Comparison of Bovine Herpesvirus 1 Vaccines for Rapid Induction of Immunity*

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1 Comprison of Bovine Herpesvirus 1 Vccines for Rpid Induction of Immunity Jmes A. Roth, DVM, PhD,b Dvid P. Crter, DVM b Deprtment of Veterinry Microbiology nd Preventive Medicine College of Veterinry Medicine Iow Stte University Ames, IA b Veterinry Resources, Inc PO Box Min Street Ames, IA This study ws supported by Meril Limited, Iselin, NJ. ABSTRACT The purpose of the study reported here ws to compre four different vccine regimens for their efficcy in protecting clves from chllenge with bovine herpesvirus 1 (BHV1) t 5 nd 14 dys fter vccintion. Nine experimentl groups of five clves ech were used to compre four vccintion regimens nd group tht received no vccine. The four vccine regimens were: BHV1 intrmusculr (IM) vccine contining both modified live virus (MLV) nd killed virus (KV); n MLV intrnsl (IN) vccine; concurrent dministrtion of MLV IN vccine nd MLV IM vccine; nd concurrent dministrtion of MLV IN vccine nd KV vccine. All vccine regimens induced solid protection ginst BHV1 chllenge t 14 dys fter vccintion. All of the vccine regimens lso induced sttisticlly significnt (P <.05) protective immunity t 5 dys fter vccintion; however, there were significnt differences (P <.05) in the degree of protection. The best protection induced by 5 dys fter vccintion ws provided by the MLV + KV combintion vccine injected IM nd the MLV vccines given IN nd IM concurrently. The MLV + KV combintion vccine dministered IM gve significntly (P <.05) better protection by 5 dys fter vccintion thn the MLV vccine dministered IN. Administering KV BHV1 vccine IM t the sme time s the MLV BHV1 IN vccine significntly (P <.05) reduced the effectiveness of the IN vccine for inducing erly protective immunity. INTRODUCTION It is sometimes very importnt to induce rpid protection ginst infection with bovine herpesvirus 1 (BHV1) fter vccintion. This is especilly importnt when vccines re used t rrivl for feedlot cttle or when they re used erly in n outbrek of infection with BHV1 in herd. 1 Shortening the intervl from vccin 220

2 J. A. Roth nd D. P. Crter TABLE 1. Experimentl Design Number Vccintion Chllenge Group of Clves Vccine Route Dy Dy 1 5 A IM A IM B IN B IN B&C IN/IM B&C IN/IM B&D IN/IM B&D IN/IM NONE N/A N/A 14 IM = Intrmusculr IN = Intrnsl See Tble 2 for description of vccines. tion to induction of protective immunity cn significntly reduce the severity of disese in the herd under these circumstnces. There re mny types of vccines licensed by the United Sttes Deprtment of Agriculture becuse they hve been shown to be sfe nd effective for preventing BHV1induced disese fter chllenge. 2 Typiclly the chllenge is dministered 2 to 5 weeks fter one or two doses of vccine is dministered. 3 8 Intrnslly dministered modified live virus (MLV) vccines hve been shown to induce mesurble protection ginst experimentl chllenge by 3 dys fter dministrtion 9 or ginst contct exposure by 48 hours fter vccintion. 10 In contrst, others 11 hve reported the filure of intrnslly dministered MLV vccines to protect ginst chllenge with BHV1 t 3 dys fter vccintion. Intrmusculrly dministered MLV vccines hve been shown to provide mesurble protection ginst contct exposure to BHV1 by 2 dys fter vccintion. 12 These studies focused on demonstrting significnt protection of vccinted nimls compred with nonvccinted nimls. The study reported here focused on demonstrting significntly improved protection when vrious vccintion regimens were compred. Veterinrins hve recommended vrious vccintion protocols in n ttempt to induce stronger immunity to BHV1 more rpidly. c These protocols hve included using vrious combintions of MLV vccines given intrmusculrly (IM) nd/or intrnslly (IN) with or without the concurrent use of killed virus (KV) vccines. The purpose of the study reported here ws to compre four vccintion regimens for their bility to induce protection from BHV1 chllenge t 5 nd 14 dys fter vccintion. MATERIALS AND METHODS Experimentl Design Fortyfive clves (250 to 300 kg) seronegtive for ntibodies to BHV1 were individully identified using er tttoos nd tgs nd rndomly ssigned to nine groups of five clves ech. Tble 1 shows the groups, vccines dministered, route of dministrtion, dy of vccintion, nd dy of chllenge. A descrip c Personl communiction with Dr. Roth t veterinry meetings. 221

