Murine Cytomegalovirus 10366 Murine cytomegalovirus (MCMV) belongs to the genus cytomegalovirus (CMV) of the β-Herpesvirinae subfamily of the Herpesviridae family. CMVs are marked by strict species specificity, tropism for hematopoietic tissue and secretory glands, and a slow replication cycle. After the resolution of acute infection, CMVs establish persistent life-long infections characterized by alternate stages of virus productivity and latency. Although producing up to 200 potentially antigenic proteins during its sequential immediate early (IE), early (E) and late (L) phases of gene expression, transmission to a new host is achieved in the face of repeatedly primed antiviral immune responses. The routes used by MCMV to enter the host have not yet been clearly determined. Infectious virus can easily be detected in saliva of chronically infected mice, and productive infection is confined to glandular epithelial cells of salivary glands for a prolonged period of time, even in a fully immunocompetent host. Therefore, it is likely that the saliva is the major source of virus for horizontal spread, and the major route of entry for MCMV is the epithelium of the gastrointestinal and the upper respiratory tract. Per oral infection of newborn mice with MCMV leads to virus spread similar to that following intraperitoneal virus administration, which confirms that MCMV can enter through the epithelium of the gastrointestinal and/or respiratory tract. In addition, sexual transmission of MCMV and viral entry through the epithelium of the genitourinary tract could be an important means of its horizontal spread. After entry into the host, the virus spreads haematologically to various organs and infects many different cell types, including epithelial and endothelial cells, myocytes, brown fat adipocytes, fibrocytes, macrophages and bone marrow (BM) stromal cells (Krmpotic et al., 2003). Baboon Papio cynocephalus 9556 Bank vole Clethrionomys glareolus 447135 Bear Ursus americanus 9643 Birds Passeroidea 175121 Brown Trout Salmo trutta 8032 Buffalo Bison bison 9901 Carnivores Vulpes 9625 Cat Felis catus 9685 Catfishes Siluriformes 7995 Cattle Bos taurus 9913 Chicken Gallus gallus 9031 Chimpanzee Pan troglodytes 9598 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer Cervus elaphus 9860 Deer mouse Peromyscus maniculatus 10042 Dog Canis familiaris 9615 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Fish Hyperotreti 117565 Gerbil Gerbillina 10045 Goat Capra hircus 9925 Gray wolf Canis lupus 9612 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Macaque Macaca fascicularis 9541 Mongolian Gerbil Meriones unguiculatus 10047 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 None None Parrot Psittacidae 9224 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rainbow trout Oncorhynchus mykiss 8022 Rat Rattus 10114 Raven Corvus corax 56781 sei whale Balaenoptera borealis 9768 Sheep Ovis aries 9940 Squirrel Spermophilus richardsonii 37591 Tree shrew Tupaiidae 9393 Trouts, salmons & chars Salmoninae 504568 Turkey Meleagris gallopavo 9103 Vole Microtus ochrogaster 79684 Water buffalo Bubalus bubalis 391902 Murine cytomegalovirus DNA vaccine pcDNA-89 encoding pp89 VO_0004323 DNA vaccine Research pcDNAI/Amp [Ref2158:González et al., 1996] Intradermal injection (i.d.) Intradermal injection (i.d.) DNA vaccine construction Vector pcDNAI/Amp expressed the murine cytomegalovirus (MCMV) immediate-early gene 1 (IE1) encodes an 89-kDa phosphoprotein (pp89) (González et al., 1996). VO_0000286 In four separate trials, an average of 63% of the mice immunized with pcDNA-89 survived after lethal challenge, compared with 18% of the mice immunized with pcDNA (control) (González et al., 1996). Murine cytomegalovirus DNA vaccine pgM-pgN VO_0004526 DNA vaccine Research pcDNA3.1 [Ref2642:Wang et al., 2013] Intramuscular injection (i.m.) Intramuscular injection (i.m.) gM and gN from murine cytomegalovirus strain Smith (Wang et al., 2013) DNA vaccine construction DNA vaccine construction VO_0000286 Co-immunization with pgM and pgN three times at a high dose was able to provide mice with full protection against a lethal SG-MCMV challenge with a 100% survival rate. The protection obtained with the gM-gN co-immunization was far greater than each antigen alone (Wang et al., 2013). Murine Cytomegalovirus Inactivated Vaccine FI-MCMV - Aluminum Adjuvant Inactivated or "killed" vaccine Research Intraperitoneal injection (i.p.) Adjuvanted inactivated murine cytomegalovirus (MCMV) vaccine with aluminum induces potent and long-term protective immunity against a lethal challenge with virulent MCMV (Wang et al., 2014) Aluminum (Wang et al., 2014) Inactivated vaccine was prepared with tissue culture-derived extracellular MCMV. Briefly, 3T3 cells were infected by MCMV and incubated at 37°C for 4–5 days. When the monolayer exhibited 100% cytopathic effect, the medium from the infected cells was harvested and the cell debris removed by centrifugation at 6,500 × g at 4°C for 20 min. The total volume of crude virus stock was quantified and inactivated by mixing with 37% formalin at a 1/4000 ratio. (Wang et al., 2014) Intraperitoneal injection (i.p.) MCMV Smith strain (Wang et al., 2014) Antibody titers after the immunization boost were higher than those after priming. IgG antibody levels correlated positively with the vaccine dose. The antibody titer was greater and the elicited immune response stronger with a higher vaccine dose. When the vaccines were formulated with an adjuvant, the antibody levels increased significantly compared with adjuvant-free vaccines. No anti-MCMV IgG was detected in MF59, alum, and chitosan alone or in mock vaccine-treated mice, which suggested that neither the adjuvant-only treatment nor the Mock vaccine could induce specific anti-MCMV immune response. MF59-adjuvanted groups had the best antibody response. (Wang et al., 2014) Each experimental group had 10 mice. The inactivated vaccines, at doses of 4 μg, 1 μg, and 0.25 μg with or without an adjuvant (MF59, alum, or chitosan), were prepared for immunization by intraperitoneal (i.p.) injection. Immunizations were performed twice, 3 weeks apart. The volume of each dose was 200 μl. The adjuvant-only group of mice was administered with MF59, alum, or chitosan only and the control group of mice received PBS. In addition, one group of mice was immunized with formalin-treated mock virus preparation (Mock). To test whether the FI-MCMV vaccine could provide long-term protection, immunizations were performed with 4 μg FI-MCMV and adjuvants, three times by i.p. injection. (Wang et al., 2014) All mice in the control group (including mock vaccine) and adjuvant-alone treatment groups died within 8 days after a lethal challenge with SG-MCMV. Immunization with FI-MCMV vaccine (no adjuvant) at all three doses did not provide sufficient protection, but immunization with adjuvanted FI-MCMV significantly improved protection. At a low dose (0.25 μg), the survival rate in the FI-MCMV plus MF59 group reached 83.3%, while mice immunized with FI-MCMV plus alum or chitosan were not protected. At the 1 μg dose, all mice in the FI-MCMV plus MF59 group survived, while mice in the alum- or chitosan-adjuvanted groups were not completely protected. At the 4 μg dose, all three adjuvanted groups had full protection. These results indicated that immunizing twice with adjuvanted FI-MCMV vaccine could achieve excellent protection. (Wang et al., 2014) At 3 weeks after the second immunization, or 6 months after the third immunization, mice were challenged with a lethal dose (5 × LD50, 200 μl/mouse) of SG-MCMV by i.p. injection. (Wang et al., 2014) Murine Cytomegalovirus Inactivated Vaccine FI-MCMV - Chitosan adjuvant Inactivated or "killed" vaccine Research Intraperitoneal injection (i.p.) Adjuvanted inactivated murine cytomegalovirus (MCMV) vaccine with chitosan induces potent and