Meningococcal Serogroup B Bivalent rLP2086 Vaccine Elicits Broad and Robust Serum Bactericidal Responses in Healthy Adolescents

Meningococcal Se og oup B Bi alen LP2086 Vaccine Elici s B oad and Robus Se um Bac e icidal Responses in Heal hy Adolescen s Timo Vesika i,1La s Øs e gaa d,2Ja ie Diez-Domingo,3Jacek Wysocki,4Ca l-E ik Flodma k,5Johannes Beeslaa ,6 Joseph Eiden,7Qin Jiang,8Ka h in U. Jansen,7Thomas R. Jones,7Shannon L. Ha is,7Robe E. O’Neill,7 Lau a J. Yo k,9G aham C ow he ,6and John L. Pe ez8 1 Uni e si y o Tampe e Medical School, Finland; 2 Depa men o In ec ious Diseases, Aa hus Uni e si y Hospi al, Denma k; 3 Á ea de In es igación en Vacunas, FISABIO-Public Heal h, Uni e sidad Ca ólica de Valencia, Spain; 4 Depa men o P e en i e Medicine, Poznan´ Uni e si y o Medical Sciences, Poland; 5 Vaccine Uni , Depa men o Pedia ics, Skåne Uni e si y Hospi al, Malmo, Sweden; 6 Pfize L d, Wal on Oaks, Tadwo h, Uni ed Kingdom; 7 Pfize Vaccine Resea ch, Pea l Ri e , New Yo k; 8 Pfize Global Vaccines, College ille, Pennsyl ania, and 9 Pfize Medical and Scien ific A ai s, College ille, Pennsyl ania Co esponding Au ho : Timo Vesika i, MD, PhD, Uni e si y o Tampe e Medical School, Vaccine Resea ch Cen e , Bioka u 10, Tampe e, Finland. E-mail: imo. esika i@u a.fi. Recei ed Janua y 20, 2015; accep ed June 9, 2015; elec onically published Augus 4, 2015. Backg ound. Neisse ia meningi idis se og oup B (MnB) is a leading cause o in asi e meningococcal disease in adolescen s and young adul s. A ecombinan ac o H binding p o ein ( HBP) accine (T umenba ® ; bi alen LP2086) was ecen ly app o ed in he Uni ed S a es in indi iduals aged 10–25 yea s. Immunogenici y and sa e y o 2- o 3-dose schedules o bi alen LP2086 we e assessed in adolescen s. Me hods. Heal hy adolescen s (11 o <19 yea s) we e andomized o 1 o 5 bi alen LP2086 dosing egimens (0,1,6-mon h; 0,2,6-mon h; 0,2-mon h; 0,4-mon h; 0,6-mon h). Immunogenici y was assessed by se um bac e icidal an ibody assay using human complemen (hSBA). Sa e y assessmen s included local and sys emic eac ions and ad e se e en s. Resul s. Bi alen LP2086 was immunogenic when adminis e ed as 2 o 3 doses; he mos obus hSBA esponses occu ed wi h 3 doses. The p opo ion o subjec s wi h hSBA i e s 1:8 a e 3 doses anged om 91.7% o 95.0%, 98.9% o 99.4%, 88.4% o 89.0%, and 86.1% o 88.5% o MnB es s ains exp essing accine-he e ologous HBP a ian s A22, A56, B24, and B44, espec i ely. A e 2 doses, esponses anged om 90.8% o 93.5%, 98.4% o 100%, 69.1% o 81.1%, and 70.1% o 77.5%. Geome ic mean i e s (GMTs) we e highes among subjec s ecei ing 3 doses and simila be ween he 2- and 3-dose egimens. A e 2 doses, GMTs ended nume ically highe among subjec s wi h longe in e als be ween he fi s and second dose (6 mon hs s 2 and 4 mon hs). Bi alen LP2086 was well ole a ed. Conclusions. Bi alen LP2086 was immunogenic and well ole a ed when adminis e ed in 2 o 3 doses. Th ee doses yielded he mos obus hSBA esponse a es agains MnB s ains exp essing accine-he e ologous sub amily B HBPs. Key wo ds. bi alen LP2086; clinical ial; unc ional immunogenici y; Neisse ia meningi idis se og oup B; sa e y. Neisse ia meningi idis se og oup B (MnB) causes mos meningococcal disease in Eu ope and app oxima ely one hi d o cases in he Uni ed S a es [1,2]. MnB in ec ion is mo e common among adolescen s and young adul s han he gene al adul popula ion [2]. MnB polysaccha ide ac- cines a e poo ly immunogenic, likely due o c oss- eac