Chividzhyan, M.

Kaprelyan, G.

Rusev, V.

Saruyan, A.

Tuncheva, K.

English

Varna Medical University Press: Journals

I l l Prophylactical problems S T U D Y O F T H E A N T I G E N I C S T R U C T U R E , I M M U N O L O G I C A L A C T I V I T Y A N D SOME G E N E T I C M A R K E R S O F I N F L U E N Z A V I R U S E S I S O L A T E D I N 1987 - 1988 G. Kaprelyan, V. Rusev, M. Chividzhyan, K. Tuncheva, A. Saruyan Key-words: influenza virus — neuraminidase — hemagglutinin — hemagglutination — infectious titre Recently, molecular biology of influenza viruses advances rapidly and en-ables the solving of numerous essential problems concerning the nature of the virus and influenza pathogenesis and immunity. Antigenic instability of influenza viruses, type A presents an unique pe-culiarity (1,3). The source and mechanisms of origin of new epidemic influenza strains are one of the actual topics of modern biology and epidemiology of in-fluenza viruses. I t is assumed that there exists a successive and systemic an-tigenic variability of influenza viruses during their circulation under natural conditions (2, 5, 6). It is also considered that when new influenza viruses, type A appear (characterized by new antigens) the etiological role of previously circulating influenza virus strains decreases or even disappears (4, 11)1 Anti-genic changes of the structure of surface protein components of influenza v i . ruses characterize, probably, the clinico-epidemiological and immunologic a j findings of influenza epidemics (7, 9, 10). In the present scientific communication we report our results from the investigations of the antigenic structure and genetic markers of influenza v i -ruses, type A isolated in the region of Varna in the period 1987 — 1988. Material and methods The following biological properties and genetic markers were studied in five out of 15 isolated influenza virus strains: ability to be isolated on chick embryo model and adaptability to some cellular cultures; hemagglutination, eluation, inhibition-sensitive, pathogenic, infectious, toxic, and immunogenic properties; thermostability of hemagglutinin, neuraminidase activity under the action of the inhibitors glutathione and L-cysteine, as well as the antigenic structure of the isolated viruses. Our complex methods used have been describ-ed in detail elsewhere (10). Determination of the residual neuraminidase ac-tivity was performed after Aminoff's method (8). N 124 6- Kaprelyan, V . RuseV, M. Chividzhyan, K . Tuncheva, A . Saruyafl Results and discussion The isolated influenza virus strains in the period 1987 — 1988 demonstrat­ed a difficult isolation ability (up to 3—4 passages) on model of 9—12 chick embryos. On cellular culture model «CEK>\ however, influenza viruses isolated up to the 4 t h passage could not adapt by characteristic cytopathic alterations. The isolated influenza virus strains as well as standard ones showed various heamabsorption and hemagglutination properties towards 11 kinds of animal and human «0>> group erythrocytes at 4 ; and 18° C. Most strains studied de­monstrated a well-expressed hemabsorption and hemagglutination activity at both temperature levels towards erythrocytes from hen, rabbit, sheep, rat, hisusfc, tortoise as well as towards human «0» group erythrocytes. Our experiments indicated that these influenza viruses posessed a mani­fested absorption and different eluation activity at 37° C in saline by using of an unheated or heated inactivated virus. Some strains were outlined to pos sess a poor eluation activity, namely A/Vn 1/87 and A/Vn 1/88. We studied the inhibition-sensitive