Absence of peripheral blood mononuclear cells priming in hemodialysis patients

As a consequence of the proinflammatory environment occurring in dialytic patients, cytokine overproduction has been implicated in hemodialysis co-morbidity. However, there are discrepancies among the various studies that have analyzed TNF-alpha synthesis and the presence of peripheral blood mononuclear cell (PBMC) priming in this clinical setting. We measured bioactive cytokine by the L929 cell bioassay, and evaluated PBMC TNF-alpha production by 32 hemodialysis patients (HP) and 51 controls. No difference in TNF-alpha secretion was observed between controls and HP (859 ± 141 vs 697 ± 130 U/10(6) cells). Lipopolysaccharide (5 µg/ml) did not induce any further TNF-alpha release, showing no PBMC priming. Paraformaldehyde-fixed HP PBMC were not cytotoxic to L929 cells, suggesting the absence of membrane-anchored TNF-alpha. Cycloheximide inhibited PBMC cytotoxicity in HP and controls, indicating lack of a PBMC TNF-alpha pool, and dependence on de novo cytokine synthesis. Actinomycin D reduced TNF-alpha production in HP, but had no effect on controls. Therefore, our data imply that TNF-alpha production is an intrinsic activity of normal PBMC and is not altered in HP. Moreover, TNF-alpha is a product of de novo synthesis by PBMC and is not constitutively expressed on HP cell membranes. The effect of actinomycin D suggests a putative tighter control of TNF-alpha mRNA turnover in HP. This increased dependence on TNF-alpha RNA transcription in HP may reflect an adaptive response to hemodialysis stimuli.Universidade de São Paulo Instituto de Ciências Biomédicas Departamento de ImunologiaInstituto Butantan Laboratório de ImunogenéticaUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de MedicinaUNIFESP, EPM, Depto. de MedicinaSciEL

A priori estimation of accuracy and of the number of wells to be employed in limiting dilution assays

The use of limiting dilution assay (LDA) for assessing the frequency of responders in a cell population is a method extensively used by immunologists. A series of studies addressing the statistical method of choice in an LDA have been published. However, none of these studies has addressed the point of how many wells should be employed in a given assay. The objective of this study was to demonstrate how a researcher can predict the number of wells that should be employed in order to obtain results with a given accuracy, and, therefore, to help in choosing a better experimental design to fulfill one's expectations. We present the rationale underlying the expected relative error computation based on simple binomial distributions. A series of simulated in machina experiments were performed to test the validity of the a priori computation of expected errors, thus confirming the predictions. The step-by-step procedure of the relative error estimation is given. We also discuss the constraints under which an LDA must be performed

Antibodies to tumor necrosis factor: a component of B cell immune responses with a role in tumor/host interaction

Item does not contain fulltex

Absence of peripheral blood mononuclear cells priming in hemodialysis patients

