Interferon Matters A quarterly newsletter by PBL InterferonSource interferonsource.com May 2007, Issue 2 u What’s Inside What’s Your IFN-a Type? New Products Are all Type I IFNs created equal? Of Mice and Men page 4 & 5 Type I Interferons were the first cytokines to be The biological significance of the expression of discovered. They are a family of homologous cytokines so many different IFN-a subtypes has yet to be Rhesus/Cynomolgus originally identified for their antiviral activity but have determined. However, reports suggest that they show IFN-a2 Subtype also been shown to have both anti-proliferative and quantitatively distinct anti-viral, anti-proliferative, immunomodulatory activities. In humans and mice and killer cell-stimulatory activities. It is therefore Mouse IFN-a4 Subtype they are encoded by an intronless multigene family of importance for researchers to characterize and clustered on human chromosome 9 and murine study these IFN-a subtypes and their possible post- chromosome 4 respectively. translational modifications. Research Tools Also in both species there are multiple Interferon- The common models for IFN-a subtypes study are page 6 & 7 Alpha (IFN-a) genes and one Interferon-Beta (IFN-b) human and mouse. In 1998, Lawson et al showed that Human IFN-a gene. Other Type I Interferon genes described include there are in vivo differences in the antiviral activity of the Interferon-Omega (IFN-w) gene, murine limitin the mouse IFN-a subtypes.1 Using an experimental Multi-Subtype ELISA Kit gene, as well as the human/murine Interferon-Epilson, mouse model of IFN transgene expression in vivo, TM Interferon-Kappa, and Interferon-Tau (IFN-e, IFN-k, and the authors showed that mouse IFN-a1 has a greater iLite Human IFN-a Kit IFN-t). In humans, at least 13 IFN-a are transcribed. antiviral activity of the three IFN subtypes (a1, a4 and Two of these genes (IFNA1 and IFNA13) are identical a9) tested for MCMV infection. in coding sequences and give rise to a single protein Protocols & Tips species. The coding sequences of the rest of human Table 1: Human Interferon Alpha Subtypes Nomenclature page 8 & 9 IFN-a genes diverge up to 8% and give rise to 12 Human Interferon Alpha Molecular Gene Theoretical pI Protein: Handle with Care protein subtypes. Similarly, one IFN-b and 13 IFN-a Subtypes Nomenclature Weight genes have been described in the mouse. Table 1 lists aA a2A IFNA2 19241 5.99 ELISA Data Analysis the common Human IFN-a subtypes. It is important to a2 a2b IFNA2 19269 5.99 note that human IFN-a nomenclature is different from aB2 a8 IFNA8 19484 5.18 murine IFN-a nomenclature. aC a10 IFNA10 19406 5.70 All IFN-a subtypes and IFN-b are structurally similar aD (Val 114) a1 IFNA1 19414 5.18 Product Citations aF a21 IFNA21 19312 5.99 page 10 and share a common cell surface receptor, which aG a5 IFNA5 19695 5.46 is composed of two chains designated as IFNAR1 aH2 a14 IFNA14 19708 6.38 and IFNAR2. In humans, only one IFN-a subtype is aI a17 IFNA17 19299 5.46 N-glycosylated and a few IFN-a species have been aJ1 a7 IFNA7 19606 5.87 aK a6 IFNA6 20061 6.43 shown to be O-glycosylated. In the mouse, nearly all a1 (Ala 114) aD IFNA1 19386 5.18 of the IFN-a species are N-glycosylated. Globally, a4a aM1 IFNA4 19379 5.58 glycosylated IFN-a subtypes do not differ from non- a4b a4 IFNA4 19379 5.76 glycosylated subtypes in terms of their biological aWA a16 IFNA16 19282 5.96 activities. A variety of studies have suggested that References: these IFN-a subtypes possess overlapping but also 1. Gewart, DR, et al (1995). J Interferon Cytokine Res. 15(5):403 2. Hussain, M., et al (1997) J. Interferon Cytokine Res. 17(9):559 unique sets of distinctive biological activities as well 3. Hussain, M., et al (2000) J. Interferon Cytokine Res. 20:763 as differential expression. (continued on page 2) InterferonWhat’s Your —Bigger IFN-a Type? and Beta Than Ever PBLPBL Interferon InterferonSource...Source...promisingpromising results results (continued from page 1) Using immortalized J2E cells to induce erythroleukemia in affect treatments. More recently, Fujioka et al reported that mice, James et al showed that different murine IFN-a subtypes IFN-a subtypes induced cells to undergo apoptosis through can activate STATs and mitogen-activated protein kinase (MAPK) p53 activation.6 Using three distinct hepatocellular carcinoma distinctively.2 Activation of different subsets of STAT molecules cell (HCC) lines, the authors reported that IFN-a subtypes may due to differential recognition of the IFNAR with the IFN-a have differential inhibitory effects on HCC cell proliferation. In subtypes. Furthermore, when the authors subjected mice with particular, they found that IFN-a8 not only has a more potent of J2E-induced erythroleukemia to IFN plasmid DNA injections, cell anti-proliferative activity, it also yields greater induction