properties of the isolated influenza virus strains towards unspecific inhibitors contained in the serum of 11 kinds of animals and man. There were influenza virus strains with high and with re­latively lower reactions to unspecific serum inhibitors. Infectious titre of these viruses on chick embryo model varied within the limits of I g L D 5 0 - —2.50 and I g L D 5 0 - —7.54. The isolated influenza viruses in this period possessed a weakly expressed pathogenic, infectious, toxic and immunogenic activity on chick embryos and experimental animals (cocks and mice). The results from the cultivation of influenza viruses on chick embryos at 26°C, 34°C, and 40°C demonstrated their heterogenic reproduction activity and hemagglutinin thermostability. Neuraminidase activity and thermosensitivity at 56°C and GO'C as well as the effect of the inhibitors L-cysteine and reduced glutathione was estimated in these strains, too. By means of the reaction of inhibition of hemagglutination it was established that these influenza virus strains pertained to antigenic subtypes N1 and N2 when their neuraminidase was concerned. Influenza v i ­ruses with an antigenic formula H3N2 were related to the standard strain А/ Leningrad 360/86 (H3N2) while the strain A/Vn(l /87 (H1N1) was related to A/England/333/80 (H1N1). Enzyme thermosensitivity indicated (fig. 1) that enzyme activity decreas­ed by about 80—90 per cent during the first 5—10 min of the experiment at 56°C and 60° C in all the strains examined. Under the influence of L-cysteine and reduced glutathione there was an enzyme inhibition by approximately 50 per cent in all the standard strains used. Fig . 2 showed that local strains were inhibited to an almost equal extent in comparison with standard ones under L-cysteine action. Fig. 3 demonstrated that local strains were inhibited to a greater extent in comparison with standard ones when reduced glutathione was applied. Our data showed that glutathione was a better inhibitor than L-cysteine by 8—12 per cent as compared with the controls. The investigations of the antigenic structure of influenza virus strains iso­lated in the period 1987—1988 and the determination of the antigenic profile of their neuraminidase component indicated their belonging to influenza virus, Study.of t h e , ц . , . , 125 type A with an antigenic formula A(H1N1) and A(H3N2). Strains with an an t i ­genic formula A(H1N1) were similar to a certain extent to the standard strain A/Taiwan/1/86 (H1N1) and those with an antigenic formula A(H3N2) — w ith the standard strains A/Leningrad /360/86 (H3N2) and A/Philipines /2/82 (H3N2) (table 1 and 2). Fig . 1. Enzyme activity changes at 56°C and 60 eC for different time periods. These studies demonstrate that intensive simultaneous circulation of both antigenic variants of influenza virus type A with their corresponding genetic markers and antigenic properties goes currently on at presence. 126. а-.-Ка№1уЛ!ЪЛ: Яиьеу, М..Cliivid^yan, K-.Zuncheua, A^Sa-myan WO 50 Fig . 2. Neuraminidase Legend: activity after 0.1 M L-cysteine treatment 1 — A (Sofia) 87 /H2N2/ . 2 — A (Leningrad) 360/86 /H3N2/ 3 — A (Chile)" 1/83 / Н Ш 1 / 4 —- A (England) 333/80 / Н Ш 1 5 — A (Varna) 4/9/87 /H3N2/ 6 — A (Varna) 9/1/88 /H3N2/ • 7— A (Varna) 9 6/88 /H3N2/ 8 — A (Varna) 9/9/88 (H3N2/ 9 — A (Varna) 14/1/87 / Н Ш 1 / 100 50- Г ~ I Fig . 3. Neuraminidase activity after 0.1 M glutathione treatment. Legend — see fig. 2. Study of the. 00 oo oo > N c и & ТЗ c 00 CM _ CO 00 oo CD 00 00 5 1 00 CM > o o Я © . .o oo o o o CM CM CO o o — © o g o o o ^ CO o o cc z: z, co o o • o © o o o o © O O O CM 00 QQ —н — CO © . O © O © © © CM O O O "s" co — i * - CO oo © o o o CM CO ^ CO — - 0 0 o 2: см , o GO —« i co co i CO 2 ^ 0 0 ' .E . 