ing epi opes o MnB capsula polysaccha ide wi h polysialic acid s uc u es on human neu onal cells [3]. Vaccines p o- duced om MnB ou e memb ane esicles ha e shown clinical e ficacy in child en olde han 4 yea s, agains MnB s ains bea ing an igens homologous o hose in he accine, bu no agains s ains exp essing an igens he e ologous o he accine [4–6]. The e o e, al e na i e app oaches based on conse ed MnB p o ein an igens ha e been pu sued o p o ide p o ec ion agains di e se s ains causing endemic and epidemic in asi e MnB disease. A ecombinan , mul icomponen MnB accine (Bexse o, No a is Vaccines, Siena, I aly) has been licensed O iginal A icle Jou nal o he Pedia ic In ec ious Diseases Socie y, Vol. 5, No. 2, pp. 152–60, 2016. DOI:10.1093/jpids/pi 039 © The Au ho 2015. Published by Ox o d Uni e si y P ess on behal o he Pedia ic In ec ious Diseases Socie y. This is an Open Access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License (h p://c ea i ecommons.o g/licenses/by/4.0/), which pe mi s un es ic ed euse, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed. in he Eu opean Union, Canada, Aus alia [7–9], and, mo e ecen ly, in he Uni ed S a es [10].Ano he mul icomponen accine candida e, bi alen LP2086 (T umenba ® ), was e- cen ly app o ed in he Uni ed S a es o p e en in asi e me- ningococcal disease caused by MnB in indi iduals 10 o 25 yea s o age [11]. The a ge an igen o his accine, LP2086, was iden ified using a combined biochemical and unc ional immunologic sc eening app oach [12]. LP2086 is a bac e ial, su ace-exposed i ulence ac o ha binds human ac o H, also known as ac o H binding p o ein ( HBP) [13]. In a comp ehensi e su ey o 1837 MnB s ains isola ed om pa ien s wi h in asi e disease in he Uni ed S a es and Eu ope, he LP2086 gene was p esen in all isola es [14], al hough in a e cases, in asi e disease-causing MnB s ains may lack a unc ional LP2086 gene [15].LP2086 p o ein sequences can be g ouped in o 2 gene ically and immunologically dis inc sub amilies, des- igna ed A (30% o isola es) and B (70% o isola es) [14]. The bi alen accine con ains ecombinan lipida ed p o- eins om each LP2086 sub amily (bi alen LP2086), because his o mula ion was shown o be mos e ec i e a inducing b oadly p o ec i e se um bac e icidal an ibodies. In phase 1 and 2 s udies, immune se a elici ed by bi alen LP2086 demons a ed bac e ial killing ac i i y, measu ed in se um bac e icidal assays using human complemen (hSBAs), agains di e se MnB s ains bea ing HBP a i- an s o bo h sub amilies and he e ologous in sequence o hose in he accine [16–19]. Fo immunogenici y e alua- ions, 4 MnB es s ains we e selec ed o eflec he epide- miology, b ead h, and global dis ibu ion o MnB s ains. Each hSBA es s ain exp esses an HBP a ian ha is di - e en om accine componen s, hus allowing an objec- i e assessmen o unc ional and p o ec i e bac e icidal immune esponses agains in asi e disease-associa ed s ains in ci cula ion [20]. This s udy e alua ed he immunogenici y, ole abili y, and sa e y o bi alen LP2086 adminis e ed in 3-dose o 2-dose egimens in heal hy adolescen s aged 11 o <19 yea s a en ollmen . The immune esponse measu ed by hSBA was e alua ed 1 mon h a e dose 2 o 3 and included he pe cen age o subjec s achie ing hSBA i e s 1:8, a mo e conse a i e indica o o po en ial se op o ec ion han he ecognized co ela e o p o ec ion, an hSBA i e 1:4 [21,22]. Sa e y assessmen s included he incidence o local eac ions, sys emic e en s, and