禽脑脊髓炎病毒VP0基因的克隆与序列测定

禽脑脊髓炎（avian encephalomyelitis,AE）又名流行性震颤,是一种主要侵害1月龄内雏鸡的病毒性传染病。该病传染迅速,既可垂直传播又可水平传播,危害极大。世界许多国家和地区均有该病发生和流行的报道。我国学者张泽纪等于1980年首次发现广东有疑似此病,1982年李心平等通过病理学方法确认此病,随后在我国各地均发现该病的发生和流行,给养鸡业造成较大的损失。 　　禽脑脊髓炎病毒（AEV）是一种无囊膜的二十面体对称的病毒粒子,大小为26nm左右。Shafren和Tannock（1991）利用放射性碘标记电泳分析AEV的结构蛋白发现,它主要有3种结构蛋白即VP1、VP2和VP3。Marvil等（1999）测定了AEV疫苗株1?143株的全基因组序列,揭示出AEV结构蛋白有VP1、VP2、VP3和VP4,其中VP2和VP4又合称VP0。我们从河北地区某艾维因肉鸡群自然发病的肉雏鸡脑组织中分离鉴定出一株AE病毒-L2Z株。根据发表的AEV序列设计引物,利用反转录聚合酶链式反应（RT-PCR）技术,在体外从AEV分离株感染的SPF鸡胚病变组织中扩增出VP0基因,在该片段的5′和3′端均加上KpnI位点,成功地将VP0 cDNA插入到质粒pBssK的KpnI位点之间,获得阳性克隆,并对其进行测序。结果发现,该基因长726bp,编码242个氨基酸,与AEV 1?143株VP0基因的核苷酸与氨基酸之间的同源性分别为95.2%和97.5%,而与小RNA病毒科其它病毒属如人甲肝病毒HAS-15株相应基因核苷酸和氨基酸之间的同源性分别为50.1%和50.8%,与肠道病毒属的PV-1 Sabin株的同源性分别为20.5%和16.5%,与鼻病毒属的HRV-14株的同源性分别为25.5%和17.8%,与心病毒属的EMCV PV2株的同源性分别为25.9%和19.4%,与口蹄疫病毒属FMDV-C株的同源性分别为26.3%和18.6%,与人类肠道致细胞病变孤儿病毒ECHOV的同源性分别为24.9%和16.1%。在系统发育树上,L2Z株与AEV 1?143株之间的距离最近,其次与甲肝病毒之间距离较近,而与小RNA病毒科其它病毒属之间的距离较远。 　　Marvil等（1999）首先测定出AEV 1143株的全基因序列,发现与甲肝病毒HM-175株之间氨基酸同源性为39%,与该科病毒其它病毒属之间的氨基酸同源性为19%～21%,其中VP0基因与甲肝病毒HM-175株相应基因氨基酸之间同源性达到71.8%,认为AEV与甲肝病毒属之间关系密切。Todd等（1999）对AEV VR株的3′端869bp序列测定表明,它与甲肝病毒相应区域氨基酸之间的同源性为35.7%,均高于与该科病毒其它病毒属相应氨基酸之间的同源性,也认为AEV与甲肝病毒之间最为接近。本试验发现：AEV分离株L2Z株VP0基因与甲肝病毒相应氨基酸之间的同源性为50.8%,而与该科病毒的其它属病毒相应氨基酸之间同源性仅在16.1%～21.9%之间。从系统发育树上也进一步表明,AEV L2Z株与甲肝病毒之间同源性之间距离最短,亲缘关系最近,而与肠道病毒属之间亲缘关系较远。第6次国际病毒学分类报告中将AEV划为小RNA病毒科肠道病毒属,但从我们的实验结果与Marvil等（1999）和Todd等（1999）的报道,从基因水平上证明AEV与甲肝病毒属更接近,而与肠道病毒属亲缘关系较远,因此认为AEV是否归为小RNA病毒科肠道毒属有待进一步确定。 　　本研究成功地进行了AEV分离株VP0基因的克隆与序列测定,为下一步进行AEV基因结构研究打下了基础。     