long-term protective immunity against a lethal challenge with virulent MCMV. (Wang et al., 2014) Chitosan (Wang et al., 2014) Inactivated vaccine was prepared with tissue culture-derived extracellular MCMV. Briefly, 3T3 cells were infected by MCMV and incubated at 37°C for 4–5 days. When the monolayer exhibited 100% cytopathic effect, the medium from the infected cells was harvested and the cell debris removed by centrifugation at 6,500 × g at 4°C for 20 min. The total volume of crude virus stock was quantified and inactivated by mixing with 37% formalin at a 1/4000 ratio. (Wang et al., 2014) Intraperitoneal injection (i.p.) MCMV Smith strain (Wang et al., 2014) Antibody titers after the immunization boost were higher than those after priming. IgG antibody levels correlated positively with the vaccine dose. The antibody titer was greater and the elicited immune response stronger with a higher vaccine dose. When the vaccines were formulated with an adjuvant, the antibody levels increased significantly compared with adjuvant-free vaccines. No anti-MCMV IgG was detected in MF59, alum, and chitosan alone or in mock vaccine-treated mice, which suggested that neither the adjuvant-only treatment nor the Mock vaccine could induce specific anti-MCMV immune response. MF59-adjuvanted groups had the best antibody response. (Wang et al., 2014) Each experimental group had 10 mice. The inactivated vaccines, at doses of 4 μg, 1 μg, and 0.25 μg with or without an adjuvant (MF59, alum, or chitosan), were prepared for immunization by intraperitoneal (i.p.) injection. Immunizations were performed twice, 3 weeks apart. The volume of each dose was 200 μl. The adjuvant-only group of mice was administered with MF59, alum, or chitosan only and the control group of mice received PBS. In addition, one group of mice was immunized with formalin-treated mock virus preparation (Mock). To test whether the FI-MCMV vaccine could provide long-term protection, immunizations were performed with 4 μg FI-MCMV and adjuvants, three times by i.p. injection. (Wang et al., 2014) All mice in the control group (including mock vaccine) and adjuvant-alone treatment groups died within 8 days after a lethal challenge with SG-MCMV. Immunization with FI-MCMV vaccine (no adjuvant) at all three doses did not provide sufficient protection, but immunization with adjuvanted FI-MCMV significantly improved protection. At a low dose (0.25 μg), the survival rate in the FI-MCMV plus MF59 group reached 83.3%, while mice immunized with FI-MCMV plus alum or chitosan were not protected. At the 1 μg dose, all mice in the FI-MCMV plus MF59 group survived, while mice in the alum- or chitosan-adjuvanted groups were not completely protected. At the 4 μg dose, all three adjuvanted groups had full protection. These results indicated that immunizing twice with adjuvanted FI-MCMV vaccine could achieve excellent protection. (Wang et al., 2014) At 3 weeks after the second immunization, or 6 months after the third immunization, mice were challenged with a lethal dose (5 × LD50, 200 μl/mouse) of SG-MCMV by i.p. injection. (Wang et al., 2014) Murine Cytomegalovirus Inactivated Vaccine FI-MCMV - MF59 Adjuvant Inactivated or "killed" vaccine Research Intraperitoneal injection (i.p.) Adjuvanted inactivated murine cytomegalovirus (MCMV) vaccine with MF59 induces potent and long-term protective immunity against a lethal challenge with virulent MCMV (Wang et al., 2014) MF59 (Wang et al., 2014) Inactivated vaccine was prepared with tissue culture-derived extracellular MCMV. Briefly, 3T3 cells were infected by MCMV and incubated at 37°C for 4–5 days. When the monolayer exhibited 100% cytopathic effect, the medium from the infected cells was harvested and the cell debris removed by centrifugation at 6,500 × g at 4°C for 20 min. The total volume of crude virus stock was quantified and inactivated by mixing with 37% formalin at a 1/4000 ratio. (Wang et al., 2014) Intraperitoneal