As a consequence of the proinflammatory environment occurring in dialytic patients, cytokine overproduction has been implicated in hemodialysis co-morbidity. However, there are discrepancies among the various studies that have analyzed TNF-alpha synthesis and the presence of peripheral blood mononuclear cell (PBMC) priming in this clinical setting. We measured bioactive cytokine by the L929 cell bioassay, and evaluated PBMC TNF-alpha production by 32 hemodialysis patients (HP) and 51 controls. No difference in TNF-alpha secretion was observed between controls and HP (859 ± 141 vs 697 ± 130 U/10(6) cells). Lipopolysaccharide (5 µg/ml) did not induce any further TNF-alpha release, showing no PBMC priming. Paraformaldehyde-fixed HP PBMC were not cytotoxic to L929 cells, suggesting the absence of membrane-anchored TNF-alpha. Cycloheximide inhibited PBMC cytotoxicity in HP and controls, indicating lack of a PBMC TNF-alpha pool, and dependence on de novo cytokine synthesis. Actinomycin D reduced TNF-alpha production in HP, but had no effect on controls. Therefore, our data imply that TNF-alpha production is an intrinsic activity of normal PBMC and is not altered in HP. Moreover, TNF-alpha is a product of de novo synthesis by PBMC and is not constitutively expressed on HP cell membranes. The effect of actinomycin D suggests a putative tighter control of TNF-alpha mRNA turnover in HP. This increased dependence on TNF-alpha RNA transcription in HP may reflect an adaptive response to hemodialysis stimuli

Cationic Liposomes As Non-viral Vector For Rna Delivery In Cancer Immunotherapy

This review presents the current status in the use of liposomes as non-viral vector for nucleic acid delivery in cancer immunotherapy. Currently, cancer treatment uses surgery, radiotherapy and/or chemotherapy. The search for new strategies to improve the efficiency of conventional treatments is a challenge, and biological therapy has emerged as a promising technique. Immunotherapy is a branch of biological therapy that uses the body's immune system to detect and destroy cancer cells. One immunotherapy approach is the activation of T lymphocytes from cancer patients by dendritic cells (DCs) loaded with tumor antigens. Among different antigens, mRNA coding the tumor antigens is advantageous due to its capability to be amplified from small amounts of tumor tissue, its safety because it is easily degraded without integrating into the host genome, and it does not need to cross the nuclear barrier to exert its biological activity. Nanotechnology is an approach to deliver tumor antigens into DCs. Specially; we review the use of nanoliposomes in the field of cancer therapy because cationic liposomes can be used as non-viral vectors for mRNA delivery. Aside from the promise of liposomes, the development of scalable processes and facilities to the use this individualized therapy is still a challenge. Thus, we also present the recent techniques used for liposome production. In this context, the integration between technological knowledge in the production of cationic liposomes and immunotherapy using mRNA may contribute to the development of new strategies for cancer therapy. © 2013 Bentham Science Publishers.7299110Misra, R., Acharya, S., Sahoo, S.K., Cancer nanotechnology: Application of nanotechnology in cancer therapy (2010) Drug Discov Today, 15 (19-20), pp. 842-850Susa, M., Milane, L., Amiji, M., Hornicek, F., Duan, Z., Nanoparticles: A promising modality in the treatment of sarcomas (2011) Pharm Res, 28 (2), pp. 260-272Bangham, A.D., Standish, M.M., Watkins, J.C., Diffusion of univalent ions across the lamellae of swollen phospholipids (1965) J Mol Biol, 13 (1), pp. 238-252Sheng, W.Y., Huang, L., Cancer immunotherapy