Adnan Nasir
Nanotechnology in Vaccine Development: A Step Forward
University of North Carolina at Chapel Hill
study nanotechnology epicutaneous delivery pharmaceuticals vaccines burgeoning topically applied nanomaterials shown enter tapestripped skin reach draining lymph nodes inbred strain mice nanomaterials form plasmid dna proteins virus particles accumulate hair follicles diffuse via dendritic cells draining lymph nodes elicit antigenspecific humoral cellmediated immunity topically immunized mice also demonstrated resistance infection live virus advantages nanotechnology include uniformity reproducibility precision synthesis manufacture candidate compounds combined novel pharmacokinetics possibility targeted therapy nanotechnologybased vaccines may prove superior existing vaccines potential open therapeutic avenues treating infectious disease malignancy study nanotechnology epicutaneous delivery pharmaceuticals vaccines burgeoning topically applied nanomaterials shown enter tapestripped skin reach draining lymph nodes inbred strain mice nanomaterials form plasmid dna proteins virus particles accumulate hair follicles diffuse via dendritic cells draining lymph nodes elicit antigenspecific humoral cellmediated immunity topically immunized mice also demonstrated resistance infection live virus advantages nanotechnology include uniformity reproducibility precision synthesis manufacture candidate compounds combined novel pharmacokinetics possibility targeted therapy nanotechnologybased vaccines may prove superior existing vaccines potential open therapeutic avenues treating infectious disease malignancy history infectious disease humans cyclical tugofwar host parasite host pathogen donelson donelson 2008donelson lg donelson je review fatal sleep africas killer disease went undiscovered centuries j clin invest 1183932008google scholar cost society course history staggering epidemics ravaging whole populations time cost diminished either scope extent according world health organization top 10 causes death world 5 result infectious disease nearly surpassing ischemic heart disease cerebrovascular disease combined world health 2003world health organization world health report 20032003crossref google scholar table 1 although impact infectious disease universal effects particularly acute developing nations heaviest burden falls poor table 2table 1top causes mortality worldwidefrom world health organization 2003rankcause deaths1ischemic heart disease1262cerebrovascular disease973lower respiratory infection684hivaids495chronic obstructive pulmonary disease486diarrheal disease327tuberculosis278malaria229cancer tracheabronchuslung2210road traffic accidents21 open table new tab table 2global burden diseasefrom world health organization 2003rankdiseaseno infected millions1malaria8002filariasis2803leishmaniasis2004schistosomiasis2005trypanosomiasis13 open table new tab looking beyond mortality statistics five prevalent diseases worldwide infectious parasitic strike literacy rates low poverty epidemic resources scarce drain reserves affected individuals families rob communities nations productive citizens beyond basic sanitation infrastructure one costeffective ways manage infectious disease vaccination traditional vaccines considerable disadvantages novel approaches make vaccines versatile effective urgently needed ideal vaccine would easy manufacture inexpensive readily transported stable broad range ambient temperatures levels humidity finally vaccine easy administer needlebased vaccines require infrastructure sterile materials trained personnel generate hazardous waste glenn et al 2000glenn gm taylor dn li x frankel montemarano alving cr transcutaneous immunization human vaccine delivery strategy using patchnat med 2000 6 14031406crossref pubmed scopus 294 google scholar furthermore painful meet psychological cultural resistance making needlefree alternatives desirable one advantage epicutaneous vaccination resident antigenpresenting cells induce powerful sustained immune responses least one study allowed antigen dilution onefifth potentially reducing vaccine cost nasir 2008nasir dermatologic toxicity nanoengineered materialsarch dermatol 2008 144 253254crossref pubmed scopus 14 google scholar increasing vaccine availability nel et al 2006nel xia mdler l li n toxic potential materials nanolevelscience 2006 311 622627crossref pubmed scopus 6813 google scholar nanoparticles nanomaterials biological activity exist nature oberdorster 2005oberdorster g et alprinciples