bluetongue viruses , pathogenic only to domestic cattle and wild ruminants ( barratt - boyes and maclachlan , 1995 ; tsuboi and imada , 1997 ) , are non - pathogenic to humans ( xiao et al . , 2004 ; hu et al . , 2008 ) and thus , humans do not have pre - existing antibodies to btvs ( jeggo et al . , 1983 ; barratt - boyes and maclachlan , 1995 ; dong et al . , 1998 )
not only can btv be grown easily in vitro ( jameson and grossberg , 1981 ; du , 1985 ; samal et al . , 1985 ; sundin and mecham , 1989 ; dong et al . , 1998 ; wechsler and mcholland , 1998 ; chen et al . , 1999 ; prasad et al .
, 1999 ; xue , 2001 ; lei et al . , 2004 ; xiao et al . , 2004 ; liang et al . , 2006 ;
hu et al . , 2008 ) , it has powerful oncolytic activity against many different in vitro cultured cancer cell lines ( chen et al .
2008 ) , such as a498 human kidney cells ( atcc : htb-4 ) , hep - g2 human liver cells ( atcc : hb-8065 ) , a549 human lung epithelial cells ( atcc : ccl-185 ) , baby hamster kidney cells ( bhk-21 ) , monkey kidney ( vero ) cells ( ramig et al . , 1989 ) , mouse fibroblast cells ( nih 3t3 ) , spc - a-1 cells ( lei et al . , 2004 ) , ma 782 cells ( liang et al . , 2006 ) , and in mouse models .
no normal human cells such as the primary human embryo lung fibroblast ( hel ) and primary murine embryos fibroblast ( mef ) have ever been successfully infected by btv ( dong et al . , 1998 ; chen et al . , 1999 ;
, 2004 ; liang et al . , 2006 ; hu et al . , 2008 ) and thus , humans are normally sero - negative .
madin  darby canine kidney ( mdck ) which is sensitive to reovirus apoptosis also proved resistant to btv infection except in high moi ( unpublished data ) .
btvs can also infect cultured insect cells ( king and alders , 1985 ; samal et al .
, 1987 ; guirakhoo et al . , 1995 ; mullens et al . , 1995 ; mertens et al . , 1996
; xu et al . , 1997 ; tan et al . , 2001 ) .
in the last decade , our lab and collaborators have used both cell culture systems ( dong et al . , 1998 ;
xiao et al . , 2004 ; liang et al . , 2006 ; hu et al . ,
2008 ) and mice ( manuscript in preparation ) to investigate and determine the oncolytic activities and capacities of btvs and their selective degradation of human cancer cells as potential treatment of human cancers .
human cancer cell lines such as hep-3b and hep - g2 , a549 , a498 , spc - a-1 , and ma 782 cells infected with btv 10 , 11 , 13 , 17 , and hbc3 at various mois exhibited the following easily detected characteristics 36  h post - infection ( pi ) but not in hel and mef cells in multiple determinations ( xiao et al . , 2004 ; hu et al . , 2008 ) .
briefly : between 70 and 90% cytopathic effects ( cpe ) were readily observed and detected along with cellular apoptosis and necrosis in many human cancer cell lines .
various degrees of morphological and cellular damage could be visualized by either light , confocal , or electron microscopy ( em ) .
cellular dilated endoplasmic reticulum , nuclear chromatin condensation , and cytoplasmic shrinking were readily detected and easily observed in em .
btv viral genomic ds - rna fragments and viral mrnas could be found in btv-10 , 13 , 17 , and hbc3 infected human cancer cells , but not in normal hel or mef cells by gel electrophoresis , elisa , rip , western blots , pcr , and qrt - pcr ( tiwari et al . , 2000 ) .
the survival rate of several human cancer cell lines was low if high moi was used .
primary hel and mef cells survived very well for 69 days independent of the moi used for testing .
human cancer cells infected with btvs had more efficient viral replication and provided higher viral progeny yields .
