Children Vaccinated with a Cold-Adapted Influenza A/HK/

Transcription

1 INFECTION AND IMMUNITY, Mr. 198, p /8/3-86/5$./ Vol. 7, No. 3 Humorl nd Cellulr Immune Responses of Seronegtive Children Vccinted with Cold-Adpted Influen A/HK/ 13/77 (HlNl) Recombinnt Virus ARYE LAZAR, NOBUHIKO OKABE, AND PETER F. WRIGHT* Deprtment of Peditrics, Vnderbilt University School ofmedicine, Nshville, Tennessee 373 Humorl nd cell-medited immune responses of young, seronegtive children were ssessed fter intrnsl vccintion with cold-dpted influen, A/HK/ 77 (HlNl) CR 35 recombinnt virus. Vccinees shedding influen virus experienced rise in hemgglutinin-inhibition ntibody 15 to 3 dys fter vccintion. Vccinees showed low but significnt lymphocyte trnsformtion to A/USSR (HlNl) by dy 8 fter vccintion, which decresed to prevccintion levels t 3 to 3 dys. The lymphocyte trnsformtion response occurred before serum ntibody rises were detected by hemgglutinin-inhibition ssy. No chnge in lymphocyte responsiveness ws observed fter vccintion s mesured by phytohemgglutinin stimultion. Lymphocytes responded to in vitro incubtion with inctivted influen (HlNl) virus by producing interferon. The interferon produced ws of type I nd ws observed in vccinees nd nonvccinees both before nd fter vccintion. The use of cold-dpted vrints of influen trils on the bsis of seronegtivity by hemgglutintion inhibition (HAI) to influen A/USSR (HlNl) viruses s donors of genes to confer ttenution to wild types hs provided new tool in ttempts virus. Children were enrolled in the Vnderbilt Peditric Vccine Clinic, Nshville, Tenn. A totl of 15 to control influen epidemics. The bility of those recombinnts to grow t low tempertures children prticipted in four studies. The children (5C) serves s n in vitro mrker for ttenution which cn be ssessed in tissue culture influen A/HK/13/77 (HlNl) clone, CR 35 (Flow were vccinted intrnslly s previously described (). A totl of.5 ml of 1-fold dilution of stock systems before evlution in humns. Recombinnts of wild-type strins with the cold- ws clculted to give totl of 165 5% tissue culture Lbortories, Rockville, Md.) ws dministered. This dpted A/Ann Arbor/6/6 strin developed by infective doses per vccine dose. One child in ech Mssb (1, 15) hve been shown to be ttenuted nd immunogenic in humns when tested control. Nsl wshes were obtined dily for virl group ws not vccinted nd served s plcebo in open popultions (1, 3, 13). Young seronegtive children re prime cndidtes for protection before vccintion nd continuing until dy 15 fter isoltions nd interferon determintion strting 3 dys from such vccines nd represent the most sensitive popultion for testing ttenution, nti- fterwrd. The blood smples were used for HAI n- vccine dministrtion. Blood smples were obtined 1 dy before vccintion nd 1, 8, 15, nd 3 to 3 dys genicity, nd genetic stbility of such live respirtory virl vccines (1, ). Virus isoltion. Dily nsl wshes collected in 1 tibody determintion nd lymphocyte studies. The cell-medited immune response of seronegtive children fter vccintion with live inen isoltion. Cultures were inoculted dily without ml of phosphte-buffered sline were used for influfluen virus is incompletely investigted. Vccinted infnts might be expected to differ from (RMK) nd continuous cnine kidney cell line freeing onto primry rhesus monkey kidney cells nturl infection in dults due to reltive immturity of the immune system, primry expo- (MDCK). Cultures were inoculted t two tempertures: 3C, permissive for growth of the cold-dpted virus, nd sure to influen virus, nd reltive ttenution 39C, temperture restrictive to growth of the vccine strin. Tubes were hemdsorbed 5 nd 1 of the vccine strin. dys fter inocultion with.1% guine pig erythrocytes. All positive tubes were pssged t 3C nd The present study exmines these vribles by describing the humorl nd cell-medited identified s influen A/USSR (HlNl) by HAI ssy immune responses of young, seronegtive children fter vccintion with cold-dpted retiserum. All originl isoltes contining influen virus (3) with n influen A/USSR/77-specific chicken ncombinnt of influen A/HK/13/77 (HlNl) were subsequently titered on MDCK cells to determine the titer of virus being shed. virus. Other procedures. All procedures for lymphocyte MATERIALS AND METHODS collection nd culture, preprtion of mitogens, mesurement of trnsformtion, nd interferon ssy re s Ptient popultion. Vccinees nd controls were children ged 13 to months selected for vccine described in the compnion pper. 86 Downloded from on July 6, 18 by guest