characterizing potential human health effects exposure nanomaterials elements screening strategyparticle fibre toxicol 2005 2pubmed google scholar viruses one salient examples viruses induce natural immunity clinical subclinical infection via epicutaneous respiratory gastrointestinal routes induction artificial immunity using nanomaterials recent phenomenon smallpox existed millennia well documented history clinical examples skin lesions identified mummy ramses v hornung 1997hornung e pharaoh inin donadoni bianchi r egyptians university chicago press chicago il1997google scholar smallpox descriptions taper late roman early christian texts suggesting disease dormant centuries making resurgence medieval europe asia earliest description process inoculating naive individual viral nanoparticles may given 1549 wu quan temple 1986temple r genius china 3000 years science discovery invention simon schuster new york1986google scholar likely therapy smallpox developed time although china scholars contend origins traced song dynasty 9601279 ad needham 1999needham j science civilization chinabiology biological technology part 6 medicine 6 cambridge university press cambridge uk1999google scholar wu quan described powder made crust smallpox lesions blown nasal passages vaccinees time understood inoculation live material patients fullblown disease variola major led high mortality procedure therefore banned lesional crust incorporation powder could harvested subjects crusts proxy variola minor practice inoculation became widespread central asia south asia asia minor wujastyk 1995wujastyk medicine indiain oriental medicine illustrated guide asian arts healing 19 serindia london1995 38google scholar later varied include scarification skin introduced england lady mary wortley montagu 1717 practiced anecdotally farmers milkmaids subsequently perfected edward jenner 1796 bray roy 2004bray roy cj antiviral prophylaxis smallpoxj antimicrob chemother 2004 54 15crossref pubmed scopus 24 google scholar predecessor vaccination methods costeffective however indicated mortality due inappropriate administration vaccines eg variola major methods nonspecific imprecise advent nanotechnology enabled precise manufacture materials desirable properties removal undesirable features table 3 selectivity combined compactness nanoscale allows development wide range useful agents safety issues notwithstanding potential nanotechnology vaccine development significant castanedotardan et al 2007castanedotardan mp nasir jacob se better understanding chemicals surround usskin aging 2007httpwwwskinandagingcomarticle7428google scholar nasir 2008nasir dermatologic toxicity nanoengineered materialsarch dermatol 2008 144 253254crossref pubmed scopus 14 google scholar nasir gaspari 1996nasir gaspari aa contact dermatitis clinical perspectives basic mechanismsclin rev allergy immunol 1996 14 151184pubmed google scholar nel et al 2006nel xia mdler l li n toxic potential materials nanolevelscience 2006 311 622627crossref pubmed scopus 6813 google scholar oberdorster 2005oberdorster g et alprinciples characterizing potential human health effects exposure nanomaterials elements screening strategyparticle fibre toxicol 2005 2pubmed google scholar warheit et al 2007warheit db borm pj hennes c lademann j testing strategies establish safety nanomaterials conclusions ecetoc workshopinhal toxicol 2007 19 631643crossref pubmed scopus 167 google scholartable 3sources nanomaterialsadapted oberdorster et al 2005anthropogenicnaturalincidentalengineeredfryingvolcanic ashforest firescarbon nanotubescookingocean sprayplasmaquantum dotssandblastingbiomagnetitemeteoritessunscreenminingferritinmineral compositessemiconductor wiresmetalworkvirusfullerenesdiesel exhaustclouds open table new tab skin active passive barrier defenses retard entry pathogens successful nanomaterials must bypass defenses variety studies demonstrated nanomaterials penetrate skin penetration typically follicular route diminishes increasing nanomaterial size mortensen et al 2008mortensen lj oberdrster g pentland ap delouise la vivo skin penetration quantum dot nanoparticles murine model effect uvrnano lett 2008 8 27792787crossref pubmed scopus 237 google scholar vogt 2006vogt et alcd1a cells transcutaneous application human skin 40 nm 750 1500 nm particles enter epidermalj invest dermatol 2006 126 13161322abstract full text full text pdf pubmed scopus 278 google scholar particles larger 100 nm likely penetrate intact skin however skin damaged flexed otherwise