selective cell cycle arrest at sub - g1 peak in several human cancer cells infected with btv-10 ( hu et al . , 2008 ) and hbc3 similar to the reovirus - induced g2/m cell cycle arrest ( poggioli et al . , 2000
after tumor cells were grown in culture and then injected as xenografts into mice ( eight mice / sample ) , tumors developed and all these mice died in less than 8 days ( preliminary data ) .
when 0.1  ml of btvs with different mois in sterile phosphate - buffered saline ( pbs ) was injected at three different sites of each xenograft , the sizes of tumors in mice were reduced 6085% .
most mice with the reduced tumors survived more than 35 days before they were sacrificed for tissue and organ samples ( kidney , lung , liver , heart  . etc .
thus , btvs selective cytotoxic effects to human tumor cells in vitro and partially in vivo have been clearly indicated and demonstrated . further in vivo research and clinical studies
are now in progress to document and investigate this unique oncolyitc potential of btvs which will provide us better and safer application in humans either in monotherapy or combination therapy in the near future .
bluetongue viruses administration has been shown to cause cell death and apoptosis in several cell lines as well as in several human tumor cell lines ( wechsler and mcholland , 1998 ; xiao et al . , 2004
; liang et al . , 2006 ; hu et al . , 2008 ) .
however , in experiments with cultured human cancer cells , the cells detached and died progressively throughout btv infection .
all btv genes are transcribed and btv proteins are expressed during infection . in the analysis of secreted analyte expression , cytokine and chemokine levels from mock- and btv-17 infected cells
were determined using multiplex immunoassay kits and reagents supplied from quansys biosciences ( logan , ut , usa ) , according to the supplier 's protocol .
twenty - five analytes were tested and these included il-1a , il-1b , il-2 , il-4 , il-5 , il-6 , il-8 , il-10 , il-12p40 , il-12p70 , il-13 , ifn , tnf , tnf , tgf - b1 , mcp-1 , rantes , ang2 , hgf , timp , tpo , vegf , pdgf , fgf - basic , and crp ( manuscript in preparation ) .
of the 25 analytes tested for during infection , only seven were found to be expressed from five cell lines : a498 human kidney cells , hep - g2 human liver cells , a549 human lung epithelial cells ( atcc : htb-4 , hb-8065 , ccl-185 , respectively ) , bhk-21 cells , or vero cells .
of these seven analytes , five were found to be consistently elevated upon btv infection , with the expression pattern consistent among cell lines .
these are il-6 , il-8 , mcp-1 , rantes , and fgf. the levels of vegf and timp were similar in control and btv - infected cells . however , a498 kidney cells responded to infection with the greatest secretion of cytokines , followed by a549 lung cells while hep - g2 liver cells secreted the least amount of cytokines .
polyinosine : polycytosine ( poly i : c ) is a synthetic ds - rna that has been used to assess the role of ds - rna segments of many ds - rna viruses in vitro since it can induce differential interferon and cytokine production ( der et al . , 1998 ;
, 2003 ; chiang et al . , 2006 ) . however , poly i : c alone did not cause cell death or have a significant impact on cytokine or chemokine expression in btv - infected human cancer cell lines . only il-6 and il-8 were found to be significantly elevated during poly i : c treatment .
increases in il-6 and il-8 were observed only in kidney cells ( a498 and vero ) and were most prevalent in samples treated with 1  g / ml
the other cytokines mcp-1 , rantes , and fgf were all at expression levels comparable to the placebo treated cells . with the identification of some of the antigenic epitopes of various btv proteins ( wang et al .
gould et al . , 1988a , 1994 ; grieder and schultz , 1989 ; geysen , 1990 ; li and yang , 1990 , 1992 ; marshall and roy , 1990 ; mecham and jochim , 1990 ; eaton et al . , 1991 ; rossitto and maclachlan , 1992 ; du plessis et al . , 1994 , 1995 ;
, 1997 ; nagesha et al . , 2001 ) , btv immunity has only been studied partially in the last two decades because of lack of supporting funding and reliable immune assays ( jeggo and wardley , 1982a , b ; gibbs et al . , 1983 ;
jeggo et al . , 1983 ; campbell and grubman , 1985 ; stott et al . , 1985 ; fukusho et al .