2 o <* C ' a f l ^- vz J w u СЛ < < < < ins I * -/) isei An > a C o o o © o ю Ю Ю Й Ю O O O © © io to to to ю o-o o o o to to tO Ю tO-© o o o o Ю Ю Ю i C lO 2 O O O O © — £g i p 00 tO tO tO § 0 0 0 0 0 - § • © to o-o-O" •••loo- '••'.00 .-oo.. 1̂ i o o © o см . • CO § § © 8 2 o © o © © S |Д й w w й 'Л N 00 X 00. 00 =3 00 ^ ,00 00 0O a/, a> 5 $ 5 ° * - и Ф a Ф 0 $~ " G Is of>>>> X < < • «ej < 128 G - Kaprelyan, V . Rusev, M. Chividzhyan, K . Tuncheva, A- Saruya n • " i ? - r ^ — t o * * — I M I , R E F E R E N C E S I . A h t o н o в а, В . I I . , и др. Хрон. B03, 33, 1979, № 4, 192—194. — 2. Г о л у -б е в, Д . Б . Йтоги науки и техники, сер. Вирусология, 185, № И , 156 с. — 3 . 3 а к с т е л ь-е к а я , Л . Я-» 3. И и к о л о в а, М. Я х н о, и др. Вопр. вирусол., 1982, JS& 6.671 — 6 7 6 . - 4. К е б е д ж и е в , Г., К. К у з м о в, X . Р а д е в а , и др. Епидемиол., мй-кробиол. и инфекц. болести, 1981, № 1, 17—22. — 5. К о ц е в a, P . , П. П а н а й о т о в . Л . Ш и н д а р о в. Епидемиол., микробиол. и инфекц. болести, 1985, № 1 , 1—6. — 6. Н и к о л о в а, 3. , Р . К о ц с в а , Г. К а п р е л я н, и др. Епидемиология, микробиол. и инфекц. болести, 1985, №> 2, 1—8. — 7. Щ и л о в, А. А., Ю. В . К о з л о в , О. Ч а к а , н др. Вопр. вирцеол., 1984, Л!° 4, 419—429. — 8 . A m i n о f f, D. Biochem. 81 ' 1961, 382—384. — 9. C o 1 rn a n, P . At., C . W . W a r d. Curr. Topics Microbiol. Immunol, И4, 1985, 177—255. — 10. K a p r c l у a n, G . , L . K a r a l v . a n o v a, K . M е t o d i -c v , et a l . Jn: Internat. Congr.-Infect. Parasit . Dis . , I X . Munich, 2 0 - 2 2 . V I . 1986. Abstracts, p. 469: Poster, V—P—12. — 11. W е b s t c r, R . G . , L . E . B r o w n , \V . G . Z a v e r . Virology, 135, 1984, No 1, 30—42. ИССЛЕДОВАНИЕ А Н Т И Г Е Н Н О Й С Т Р У К Т У Р И , ИММУНОЛОГ ИЧЕСКОЙ А К Т И В Н О С Т И И H E K O T O P b l X Г Е Н Е Т И Ч Е С К И Х М А Р К Е Р О В В И Р У С О В ГРИППА, ИЗОЛ И Р О В А Н Н Ь Г Х В 1987 - 1988 Г. 4 . | Г. Капрелян, В. Русев, М. Чивидлсиян, /<. Тунчева, А. Сараян Р Е З Ю М Е С помошью совремеш-шх вирусологических, биолошческих и биохимических методов иеследовались аитнгенная структура, некоторьге биологические и нммунологическне свой­ства гриппозного вируса, изолированного в 1987—1988 г. в Варненской области. Прослеживалось изменение антигеннмх свонств поверхностньгх белковьгх компо-нентов — гемаглутинина и невраминидазьг двадцати штаммов вирусов гриппа типа А с антигенной формулой А / Н Ш 1 и A/H3N2. Б н л и изучени следующие биологические свойства изолированнмх вирусов гриппа: способность изолируемости модели куриннх змбрионок, а также гемадсорбцнонние, гемаглютинациошше, ингибиторчувствительнне, злюирующис, патогенние, инфекционние, токсическне и иммуногеннне свойства. Изучалась также тер-мостабильность поверхностннх антигенов и акгивность невраминидазьг. Б и л а установлена способность к репродукции изолнрованних штаммов вирусов гриппа при температура 2 6 ' С 34°С и 40°G. : С помбщью проведеннцх неследованин бнли усганозленьг закономерности, которьге дают более полную характеристику циркулирующих в последнее время вирусов гриппа, 

STUDY OF THE ANTIGENIC STRUCTURE, IMMUNOLOGICAL ACTIVITY AND SOME GENETIC MARKERS OF INFLUENZA VIRUSES ISOLATED IN 1987 - 1988

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STUDY OF THE ANTIGENIC STRUCTURE, IMMUNOLOGICAL ACTIVITY AND SOME GENETIC MARKERS OF INFLUENZA VIRUSES ISOLATED IN 1987 - 1988

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