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J Surg Res. 1999 Mar;82(1):78-87.
Activated natural killer cell tumor retention and cytokine production in colon tumor using a tissue-isolated model.

Ribeiro U Jr, Whiteside TL, Basse PH, Safatle-Ribeiro AV, Huneke CE, Posner MC.

University of Pittsburgh, The University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, USA.

BACKGROUND: Adoptively transferred activated natural killer (A-NK) cells are capable of selectively infiltrating solid tumors, but only at low efficiency when administered systemically. It is unclear if human A-NK cells can be retained in tumor tissue and, if so, what is their action. We investigated intratumor A-NK cell retention and in situ cytokine production, using an xenogeneic ex vivo tissue-isolated tumor model, which permits direct intraarterial infusion. MATERIALS AND METHODS: Human colon adenocarcinoma (HT-29) was implanted in the ovarian fat pad of nude rats. The tumors were perfused ex vivo 25 to 30 days postimplant with a known number of cells, and the effluent was collected over time. The number of human A-NK cells and cell surface antigen expression of cells infused and exiting the tumor were calculated, using cell counts and flow cytometry, respectively. Frozen sections were stained with Giemsa and also immunostained for the presence of interleukin-2, -4, and -10, tumor necrosis factor alpha (TNF-alpha), and interferon. RESULTS: Six perfusions with 8 x 10(6) A-NK cells were performed. The mean number of infused A-NK cells that remained in the tumor at the completion of perfusion was 4.74 x 10(6) (59.2%). No differences were noted in cellular phenotype between the infused cells and the cells exiting the tumor: expression of the markers CD45 (97.5% vs 94. 5%), CD14 (0 vs 0), CD3 (3.83% vs 2.83%), and CD56 (86% vs 83%) was unchanged, P > 0.05. Microscopic examination of tumor sections showed tumor surrounded by A-NK cells, with some tumor nests infiltrated by A-NK cells. In situ immunopositivity for interleukin-2 (2/6), interleukin-4 (3/6), interleukin-10 (2/6), and TNF-alpha (2/6) specimens was observed. Immunostaining for interferon-gamma was negative. Conclusions. The retention of A-NK cells in the transplanted human colon tumor tissue was found to be efficient (59.2 %) in this model. Although perfusion time was limited, A-NK cells were able to infiltrate the tumor and initiate cytokine production. 1999 Academic Press.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10068530&dopt=Abstract

FASEB J. 1999 Mar;13(3):495-502.
Psoriatic keratinocytes show reduced IRF-1 and STAT-1alpha activation in response to gamma-IFN.

Jackson M, Howie SE, Weller R, Sabin E, Hunter JA, McKenzie RC.

Department of Dermatology, University of Edinburgh, Scotland, UK.

Psoriasis is a chronic inflammatory dermatosis characterized by hyperproliferative keratinocytes (KC). The skin lesions are infiltrated by T cells, which secrete gamma interferon (gamma-IFN) and are believed to be necessary to maintain the psoriatic phenotype. In normal KC, gamma-IFN is a potent inhibitor of proliferation, but proliferation of KC persists in psoriatic plaques despite the presence of gamma-IFN. Immunostaining of interferon regulatory factor-1 (IRF-1) revealed that IRF-1 was localized to the basal cells of the epidermis in normal and in nonlesional psoriatic skin, but was suprabasal or completely absent in lesional psoriatic skin. This finding led to the hypothesis that abnormal signaling in the gamma-IFN pathway may occur in psoriatic KC. To test this hypothesis, we measured activation of IRF-1 and signal transducer and activator of transcription (STAT)-1alpha transcription factors in KC after stimulation with gamma-IFN. Primary cultures of KC from normal and nonlesional psoriatic skin were stimulated with gamma-IFN and subsequent transcription factor activation was measured by electrophoretic mobility shift assay. Psoriatic KC showed a reduced induction of IRF-1 and STAT-1alpha activation after stimulation with gamma-IFN, compared with normal KC. Reduced activation of IRF-1 and STAT-1alpha in response to gamma-IFN indicates a fundamental defect in the growth and differentiation control of psoriatic KC in the absence of the influence of other cell types.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10064616&dopt=Abstract

Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2367-72.
Interferon gamma expressed by a recombinant respiratory syncytial virus attenuates virus replication in mice without compromising immunogenicity.

Bukreyev A, Whitehead SS, Bukreyeva N, Murphy BR, Collins PL.

Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Building 7, Room 100, 7 Center Drive, Bethesda, MD 20892-0720, USA.

