In order to surpass the problem of genetic variability of hepatitis C virus envelope proteins during vaccine development, we used the so-called "reverse vaccinology"approach--"from genome to vaccine". Database of HCV protein sequences was designed, viral genome analysis was performed, and several highly conserved sites were revealed in HCV envelope proteins in the framework of this approach. These sites demonstrated low antigenic activity in full-size proteins and HCV virions: antibodies against these sites were not found in all hepatitis C patients. However, two sites, which contained a wide set of potential T-helper epitope motifs, were revealed among these highly conserved ones. We constructed and prepared by solid-phase peptide synthesis several artificial peptide constructs composed of two linker-connected highly conserved HCV envelope E2 protein sites; one of these sites contained a set of T-helper epitope motifs. Experiments on laboratory animals demonstrated that the developed peptide constructs manifested immunogenicity compared with one of protein molecules and were able to raise antibodies, which specifically bound HCV envelope proteins. We succeeded in obtaining antibodies reactive with HCV from hepatitis C patient plasma upon the immunization with some constructs. An original preparation of a peptide vaccine against hepatitis C is under development on the basis of these peptide constructs.
Chtoby oboĭti problemu geneticheskoĭ variabel'nosti i izmenchivosti obolochechnykh belkov VGS pri sozdanii vaktsiny, nami byl ispol'zovan podkhod, nazyvaemyĭ “obratnaia vaktsinologiia” – “ot genoma k vaktsine”. V ramkakh étogo podkhoda byla sozdana baza dannykh posledovatel'nosteĭ belkov VGS, proveden analiz genoma virusa i vyiavleny neskol'ko vysokokonservativnykh uchastkov v sostave obolochechnykh belkov VGS. V sostave tselykh belkov i virusnykh chastits éti uchastki proiavliali nevysokuiu antigennuiu aktivnost': antitela protiv dannykh uchastkov obnaruzhivalis' daleko ne u vsekh bol'nykh gepatitom S. Odnako sredi vysokokonservativnykh uchastkov byli vyiavleny dva takikh, kotorye soderzhali shirokiĭ nabor potentsial'nykh motivov T-khelpernykh épitopov. Nami byli razrabotany i polucheny metodom tverdofaznogo peptidnogo sinteza neskol'ko iskusstvennykh peptidnykh konstruktsiĭ, sos toiashchikh iz dvukh vysokokonservativnykh uchastkov obolochechnogo belka E2 VGS, soedinennykh linkerom; odin iz étikh uchastkov soderzhal nabor T-khelpernykh épitopnykh motivov. V opytakh na laboratornykh zhivotnykh poluchennye peptidnye konstruktsii proiavliali immunogennost', sravnimuiu s immunogennost'iu belkovykh molekul, i byli sposobny vyzyvat' obrazovanie antitel, spetsifichno sviazyvaiushchikh obolochechnye belki VGS. Pri immunizatsii nekotorymi konstruktsiiami udalos' poluchit' antitela, sviazyvaiushchie VGS iz plazmy krovi bol'nykh gepatitom S. Na osnove ukazannykh peptidnykh konstruktsiĭ razrabatyvaetsia original'nyĭ preparat sinteticheskoĭ peptidnoĭ vaktsiny protiv gepatita S.
Keywords: antigens; envelope proteins; hepatitis C; peptides; vaccine; virus.