and nanomedicine (2011) Pharm Res, 28 (2), pp. 200-214Sullenger, B.A., Gilboa, E., Emerging clinical applications of RNA (2002) Nature, 418 (6894), pp. 252-258Trevisan, J.E., Cavalcanti, L.P., Oliveira, C.L.P., de la Torre, L.G., Santana, M.H.A., Technological aspects of scalable processes for the production of functional liposomes for gene therapy (2011) Non-Viral Gene Therapy. In Tech, pp. 267-292. , In: Yuan X-b, EdsWood, G.W., (2002) Composition and Method of Cancer Antigen Immunotherapy, , US6406699Rubiolo, C., (2011) Dendritic Cells, , US20110097346Maeda, H., Greish, K., (2005) Antitumor Agent and Process For Producing the Same, , US20050208136Tomalia, D.A., Pulgam, V.R., Swanson, D.R., Huang, B., (2011) Janus Dendrimers and Dendrons, , US7977452Klimash, J.W., Brothers, H.M., Swanson, D.R., Yin, R., Spindler, R., Tomalia, D.A., Hsu, Y., Cheng, R.C., (2000) Disulfidecontaining Dendritic Polymers, , US6020457Kim, J.U., Choi, H.Y., (2010) X-ray System For Dental Diagnosis and Oral Cancer Therapy Based On Nano-material and Method Thereof, , US7771117Hirsch, A., Sagman, U., Wilson, S.R., Rosenblum, M.G., Wilson, L.J., (2008) Use of Carbon Nanotube For Drug Delivery, , US20080193490Carol, M.P., Heanue, J.A., (2009) Delivery System For Radiation Therapy, , US20090154646Zhukov, T.A., Ostapenko, S., Sutphen, R., Lancaster, J., Sellers, T.A., Zhang, J.Z., (2006) Luminescence Characterization of Quantum Dots Conjugated With Biomarkers For Early Cancer Detection, , US20060003465Bao, G., Nie, S., Nitin, N., la Conte, L., (2005) Multifunctional Magnetic Nanoparticle Probes For Intracellular Molecular Imaging and Monitoring, , US20050130167Holland, J.W., Madden, T.D., Cullis, P.R., (1999) Bilayer Stabilizing Components and Their Use In Forming Programmable Fusogenic Liposomes, , US5885613Duzgunes, N., Simoes, S., Slepushkin, V., de Lima, M.C.P., (2001) Nonligand Polypeptide and Liposome Complexes As Intracellular Delivery Vehicles, , US6245427Chancellor, M.B., Fraser, M.O., Chuang, Y.-C., de Groat, W.C., Huang, L., Yoshimura, N., (2006) Application of Lipid Vehicles and Use For Drug Delivery, , US7063860Rahman, A., (1990) Liposome-encapsulated Vinca Alkaloids and Their Use In Combatting Tumors, , US4952408Graham, R., Barbisin, M., (2006) Cationic Liposomes and Methods of Use, , US20060159738Mezei, M., Nugent, F.J., (1984) Method of Encapsulating Biologically Active Materials In Multilamellar Lipid Vesicles (MLV), , US4485054Kung, V.T., Canova-Davis, E., (1986) Liposome Immunoassay Reagent and Method, , US4622294Tournier, H., Schneider, M., Guillot, C., (1999) Liposomes With Enhanced Entrapment Capacity and Their Use In Imaging, , US5980937Ho, J.-A., Lin, Y.-C., (2011) Device For Preparation of Liposomes and Method Thereof, , US20110163468Barenholz, Y., Gabizon, A., (1990) Liposome/doxorubicin Composition and Method, , US4898735Rahman, A., (2002) Method of Administering Liposomal Encapsulated Taxane, , US6461637Rahman, A., Rafaeloff, R., Husain, S.R., (1995) Liposome Encapsulated Taxol and A Method of Using the Same, , US5424073Iga, K., Hamaguchi, N., Ogawa, Y., (1991) Liposome Composition and Production Thereof, , US5000959Esuvaranathan, K., Mahendran, R., Lawrencia, C., (2010) Methods and Compositions For Delivery of Pharmaceutical Agents, , US7709457Santana, M.H.A., Rosada, R.S., Castelo, A.A.M.C., Silva, C.L., de la Torre, L.G., (2009) Ternary Liposomal Composition Containing a Polynucleotide, , WO2009073941Stewart, B.W., Kleihues, P., (2003) World