ad e se e en s (AEs). METHODS S udy Pa icipan s and Design This phase 2, mul icen e , andomized, single-blind s udy was conduc ed be ween Ma ch 3, 2011 and Augus 30, 2013, in heal hy adolescen s 11 o <19 yea s o age om he Czech Republic, Denma k, Finland, Ge many, Poland, Spain, and Sweden. Subjec s we e judged heal hy by he in es iga o and ag eed o p ac ice an e ec i e o m o con acep ion. Key exclusion c i e ia we e p e ious accina ion wi h any MnB accine; p e ious anaphylac ic eac ion o any accine o accine- ela ed componen ; his- o y o cul u e-p o en N meningi idis o Neisse ia gono - hoeae disease; p egnancy o nu sing; cu en ch onic use o sys emic an ibio ics; bleeding dia hesis o condi ion ha would con aindica e in amuscula injec ion; known o suspec ed disease o he immune sys em o e- ceip o immunosupp essi e he apy; any neu oinflamma- o y o au oimmune condi ion; and eceip o any blood p oduc s, including immunoglobulin, wi hin 6 mon hs be- o e he fi s s udy accina ion. Subjec s we e andomized 3:3:3:2:1 co esponding o a 0,1,6-mon h, 0,2,6-mon h, 0,6-mon h, 0,2-mon h, and 0,4-mon h bi alen LP2086 dosing schedule, espec i ely (g oups 1–5) (Table 1; Figu e 1). The unequal andomiza- ion schedule eflec s he compa isons o in e es o his s udy. The fi s 3 g oups ( andomized 3:3:3) we e assigned o mee he hypo hesis es ing objec i es; he emaining 2 g oups ( andomized 2:1) we e desc ip i e and we e used o explo e di e en LP2086 dosing schedules. Subjec s we e andomized using an in e ac i e oice- o web-based esponse sys em. En ollmen was s a ified by age (11 o <14 and 14 o <19 yea s) o ensu e ha all ages we e ade- qua ely ep esen ed in he s udy. Each subjec ecei ed 4 s udy injec ions, wi h adminis a ion o 2 o 3 doses o bi alen LP2086 acco ding o he a o emen ioned dosing schedule; saline was adminis e ed when bi alen LP2086 was no scheduled. Pa icipan s we e blinded o hei assigned dose egimen. All labo a o y s a we e blinded o subjec , isi , and ea men (ie, andomiza ion g oup), bu in es iga o s and sponso knew he alloca ion o sub- jec s h oughou he s udy. This s udy was conduc ed in acco dance wi h he Decla a ion o Helsinki [23] and he In e na ional Con e ence on Ha monisa ion Guidelines o Good Clinical P ac ice [24] (ClinicalT ials.go : NCT01299480). Vaccine and Placebo Subjec s ecei ed 1 in amuscula dose o bi alen LP2086 o saline in he uppe del oid muscle. Each 0.5-mL dose o bi alen LP2086 con ained 60 µg each o LP2086 A (A05) and B (B01) sub amily p o eins p o- duced in Esche ichia coli and o mula edwi haluminum phospha e in an iso onic bu e in p efilled sy inges. Saline (0.9% sodium chlo ide) was adminis e ed as a 0.5-mL dose. Bi alen LP2086 in Adolescen s 153 Immunogenici y Assessmen s Blood samples o hSBA we e ob ained a 0, 2, 3, and 7 mon hs (Table 1). Func ional bac e icidal an ibodies we e assessed agains 4 MnB es s ains exp essing HBP a i- an s ha a e he e ologous in sequence compa ed wi h he accine componen s; 2 s ains exp ess HBP sub amily A a ian s (A22 and A56), and 2 s ains exp ess HBP sub- amily B a ian s (B24 and B44). The HBP a ian s ep e- sen 4 o he 6 majo HBP a ian subg oups ha accoun o >90% o in asi e meningococcal disease isola es ci cu- la ing in he Uni ed S a es and Eu ope combined [20]. As an indica ion o he di e si y o hese 4 MnB es s ains, he mul ilocus sequence ype clonal complex (ie, he gene ic backg ound) o each s ain ep esen s 1 o he 4 mos p e alen clonal complexes in ci cula ing