3 TABLE 2. Vccines Used Vccine Description 2 A Modifiedlive nd killed forms of BHV1 nd modified live BVDV, PI3, nd killed BRSV. B Modifiedlive temperturespecific BHV1 nd PI3. C Chemicllyltered modified live BHV1 nd PI3 viruses, modified live BRSV, nd killed cytopthic nd noncytopthic BVDV. D Killed BHV1, BVDV I & II, BRSV, nd PI3. IBR PLUS TM 4wy; Meril Limited, Iselin, NJ. TSV2 ; Pfizer Animl Helth, Exton, PA. Cttlemster 4 ; Pfizer Animl Helth, Exton, PA. Virshield 5; Grnd Lbortories, Lrchwood, IA. tion of the vccines is found in Tble 2. The clves were treted with vitmin A nd D, vitmin Eselenium, vitmin B complex, nd ivermectin upon rrivl. Decoquinte (Deccox, Alphrm, Fort Lee, NJ) ws plced in the feed for the durtion of the study. The nimls were rested nd cclimted to their environment for 2 weeks before experimentl procedures were strted. From dy 14 to dy 0 of the study ll clves were housed together. From dy 0 to dy 9 the clves in the nine tretment groups were housed in four seprte pens s follows: pen 1 (group 1); pen 2 (group 9); pen 3 (groups 3, 5, nd 7); pen 4 (groups 2, 4, 6, nd 8). From dys 9 to 24 the clves in the nine tretment groups were housed in four seprte pens s follows: pen 1 (groups 1 nd 2); pen 2 (group 9); pen 3 (groups 3, 5, nd 7); pen 4 (groups 4, 6, nd 8). All clves were chllenged intrnslly with the United Sttes Deprtment of Agriculture (USDA) recommended chllenge dose of BHV1 ( TCID 50 of the Cooper s strin of BHV1; lot number 88 9) obtined from the USDA, Animl nd Plnt Helth Inspection Service (APHIS), Ntionl Veterinry Services Lbortories (Ames, IA). Clinicl Observtions Clinicl signs evluted included dyspne, coughing, nsl dischrge, nd pthy. Ech clinicl sign ws evluted independently (on dys 14 through 24) by two individuls who were unwre of the vccines ech niml received nd who grded ech clinicl sign using the following scle: 0 (norml); 1 (slight); 2 (mild); 3 (moderte); or 4 (severe). The two scores for ech clinicl sign were verged for ech dy. A composite clinicl score for ech niml for ech dy ws clculted by dding the score for ech of the clinicl signs nd the rectl temperture. BHV1 Titrtion in Nsl Secretions Nsl secretions were collected by suction from ech ventrl nsl metus through modified pipette into trpping tube. 13 The nsl secretions were stored frozen t 20 C until tested s previously described. 14 BHV1 in nsl secretions ws quntified by thwing, sonicting, nd centrifuging (15,600 grvity for 3 minutes) the nsl secretions nd inoculting two secondry bovine turbinte cell monolyers with 0.2 ml of tenfold seril dilutions of the superntnt fluid. After 1 hour dsorption, the cells were overlid with MEM (GIBCO Lbortories, Githersburg, MD) contining 7.0 g grnulted gr (DIFCO Lbortories, Detroit, MI), 30 ml hetincti 222