Cancer Report, , IARC Press: Lyon, France(2011) Are the number of cancer cases increasing or decreasing in the world?, , http://www.who.int/features/qa/15/en/index.html, (WHO) WHO Available at, Accessed on: July 16Morgan, G., Ward, R., Barton, M., The contribution of cytotoxic chemotherapy to 5-year survival in adult malignancies (2004) Clin Oncol, 16 (8), pp. 549-560Pastor, F., Kolonias, D., Giangrande, P.H., Gilboa, E., Induction of tumour immunity by targeted inhibition of nonsense-mediated mRNA decay (2010) Nature, 465 (7295), pp. 227-230Folkman, J., Tumor angiogenesis: Therapeutic implications (1971) N Engl J Med, 285 (21), pp. 1182-1186Pang, R.W., Poon, R.T., Clinical implications of angiogenesis in cancers (2006) Vasc Health Risk Manag, 2 (2), pp. 97-108Bertino, J.R., Hait, W., Princípios do tratamento do câncer (2005) Cecil, Tratado De Medicina Interna. Rio De Janeiro, pp. 1316-1330. , In: Ausiello D, Goldman L, Eds., Brazil: Elsevier(2011) Biological Therapy, , http://www.cancer.gov/cancertopics/treatment/biologicaltherapy, (NCI) NCI, Available at, Accessed on: August 28Hanahan, D., Weinberg, R.A., Hallmarks of Cancer: The Next Generation (2011) Cell, 144 (5), pp. 646-674Coley, W.B., The Treatment of Inoperable Sarcoma by Bacterial Toxins (the Mixed Toxins of the Streptococcus erysipelas and the Bacillus prodigiosus) (1910) Proc R Soc Med, 3, pp. 1-48(2007) Understanding the Immune System - How it Works, pp. 1-54. , National Institute of Allergy and Infectious Diseases, U.S. Departament of Healthy and Human Services National Institutes of Health, Eds. USA: NIH PublicationPrestwich, R.J., Errington, F., Hatfield, P., Merrick, A.E., Ilett, E.J., Selby, P.J., The immune system - is it relevant to cancer development, progression and treatment? (2008) Clin Oncol, 20 (2), pp. 101-112Pulendran, B., Banchereau, J., Maraskovsky, E., Maliszewski, C., Modulating the immune response with dendritic cells and their growth factors (2001) Trends Immunol, 22 (1), pp. 41-47Markovic, S.N., Celis, E., Antibodies and vaccines as novel cancer therapeutics (2006) Novel Anticancer Agents, pp. 207-221. , In: Alex AA, John KB, Eds., Burlington: Academic PressScanlan, M.J., Gure, A.O., Jungbluth, A.A., Old, L.J., Chen, Y.-T., Cancer/ testis antigens: An expanding family of targets for cancer immunotherapy (2002) Immunol Rev, 188 (1), pp. 22-32Dunn, G.P., Bruce, A.T., Ikeda, H., Old, L.J., Schreiber, R.D., Cancer immunoediting: From immunosurveillance to tumor escape (2002) Nat Immunol, 3 (11), pp. 991-998Steinman, R.M., Dendritic cells: Understanding immunogenicity (2007) Eur J Immunol, 37 (S1), pp. S53-S60Banchereau, J., Steinman, R.M., Dendritic cells and the control of immunity (1998) Nature, 392 (6673), pp. 245-252Baleeiro, R.B., Anselmo, L.B., Soares, F.A., Pinto, C.A.L., Ramos, O., Gross, J.L., High frequency of immature dendritic cells and altered in situ production of interleukin-4 and tumor necrosis factor-α in lung cancer (2008) Ancer Immunol Immunother, 57 (9), pp. 1335-1345Sallusto, F., Lanzavecchia, A., Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/ macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha (1994) J Exp Med, 179 (4), pp. 1109-1118Barbuto, J.A.M., Ensina, L.F.C., Neves, A.R., Bergami-Santos, P.C., Leite, K.R.M., Marques, R., Dendritic cell-tumor cell hybrid vaccination for metastatic cancer (2004) Cancer Immunol Immunother, 53 (12), pp. 1111-1118Gilboa, E., Vieweg, J., Cancer