2 VOL. 7, 198 RESULTS Virus shedding. Eight of eleven vccinted children shed influen virus for n verge of 8. dys. Virus shedding begn on dy 1 or fter vccintion nd continued in some vccinees until dy 13 (Fig. 1). The mximl level of virl repliction occurred on dy 6 fter dministrtion of the virus, followed by stedy decrese in titer. In one tril two vccinees shed nturlly occurring A/Bril/78, beginning before vccintion in one child nd in contct on dy 8. All A/USSR/77 isolted retined their temperture sensitivity s shown by lck of growth t restrictive temperture (39C). Antibody response. At the beginning of the trils, both vccinees nd plcebos hd serum HAI ntibody titer of 1:8 or lower. This titer ws not chnged 8 dys fter vccine dministrtion (Fig. ). By weeks, HAI ntibody titer rose in eight virus shedders by men of 6.9-fold nd remined t this level when mesured 3 to 3 dys postvccintion. Four plcebo nd the three nonshedding vccinees remined seronegtive. Lymphocyte trnsformtion (LTF) response of vccinees to influen A/USSR virus. The trnsformtion responses of children's lymphocytes s response to in vitro stimultion by influen A/USSR-inctivted virus were mesured 1 dy before vccine dministrtion (dy -1) nd on dys 1, 8, 15, nd 3 to 3 IMMUNE RESPONSES TO INFLUENZA VACCINATION 863 therefter. For sttisticl nlysis, the three children who neither shed influen virus nor hd n HAI response fter vccintion were dded w I- co I- n or w w cr S. *- *m B yo=r 1 1 * -I 8 15 * *-L e 3-3 FIG.. Serum HAI ntibody response in children fter intrnsl dministrtion of influen A/HK/ 13/77 (HINI) cold-dpted virus. Horiontl brs indicte rithmetic mens. Symbols:, children shedding influen HiN1 virus;, children not shedding influen HINM virus nd plcebos. A i Downloded from Cl) -J I -J E 6 3 o o To on July 6, 18 by guest - w en f -J FIG. 1. InfluenHINI cold-dpted virussheddingfterintrnsl dministrtion of vccine. Horiontl brs indicte rithmetic mens.

3 86 LAZAR, OKABE, AND WRIGHT to the four plcebo controls, nd the LTF response of this pprently uninfected group ws men levels of interferon being between 15 nd fected children during the study period with compred with the eight children who shed influen A/USSR (infected group). No significnt of both groups ws resistnt t ph nd stble 5 U. The interferon produced by lymphocytes trnsformtion response ws noted on dys -1 fter 56C incubtion for 6 min nd showed or 1 (Fig. 3). On dy 8, consistent low, but species specificity by lck of inhibition of vesiculr stomtitis virus plque formtion on mouse significnt, response ws observed in children who shed influen virus when compred with cells. These tests indicte tht the interferon the noninfected group (P =.7, Student's t produced ws type I nd reflected in vitro lymphocyte response to n interferon inducer. test). This response ws decresed by dy 15 but ws still significnt (P =.1). The LTF response returned to prevccine levels when mes- mesurble interferon could be detected in nsl Interferon in nsl wshes. In vccinees no ured 3 to 3 dys postvccintion. The lck of wshes fter vccintion when dily smples ny chnge in the LTF response of the nonshedders supports their inclusion in the noninfected experienced nturl infection with A/Bril/78 were exmined. In contrst, two children tht group. (HiN1) during the trils hd pek interferon LTF response of vccinees to phytohemgglutinin. Lymphocyte cultures of vcci- virus shedding. titers of 83 nd 79 U t the time of mximum nted children were incubted with phytohemgglutinin fter virl dministrtion to deter- DISCUSSION InsultAd st~+1%.