in asi e MnB s ains (Table 2)[20]. Each o hese 4 MnB es s ains ha e de ec able HBP su ace exp ession le els ep esen a- i e o hei espec i e HBP a ian g oup, as measu ed in aflow cy ome y assay adap ed om McNeil e al [25] using he an i- HBP b oadly c oss- eac i e monoclonal an- ibody MN86-994-11-1. The p ima y immunogenici y endpoin s we e he p o- po ion o subjec s ecei ing 3 doses o bi alen LP2086 ( accina ion a 0, 1, 6 and 0, 2, 6 mon hs) who achie ed hSBA i e 1:8 o each o he 4 MnB es s ains 1 mon h a e he hi d dose o bi alen LP2086. An hSBA i e 1:8 is a mo e conse a i e indica o o se op o ec- ion han a i e 1:4, which is he ecognized co ela e Table 1. S udy Design LP2086, mo Visi 1* (Mon h 0) Visi 2 (Mon h 1) Visi 3* (Mon h 2) Visi 4* (Mon h 3) Visi 5 (Mon h 6) Visi 6* (Mon h 7) Visi 7 (Mon h 12) Injec ion 1 Injec ion 2 Injec ion 3 Blood D aw Only Injec ion 4 Blood D aw Only Telephone Con ac Only 0,1,6 LP2086 LP2086 Saline LP2086 0,2,6 LP2086 Saline LP2086 LP2086 0,2 LP2086 Saline LP2086 Saline 0,4 Saline Saline LP2086 LP2086 0,6 LP2086 Saline Saline LP2086 Abb e ia ions: HBP, ac o H binding p o ein; hSBA, se um bac e icidal assay using human complemen ; LP2086, bi alen LP2086 accine. *Blood d aw a isi s 1, 3, 4, and 6 o hSBA pe o med wi h s ains exp essing HBP a ian s A22, A56, B24, and B44. Figu e 1. Subjec disposi ion. *Values in his ow used as denomina o s o pe cen ages. 154 Vesika i e al o p o ec ion agains meningococcal disease [21,22]and add esses he abili y o he hSBA o de e mine p ecisely and accu a ely a posi i e om a nega i e se o esponse. The hypo hesis- es ing seconda y endpoin was he p o- po ion o subjec s ecei ing 2 doses o bi alen LP2086 a 0,6 mon hs who achie ed hSBA i e 1:8 o each o he 4 MnB es s ains 1 mon h a e he second dose o bi- alen LP2086. O he seconda y immunogenici y end- poin s included assessmen o geome ic mean i e s (GMTs) o each o he 4 MnB es s ains and he p opo - ion o esponde s wi h hSBA i e s 1:8 o all g oups a each sampling poin . Responses a e 1 dose o bi alen LP2086 we e only a ailable o he 0,4-mon h dosing g oup. Sa e y Assessmen s Subjec s we e obse ed o 20 minu es (o longe pe local p ac ice) a e accina ion o immedia e eac ions, which we e documen ed as AEs. Reac ogenici y da a we e collec ed by elec onic dia y (e-dia y) o 7 days a e each injec ion and included solici ed local eac ions and sys emic e en s and use o an ipy e ic medica ions. Injec ion-si e edness and swelling we e measu ed by sub- jec s using calipe s p o ided, and measu emen s we e sub- sequen ly g aded as mild, mode a e, o se e e as defined p ospec i ely by s udy p o ocol. Injec ion-si e pain and sys- emic symp oms o headache, a igue, chills, omi ing, dia hea, muscle pain, and join pain we e g aded by he subjec s as mild, mode a e, o se e e. Tempe a u e was measu ed a bed ime; e e was defined as empe a u e 38.0°C (100.4°F). Only he highes e e measu emen eco ded among measu emen s aken du ing any single 24-hou pe iod was epo ed. Unsolici ed AEs we e collec - ed om signing o in o med consen h ough 1 mon h a e he las injec ion (bi alen LP2086 o saline). Se ious AEs (SAEs) we e collec ed h oughou he s udy. S a is ical Analysis The e aluable immunogenici y popula ion included sub- jec s who ecei ed all doses o bi alen LP2086 as an- domized, had no majo p o ocol iola ion o use o p ohibi ed accines, had blood d awn be o e dose 1 and 1 mon h a e he las dose o bi alen LP2086, and had a alid and de e