immunotherapy with mRNAtransfected dendritic cells (2004) Immunol Rev, 199, pp. 251-263Banchereau, J., Briere, F., Caux, C., Davoust, J., Lebecque, S., Liu, Y.J., Immunobiology of dendritic cells (2000) Annu Rev Immunol, 18, pp. 767-811Ashley, D.M., Faiola, B., Nair, S., Hale, L.P., Bigner, D.D., Gilboa, E., Bone marrow-generated dendritic cells pulsed with tumor extracts or tumor RNA induce antitumor immunity against central nervous system tumors (1997) J Exp Med, 186 (7), pp. 1177-1182Boczkowski, D., Nair, S.K., Nam, J.H., Lyerly, H.K., Gilboa, E., Induction of tumor immunity and cytotoxic T lymphocyte responses using dendritic cells transfected with messenger RNA amplified from tumor cells (2000) Cancer Res, 60 (4), pp. 1028-1034Boczkowski, D., Nair, S.K., Snyder, D., Gilboa, E., Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo (1996) J Exp Med, 184 (2), pp. 465-472Granstein, R.D., Ding, W., Ozawa, H., Induction of anti-tumor immunity with epidermal cells pulsed with tumor-derived RNA or intradermal administration of RNA (2000) J Invest Dermatol, 114 (4), pp. 632-636Koido, S., Kashiwaba, M., Chen, D., Gendler, S., Kufe, D., Gong, J., Induction of antitumor immunity by vaccination of dendritic cells transfected with MUC1 RNA (2000) J Immunol, 165 (10), pp. 5713-5719Nair, S.K., Heiser, A., Boczkowski, D., Majumdar, A., Naoe, M., Lebkowski, J.S., Induction of cytotoxic T cell responses and tumor immunity against unrelated tumors using telomerase reverse transcriptase RNA transfected dendritic cells (2000) Nat Med, 6 (9), pp. 1011-1017Zhang, W., He, L., Yuan, Z., Xie, Z., Wang, J., Hamada, H., Enhanced therapeutic efficacy of tumor RNA-pulsed dendritic cells after genetic modification with lymphotactin (1999) Hum Gene Ther, 10 (7), pp. 1151-1161Heiser, A., Dahm, P., Yancey, D.R., Maurice, M.A., Boczkowski, D., Nair, S.K., Human dendritic cells transfected with RNA encoding prostate-specific antigen stimulate prostate-specific CTL responses in vitro (2000) J Immunol, 164 (10), pp. 5508-5514Heiser, A., Maurice, M.A., Yancey, D.R., Coleman, D.M., Dahm, P., Vieweg, J., Human dendritic cells transfected with renal tumor RNA stimulate polyclonal T-cell responses against antigens expressed by primary and metastatic tumors (2001) Cancer Res, 61 (8), pp. 3388-3393Heiser, A., Maurice, M.A., Yancey, D.R., Wu, N.Z., Dahm, P., Pruitt, S.K., Induction of polyclonal prostate cancer-specific CTL using dendritic cells transfected with amplified tumor RNA (2001) J Immunol, 166 (5), pp. 2953-2960Nair, S.K., Boczkowski, D., Morse, M., Cumming, R.I., Lyerly, H.K., Gilboa, E., Induction of primary carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocytes in vitro using human dendritic cells transfected with RNA (1998) Nat Biotech, 16 (4), pp. 364-369Sæbøe-Larssen, S., Fossberg, E., Gaudernack, G., mRNA-based electrotransfection of human dendritic cells and induction of cytotoxic T lymphocyte responses against the telomerase catalytic subunit (hTERT) (2002) J Immunol Methods, 259 (1-2), pp. 191-203Strobel, I., Berchtold, S., Götze, A., Schulze, U., Schuler, G., Steinkasserer, A., Human dendritic cells transfected with either RNA or DNA encoding influenza matrix protein M1 differ in their ability to stimulate cytotoxic T lymphocytes (2000) Gene Ther, 7 (23), pp. 2028-2035Su, Z., Peluso, M.V., Raffegerst, S.H., Schendel, D.J., Roskrow, M.A., The generation of LMP2a-specific cytotoxic T lymphocytes for the treatment of patients with