基于宏病毒组学技术挖掘猪腹泻粪便的病毒群落组成

为系统鉴定现阶段上海地区仔猪腹泻样品中的病毒群落组成,本研究利用宏病毒组学技术对临床采集的猪腹泻粪便进行了宏病毒高通量测序。结果显示,测序获得了1,676,726个RNAcontig和95,111个DNAcontig,RNA病毒和DNA病毒序列分别占比38.58%和3.10%。其中星状病毒科、杯状病毒科和小RNA病毒科是病毒总群落中占比最高的前3个科。虽然DNA病毒在病毒总群落中的占比较低,但其单组分环状DNA (CRESS-DNA)病毒种类丰富多样,包含有18种CRESS-DNA病毒。经基因组序列深度拼接,共获得了46条病毒全基因组序列,包括2株猪星状病毒和3株类似星状病毒、2株札幌病毒、16株小双节RNA病毒和23株CRESS-DNA病毒。其中1株札幌病毒归属于GII/3亚型,与人札幌病毒亲缘性较近。16株小双节RNA病毒中有4株归属于GGI亚型,其余12株归属于GGII亚型。23株CRESS-DNA病毒中有1株归属于类双生病毒科、12株CRESS-DNA病毒归属于小环状DNA病毒科,另外10株CRESS-DNA病毒未分类。本研究揭示现阶段猪腹泻粪便中以RNA病毒为主导,且存在宿主...     

家蚕传染性软化病病毒（桐乡株）VP1基因片段的克隆及序列分析

以首株在中国分离到的家蚕传染性软化病病毒（Bombyx mori infectious flacherie virus,BmIFV）BmIFV-CHN001基因组为模板,扩增了编码主要结构蛋白的VP1基因。克隆测序后得到VP1基因片段906 bp。该序列与已发表的日本毒株相比,核苷酸序列的相似性为99.3%,编码氨基酸的相似性为100%,证明该毒株与家蚕传染性软化病病毒日本株的同源性较高。把BmIFV-CHN001的VP1序列与同属的另外6个昆虫小RNA病毒的结构蛋白进行序列比对,构建系统发育树,对其进化关系进行了初步分析,结果显示这7种病毒具有相近的亲缘关系,而BmIFV-CHN001与蜜蜂囊雏病毒的亲缘关系最近。     

云南蝙蝠轮状病毒的分离与鉴定

蝙蝠是携带人兽共患病毒最多的一种哺乳动物,调查蝙蝠携带病毒的病原生态学本底对防范蝙蝠病毒威胁人类健康具有重要意义。本研究采集云南省部分地区蝙蝠进行病毒宏基因组学分析,在一组食虫蝙蝠样品中发现了A群轮状病毒(Group A rotaviruses,RVA)序列,经过进一步RT-PCR筛查验证及病毒的分离鉴定,最终从勐远县的1只三叶蹄蝠中分离出一株轮状病毒。扩增该毒株的VP7与VP4基因进行分型与系统进化分析,结果表明其VP7基因为G3型,与来自阿根廷的1株马轮状病毒的同源性最高,为93%;其VP4基因为P[10]型,与来自泰国的1株人轮状病毒同源性最高,为94.8%。通过与本实验室之前分离的首株蝙蝠G3P[3]型轮状病毒MSLH14的比较,确定该毒株为一株新的蝙蝠轮状病毒,命名为RVA/Bat-tc/MYAS33/2013/G3P[10],简称MYAS33。本研究结果进一步证明了蝙蝠轮状病毒的多样性,突显了蝙蝠作为轮状病毒潜在宿主的重要性。     

桂越边境地区蝙蝠病毒组研究

桂越地区包括越南和我国广西,地理位置的战略意义突出,该地区蝙蝠资源丰富,且越南境内的蝙蝠携带多种人兽共患病毒.为了监测桂越边境地区蝙蝠病毒的跨境传播,防范蝙蝠病毒引发新发传染病,掌握该地区蝙蝠携带病毒的病原本底和重要病毒遗传进化特征具有重要意义.本研究在桂越边境采集蝙蝠样本并对其进行病毒宏基因组学分析,结果发现了49个科的病毒,包括脊椎动物病毒、植物病毒、昆虫病毒和噬菌体.根据病毒宏基因组学结果,对注释到的嗜肝DNA病毒、博卡细小病毒、A型轮状病毒、星状病毒、弹状病毒和冠状病毒进行检测和进化分析,结果发现星状病毒广泛分布,且具有丰富的遗传多样性;其他病毒则呈局部分布,其中嗜肝DNA病毒、冠状病毒、轮状病毒与各自参考毒株进化关系较近,为这些病毒的变异毒株;而博卡细小病毒和弹状病毒与已知病毒具有很明显的遗传差异性,可能为病毒新种;值得注意的是,发现的蝙蝠轮状病毒与广西当地人轮状病毒具有很高的同源性,提示该病毒的跨种传播和基因重排.本研究获得了桂越边境地区蝙蝠携带病毒的本底数据以及部分重要病毒在该地区蝙蝠中的流行情况和遗传进化特征,为监测蝙蝠病毒的跨境传播、防范新发病毒病提供了重要的基础数据.     