mina e assay esul o he p oposed anal- ysis. The sa e y popula ion included all pa icipan s who ecei ed a leas 1 injec ion (bi alen LP2086 o saline) and had any sa e y da a a ailable. Immunogenici y endpoin s, defined as he p opo ion o subjec s esponding wi h an hSBA i e 1:8, we e summa- ized along wi h he exac 2-sided 95% confidence in e al ([CI] o Cloppe -Pea son confidence limi ) o he p opo - ion; he exac CI o he p opo ion was compu ed using he F dis ibu ion. The esponse a es in subjec s ecei ing 3 e sus 2 doses o bi alen LP2086 we e compa ed wi h 50% a a significance le el o 1.25%, using a 1-sided exac es based on binomial dis ibu ion. GMTs we e compu ed wi h 2-sided 95% CIs. RESULTS Subjec s A o al o 1714 subjec s we e en olled and 1713 we e an- domized; 1 subjec ecei ed saline wi hou andomiza ion and wi hd ew om he s udy (Figu e 1). A o al o 1712 subjec s ecei ed a leas 1 dose o bi alen LP2086 o sa- line, and 1550 (90.5%) comple ed he s udy. Mos subjec s we e whi e (99.0%) and non-Hispanic (98.5%). A he ime o fi s injec ion, 63.4% o subjec s we e 14–18 yea s o age; mean age o he o al popula ion was 14.4 yea s (Table 3). Demog aphic cha ac e is ics we e simila ac oss dosing egimens. Immunogenici y A o al o 1450 subjec s comp ised he e aluable immuno- genici y popula ion. One mon h a e dose 3, he p opo - ion o subjec s wi h hSBA i e s 1:8 a e 3 doses o bi alen LP2086 adminis e ed a 0,1,6 mon hs was 91.7%, 99.4%, 89.0%, and 88.5% o MnB es s ains exp essing accine-he e ologous HBP a ian s A22, A56, B24, and B44, espec i ely. Fo subjec s accina ed a 0,2,6 mon hs, 95.0%, 98.9%, 88.4%, and 86.1% had hSBA i e s 1:8 (Figu e 2). The e we e no significan di - e ences in immunogenici y among subjec s who ecei ed bi alen LP2086 on he 0,1,6-mon h and 0,2,6-mon h schedules. Table 2. Geno ypic Cha ac e is ics o N meningi idis se og oup B Tes S ains S ain ID HBP Va ian ( HBP Pep ide ID a ) Homology, b % HBP Subg oup ST Clonal Complex Po A Sub ype PMB2001 A56 (187) 98.1 N1C2 cc213 P1.22,14 PMB2707 B44 (15) 91.6 N4/N5 cc269 P1.19-1,10-4 PMB80 A22 (19) 88.9 N2C2 cc41/44 P1.21,16 PMB2948 B24 (1) 86.2 N6 cc32 P1.12-1,13-1 Abb e ia ions: HBP, ac o H binding p o ein; ID, iden i ica ion; ST, sequence ype. a HBP pep ide ID is om h p://pubmls .o g/neisse ia/ Hbp/. b Amino acid iden i y o he HBP sub amily ma ched accine componen : A05 o he s ains exp essing HBP sub amily A a ian s, o B01 o he s ains exp essing HBP sub amily B a ian s. Wi hin HBP sub amily, he pe cen age amino acid iden i y is >83%. Bi alen LP2086 in Adolescen s 155 Fo HBP sub amily A s ains, among hose ecei ing 2 doses o bi alen LP2086, >90% o subjec s had hSBA i- e s 1:8 o A22 and >98% o subjec s had hSBA i e s 1:8 o A56 1 mon h a e he second dose. Fo HBP sub amily B s ains, >69% o subjec s had hSBA i e s 1:8 o B24 and >70% o B44 (Figu e 2). The s udy de- sign allowed assessmen o hSBA esponses a 1 mon h a e 1 dose o bi alen LP2086 (0,4-mon h g oup only). A e he fi s dose, 55.9%, 67.6%, 56.9%, and 23.8% o subjec s had hSBA i e s 1:8 o A22, A56, B24, and B44, espec i ely. P e accina ion hSBA esponses o all 4 MnB es s ains we e low, anging om 5.2% (B44) o 28.1% (A22). Pos accina ion GMTs inc eased wi h each dose o bi a- len LP2086 and we e highes among subjec s ecei ing 3 doses o bi alen LP2086. Fo subjec s accina ed a 0,1,6 mon hs, GMTs 1 mon h a e dose 3 we e 55.1, 152.9, 29.1, and 40.3 o A22, A56, B24, and B44, espec i ely. Fo subjec s accina ed a 0,2,6 