of it was found that IFN-a6 and IFN-a9 were the most effective apoptosis when compared to other IFN-a subtypes in the cell subtypes in extending the survival period of the mice. line PLC/PRF/5. Similar findings have also been reported in humans. Early studies carried out by Meager et al had shown that antisera Do you know what’s your type? raised against human IFN-a1 and human IFN-a2 exhibited Although all Type I IFNs, including all IFN-a subtypes, bind to a strong neutralizing activity only to their homologous subtypes common receptor (composed of two subunits IFNAR-1 and IFNAR- and was less active against heterologous IFN-a subtypes.3 2), the variations in biological activities among IFN-a subtypes Evaluation of anti-viral activity of IFN-a subtypes in MT-2 suggest that receptor affinities may not be the only explanation cells done by Pestka et al showed significant anti-HIV activity for the diverse effects of IFN-a subtypes. It may be possible that among different IFN-a subtypes (Figure 1).4 In 2003, Kerr et al IFN-a subtype-specific signaling pathways contribute to the studied the signaling and transcriptional responses of JAK/STAT differences between IFN-a subtypes. Elucidating the functional pathways to different IFN-a subtypes. The signaling pathways significances of various IFN-a subtypes not only can lead to a and gene expression profiles were analyzed for human dendritic better understanding on the role interferons have in the immune cells and T cells treated with different IFN-a subtypes.5 They system, but also to the refinement of current interferon based reported differentials in IFN-a subtype/cell-type responses therapeutic applications. and expression in IFN-stimulated genes (ISGs). Interestingly, PBL InterferonSource offers the largest selection of interferons, they found that IFN-a2 has a higher induction of chemokine IP- including alpha subtypes from different animal species and 10 when compare to IFN-a1. IP-10 is an IFN-g-induced protein assay kits to analyze them. This issue of Interferon Matters that plays an important role in recruiting Th-1 cells into sites of features some of these products. For more information on these inflammation. The authors surmised that this may be of clinical products, please contact us at [email protected] or importance in cases where Th-1-related side effects adversely visit www.interferonsource.com. Figure 1: Varying Effectiveness of Alpha Interferon Subtypes References: 1) Lawson et al. (1998) J. Immunol. 60:2932 2) James et al. (2003) Blood 101:2727 3) Meager et al. (1984) J. Gen. Virol. 65:2277 4) Pestka et al. (1992) J. Interferon Res. 12:363 5) Kerr et al. (2003) J. Immunol. 171:5255 6) Fujioka et al. (2006) Biomed. Res. 27(5):219 2 Toll Free: 1 877-PBL-8881 Fax: +1 732-777-9141 www.interferonsource.com [email protected] Mouse TLR-3Interferon ReadyTM Alpha IFN-b Subtypes ELISA Kit PBL InterferonSource...promising results Human Interferons Alpha Sampler Pack (PBL Cat # 11001-1) PBL‘s Human Interferon Alpha Sampler Pack (Cat # 11001-1) includes 12 individual subtypes including Human IFN-aA, B2, C, D, F, G, H2, I, J1, K, 4b, and WA. The Human Interferon Sampler Pack can help you find the right alpha interferon for your research. Each Pack includes 20,000 units of each of the subtypes. For more background infor- mation on the different interferon subtypes, please contact PBL’s technical support at [email protected] or visit our web site at www.interferonsource.com. GenBank Interferon Alpha Protein Specific Activity (U/mg) Size Product # Accession # Human IFN-aA (a2a) 2.8 x 108 * VOO549 5 x 106 U 11100-1 Human IFN-a2 (a2b) 2.8 x 108 * VOO548 1 x 105 U 11105-1 Human IFN-aB2 (a8) 4.6 x 108 * XO3125 1 x 105 U 11115-1 Human IFN-aC (a10) 2.3 x 108 * XO2961 1 x 105 U 11120-1 Human IFN-aD (a1) 7.5 x 107 * VOO538 1 x 105 U 11125-1 Human IFN-aF (a21) 6.4 x 108 * VOO540 1 x 105 U 11130-1 Human IFN-aG (a5) 2.3 x 108 * XO2956 1 x 105 U 11135-1 Human IFN-aH2 (a14) 1.1 x 108 * XO2959 1 x 105 U 11145-1 Human IFN-aI (a17) 1.4 x 108 * VOO532 1 x 105 U 11150-1 Human IFN-aJ1 (a7) 2.6 x 108 * XO2960 1 x 105 U 11160-1 Human IFN-aK (a6) 1.5 x 108 * XO2958 1 x 105 U 11165-1 Human IFN-a1 (aD) 1.4 x 108 * JOO210 1 x 105 U 11175-1 Human IFN-a4a (aM1) 2.1 x 108 * NM_021068 1 x 105 U 11177-1 Human IFN-a4b (a4) 1.8 x 108 * XO2955 1 x 105 U 11180-1 Human IFN-aWA (a16) 2.4 x 108 * XO2957 1 x 105 U 11190-1 Human IFN Alpha Sampler Pack Call for more inofrmation n/a 2 x 104 U of each 11001-1 New Rhesus/Cynomolgus IFN a2 4.0 x 108 * XM_001107516 1 x 105 U 14110-1 New Mouse IFN-a4 5.0 x 107 * NM_010504 1 x 105 U 12110-1 * Biological activity for recombinant proteins measurerd on bovine MDBK cells with vesicular stomatitis virus (VSV) may vary Universal IFN-aA/D (Bgl ll) Interferon (PBL Cat # 11200) Also available from PBL is the Universal Type I Interferon (Cat # 11200).