三株牛源亚洲1型口蹄疫病毒VP1基因的克隆与序列分析

以3株国内分离的亚洲1型口蹄疫病毒(分别命名为F1、F2、F3)为研究目标,根据GenBank中注册的FMDV VP1基因的序列设计1对引物,采用RT-PCR方法成功地扩增出含有VP1全基因的片段,并测定了3个毒株VP1基因的序列.结果表明,3株亚洲1型FMDV毒株VP1基因长度均为633 bp,编码211个氨基酸.3株毒株彼此之间的核苷酸序列同源性在82.8% ～99.1%之间,推导氨基酸序列同源性在89.1% ～99.1%之间.从系统发生树看,F1株与我国香港2005年牛毒株序列同源性99.5%,属同一遗传谱系,F2株、F3株与2005年引起河北省万全县、北京市延庆县、甘肃静宁县疫情的毒株分属同一个基因群.     

三株牛源亚洲1型口蹄疫病毒聊基因的克隆与序列分析

以3株国内分离的亚洲1型口蹄疫病毒（分别命名为F1、F2、F3）为研究目标,根据GenBank中注册的FMDV VP1基因的序列设计1对引物,采用RT-PCR方法成功地扩增出含有VP1全基因的片段,并测定了3个毒株VP1基因的序列。结果表明,3株亚洲1型FMDV毒株VP1基因长度均为633bp,编码211个氨基酸。3株毒株彼此之间的核苷酸序列同源性在82．8％～99．1％之间,雅导氨基酸序列同源性在89．1％～99．1％之间。从系统发生树看,F1株与我国香港2005年牛毒株序列同源性99．5％,属同一遗传谱系,F2株、F3株与2005年引起河北省万全县、北京市延庆县、甘肃静宁县疫情的毒株分属同一个基因群。     

鸭肝炎病毒基因组3′末端序列的克隆和分析

用3′RACE和RT-PCR扩增并克隆鸭肝炎病毒(Duck hepatitis virus,DHV)Ⅰ型毒株C80和Ⅰ型变异株E63的3′末端序列。分析结果显示,C80株和E63株基因组3′末端均包含1 359 nt的3D、终止密码子TGA、长314nt的3′UTR,而poly(A)尾分别含18个A和19个A。由2株DHV 3D核苷酸序列所推导的3D蛋白均含453个氨基酸,均包含KDELR、DxxxxD、GxxCSGxxxTxxxNS、YGDD和FLKR等小RNA病毒RNA聚合酶的特征基序,该结果进一步证实Ⅰ型DHV属于小RNA病毒科的成员。两株DHV与小RNA病毒科9个已知属之间3D蛋白的氨基酸序列同源性为16%~37%,介于属间3D蛋白的氨基酸序列同源性范围(18%~60%)之内;此外,Ⅰ型DHV的3′UTR在小RNA病毒科是最长的。用3D蛋白进行进化分析的结果表明,Ⅰ型DHV可能属于小RNA病毒科的一个独立的病毒属。     

鸭肝炎病毒基因组3'末端序列的克隆和分析

用3'RACE和RT-PCR扩增并克隆鸭肝炎病毒(Duck hepatitis virus,DHV)Ⅰ型毒株C80和Ⅰ型变异株E63的3'末端序列.分析结果显示,C80株和E63株基因组3'末端均包含1 359 nt的3D、终止密码子TGA、长314nt的3'UTR,而poly(A)尾分别含18个A和19个A.由2株DHV 3D核苷酸序列所推导的3D蛋白均含453个氨基酸,均包含KDELR、DxxxxD、GxxCSGxxxTxxxNS、YGDD和FLKR等小RNA病毒RNA聚合酶的特征基序,该结果进一步证实Ⅰ型DHV属于小RNA病毒科的成员.两株DHV与小RNA病毒科9个已知属之间3D蛋白的氨基酸序列同源性为16%～37%,介于属间3D蛋白的氨基酸序列同源性范围(18%～60%)之内;此外,Ⅰ型DHV的3'UTR在小RNA病毒科是最长的.用3D蛋白进行进化分析的结果表明,Ⅰ型DHV可能属于小RNA病毒科的一个独立的病毒属.     