+t TVP1 I+QV-iQlMrA Ni r reponse w itereu Cold-dpted influen A/HK (H1Ni) is duri rg vccintion. No significnt chnges in the cliniclly ttenuted vccine tht is geneticlly LTF response of either infected or noninfected stble in spite of considerble repliction in the child.en tht were noted fter vccintion, suggesting upper respirtory trct. Eqully importnt to the vccine virus used in this study ws not evlute re the effects of live virl vccine on iunmunosuppressive s determined by this ssy. the host immune system. Interferon production by vccines' lym- In the current phon study the in vitro trnsformcytes. Culture medi of in vitro-stimulted tion tlonresons response ofstimulted y lymphocytes ws lymy.hocytes from vccinted children were used noted in vccinted children who shed influen for iinterferon determintion. Some children's ~virus nd exhibited n HAI response. This sug- showed consistently high level of gets tht the cell-medited immune system is lymiphocytes inteirferon production, wheres some showed low involved in the immune rection to primry levelis. No sttisticl differences in interferon i leve] Is wre fund etwen ifectd n uniifection with live ttenuted influen virus. : UI) 3 compred with nturlly infected children nd FIXG. 3. Lymphocyte trnsformtion response to in- dults with H1N1 virus. flueni A/USSR (HJNJ) virus in children fter in- Suppression of the immune system due to trncesl dministrtion of influen A/HK/13/77 virl infection is well-recognied phenomenon (HlA press INFECT. IMMUN. The erly detection of LTF fter vccintion is consistent with the observtions of Ruben et l. (), Dolin et l. (5), nd Chow et l. (), who found elevted blstogenic responses 1 to 1 dys fter vccintion with inctivted influen virus. Since blood smples in the present study could not be obtined dily, the point of mximl ;T; g trnsformtion response fter vccintion ws not fully defined. But clerly the blstogenic response occurred erlier thn serum ntibody rises were detected by HAI ssy. The return of the LTF to bse line within 3 dys fter vccintion nd the low LTF even t 1 to weeks 8 I suggest diminished lymphocyte response when 1) cold-dpted virus. Trnsformtion exwhich must be considered when evluting new 3ed s stimultion rtio derived by dividing the live virl vccines, (19, 3). The degree of im- metrt counts per minute incorported in the presence. ' ' of viirl ntigens by the men counts per minute munosuppression resultnt from influen virus incoirported in the bsence of virus. Dt re given infection is controversil. Some investigtors s m mens (brs) + stndrd error (brckets). Striped hve described reduced lymphocyte trnsforrepresent infected children (n = 8); open brs mtion during influen infection (6, 9, 1), brs reprosent uninfected children (n = 7). wheres others hve filed to demonstrte such Downloded from on July 6, 18 by guest

4 VOL. 7, 198 immunosuppression (11, 1). These conflicting observtions could be explined by vribles such s severity of infection (11) or differences in the immune response to different influen types. Reduced immune responsiveness hs not been demonstrted during vccintion with inctivted influen virus (5, 1). One live ttenuted vccine (A/H3N/Englnd/8/68), on the other hnd, hs been shown to be immunosuppressive (9). The lck of immunosuppression s mesured by phytohemgglutinin stimultion with the cold-dpted vccine is ressuring. Interferon production by stimulted lymphocytes during influen infection hs been investigted to only limited extent. In the present study, the interferon produced ws of the nonimmune type (type I) nd ws detected in vccinees nd nonvccinees s well. These results re consistent with nonspecific response of lymphocytes to n interferon inducer rther thn specific cell-medited immune response. However, since type I interferon hs been detected in ser nd nsophryngel wshings of ptients fter influen infection (7, 8, 16, 18), it my ply n importnt role in recovery from the disese. Vccine infection did not induce mesurble interferon in nsl wshes in the present study. The mcrophge-lymphocyte technique, useful for detection of immune interferon, requires two specimens of 15 nd 5 ml of blood () nd is, therefore, unsuitble for studies in young children. Live, ttenuted influen vccine induces lymphocyte stimultion tht is of shorter durtion thn tht seen with nturl infection perhps relted to more limited virus repliction. However, lymphocyte blstogenesis is detected s erly s 8 dys fter vccintion. The possible importnce of cell-medited immunity s the erliest documented event in host's response to influen virl infections requires further study. ACKNOWLEDGMENTS We thnk Mimi Kervin, Judi Thompson, nd Robert Bskette for technicl ssistnce nd to Judi Tinnin for secretril ssistnce. This work ws supported by Public Helth Service contrct NO1-A1-57 with the Development nd Applictions Brnch, Ntionl Institute for Allergy nd Infectious Diseses. LITERATURE CITED 1. Bere, A. S., H. F. Mssb, D. A. J. Tyrrell, A. N. Slepushkin, nd T. S. Hll A comprtive study of ttenuted influen viruses. Bull. W.H.O. : Chow, T. C., K. R. Beutner, nd P. L. Ogr Cellmedited immune responses to the hemgglutinin nd neurminidse ntigens of influen A virus fter immunition in humns. Infect. Immun. 