mon hs, GMTs we e 56.3, 155.6, 25.6, and 35.0 (Figu e 3). Fo HBP sub amily A s ains, among subjec s ecei ing 2 doses o bi alen LP2086, GMTs anged om 37.1 o 48.4 o A22, and 104.9 o 125.6 o A56 one mon h a e he second dose. Fo he HBP sub amily B s ains, GMTs anged om 14.7 o 20.6 o B24 and 17.8 o 22.5 o B44. A e 2 doses o bi alen LP2086, GMTs ended nume ically highe among subjec s wi h a longe in e al be ween doses; subjec s wi h a 6-mon h in e al be ween doses (0,6- mon h g oup) had highe GMTs o all 4 MnB es s ains compa ed wi h subjec s ha ing a 4-mon h (0,4-mon h), 2-mon h (0,2- and 0,2,6-mon h), o 1-mon h (0,1,6- mon h) in e al be ween doses (Figu e 3). A e 1 dose o bi alen LP2086 (0,4-mon h g oup), GMTs we e 16.0, 26.8, 12.6, and 6.8, espec i ely, o A22, A56, B24, and B44. Sa e y The equency o local eac ions was highe a e bi alen LP2086 adminis a ions compa ed wi h saline; pain a he injec ion si e was he mos common local eac ion (Figu e 4). Ac oss dosing schedules, mos cases o pain a e bi alen LP2086 and saline adminis a ion we e mild o mode a e. Se e e pain was epo ed by 9.9% o Figu e 2. Pe cen age o subjec s wi h hSBA i e s 1:8 agains N meningi idis se og oup B es s ains A22, A56, B24, and B44 a baseline and 1 mon h a e injec ion wi h bi alen LP2086 o saline. E o s shown a e 95% confidence in e - als. hSBA, human se um bac e icidal an ibody assay using human complemen . Table 3. Demog aphics, Sa e y Popula ion Bi alen LP2086 Dosing Schedule 0,1,6 mo n = 426 0,2,6 mo n = 414 0,6 mo n = 451 0,2 mo n = 277 0,4 mo n = 144 To al N = 1712 Sex, n (%) Female 212 (50) 217 (52) 227 (50) 135 (49) 79 (55) 870 (51) Male 214 (50) 197 (48) 224 (50) 142 (51) 65 (45) 842 (49) Race, n (%) Whi e 423 (99) 408 (99) 448 (99) 272 (98) 144 (100) 1695 (99) Age a i s injec ion, y, n (%) 11 o <14 155 (36) 154 (37) 164 (36) 101 (37) 53 (37) 627 (37) 14 o <19 271 (64) 260 (63) 287 (64) 176 (64) 91 (63) 1085 (63) Mean (SD) 14.4 (2.22) 14.4 (2.23) 14.4 (2.17) 14.4 (2.25) 14.3 (2.11) 14.4 (2.20) Abb e ia ion: SD, s anda d de ia ion. 156 Vesika i e al subjec s who ecei ed bi alen LP2086 and 0.3% sub- jec s who ecei ed saline, o each o he 4 injec ions. O he common local eac ions included edness and swell- ing, which we e mild o mode a e in se e i y (Figu e 4). The mean du a ion o all local eac ions a e each dose o bi alen LP2086 was 2.1 o 3.2 days; 4 local eac ions, all o which we e pain a he injec ion si e, had a du a ion g ea e han 14 days, bu no subjec s wi hd ew om he s udy due o pain. Ac oss all injec ions, he mos common sys emic e en s we e headache and a igue. The majo i y o sys emic e en s we e mild o mode a e in se e i y. Se e e headache was e- po ed by 1.6% o subjec s a e ei he bi alen LP2086 o saline. Se e e a igue was epo ed by 3.6% o subjec s a e ei he injec ion (Figu e 5). In o al, 3 subjec s wi h- d ew due o a sys emic e en . Fe e 38°C wi hin 7 days o accina ion was in e- quen , epo ed by 1.7%–4.3% and 1.5%–2.1% o bi alen LP2086 o saline ecipien s, espec i ely. Fe e 39°C was in equen (<1% ac oss g oups). The median du a ion o e e was 1 day a e bi alen LP2086 o saline adminis a ion. Ac oss all 4 injec ions, 13.6%–16.2% and 7.5%–9.1% o subjec s used an ipy e ic medica ion a e bi alen LP2086 and saline, espec i ely. Du ing he accina ion phase, 35.5%–37.5% o subjec s epo ed 1 AE ac oss he 5 dosing g oups. Mos e en s we e mild o mode a e in se e i y. The mos common AE was nasopha yngi is (5.5% −10.1%). Ele en subjec s