, 1987a ; marshall and roy , 1990 ; martyn et al . , 1991 ;
; lin and zhou , 1996 ; wade - evans et al . , 1996 ; odeon et al . , 1999 ; prasad and minakshi , 1999 ; demaula et al . , 2000 ; roy , 2003 ) . with the recent development of more reliable immune assays and microarrays
, we have found that btv can induce inflammatory cytokine expression in immortalized human kidney , lung , and liver cells and subsequently kill the infected cell with cell line - dependent severity .
analysis of results from the cytokine profiles and expression levels in these five cell lines indicated that human tumor cells and tumor reduction might be a result of the combined effects of direct viral induced cell death and recruitment of t - lymphocytes ( jeggo and wardley , 1982a , b ; jones et al .
, 1996 , 1997 ) to the tumor through elevated cytokine / chemokine production ( shmulevitz et al . , 2005 , 2010 ) .
very high levels of pro - inflammatory cytokines were secreted from btv - infected cells and cytokine levels secreted from each of the seven cell types were directly proportional to the measured death of infected cells .
thus , we hypothesize that the cell death and cytokine expression are related and may be caused by shared signaling pathways or events .
if these same events occurred in human tumors , btv would be expected to reduce tumor size in infected patients by directly causing infected human cancer cell death and by inducing cytokine expression from the tumor that would lead to lymphocyte recruitment to the site of the tumor .
thus , btv has the therapeutic potential in cancer because of its selective ability to cause preferential apoptosis of various human cancer cell lines and tumors .
ideal oncolytic viruses should have essential survival data , systemic effect , resist immune system clearance , targeted delivery , and distribution , be highly mobile for intravenous spread , travel to and kill distant metastases , have a high rate of replication to stay ahead of immune system clearance , tolerable toxicity , and side effects , etc .
systemic delivery makes sense in cancer where primary tumors and metastases are dispersed . however , systemic approach in cancer is a big problem because we do not have a good target specific delivery device that would facilitate uptake specifically for certain cells .
if this can be achieved , then it would be better than the current and conventional therapies .
however , it is very difficult to get enough control patients in clinical trials and to get biopsies from different tissues , organs and tumors after the oncolytic virus treatments to determine the efficacy or survival of most oncolytic viruses .
oncovirologists consider oncolytic virus as a dark horse treatment but not a dead one since they know that many anti - cancer agents appear to be actively effective in lab cultures and animal models but failed in human trials .
many oncolytic viruses ( norman and lee , 2005 ; alain et al . , 2006 , 2007 ; kim et al .
, 2006 ; shen and nemunaitis , 2006 ; burroughs et al . , 2007 ; qi et al . , 2007
, 2009 ; ou and yen , 2010 ) that are currently in phase i / ii / iii human clinical trial status include adenovirus , vaccinia virus , coxsackie virus , reovirus , newcastle disease virus , herpes simplex virus 1 , measles virus , and seneca valley virus ( burroughs et al . , 2007 ) . in 2007 , the only oncolytic virus approved for phase iii clinical trials in china was the adenovirus ( h101 ) with e1b deletion produced jointly by the sunway biotech of china and onyx pharmaceuticals of emeryville , ca , usa ( privilege communication ) .
since mid-2005 to the end of 2010 , several other oncolytic viruses have also been approved for phase iii clinical trials primarily outside us as briefly described below .
thus , cancer therapy with oncolytic viruses has survived and revived , and this unique approach is poised for a comeback ( aghi and martuza , 2005 ; norman and lee , 2005 ; alain et al .
the results of the last 1015 years of investigation generated by researchers have increased interest in oncolytic viruses .
investigators have generated and obtained some genetically modified viruses with greater oncolytic potentials ( aghi and martuza , 2005 ; norman and lee , 2005 ; alain et al .
a significant effort is underway to understand the genetic and mechanistic basis of cancers , selective degradation of cancer cells by these oncolytic viruses , and their effects on downstream cellular events and pathways ( strong et al .
vorburger et al . , 2004 ; aghi and martuza , 2005 ; norman and lee , 2005 ; shmulevitz et al . , 2005 , 2010 ; alain et al . , 2006 ; liu , 2006 ; kim et al . , 2007 ; liu et al . , 2007 ;
2009 ; hill and lee , 2010 ; ou and yen , 2010 ; thirukkumaran et al . , 2010 ) .