鸡传染性法氏囊病病毒基因组B片段的克隆与序列分析

鸡传染性法氏囊病病毒(IBDV)属双链双节段RNA病毒科,禽双链RNA病毒属.IBDV基因组由A、B两个RNA片段组成.VP2是最主要的IBDV结构蛋白,由A片段编码,它的变异最有可能导致IBDV血清型变异.最近的研究表明:B片段对病毒的毒力也有一定的影响.而我国对B片段的研究还未见报道.为此,我们克隆了我国甘肃地区IBDV分离强毒株Ts毒株的B片段全序列,并与报道的序列进行了比较分析.     

Neutralizing monoclonal antibodies against three serotypes of porcine rotavirus.

Using three serotypes (four strains) of cultivable porcine rotavirus as immunizing antigens, 10 neutralizing monoclonal antibodies were characterized. One VP4-specific monoclonal antibody directed against porcine rotavirus BEN-144 (serotype G4) neutralized human rotavirus strain ST-3 in addition to the homologous porcine virus. All nine VP7-specific monoclonal antibodies were highly specific for viruses of the same serotype as the immunizing rotavirus strain. One exception was the VP7-specific monoclonal antibody C3/1, which neutralized both serotype G3 and G5 rotaviruses. However, this monoclonal antibody did not neutralize the porcine rotavirus AT/76, also of serotype G3, nor mutants of SA-11 virus (serotype G3) which were selected with monoclonal antibody A10/N3 and are known to have mutations affecting the C antigenic region.     

G5型人A组轮状病毒LL36755株的VP4、VP6和NSP4编码基因的分子特征

最近在亚洲首次发现并报道了感染人的G5型人A组轮状病毒LL36755株,为进一步探讨其进化来源,克隆了G5型人A组轮状病毒LL36755株的VP4、VP6、NSP4编码基因,并分析其基因序列的分子特征。结果发现卢龙株LL36755为罕见的G5P[6]型,其VP6的亚群为SGⅡ型,NSP4的基因型为B型。系统进化树分析表明,卢龙株LL36755的VP7、VP4编码基因与猪来源的毒株关系密切,而VP6、NSP4编码基因与人来源的毒株紧密相联系。可以推断新的人腹泻A组轮状病毒LL36755株是猪的VP7,VP4编码基因与人的VP6,NSP4编码基因的自然重组;而且该毒株不是G5的原型,很可能是人类轮状病毒与猪轮状病毒毒株的自然重组后逐步进化而来。     

Infection immunity of piglets to either VP3 or VP7 outer capsid protein confers resistance to challenge with a virulent rotavirus bearing the corresponding antigen.

A single-gene substitution reassortant 11-1 was generated from two porcine rotaviruses, OSU (serotype 5) and Gottfried (serotype 4). This reassortant derived 10 genes, including gene 4 encoding VP3, from the OSU strain and only gene 9, encoding a major neutralization glycoprotein (VP7), from the Gottfried strain and was thus designated VP3:5; VP7:4. Oral administration of this reassortant to colostrum-deprived gnotobiotic newborn pigs induced a high level of neutralizing antibodies not only to Gottfried VP7 but also to OSU VP3, thus demonstrating that VP3 is as potent an immunogen as VP7 in inducing neutralizing antibodies during experimental oral infection. Gnotobiotic piglets infected previously with the reassortant were completely resistant to oral challenge with the virulent Gottfried strain (VP3:4; VP7:4), as indicated by failure of symptoms to develop and lack of virus shedding. Similarly, prior infection with the reassortant induced almost complete protection against diarrhea and significant restriction of virus replication after oral challenge with the virulent OSU strain (VP3:5; VP7:5). Thus, it appears that (i) the immune system of the piglet responds equally well to two rotavirus outer capsid proteins, VP3 and VP7, during primary enteric rotavirus infection; (ii) antibody to VP3 and antibody to VP7 are each associated with resistance to diarrhea; and (iii) infection with a reassortant rotavirus bearing VP3 and VP7 neutralization antigens derived from two viruses of different serotype induces immunity to both parental viruses. The relevance of these findings to the development of effective reassortant rotavirus vaccines is discussed.     