4 J. A. Roth nd D. P. Crter vted fetl bovine serum, 100 mg gentmicin (Schering Corp, Kenilworth, NJ), nd 50,000 units of mycosttin (E.R. Squibb nd Sons, Inc, Princeton, NJ) per liter nd incubted t 37 C in 5% CO 2 tmosphere for 5 dys. The gr ws removed, the cell lyer ws stined with crystl violet, nd the virl plques were enumerted. Sttisticl Anlysis Clinicl signs of disese were most prominent on dys 3 through 6 fter chllenge with BHV1 in the nonvccinted group. Therefore, to determine if there were significnt differences between tretment groups, the composite clinicl scores for dys 3 through 6 were compred using n nlysis of vrince procedure with dy fter infection s blocking fctor using SAS softwre (SAS, Cry, NC). Lest significnt difference t P <.05 ws then used to determine sttisticlly significnt differences between groups. RESULTS The results for body temperture nd composite clinicl score for the groups vccinted 14 dys before chllenge re found in Figures 1A nd 1B. Results for the groups vccinted 5 dys before chllenge re found in Figures 2A nd 2B. The composite clinicl score for dys 3 through 6 provides the best summry of overll level of protection by the vccines nd is therefore presented grphiclly in Figures 3A nd 3B. All four vccine combintions used provided solid protection from BHV1 chllenge when dministered 14 dys before chllenge. There ws no significnt difference between them (P >.05). All four vccines used provided significnt (P <.05) protection compred with the nonvccinted group when given 5 dys before chllenge; however, some provided better protection thn others. Vccine A provided significntly (P <.05) better protection thn vccine B or vccines B plus D when given 5 dys before chllenge. Vccine B nd vccines B plus C provided significntly (P <.05) better protection thn vccines B plus D. Vccine A nd vccines B plus C provided essentilly equivlent protection t 5 dys nd 14 dys fter vccintion (Figures 3A nd 3B). Vccine B nd vccines B plus D provided significntly (P <.05) better protection t 14 dys fter vccintion thn t 5 dys fter vccintion. The results of BHV1 isoltion from nsl secretions re found in Figures 1C nd 2C. The nonvccinted clves (group 9) hd detectble virus in the nsl secretions from dys 2 through 10 fter chllenge with pek shedding on dy 4. All of the vccinted groups of clves hd less virl shedding thn the nonvccinted group (group 9). All of the vccines, when dministered 14 dys before chllenge, nerly eliminted virus shedding in the nsl secretions. The vccines were somewht less effective t preventing virl shedding when given 5 dys before chllenge, especilly vccines B plus D. DISCUSSION All of the vccintion regimens used induced solid protective immunity t 14 dys fter vccintion with no significnt difference between them. All of the vccintion regimens used lso induced sttisticlly significnt protective immunity t 5 dys fter vccintion; however, there were significnt differences in the degree of protection fforded by the vrious vccintion protocols. The best protection induced by 5 dys fter vccintion ws provided by the MLV + KV combintion vccine (vccine A) injected IM nd the MLV vccines given IN + IM concurrently (vccines B + D). The MLV + KV combintion vccine (vccine A) dministered IM gve significntly better protection thn the MLV vccine dministered IN (vc 223

5 Degrees Fhrenheit Figure 1A Body Tempertures = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Score Composite Clinicl Score = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Figure 1B Log 10 TCID BHV1 Isoltion = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Figure 1C Figure 1. Men rectl temperture (1A), composite clinicl score (1B), nd isoltion of BHV1 in nsl secretions (1C), from clves chllenged 14 dys fter vccintion. 224

6 J. A. Roth nd D. P. Crter Degrees Fhrenheit Figure 2A Body Tempertures = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Score Composite Clinicl Score = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Figure 2B Log 10 TCID BHV1 Isoltion = Group 1 = Group 3 = Group 5 = Group 7 = Group 9 Figure 2C Figure 2. Men rectl temperture (2A), composite clinicl score (2B), nd isoltion of BHV1 in nsl secretions (2C), from clves chllenged 5 dys fter vccintion. 225

7 Men Composite Clinicl Scores for Dys 3 Through 6 After Chllenge 110 c 108 Score ,b Tretment Group Figure 3A Men Composite Clinicl Scores for Dys 3 Through 6 After Chllenge 110 c Score b,b d Tretment Group Figure 3B Figure 3. Men (+ SEM) composite clinicl scores for dys 3 through 6 fter chllenge for groups chllenged t 14 dys fter vccintion (3A) or 5 dys fter vccintion (3B). Mens with the sme letter re not significntly different ( P <.05). cine B). Administering KV BHV1 vccine (vccine D) IM t the sme time s the MLV BHV1 vccine (vccine B) IN reduced the effectiveness of the IN vccine for inducing erly protection (Figures 3A nd 3B). Erly onset of protection fter vccintion ginst BHV1 cn be cused by combintion of induction of cytokines (including interferon), induction of systemic or intrnsl ntibody, ctivtion of T lymphocyte subsets, ctivtion of nturl killer lymphocytes, nd perhps other mechnisms. The mechnisms responsible for the more rpid induction of immunity with the vrious vccintion regimens used is unknown. The best protection t 5 dys fter vccintion ws fforded by the two vccines tht included MLV vccine dministered IM. One of the two vccines lso included killed BHV1 in the vccine formultion IM; the other vccintion regimen included concurrent dministrtion of MLV IN vccine. Becuse n MLV vccine dministered IM by itself ws not test 226