increasing attention is being paid to protein families like proteases , caspases , and kinases which play critical roles in cancer .
current therapeutic approaches for human cancer include the regimen of selective surgery , high dose but cytotoxic chemotherapy with two to three drugs and radiation .
these approaches are not very effective to combat the increasing numbers of human cancers . using oncolytic viruses generates a paradigm shift from current cancer treatment using chemotherapy and radiation which typically destroy both normal and cancer tumor cells .
as we learn more about cancer biology and oncolytic viruses , new strategies for treating cancers are rapidly evolving .
uses of oncolytic virus alone or with synergistic anti - cancer drugs or irradiation in human clinical trials are currently under consideration or in progress .
reovirus leads the field of oncolytic viruses and positive clinical trials are very encouraging ( strong et al . , 1998 ; norman et al . , 2004 ; norman and lee , 2005 ; shmulevitz et al . , 2005 , 2010 ; alain et al . , 2006 ; kim et al .
2009 ; hill and lee , 2010 ; thirukkumaran et al . , 2010 ) .
other companies and institutes are not far behind . in mid - may 2010 , researchers of the university of helsinki and oncos therapeutics have treated 200 patients with their  modified  gmcsf armed oncolytic adenovirus which exhibited anti - tumor immunity that recruited natural killer cells and induced tumor - specific cytotoxic t - cells with strong efficacy and safety .
the media report indicated that its clinical benefit is about 45% according to the recist criteria with no grade four to five side effects detected .
this represents an extension of their initial investigation in 2008 . with these successful applications and those from other oncolytic viruses
, we can now add the simple but oncolytic btv as a key component to the armamentarium of oncologists to combat and destroy the insidious cancer cells since btvs have similar characteristics and additional subtle advantages over reovirus and other oncolytic viruses .
bluetongue viruses and reovirus are more advantageous than the genetically modified oncolytic dna viruses since the integration of viral ds - rna genome into the host cell genome have not been shown or reported . since humans are sero - negative to btvs , potential future use of btv to degrade human cancer by direct intratumoral injection to degrade the injected tumors will not be inhibited by pre - existing immune components within human cancer cells .
. the current mechanisms of oncolytic reovirus toward human cancer cells have recently been summarized ( strong et al . , 1998 ; norman et al . , 2004 ; norman and lee , 2005 ; shmulevitz et al . , 2005 , 2010 ; alain et al . , 2006 ; kim et al .
, 2009 ; hill and lee , 2010 ; thirukkumaran et al . , 2010 ) .
genetically modified reovirus serotype 3 , but not types 1 and 2 , is a proprietary product called reolysin produced by reolytics biotech , inc .
data for cancer treatment with this agent from the last decade in animal models and lately in phase i / ii human clinical trials have shown encouraging indications of its potential oncolytic activities in cancer cells bearing an activated ras pathway .
phase iii human clinical trials have been approved and initiated in 2009 . however , the pre - existing anti - reovirus sera in humans worldwide can trigger greater immune responses against this virus , reducing its efficacy and effectively shutting down its potential oncolytic treatment . uses of oncolytic btv have greater advantages over reovirus since no anti - btv serum is detected in most people globally .
even though btv injected into tumors can activate dendritic cells for early detection of viral infection , it will take 1014 days before adaptive immune responses can be generated against btvs which can eradicate xenografted tumor cells in mice in less than 10 days ( preliminary data ) .
oncolytics biotech at calgary , ab , canada showed the uses of reovirus ( reolysin ) plus cisplatin , a standard chemotherapy agent in both cultured melanoma cells and a mouse model .
the combined uses of reolysin and cisplatin were much better than either of these two agents alone since they appeared to work synergistically ( abstract of the 4th international symposium on the  treatment of cancers with oncolytic viruses  at scottsdale , arizona march 1517 , 2007 ) .
