genomic organization, sequence analysis and transcriptional regulation of the human MCP-4 chemokine gene (SCYA13) in dermal fibroblasts: a comparison to other eosinophilic beta-chemokines.

Hein H, Schluter C, Kulke R, Christophers E, Schroder JM, Bartels J.

Clinical Research Unit, Department of Dermatology, University of Kiel, Germany.

The eosinophil chemotactic beta-chemokine MCP-4 is assumed to be involved in the accumulation of eosinophils characteristic for eosinophilic inflammatory diseases. We here describe the genomic organisation (3 exons of 138, 115 and 578 bp, 2 introns of 867 and 437 bp and 1.4 kb of regulatory sequences from the immediate 5' upstream region), sequence (genomic and transcribed) and mRNA expression of the human MCP-4 gene in dermal fibroblasts. Among the promoter elements potentially regulating MCP-4 gene expression and/or mediating the effects of anti-inflammatory drugs we identified consensus sequences known to interact with nuclear factors like NF-IL6, AP-2, a NF-kappaB like consensus sequence, gamma-interferon- response and YY-1 elements as well as glucocorticoid response elements. Like MCP-3, MCP-4 mRNA expression in dermal fibroblasts is upregulated by TNF-alpha, IL-1alpha, IFN-gamma or IL-4 and differs from RANTES and eotaxin mRNA expression in its response to IFN-gamma and/or IL-4. 1999 Academic Press.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10049733&dopt=Abstract

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