ex- pe ienced se e e AEs conside ed o be ela ed o bi alen LP2086, including headache, injec ion-si e pain, py exia, omi ing, and injec ion-si e swelling. Two subjec s epo - ed SAEs conside ed o be ela ed o bi alen LP2086; 1 subjec expe ienced e igo, chills, and headache a e dose 3, and ano he expe ienced py exia and omi ing a e dose 1. O e all, he e we e no di e ences in he inci- dence o SAEs be ween bi alen LP2086 and saline ecip- ien s o be ween he 3-dose and 2-dose schedules and no inc ease in AEs wi h subsequen dosing. Nine een subjec s (1.1%) wi hd ew due o an AE. O hese, 9 AEs we e con- side ed o be ela ed o s udy accina ion and included injec ion-si e pain (n = 4), headache (n = 2), mig aine, a igue, and e igo (n = 1 each). No dea hs we e epo ed. DISCUSSION Meningococcal B in ec ion can lead o se e e and debili a - ing disease. Vaccina ion agains non-B meningococcal se og oups (ACWY) has p o en e ficacious in all age g oups, pa icula ly in adolescen s who a e a high isk o meningococcal disease [26]. The need o an e ec i e MnB accine is unde sco ed by ecen ou b eaks on US college campuses and by endemic in ec ions wo ldwide [27–29]. The low basal immuni y o meningococcal B s ains in his s udy emphasizes he ulne abili y o adoles- cen s o in ec ion and disease caused by Nmeningi idis se og oup B. In his s udy, all 5 bi alen LP2086 accina ion egi- mens, ega dless o schedule o equency, we e immuno- genic and well ole a ed in heal hy adolescen s. The 2-dose and 3-dose egimens elici ed obus immune e- sponses. Conside ing ha hSBA i e s 1:4 a e an accep ed co ela e o p o ec ion agains in asi e meningococcal disease [21,22] and ha se op o ec i e esponses in his s udy we e defined using mo e s ingen c i e ia o hSBA i- e s 1:8, bi alen LP2086 has demons a ed a subs an- ial and b oad immune esponse a e 2 doses and a mo e obus esponse a e 3 doses, when adminis e ed ac oss a ange o dosing schedules. This s udy also allowed assess- men o hSBA esponses a e 1 dose o bi alen LP2086. Figu e 3. Geome ic mean i e s (GMTs) agains N meningi idis se og oup B es s ains A22, A56, B24, and B44 a baseline and 1 mon h a e injec ion wi h bi alen LP2086 o saline. Bi alen LP2086 in Adolescen s 157 Figu e 4. Local injec ion-si e eac ogenici y ( eco ded by elec onic dia y [e-dia y]). Da a ha e been agg ega ed ac oss g oups o show eac ogenici y a e each dose. Figu e 5. Sys emic eac ogenici y ( eco ded by elec onic dia y [e-dia y]). Da a ha e been agg ega ed ac oss g oups o show eac ogenici y a e each dose. *Fe e : 38.0–38.4°C = mild; 38.5–38.9°C = mode a e; and 39°C = se e e. 158 Vesika i e al A e a single accina ion, inc eased se op o ec i e esponses compa ed wi h baseline agains all MnB es s ains we e obse ed. Among ecipien s ecei ing 3 doses o bi alen LP2086, he iming o he second accina ion, whe he 1 o 2 mon hs a e ini ial accina ion, had no disce nable e ec on immunogenici y. None heless, he abili y o elici obus esponses a e 2 doses 1 mon h apa may be aluable du - ing ou b eaks when imely p o ec ion is c i ical. Immune esponses a e 2 doses o bi alen LP2086 we e subs an- ial and indica i e o a b oad immune esponse o he MnB es s ains ha we e selec ed o ep esen epidemiological- ly and an igenically ele an in asi e MnB s ains. Immunogenici y gene ally inc eased in he 2-dose sched- ules when he e was a longe in e al be ween he fi s and second dose, as seen by he highe GMTs and gene ally highe p opo ion o subjec s wi h hSBA i e