furthermore , they had also showed that simultaneous and combined uses of reolysin and gemcitabine could completely eradicate the transplanted colon cancer in four of the five mice ( abstract of the american association for cancer research ( aacr ) annual meeting in april 2007 ) . since mid - april , 2007
, oncolytics biotech has demonstrated some success in multiple phase ii trials with several medical centers in the uk and us , administering intravenous reolysin to patients with sarcomas that have metastasized to the lung .
multiple clinical trials for other tumors with the us national cancer institute ( nci ) were conducted in late 2007 .
this will help to determine whether reolysin can migrate systemically to metastasized tumor sites and exhibit its oncolytic activity .
oncolytics biotech has expended more than 65 million dollars and a decade of genetic modifications of reolysin before human clinical trials were approved .
similar and greater expenditures have also been made by other companies and institutes in the last 1215 years to develop different oncolytic viruses with tremendous personnel efforts before some clinical trials are approved and conducted .
we strongly believe that btv is the front runner of the next generation oncolytic viruses since it needs no genetic modifications and attenuation , and carries no payloads .
it can grow fast and spread easily within many human cancer cells . since there is no pre - existing neutralizing antibody against btv in humans , they should be undetected nor immunologically inhibited in the initial injection alone or with anti - cancer agents .
in addition to the comparison of the oncolysis of human cancer cells by reovirus and btv as shown in table 2 , btv has additional advantages .
briefly : since btv does not infect normal human cells , we do not have to suppress the immune system before use
btv is very stable since it can be stored at ph 8.28.4 at 4c for over 20 years with only 510% loss of infectivity ( unpublished data ) .
thus , it can be shipped by air - mail globally without significant loss of its infectivity and oncolytic activities ( unpublished data ) .
2-d gel electrophoresis and the 3-d assay ( lam et al . , 2007 ) for the measurement of btv - induced oncolysis are currently under optimization and further development and adaptation .
btv infection of many cultured human tumor cells has led to the production of many  inflammatory  cytokines such as tnf - alpha and il-6 which strongly activate the kinase activity of p38 mapk and erk1/2 .
these cytokines also assist the subsequent activation of p38-dependent mapk and pkr pathways which can bring about the necrosis and apoptosis of cancer cells .
this strongly suggests that btv can prime and direct the immune system of cancer patients against certain solid tumors such as breast and prostate cancer .
repeat screening of over 500 human cytokines using biotin label - based cytokine antibody array is currently in progress and data are under comparative analysis .
construction and rescue of recombinant infectious btv ( boyce and roy , 2007 ) expressing the enhanced green fluorescent protein ( egfp ) , using a protocol similar to one recently published by our lab for human parainfluenza virus type 3 ( roth et al .
comparison of oncolytic reovirus and bluetongue virus . * reovirus references : strong et al .
an activated ras / ralgef / p38 pathway is potentially involved with the permissiveness of host cells to reovirus infection ( norman et al . , 2004 ; shmulevitz et al . , 2005 ; alain et al .
, 2007 ; kim et al . , 2007 ; marcato et al . , 2007 ) .
it has been shown that the c - k - ras gene inside the lung carcinoma cell line a549 we used has mutation at codon 12 ( valenzuela and groffen , 1986 ) .
since the ras / ralgef / p38 signal pathway inside the a549 cell has been activated , this might allow btv to degrade human tumor cells efficiently similar to reovirus ( stott et al .
, 1985 ; shmulevitz et al . , 2005 , 2010 ; kim et al . , 2007 ; marcato et al . , 2007
we have further hypothesized that the genetic alterations / mutations of different human cancer cells , such as signals within a ras - activated pathway , play a key role in the susceptibility of different oncolytic viruses in addition to specific viral receptors which might be present in human cancer cells but not in normal human cells .
identification of btv receptor(s ) on the surface of human cancer cells is also currently in progress .
comparison of oncolytic potentials between reovirus and btv is summarized in table 2 . with regard to the potential cytotoxic effect mechanisms of btvs , the gp5 protein of btv identified by our lab ( li and yang , 1992 ; yang and li , 1992 , 1993 ; yang et al .