Epstein-Barr virus-positive Hodgkin disease (2001) Eur J Immunol, 31 (3), pp. 947-958Thornburg, C., Boczkowski, D., Gilboa, E., Nair, S.K., Induction of cytotoxic T lymphocytes with dendritic cells transfected with human papillomavirus E6 and E7 RNA: Implications for cervical cancer immunotherapy (2000) J Immunother, 23 (4), pp. 412-418van Tendeloo, V.F.I., Ponsaerts, P., Lardon, F., Nijs, G., Lenjou, M., van Broeckhoven, C., Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: Superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells (2001) Blood, 98 (1), pp. 49-56Weissman, D., Ni, H., Scales, D., Dude, A., Capodici, J., McGibney, K., HIV gag mRNA transfection of dendritic cells (DC) delivers encoded antigen to MHC class I and II molecules, causes DC maturation, and induces a potent human in vitro primary immune response (2000) J Immunol, 165 (8), pp. 4710-4717Heiser, A., Coleman, D., Dannull, J., Yancey, D., Maurice, M.A., Lallas, C.D., Autologous dendritic cells transfected with prostatespecific antigen RNA stimulate CTL responses against metastatic prostate tumors (2002) J Clin Invest, 109 (3), pp. 409-417Bawa, R., Bawa, S.R., Maebius, S.B., Flynn, T., Wei, C., Protecting new ideas and inventions in nanomedicine with patents (2005) Nanomed Nanotechnol Biol Med, 1 (2), pp. 150-158Moghimi, S.M., Hunter, A.C., Murray, J.C., Nanomedicine: Current status and future prospects (2005) FASEB Journal: Official Publication of the Federation of American Societies For Experimental Biology, 19 (3), pp. 311-330Ferrari, M., Cancer nanotechnology: Opportunities and challenges (2005) Nat Rev Cancer, 5 (3), pp. 161-171Ferrari, M., Nanovector therapeutics (2005) Curr Opin Chem Biol, 9 (4), pp. 343-346Fredika, R., Mauro, F., Introduction and rationale for nanotechnology in cancer therapy (2006) Nanotechnology For Cancer Therapy, pp. 3-10. , In: Eds, CRC PressShigeru, K., Mitsuru, H., Yuriko, H., Pharmacokinetics of Nanocarrier-Mediated Drug and Gene Delivery (2006) Nanotechnology For Cancer Therapy, pp. 43-58. , In: Amiji MM, Eds., CRC PressFenart, L., Casanova, A., Dehouck, B., Duhem, C., Slupek, S., Cecchelli, R., Evaluation of effect of charge and lipid coating on ability of 60-nm nanoparticles to cross an in vitro model of the bloodbrain barrier (1999) J Pharmacol Exp Ther, 291 (3), pp. 1017-1022Furumoto, K., Nagayama, S., Ogawara, K.-I., Takakura, Y., Hashida, M., Higaki, K., Hepatic uptake of negatively charged particles in rats: Possible involvement of serum proteins in recognition by scavenger receptor (2004) J Controlled Release, 97 (1), pp. 133-141Oberdürster, G., Toxicology of ultrafine particles: In vivo studies. Philosophical Transactions of the Royal Society of London. Series A: Mathematical (2000) Physical and Engineering Sciences, 358 (1775), pp. 2719-2740Ogawara, K.-I., Yoshida, M., Higaki, K., Toshikiro, K., Shiraishi, K., Nishikawa, M., Hepatic uptake of polystyrene microspheres in rats: Effect of particle size on intrahepatic distribution (1999) J Controlled Release, 59 (1), pp. 15-22Ogawara, K.-I., Yoshida, M., Kubo, J.