Identification and Characterization of a Novel Recombinant Porcine Astrovirus from Pigs in Anhui,China

Porcine astroviruses (PAstVs) have wide distribution in swine herds worldwide. At present, five porcine astrovirus genotypes have been identified. In this study, using viral metagenomics, a novel PAstV strain (designated as Ahast) was identified in fecal samples from pigs in Anhui of China, and the complete genomic sequence of Ahast was obtained by assembling and PCR amplification. Genomic structural analysis indicated that Ahast had a typical ribosomal frameshifting signal, and some conserve amino acid motifs were also found in virally encoded proteins. Phylogenetic analysis and sequence comparison indicated that this virus belonged to porcine astrovirus genotype 4 (PAstV4), which formed a clade clustered with other PAstV4. Multiple recombinant events were confirmed by recombination analysis and indicated that Ahast was a potential recombinant. Epidemiological investigation indicated that PAstV4 has a 10.7% prevalence in this pig farm. The new recombinant identified in this study will be beneficial to comprehend the origin, genetic diversity, and evolution of porcine astroviruses in Anhui of China.Key words: porcine astroviruses, viral metagenomics, genome recombination     

Purification and characterization of adult diarrhea rotavirus: identification of viral structural proteins.

Adult diarrhea rotavirus (ADRV) is a newly identified strain of noncultivable human group B rotavirus that has been epidemic in the People's Republic of China since 1982. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western (immuno-) blot analysis to examine the viral proteins present in the outer and inner capsids of ADRV and compared these with the proteins of a group A rotavirus, SA11. EDTA treatment of double-shelled virions removed the outer capsid and resulted in the loss of three polypeptides of 64, 61, and 41, kilodaltons (kDa). Endo-beta-N-acetylglucosaminidase H digestion of double-shelled virions identified the 41-kDa polypeptide as a glycoprotein. CaCl2 treatment of single-shelled particles removed the inner capsid and resulted in the loss of one polypeptide with a molecular mass of 47 kDa. The remaining core particle had two major structural proteins of 136 and 113 kDa. All of the proteins visualized on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were antigenic by Western blot analysis when probed with convalescent-phase human and animal antisera. A 47-kDa polypeptide was most abundant and was strongly immunoreactive with human sera, animal sera raised against ADRV and against other group B animal rotaviruses (infectious diarrhea of infant rat virus, bovine and porcine group B rotavirus, and bovine enteric syncytial virus) and a monoclonal antibody prepared against infectious diarrhea of infant rat virus. This 47-kDa inner capsid polypeptide contains a common group B antigen and is similar to the VP6 of the group A rotaviruses. Human convalescent-phase sera also responded to a 41-kDa polypeptide of the outer capsid that seems similar to the VP7 of group A rotavirus. Other polypeptides have been given tentative designations on the basis of similarities to the control preparation of SA11, including a 136-kDa polypeptide designated VP1, a 113-kDa polypeptide designated VP2, 64- and 61-kDa polypeptides designated VP5 and VP5a, and several proteins in the 110- to 72-kDa range that may be VP3, VP4, or related proteins. The lack of cross-reactivity on Western blots between antisera to group A versus group B rotaviruses confirmed that these viruses are antigenically quite distinct.     

Similarity of the outer capsid protein VP4 of the Gottfried strain of porcine rotavirus to that of asymptomatic human rotavirus strains.