, 1992a ) has recently been shown to induce or trigger apoptosis in mammalian cells ( mortola et al . , 2004 ) .
furthermore , the non - structural btv protein , ns-3 , has been shown to be a 
viroporin ,  the polymerization of which forms pores on the surface of btv - infected cells , leading to severe cellular leakage , and potential cell death ( han and harty , 2004 ) .
we hypothesize that these two btv proteins and other(s ) are involved with the cytotoxic effect of human cancer cells after btv infection .
individual btv gene  silencing  with rnai to determine the contribution of each of the btv gene products to onclysis of human cancer cells are also in progress .
the protein toll ( means  weird  in german ) was discovered in the fruit fly drosophila melanogaster in 1980s .
however , if mutations in toll occurred in the fruit fly , the infection by aspergillus fumigates ( a fungus ) is lethal .
toll - related molecules involved with human innate immunity and coined as toll - like receptors ( tlrs ) were then discovered in mid-1990s by ruslan medzhitov and charles janeway ( medzhitov and janeway , 2002 ; norman et al . , 2004 ) .
tlrs are molecular pattern recognition receptors and can recognize molecules that are broadly shared by many pathogens .
thus , tlrs pathways play a key role in both innate immunity and normal immune physiology that can sense different pathogens and cancer cells .
to determine whether mutated tlrs are involved with the recognition of btvs , we have initiated the  silencing  of each 1 of the 10 known human tlrs tlr- 110 using rnai .
human cancer cell lines , a498 and hek 239 , are transfected with dna plasmids containing sirna sequences against human tlrs ( invivogen , ca , usa ) .
preliminary data indicated that endosome - located tlr-3 which specifically recognizes ds - rna is potentially involved since cells transfected with this tlr-3 rnai plasmid produced substantially less btv 17 and 10 progeny .
cells transfected with tlr-2 , 4 , 7 , 8 , and 9 rnai plasmids produced slightly less viral progeny in these two human cancer cell lines .
experiments are now underway to determine what types of mutation are in the tlr-3 in these four human cancer cell lines and this might provide a potential explanation of why normal human cells are not infected by btv because the regular tlr-3 has not been modified nor mutated .
several signal pathways related to tlrs are currently under screening using pathway screen arrays ( qiagen / sabiosciences ) .
the protein toll ( means  weird  in german ) was discovered in the fruit fly drosophila melanogaster in 1980s .
however , if mutations in toll occurred in the fruit fly , the infection by aspergillus fumigates ( a fungus ) is lethal .
toll - related molecules involved with human innate immunity and coined as toll - like receptors ( tlrs ) were then discovered in mid-1990s by ruslan medzhitov and charles janeway ( medzhitov and janeway , 2002 ; norman et al . , 2004 ) .
tlrs are molecular pattern recognition receptors and can recognize molecules that are broadly shared by many pathogens .
thus , tlrs pathways play a key role in both innate immunity and normal immune physiology that can sense different pathogens and cancer cells .
to determine whether mutated tlrs are involved with the recognition of btvs , we have initiated the  silencing  of each 1 of the 10 known human tlrs tlr- 110 using rnai .
human cancer cell lines , a498 and hek 239 , are transfected with dna plasmids containing sirna sequences against human tlrs ( invivogen , ca , usa ) .
preliminary data indicated that endosome - located tlr-3 which specifically recognizes ds - rna is potentially involved since cells transfected with this tlr-3 rnai plasmid produced substantially less btv 17 and 10 progeny .
cells transfected with tlr-2 , 4 , 7 , 8 , and 9 rnai plasmids produced slightly less viral progeny in these two human cancer cell lines .
experiments are now underway to determine what types of mutation are in the tlr-3 in these four human cancer cell lines and this might provide a potential explanation of why normal human cells are not infected by btv because the regular tlr-3 has not been modified nor mutated .
several signal pathways related to tlrs are currently under screening using pathway screen arrays ( qiagen / sabiosciences ) .
human cancers can be divided into two major types , 90 and 10% of which are solid ( carcinoma and sarcoma ) or liquid ( leukemia ) tumors , respectively .
recent data and epidemiological analysis of many tumors reveal that most fda - approved cancer therapeutic drugs can only provide small incremental improvement and survival to cancer patients .