altered may permeable particles patients atopic dermatitis demonstrate slightly greater vulnerability cutaneous viral infections example molluscum contagiosum herpes simplex bork brauninger 1988bork k brauninger w increasing evidence eczema herpeticum analysis seventyfive casesj acad dermatol 1988 19 10241029abstract full text pdf pubmed scopus 130 google scholar erdmann et al 2004erdmann sm rubben frank j pobletegutierrez p mollusca contagiosa infant atopic eczema therapeutic challengehautarzt 2004 55 991994crossref pubmed google scholar kakourou et al 2005kakourou zacharides anastasiou et almolluscum contagiosum greek children case seriesint j dermatol 2005 44 221223crossref pubmed scopus 31 google scholar wollenberg et al 2003wollenberg wetzel burgdorf w haas j viral infections atopic dermatitis pathogenic aspects clinical managementj allergy clin immunol 2003 112 667674abstract full text full text pdf pubmed scopus 202 google scholar furthermore nanomaterials vehicles may modified gain purchase skin enter via follicular interfollicular routes issue mahe et al 2009mahe b vogt liard c duffy abadie v bonduelle et alnanoparticlebased targeting vaccine compounds skin antigenpresenting cells hair follicles transport micej invest dermatol 2009 129 11561164abstract full text full text pdf pubmed scopus 102 google scholar report mouse model testing nanomaterial penetration immunogenicity authors increase potential nanomaterial penetration three means first skin antigen applied tapestripped tapestripping may decrease barrier function also may increase expression immunostimulatory cytokines surface markers keratinocytes second mice anesthetized 1 hour allow undisturbed penetration applied material third authors selected c57bl6 inbred mouse strain unique immunological favor th1 response behaviorial tend groom barber form dominance properties sarna et al 2000sarna jr dyck rh whishaw iq dalila effect c57bl6 mice barber whiskers pluckingbehav brain res 2000 108 3945crossref pubmed scopus 90 google scholar von stebut udey 2004von stebut e udey mc requirements th1dependent immunity infection leishmania majormicrobes infect 2004 6 11021109crossref pubmed scopus 73 google scholar series elegant experiments authors found nanomaterials penetrate tapestripped skin used several methods demonstrate penetration figure 1 fluorescent 40 200nm particles called fluorospheres emit yellowgreen fluorescence 515 nm vivo confocal laser scanning microscopy demonstrated accumulation particles round aggregates depth 80 corresponding location hair follicles initial distribution compact bright 5 hours gradually diffused 24 hours suggesting skin uptake epidermal cells harvested 3 hours application sorted antigenpresenting cells cd11c cd207 nearly onethird contained fluorospheres 4 24 48 hours application authors harvested draining lymph nodes compared nondraining lymph nodes presence fluorospheres able demonstrate accumulations early 4 hoursand peak 40 nm fluorospheres 24 hoursin draining lymph nodes absence fluorescence nondraining lymph nodes results 200nm fluorospheres showed smaller peak 4 hours approximately onethird 40nm peak subsequent diminution 24 48 hours cd11c cells purified draining lymph nodes found contain fluorospheres studies suggest rapid accumulation nanoparticles hair follicles attachment uptake dendritic cells migration fluorosphereladen dendritic cells ipsilateral lymph node within 424 hours absence fluorescence nondraining nodes indicates cutaneouslymphatic route migration rather cutaneouscirculatory routenanomaterials penetrate tapestripped skin effectively next authors tested ability nanoparticulate antigens generate immune response topical application antigens selected plasmid dna encoding ovalbumin ova ova protein ova conjugated cholera toxin based calculations current literature particles measure 2070 5 55 nm respectively braga ricci 2004braga pc ricci atomic force microscopy biomedical methods applications humana press totowa nj2004google scholar experiments host mice first intravenously administered 4 million prelabeled naive cd8ot1 cells strain mice specific immune response peptides 257264 siinfekl ova cells lost intensity label cytoplasmic dilution underwent proliferation division cellmediated responses topical application antigen compared pbs controls measured day 4 flow cytometry fluorescent labeling cd8 cells draining nondraining lymph nodes authors found cd8ot1 proliferation greatest response ova protein 83 proliferating cells draining lymph nodes vs 68 