Genomic segment 4 of the porcine Gottfried strain (serotype 4) of porcine rotavirus, which encodes the outer capsid protein VP4, was sequences, and its deduced amino acid sequence was analyzed. Amino acid homology of the porcine rotavirus VP4 to the corresponding protein of asymptomatic or symptomatic human rotaviruses representing serotypes 1 to 4 ranged from 87.1 to 88.1% for asymptomatic strains and from 77.5 to 77.8% for symptomatic strains. Amino acid homology of the Gottfried strain to simian rhesus rotavirus, simian SA11 virus, bovine Nebraska calf diarrhea virus, and porcine OSU strains ranged from 71.5 to 74.3%. Antigenic similarities of VP4 epitopes between the Gottfried strain and human rotaviruses were detected by a plaque reduction neutralization test with hyperimmune antisera produced against the Gottfried strain or a Gottfried (10 genes) x human DS-1 rotavirus (VP7 gene) reassortant which exhibited serotype 2 neutralization specificity. In addition, a panel of six anti-VP4 monoclonal antibodies capable of neutralizing human rotaviruses belonging to serotype 1, 3, or 4 was able to neutralize the Gottfried strain. These observations suggest that the VP4 outer capsid protein of the Gottfried rotavirus is more closely related to human rotaviruses than to animal rotaviruses.     

VP7 mediates the interaction of rotaviruses with integrin alphavbeta3 through a novel integrin-binding site

Rotavirus entry is a complex multistep process that depends on the trypsin cleavage of the virus spike protein VP4 into polypeptides VP5 and VP8 and on the interaction of these polypeptides and of VP7, the second viral surface protein, with several cell surface molecules, including integrin alphavbeta3. We characterized the effect of the trypsin cleavage of VP4 on the binding to MA104 cells of the sialic acid-dependent virus strain RRV and its sialic acid-independent variant, nar3. We found that, although the trypsin treatment did not affect the attachment of these viruses to the cell surface, their binding was qualitatively different. In contrast to the trypsin-treated viruses, which initially bound to the cell surface through VP4, the non-trypsin-treated variant nar3 bound to the cell through VP7. Amino acid sequence comparison of the surface proteins of rotavirus and hantavirus, both of which interact with integrin alphavbeta3 in an RGD-independent manner, identified a region shared by rotavirus VP7 and hantavirus G1G2 protein in which six of nine amino acids are identical. This region, which is highly conserved among the VP7 proteins of different rotavirus strains, mediates the binding of rotaviruses to integrin alphavbeta3 and probably represents a novel binding motif for this integrin.     

在中国卢龙县发现G5型人A组轮状病毒

轮状病毒是引起我国儿童重症腹泻的主要病原。按照WHO轮状病毒监测方案,对2003年间河北省卢龙县开展了以医院和社区为基础的小于5岁儿童轮状病毒腹泻的监测,发现一株新型轮状病毒。该病毒用传统分型引物(G1、G2、G3、G4)扩增不出条带,对其VP7基因全序列测定和分析后确定该毒株为G5型。这是我们在亚洲首次发现的人类G5型轮状病毒。该毒株与猪的G5型C134毒株核苷酸和氨基酸序列同源性分别为88·6%和95·4%,与非洲发现的人的G5型毒株MRC3105核苷酸和氨基酸的同源性为89·9%和94%,与巴西发现的IAL-28毒株核苷酸和氨基酸的同源性为87·2%和93·3%。系统发生树分析表明:卢龙毒株LL36755与其他已经报告的两种猪和一种人类的G5型毒株可能具有相同的起源。这是人轮状病毒G5型首次在亚洲国家发现,而且该毒株可能是由人类轮状病毒与动物轮状病毒毒株自然重组产生。     

Expression of the OSU rotavirus outer capsid protein VP4 by an adenovirus recombinant.

Full-length cDNA of the VP4 gene of porcine rotavirus strain OSU was cloned into adenovirus type 5 (Ad5) downstream of the E3 promoter. The plaque-purified recombinant (Ad5-OSU VP4) expressed apparently authentic VP4 rotavirus outer capsid protein. The protein had the same molecular size (85 kDa) and electrophoretic mobility as did native OSU VP4 and was immunoprecipitated by a polyclonal antiserum raised to OSU VP4. Cotton rats that possessed prechallenge rotavirus antibodies that may have been acquired either passively or actively developed neutralizing antibodies against the OSU strain following intranasal administration of the live Ad5-OSU VP4 recombinant. The neutralizing activity was enhanced by a parenteral booster injection with baculovirus-expressed OSU VP4 antigen. In addition, a high titer of neutralizing antibodies was induced by parenteral administration of the latter antigen and subsequent intranasal administration of the Ad5-OSU VP4 recombinant. These observations indicate that the VP4 outer capsid protein of a rotavirus strain can be expressed by a recombinant adenovirus vector. This approach warrants further exploration for immunization against rotavirus disease.