s 1:8 a e dose 2 in he 0,6-mon h dosing egimen compa ed wi h he o he dosing schedules. Simila obse a ions we e desc ibed o quad i alen human papilloma i us accina- ion [30,31]. In hese s udies, inc easing he in e al be- ween doses 2 and 3 om 4 o 10 mon hs esul ed in highe an ibody i e s agains all 4 human papilloma i us ypes examined. Simila o o he clinical s udies o bi alen LP2086 in adolescen s and adul s [18,19,32,33], he majo i y o sa e- y e en s obse ed in his s udy we e local eac ions and sys emic e en s ha we e mild o mode a e in se e i y, an- sien , and wi hou po en ia ion a subsequen dosing. App oxima ely wo hi ds o subjec s did no epo any AEs du ing he s udy. O e all, 90.6% o subjec s we e able o comple e hei accina ion se ies, indica ing ha accina ions we e well ole a ed in adolescen s and ha any sa e y e en s, such as injec ion-si e pain, we e no an impedimen o accina ion. Sa e y e en s we e no in- c eased in subjec s ecei ing a hi d dose o bi alen LP2086. CONCLUSIONS In summa y, 2 o 3 doses o bi alen LP2086 we e immu- nogenic and well ole a ed. The 2-dose egimens p o ided subs an ial hSBA esponses agains di e se MnB s ains ex- p essing HBP a ian s he e ologous o accine an igen. Howe e , he 3-dose egimens yielded he highes se ocon- e sion a es agains sub amily B s ains and a highe le el o hSBA an ibodies as measu ed by GMTs. The con e- nience o a 2-dose schedule and he po en ial benefi o highe le el o p o ec i e an ibodies a e a 3-dose schedule should be ca e ully conside ed o u u e MnB immuniza- ion schedules. Acknowledgmen s We hank all he pa en s and gua dians o all pa icipan s and he many in es iga o s and s udy s a o hei indi idual con ibu ions o his s udy. Debo ah M. Campoli-Richa ds, BSPha m, RPh, and Susan E. DeRocco, PhD (Comple e Heal hca e Communica ions, Inc.) p o ided assis ance in p epa ing and edi ing he manusc ip . The s udy was egis e ed wi h ClinicalT ials.go , numbe NCT00657709. Financial suppo . This s udy was unded by Pfize Inc. Po en ial conflic s o in e es . T. V. ecei ed consul ing ees and suppo o mee ings, a el, o accommoda ion expenses om GlaxoSmi hKline; and he is a consul an and speake o Me ck, SanofiPas eu -Me ck Sha p & Dohme (SPMSD), MedImmune, No a is, and Pfize . J. D.-D. pa icipa ed in ad iso y boa ds and speake bu eaus o GlaxoSmi hKline, SPMSD, and Pfize , o which paymen is ecei ed; and he was a p incipal in es iga o in clin- ical ials o GlaxoSmi hKline and SPMSD. J. W. was he p incipal in es iga o o clinical ials sponso ed by GSK, No a is, Wye h, and Pfize ; and he has ecei ed a el g an s o pa icipa e in scien ific con e ences and was paid o lec u es. J. B., J. E., Q. J., K. U. J., T. R. J., S. L. H., R. E. O., L. J. Y., J. L. P., and G. C. a e ull- ime employees o Pfize Inc. L. O. has se ed as a p incipal in es iga o o ials spon- so ed by Pfize , GSK, and Sanofi-Pas eu MSD. C.-E. F. has been p in- cipal in es iga o o clinical ials o GlaxoSmi hKline, SPMSD, Wye h, Pfize , Smi hKline Beecham, and MSD. All au ho s ha e submi ed he ICMJE Fo m o Disclosu e o Po en ial Conflic s o In e es . Conflic s ha he edi o s conside ele an o he con en o he manusc ip ha e been disclosed. Re e ences 1. EU-IBIS Ne wo k. In asi e Neisse ia meningi idis in Eu ope 2006. A ailable a : h p://www.hpa-bioin o ma ics.o g.uk/ euibis/documen s/2006_meningo.pd . Accessed 27 Ma ch 2015. 2. Cen e s o Disease Con ol and P e en ion. 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