nondraining lymph nodes compared ova plasmid dna 21 proliferating cells draining lymph nodes vs 15 nondraining lymph nodes efficacy plasmid dna roughly equivalent whether applied topically injected tibialis anterior muscle humoral responses assessed measuring antiova igg day 14 whereas phosphatebuffered saline pbs ova protein elicited negligible antibody responses ova protein cholera toxin ova plasmid dna elicited vigorous nearly equivalent responses authors subsequently determined whether modified vaccinia ankara mva virus could penetrate skin elicit immune responses virus particle modified green fluorescent protein allow visualization approximately 290 nm diameter authors able detect particles situ using confocal fluorescent microscopy antimva antibodies demonstrated intense follicular localization mva diffusion loss 2448 hours mva antigen could seen cd207 cells particularly around perifollicular dendrites mva antigen could also detected draining lymph nodes within 4 hours topical application nondraining lymph nodes application 15 106 particleforming units pfu mva topically elicited vigorous production ifn cd4 cd8 cells day 14 similarly vigorous elevation neutralizing antibody titer noted day 28 immunized animals compared pbs controls finally mahe et al immunized mice topically 5 106 pfu mva pbs control two months later mice challenged intranasally 5 106 pfu live vaccinia virus animals measured weight loss percentage body weight topically immunized mice lost peak 5 body weight study whereas control mice lost nearly 11 suggesting modest vivo protection live virus challenge authors nicely demonstrated model system nanoparticles 40 200nm range effectively penetrate tapestripped skin enter lymph nodes via dendritic cells langerhans cells demonstrated proteins plasmid dna traffic draining lymph nodes mechanism elicit cellmediated humoral immunity authors shown vaccinia virus particle approximately 290 nm size penetrate tapestripped skin migrate within dendritic cells draining lymph nodes stimulate tcell cytokine production neutralizing antibody production thereby conferring relative immunity live virus challenge several interesting questions remain applied nanomaterials localize solely hair follicles nanomaterials nonfollicular skin groomed rubbed away accumulate aggregate follicles funnel drain effect properties infundibulofollicular epithelium example charge ph lipids commensal organisms permissive nanoparticle accumulation nanoparticles accumulate follicle require threshold density concentration penetrate skin density lacking interfollicular skin would occlusion enhance epicutaneous penetration even interfollicular skin form nanoparticles dendritic cells prefer uptake dispersed aggregated bulk surface properties nanoparticles favor dendritic cell uptake 2007he lz antigenic targeting human mannose receptor induces tumor immunityj immunol 2007 178 62596267crossref pubmed scopus 111 google scholar ramakrishna et al 2007ramakrishna v vasilakos jp tario jd berger wallace pk keler tolllike receptor activation enhances cell mediated immunity induced antibody vaccine targeting human dendritic cellsj translat med 2007 5crossref pubmed scopus 46 google scholar example models nanoparticles readily taken cells spherical rather cylindrical whereas models gratton et al 2008gratton sea ropp pa pohlhaus pd luft jc madden vj napier et althe effect particle 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development long tangled unpredictable countless investigator years solid supportive preliminary data led one largest costliest hiv vaccine clinical trials history yet resultincreased infection rates treated individualswas puzzling profoundly disappointing complexity population studied prior history nonhiv viral infections difficulty reaching necessary strength humoral cellmediated arms immunity likely contributed failure cohen 2007cohen j mercks failed hiv vaccine cause harmscience 2007 318 10481049crossref pubmed scopus 90 google scholar sekaly 2008sekaly rp failed hiv merck vaccine study step back launching point future vaccine developmentj exp med 2008 205 712crossref pubmed scopus 320 google scholar advent nanotechnology tools becoming available develop next generation vaccines featuring easy application transportability stability compact size specific targeting enhanced effectiveness results reported mahe coworkers represent notable step direction author states conflict interest author thanks anthony gaspari support mentorship encouragement lowell goldsmith guidance