companies that make many unrealistic claims for human cancer cures in scientific tabloid media do so strictly for financial reasons since these expensive cancer drugs can generate great revenues via physicians who make drug recommendation and to whom drug companies provide substantial and financial 
however , the 90% of deaths of cancer patients are primarily caused by those cancer stem cells that have metastasized to other regions to form metastatic tumors .
humans are sero - negative toward btvs since btvs do not infect normal human cells .
infection and selective degradation of several human cancer cell lines , but not normal cells , in vitro by five us btv serotypes have been investigated by our lab for the last few years when btv oncolytic potential was discovered .
we have also recently found that direct single injection of oncolytic btv into the human tumor xenografts in mice , have led to cytotoxic destruction and preferential lysis of cancer cells within the xenograft but not in normal healthy human or mouse cells ( preliminary data ) .
thus , oncolytic btvs are safer and more effective than most current cancer treatments . however , we believe that oncolytic btv is not a  stand - alone  therapeutic and that simultaneous combination of btv with either radiation or chemotherapy can obliterate the tumors more rapidly and extensively .
cancer patients should have fewer side effects since small dosage radiation and anti - cancer drugs are used for this combined treatment .
related to the oncolytic btvs , the following questions have been frequently asked : is widely curing cancer by oncolytic viruses possible or just hype ?
big  is the gap between in vitro / in vivo studies and human application ?
is oncolytic btv a vital alternative for cancer therapy ? what are the pathways btv used to carry out this task ?
overall , we believe that the oncolytic ability of btv represents a significant potential to selectively degrade human cancer cells alone and would even be greater when combined with simultaneous anti - cancer drug and radiation treatment .
our group and collaborators are currently engaged in several types of investigations with these goals : ( 1 ) study the mechanistic basis of lung , kidney , and liver cancer destruction by btvs , ( 2 ) examine and determine biological profiles of three types of solid tumors after btv infection , ( 3 ) correlate the responses of these human cancer cells to changes at the cellular dna , rna and protein level , and ( 4 ) construction and rescue of an infectious recombinant btv 17 expressing the enhanced egfp using the method that we have developed for the human parainfluenza virus type 3 ( roth et al . , 2010 ) with btv ds - rna fragments ( kowalik et al . , 1990b ; boyce and roy , 2007 ) , the success of which can further reveal the oncolytic pathways used by btvs .
taking the next step in the investigation of btv oncolysis is very challenging since no one can individually excel in all the technical , scientific , and clinical avenues involved in this endeavor .
our ongoing in vitro work with different human cancer cell lines will assist us in elucidating the potential mechanisms of the btv oncolytic activities .
no cpe was detected nor viral progeny obtained when several primary human cell lines were infected by btv .
when xenografts implanted in mice were injected directly with btv , no pathological damages were detected in any tissue or organs 721 days post - infection ( preliminary data ) .
thus , the potential risks of btv oncotherapy is substantially lower than risks associated with other oncolytic viruses . however
, this author sincerely hopes that expert advice and suggestions will be forthcoming from different  champions  after they have read this brief review .
the author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest .
 humans are sero - negative toward bluetongue viruses ( btvs ) since btvs do not infect normal human cells . 
 infection and selective degradation of several human cancer cell lines but not normal ones by five us btv serotypes have been investigated . 
 we determined the susceptibilities of many normal and human cancer cells to btv infections and made comparative kinetic analyses of their cytopathic effects , survival rates , ultra - structural changes , cellular apoptosis and necrosis , cell cycle arrest , cytokine profiles , viral genome , mrnas , and progeny titers . 
 the wild - type us btvs , without any genetic modifications , could preferentially infect and degrade several types of human cancer cells but not normal cells . 
 their selective and preferential btv - degradation of human cancer cells is viral dose  
 dependent , leading to effective viral replication , and induced apoptosis . 
 xenograft tumors in mice were substantially reduced by a single intratumoral btv injection in initial in vivo experiments . 
 thus , wild - type btvs , without genetic modifications , have oncolytic potentials . 
 they represent an attractive , next generation of oncolytic viral approach for potential human cancer therapy combined with current anti - cancer agents and irradiation . 