5: Dvenport, F. M., A. V. Hennessey, H. F. Mssb, E. Minuse, L. C. Clrk, G. D. Abrms, nd J. R. IMMUNE RESPONSES TO INFLUENZA VACCINATION 865 Mitchell Pilot studies on recombinnt colddpted live type A nd B influen virus vccine. J. Infect. Dis. 136: Dvenport, F. AL, nd E. Minuse Influen viruses, p In E. H. Lennette nd N. J. Schmidt (ed.), Dignostic procedures for virl nd rickettsil diseses, 3rd ed. Americn Public Helth Assocition, Inc., New York. 5. Dolin, R., B. R. Murphy, nd E. A. Cpln Lymphocyte blstogenesis response to influen virus ntigen fter influen infection nd vccintion in humns. Infect. Immun. 19: Dolin, R., D. Richmn, B. R. Murphy, nd A. S. Fuci Cell-medited immune responses following induced influen in humns. J. Infect. Dis. 177: Hll, C. B., G. Dougls, R. L Simons, nd J. M. Geimn Interferon production in children with respirtory syncytil, influen nd prinfluen virus infections. J. Peditr. 93: Jo, R. L, E. F. Wheelock, nd G. G. Jckson Production of interferon in volunteers with Asin influen. J. Infect. Dis. 11: Knler, G. B., S. F. Luteri, C. F. Cusumno, J. D. Lee, nd R H. Gnguly Immunosuppression during influen virus infection. Infect. Immun. 1: Kuffmn, C. A., C. C. Linnemnn, Jr., G. M. Schiff, nd J. P. Phir Effect of virl nd bcteril pneumonis on cell-medited immunity in humns. Infect. Immun. 13: Kuffmn, C. A., C. C. Linnemnn, Jr., J. S. Tn, G. M. Schiff, nd J. P. Phir Cell-medited immunity in humns during virl infection: derml hypersensitivity nd in vitro lymphocyte prolifertion during mild virl respirtory infections. Infect. Immun. 1: Kim, H. W., J.. Arrobio, C. D. Brndt, P. Wright, D. Hodes, R. M. Chnock, nd R H. Prrott Sfety nd ntigenicity of temperture sensitive (ts) mutnt respirtory syncytil virus (RSV) in infnts nd children. Peditrics 5: Kitym, T., Y. Togo, R. B. Hornick, nd W. T. Friedwld Low-temperture-dpted influen A/AA6/6 virus vccine in mn. Infect. Immun. 7: Mssb, H. F Adpttion nd growth chrcteristics of influen virus t 5'C. Nture (London) 13: Mssb, H. F Biologic nd immunologic chrcteristics of cold-dpted influen virus. J. Immunol. 1: McIntosh, K Interferon in nsl secretions from infnts with virl respirtory trct infections. J. Peditr. 93: Merign, T. C A plque inhibition ssy for humn interferon employing humn neonte skin fibroblst monolyers nd bovine vesiculr stomtitis virus, p In B. R. Bloom nd P. R. Glde (ed.), In vitro methods in cell-medited immunity. Acdemic Press Inc., New York. 18. Murphy, B. R., S. Bron, E. G. Ghlhub, C. P. Uhlendorf, nd R. M. Chnock Temperture-sensitive mutnts of influen virus: IV, Induction of interferon in the nsophrynx by wild-type nd temperture-sensitive recombinnt virus. J. Infect. Dis. 18: Notkins, A. L., S. E. Mergenhgen, nd R. J. Howrd Effect of virus infections on the function of the immune system. Annu. Rev. Microbiol. : Rsmussen, L. E., G. W. Jordn, D. A. Stevens, nd T. C. Merign Lymphocyte interferon production nd trnsformtion fter herpes simplex infections in humns. J. Immunol. 11: Downloded from on July 6, 18 by guest

5 866 LAZAR, OKABE, AND WRIGHT 1. Reed, W. P., J. W. Olds, nd A. L. Kisch Decresed skin-test rectivity ssocited with influen. J. Infect. Dis. 15: Ruben, F. L, G. G. Jckson, nd S. P. Gotoff Humorl nd cellulr response in humns fter immunition with influen vccine. Infect. Immun. 7: INFECT. IMMUN. 3. Wheelock, E. F., nd S. T. Toy Prticiption of lymphocytes in virl infections. Adv. Immunol. 16: Wright, P. F., S. H. Sell, T. S. Shinoki, J. Thompson, nd D. T. Kron Sfety nd ntigenicity of influen A/Hong Kong/68-ts-1 E (H3N) vccine in young seronegtive children. J. Peditr. 87: Downloded from on July 6, 18 by guest