8 J. A. Roth nd D. P. Crter ed in this experiment, the uthors cnnot be certin tht the other two components contributed to more rpid onset of protective immunity. It is possible tht n MLV IM vccine dministered by itself would provide equivlent protection t 5 dys fter vccintion. It is interesting to note tht one of the vccines tht induced the best protection t 5 dys fter vccintion contined both MLV nd KV BHV1 dministered IM, wheres KV BHV1 vccine significntly (P <.05) reduced the efficcy of concurrently dministered MLV IN vccine. This difference could be becuse of the difference in route of dministrtion of the MLV vccines (IM versus IN) or to differences in djuvnts or other properties of the different KV vccines used. The clves used in this experiment were seronegtive for BHV1 t the time of vccintion. Therefore, these results re vlid for clves tht do not hve mternl ntibody ginst BHV1. Clves with mternl ntibody cn respond differently to the vccines; however, clves with mternl ntibody lredy hve some protection ginst BHV1 1 nd do not require rpid onset of ctive immunity in order to be protected during the first few dys fter vccintion. CONCLUSION All vccine regimens in this study induced solid protection ginst BHV1 chllenge t 14 dys fter vccintion. All of the vccine regimens lso induced sttisticlly significnt (P <.05) protective immunity t 5 dys fter vccintion; however, there were significnt differences (P <.05) in the degree of protection. The best protection induced by 5 dys fter vccintion ws provided by the MLV + KV combintion vccine injected IM nd the MLV vccines given IN nd IM concurrently. The MLV + KV combintion vccine dministered IM gve significntly (P <.05) better protection by 5 dys fter vccintion thn the MLV vccine dministered IN. Administering KV BHV1 vccine IM t the sme time s the MLV BHV1 IN vccine significntly (P <.05) reduced the effectiveness of the IN vccine for inducing erly protective immunity. REFERENCES 1. Hjerpe CA: Bovine vccines nd herd vccintion progrms, in Smith B, ed. Lrge Animl Internl Medicine. St. Louis, MO, CV Mosby Co, 1990, pp Compendium of Veterinry Products, ed. 5. Port Huron, MI, N Americn Compendiums, 1999, pp McKercher DG, Crenshw GL: Comprtive efficcy of intrnslly nd prenterlly dministered infectious bovine rhinotrcheitis vccines. JAVMA 159: , vndrunenlittelvndenhurk S, Prker MD, Mssie B, et l: Protection of cttle from BHV1 infection by immuniztion with recombinnt glycoprotein giv. Vccine 11(1):25 34, Gerber JD, Mrron AE, Kucer CJ: Locl nd systemic cellulr nd ntibody immune responses of cttle to infectious bovine rhinotrcheitis virus vccines dministered intrnslly or intrmusculrly. Am J Vet Res 39(5): , Strub OC, Mwhinney IC: Vccintion to protect clves ginst infectious bovine rhinotrcheitis. Vet Rec 122: , Mnn DD, Buening GM, Thorne JG: Efficcy of erosol, intrnsl nd intrmusculr vccintion ginst selected bovine virl diseses. Cornell Vet 73: , Kucer CJ, White RG, Beckenhuer WH: Evlution of the sfety nd efficcy of n intrnsl vccine contining temperturesensitive strin of infectious bovine rhinotrcheitis virus. Am J Vet Res 39(4): , Todd JD, Volenec FJ, Pton IM: Intrnsl vccintion ginst infectious bovine rhinotrcheitis: Studies on erly onset of protection nd use of the vccine in pregnnt cows. JAVMA 159(11): , Kucer CJ, Beckenhuer WH: Time required to stimulte protection with intrnsl dministrtion of temperturesensitive, infectious bovine rhinotrcheitis virus vccine. Vet Med/Smll Anim Clin Jnury 1978: Svn M, Angulo AB, Derbyshire JB: Interferon, ntibody responses nd protection induced by n in 227

9 trnsl infectious bovine rhinotrcheitis vccine. Cn Vet J 20: , Sutton M: Rpid onset of immunity in cttle fter intrmusculr injection of modifiedlive virus. Vet Med/Smll Anim Clin September 1980: Mrshll RG, Frnk GH: Clinicl nd immunologic responses of clves with colostrlly cquired mternl ntibody ginst prinfluenz3 virus to homologous virl infection. Am J Vet Res 36: , Frnk GH, Mrshll RG, Smith PC: Clinicl nd immunologic responses of cttle to infectious bovine rhinotrcheitis virus fter infection by virl erosol or intrmusculr inocultion. Am J Vet Res 38: ,

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