Interferon gamma (IFN-gamma) has pleiotropic biological effects, including intrinsic antiviral activity as well as stimulation and regulation of immune responses. An infectious recombinant human respiratory syncytial virus (rRSV/mIFN-gamma) was constructed that encodes murine (m) IFN-gamma as a separate gene inserted into the G-F intergenic region. Cultured cells infected with rRSV/mIFN-gamma secreted 22 microg mIFN-gamma per 10(6) cells. The replication of rRSV/mIFN-gamma, but not that of a control chimeric rRSV containing the chloramphenicol acetyl transferase (CAT) gene as an additional gene, was 63- and 20-fold lower than that of wild-type (wt) RSV in the upper and lower respiratory tract, respectively, of mice. Thus, the attenuation of rRSV/mIFN-gamma in vivo could be attributed to the activity of mIFN-gamma and not to the presence of the additional gene per se. The mice were completely resistant to subsequent challenge with wt RSV. Despite its growth restriction, infection of mice with rRSV/mIFN-gamma induced a level of RSV-specific antibodies that, on day 56, was comparable to or greater than that induced by infection with wt RSV. Mice infected with rRSV/mIFN-gamma developed a high level of IFN-gamma mRNA and an increased amount of interleukin 12 p40 mRNA in their lungs, whereas other cytokine mRNAs tested were unchanged compared with those induced by wt RSV. Because attenuation of RSV typically is accompanied by a reduction in immunogenicity, expression of IFN-gamma by an rRSV represents a method of attenuation in which immunogenicity can be maintained rather than be reduced.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10051648&dopt=Abstract

J Pediatr. 1999 Mar;134(3):324-32.
Interferons and cerebral palsy.

Grether JK, Nelson KB, Dambrosia JM, Phillips TM.

California Birth Defects Monitoring Program, California Department of Health Services, Emeryville, California, USA.

OBJECTIVE: To explore the association of neonatal interferons (IFNs) with spastic cerebral palsy (CP) and with other measured substances. STUDY DESIGN: Assays of archived neonatal blood of 31 predominantly term children with CP and 65 children in a control group were obtained by recycling immunoaffinity chromatography with laser-enhanced fluorescence and chemiluminescence detection. RESULTS: Fourteen of 31 children with spastic CP had concentrations of IFNs-alpha, beta, and gamma exceeding any control. Levels of interleukins-1, 6, 8, tumor necrosis factor-alpha, chemokines, colony stimulating factors, transforming growth factor-beta, complement components and regulators, certain neuropeptides, and thyroid hormones also differed from control levels in these 14 children. The 17 children with CP whose IFN concentrations were within the control range had levels of inflammatory cytokines higher than but near to control values; 13 of these 17 had values for coagulation factors that exceeded control values. Seven of 9 children with spastic diplegia had high IFNs, and 8 of 10 hemiplegic children had normal IFNs. CONCLUSION: Neonatal IFNs exceeding control concentrations were associated with other biochemical and clinical indicators of inflammation and with spastic diplegia. In these children with CP, IFNs within the control range were associated with concentrations of other inflammatory markers that were near to control values and with spastic hemiplegia.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10064670&dopt=Abstract

J Infect Dis. 1999 Apr;179(4):1038-41.
Comparison of interferon-gamma, granulocyte colony-stimulating factor, and granulocyte-macrophage colony-stimulating factor for priming leukocyte-mediated hyphal damage of opportunistic fungal pathogens.

Gaviria JM, van Burik JA, Dale DC, Root RK, Liles WC.

Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, WA 98195, USA.

Proinflammatory cytokines have been proposed as adjunctive therapeutic agents to enhance the host immune response during infections caused by opportunistic fungi. The study compared the differential in vitro priming effects of interferon-gamma (IFN-gamma), granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) on hyphal damage of opportunistic fungi mediated by isolated neutrophils (polymorphonuclear leukocytes, PMNL) and buffy coat cells (polymorphonuclear leukocytes/peripheral blood mononuclear cells, PMNL/PBMC) from healthy donors. IFN-gamma (1000 U/mL) effectively primed both PMNL and PMNL/PBMC for enhanced hyphal damage of Aspergillus fumigatus, Fusarium solani, and Candida albicans. G-CSF (100 ng/mL) increased hyphal damage mediated by both PMNL and PMNL/PBMC against F. solani, and GM-CSF (100 ng/mL) augmented the antifungal activity of PMNL/PBMC against hyphal forms of both F. solani and C. albicans. IFN-gamma may be superior to G-CSF or GM-CSF for enhancing the microbicidal activity of PMNL and PMNL/PBMC against opportunistic fungi.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10068606&dopt=Abstract

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