-I., Nishikawa, M., Takakura, Y., Hashida, M., Mechanisms of hepatic disposition of polystyrene microspheres in rats: Effects of serum depend on the sizes of microspheres (1999) J Controlled Release, 61 (3), pp. 241-250Hamaguchi, T., Matsumura, Y., Suzuki, M., Shimizu, K., Goda, R., Nakamura, I., NK105, a paclitaxel-incorporating micellar nanoparticle formulation, can extend in vivo antitumour activity and reduce the neurotoxicity of paclitaxel (2005) Br J Cancer, 92 (7), pp. 1240-1246Davis, M.E., Chen, Z., Shin, D.M., Nanoparticle therapeutics: An emerging treatment modality for cancer (2008) Nature Reviews Drug Discovery, 7, pp. 771-782Brigger, I., Dubernet, C., Couvreur, P., Nanoparticles in cancer therapy and diagnosis (2002) Adv Drug Deliv Rev, 54 (5), pp. 631-651Randall, M., Active Targeting Strategies in Cancer with a Focus on Potential Nanotechnology Applications (2006) Nanotechnology For Cancer Therapy, pp. 19-42. , In: Amiji MM, Eds., CRC PressDuncan, R., Nanomedicine gets clinical (2005) Mater Today, 8 (8 SUPPL.), pp. 16-17Shimab, S.A.K., Anwar, S., Jain, P., Nano structure based drug delivery system: An approach to treat cancer (2012) Int J Drug Develop Res, 4 (2), pp. 394-407Antunes, F.E., Marques, E.F., Miguel, M.G., Lindman, B., Polymervesicle association (2009) Adv Colloid Interface Sci, 147-148, pp. 18-35Filipe, E.J.M., Quando as moléculas se auto-organizam: Micelas e outras estruturas supremoleculares (1996) Revista De Cultura Científica, 18, pp. 25-38Park, J.W., Benz, C.C., Martin, F.J., Future directions of liposome-and immunoliposome-based cancer therapeutics (2004) Semin Oncol, 31 (13 SUPPL.), pp. 196-205Evans, D.F., Wennerström, H., (1999) The Colloidal Domain - Where Physics, Chemistry, Biology, and Technology Meet, , 2nd ed. Wiley-vch: USAIsraelachvili, J.N., (1992) Intermolecular and Surface Forces, , 2nd ed. Academic Press: California, USALorenz, R.M., Edgar, J.S., Jeffries, G.D.M., Zhao, Y., McGloin, D., Chiu, D.T., Vortex-trap-induced fusion of femtoliter-volume aqueous droplets (2006) Anal Chem, 79 (1), pp. 224-228Yagi, K., (1986) Medical Application of Liposomes, , Japan Scientific Societies PressHiemenz, P.C., Rajagopalan, R., (1997) Principles of Colloid and Surface Chemistry, , third edition, revised and expanded. Taylor & FrancisLasic, D.D., (1993) Liposomes: From Physics to Applications, , Elsevier Science Publishers B.V.: AmsterdamPortney, N., Ozkan, M., Nano-oncology: Drug delivery, imaging, and sensing (2006) Anal Bioanal Chem, 384 (3), pp. 620-630Forssen, E.A., Tokes, Z.A., Use of anionic liposomes for the reduction of chronic doxorubicin-induced cardiotoxicity (1981) Proc Natl Acad Sci USA, 78 (3), pp. 1873-1877Robert, N.J., Vogel, C.L., Henderson, I.C., Sparano, J.A., Moore, M.R., Silverman, P., The role of the liposomal anthracyclines and other systemic therapies in the management of advanced breast cancer (2004) Semin Oncol, 31 (13 SUPPL.), pp. 106-146Straubinger, R.M., Lopez, N.G., Debs, R.J., Hong, K., Papahadjopoulos, D., Liposome-based therapy of human ovarian cancer: Parameters determining potency of negatively charged and antibody-targeted liposomes (1988) Cancer Res, 48 (18), pp. 5237-5245Campbell, R.B., Balasubramanian, S.V., Straubinger, R.M., Influence of cationic lipids on the stability and membrane properties of paclitaxel-containing liposomes (2001) J Pharm Sci, 90 (8), pp. 1091-1105Rowinsky, E.K., Donehower, R.C., Paclitaxel (Taxol) (1995) N Engl J Med, 333 (1), pp. 1004-1014Dye, D., Watkins, J., Suspected anaphylactic reaction to Cremophor EL (1980) Br Med J, 280 (6228), p. 1353Lorenz, W., Reimann, H.J., Schmal, A., Dormann, P., Schwarz, B., Neugebauer, E., Histamine release in dogs by Cremophor E1 and its derivatives: Oxethylated oleic acid is the most effective constituent (1977) Agents Actions, 7 (1), pp. 63-67Yoshizawa, Y., Kono, Y., Ogawara, K.-I., Kimura, T., Higaki, K., PEG liposomalization of paclitaxel improved its in vivo disposition and anti-tumor efficacy (2011) Int J Pharm, 412 (1-2), pp. 132-141Sapra, P., Allen, T.M., Ligand-targeted liposomal anticancer drugs (2003) Prog Lipid Res, 42 (5), pp. 439-462Sorgi, F.L., Huang, L., Large scale production of DC-Chol cationic liposomes by microfluidization (1996) Int J Pharm, 144 (2), pp. 131-139New, R.R.C., (1994) Liposomes: A Practical Approach, , IRL Press: Oxford University PressTorre, L.G., Carneiro, A.L., Rosada, R.S., Silva, C.L., Santana, M.H.A., A mathematical model describing the kinetic of cationic liposome production from dried lipid films adsorbed in a multitubular system (2007) Braz J Chem Eng, 24, pp. 477-486Barnadas-Rodriguez, R., Sabes, M., Factors involved in the production of liposomes with a high-pressure homogenizer (2001) Int J Pharm, 213 (1-2), pp. 175-186Jahn, A., Vreeland, W.N., de Voe, D.L., Locascio, L.E., Gaitan, M., Microfluidic Directed Formation of Liposomes of Controlled Size (2007) Langmuir, 23 (11), pp. 6289-6293Su, Z., Dannull, J., Heiser, A., Yancey, D., Pruitt, S., Madden, J., Immunological and clinical responses in metastatic renal cancer patients vaccinated with tumor rna-transfected dendritic cells (2003) Cancer Res, 63 (9), pp. 2127-2133Bonehill, A., van Nuffel, A.M., Corthals, J., Tuyaerts, S., Heirman, C., Francois, V., Single-step antigen loading and activation of dendritic cells by mRNA electroporation for the purpose of therapeutic vaccination in melanoma patients (2009) Clin Cancer Res, 15 (10), pp. 3366-3375Grunebach, F., Muller, M.R., Nencioni, A., Brossart, P., Delivery of tumor-derived RNA for the induction of cytotoxic T-lymphocytes (2003) Gene Ther, 10 (5), pp. 367-374Milazzo, C., Reichardt, V.L., Müller, M.R., Grünebach, F., Brossart, P., Induction of myeloma-specific cytotoxic T cells using dendritic cells transfected with tumor-derived RNA (2003) Blood, 101 (3), pp. 977-982Müller, M.R., Grünebach, F., Nencioni, A., Brossart, P., Transfection

Thioglycollate-elicited murine macrophages are cytotoxic to Mycoplasma arginini-infected YAC-1 tumor cells

Macrophages are important components of natural immunity involved in inhibition of tumor growth and destruction of tumor cells. It is known that these cells can be activated for tumoricidal activity by lymphokines and bacterial products. We investigated whether YAC-1 tumor cells infected with Mycoplasma arginini stimulate nitric oxide (NO) release and macrophage cytotoxic activity. Thioglycollate-elicited macrophages from male BALB/c mice were co-cultured for 20 h with YAC-1 tumor cells infected or not with Mycoplasma arginini. The cytotoxic activity was evaluated by MTT assay and nitrite levels were determined with the Griess reagent. Thioglycollate-elicited macrophages co-cultured with noninfected YAC-1 cells showed low cytotoxic activity (34.7 ± 8.6%) and low production of NO (4.7 ± 3.1 µM NO2-). These macrophages co-cultured with mycoplasma-infected YAC-1 cells showed significantly higher cytotoxic activity (61.4 ± 9.1%; P<0.05) and higher NO production (48.5 ± 13 µM NO2-; P<0.05). Addition of L-NAME (10 mM), an inhibitor of NO synthesis, to these co-cultures reduced the cytotoxic activity to 37.4 ± 2% (P<0.05) and NO production to 3 ± 4 µM NO2- (P<0.05). The present data show that Mycoplasma arginini is able to induce macrophage cytotoxic activity and that this activity is partially mediated by NO