injection (i.p.) MCMV Smith strain (Wang et al., 2014) Antibody titers after the immunization boost were higher than those after priming. IgG antibody levels correlated positively with the vaccine dose. The antibody titer was greater and the elicited immune response stronger with a higher vaccine dose. When the vaccines were formulated with an adjuvant, the antibody levels increased significantly compared with adjuvant-free vaccines. No anti-MCMV IgG was detected in MF59, alum, and chitosan alone or in mock vaccine-treated mice, which suggested that neither the adjuvant-only treatment nor the Mock vaccine could induce specific anti-MCMV immune response. MF59-adjuvanted groups had the best antibody response. (Wang et al., 2014) Each experimental group had 10 mice. The inactivated vaccines, at doses of 4 μg, 1 μg, and 0.25 μg with or without an adjuvant (MF59, alum, or chitosan), were prepared for immunization by intraperitoneal (i.p.) injection. Immunizations were performed twice, 3 weeks apart. The volume of each dose was 200 μl. The adjuvant-only group of mice was administered with MF59, alum, or chitosan only and the control group of mice received PBS. In addition, one group of mice was immunized with formalin-treated mock virus preparation (Mock). To test whether the FI-MCMV vaccine could provide long-term protection, immunizations were performed with 4 μg FI-MCMV and adjuvants, three times by i.p. injection. (Wang et al., 2014) All mice in the control group (including mock vaccine) and adjuvant-alone treatment groups died within 8 days after a lethal challenge with SG-MCMV. Immunization with FI-MCMV vaccine (no adjuvant) at all three doses did not provide sufficient protection, but immunization with adjuvanted FI-MCMV significantly improved protection. At a low dose (0.25 μg), the survival rate in the FI-MCMV plus MF59 group reached 83.3%, while mice immunized with FI-MCMV plus alum or chitosan were not protected. At the 1 μg dose, all mice in the FI-MCMV plus MF59 group survived, while mice in the alum- or chitosan-adjuvanted groups were not completely protected. At the 4 μg dose, all three adjuvanted groups had full protection. These results indicated that immunizing twice with adjuvanted FI-MCMV vaccine could achieve excellent protection. (Wang et al., 2014) At 3 weeks after the second immunization, or 6 months after the third immunization, mice were challenged with a lethal dose (5 × LD50, 200 μl/mouse) of SG-MCMV by i.p. injection. (Wang et al., 2014) gM Murid betaherpesvirus 1 3293854 61660210 MuHV1_gp093 GU305914 YP_214101 10366 144295 145410 - Glycoprotein M or integral membrane protein 8.98 38877.66 371 >NC_004065.1:144295-145410 Murid herpesvirus 1, complete genome CTCACTCTGCGTCGCTTCGCAGGGCGCGTATCTCTTCGTTGTCCAGGGCCAGGGCTCGATATCGCTTGTT GCGGCGCAGCAAGAACCTCACCGTACGGATGACCGAGAACGCCACGCAGAGCAGTAGCAGCACCGTGATG CCGATGTTGATGTCCCTCGCGTAGCTCGAGGCGTTCAGCGCCTCGTAGCGGATGATGGGGTACATGGCGC CGCAGACGCCCAGTATCGTCCCGATGTGGTAGCCGAACTGCACCTTCATGTAGCGCGACAGCACCGACTC GATGATCGAGAAGTACACGCACGCCACGATCGCGAAGGCGACCATGGCGCCGAACACGACGTGCGCCGTC TTCACGAAAAAGCTGTTGCCGAAGCCGAGCGCCAGCGACATGGCCAGCACCATGGTCGCGAAGCCCAAGA TCAGCTGGGTCAGGTTCACCACCGCCGTCCGATAGCGGATGGTCCCCTGCAGCTTCGGGTGGATCTTCGA CAGCGCGAAAGCGCTGCGCTCGCGCGACTCGTAGTGGGTCACGAGCGTCACCACGAAGGCCGTCAGGCAC ATAAAGTACATGCACTTGCTGAACGCGACCATGGACGGTAGGCGGAACGACAGGCTCAGCAGGAAGATCT GAAAGGTGTCCATCGTCAGCACGAAGGTGAAGCACGTCGCCGAGTCGCCCATGAAGTTGATGTCGCGGGT CGACTGGTTCACCTGCGTGCCGTGCTCGCTGCGGAAGTAGATCTGCACCCAGCACACGATGTAGTAGCTC AGCACGCAGAAGAAGATCAGCTCCGTGAAGATCACGTACACGATGAGCTGCACGGGGTCCAGGAACAGCT GCGGCGTCAGATGGTGGATCGCGTTGTACATCGTCAGGTTCATGTTGTCGAAGTCGATGATGCGCGGGTA ATAGCACGGAAAACCGATGCCGGGGAAGTGCGCCGCCACCGAGAACACCACGATGTTCACGAACGACAGC AGGCAGCACGCGATGGCCATCGTCCACGTCCGCAGGTTCATGCGGTCCACGTGGGAGAGCGTCATGACGC CCGCTTTCGCCATCGTGCAGGTGTCTCGTGTCCGCGGTCGACGGGGCGGGTCAAATAGTGAGAGCA >YP_214101.1 Glycoprotein M or integral membrane protein [Murid betaherpesvirus 1] MLSLFDPPRRPRTRDTCTMAKAGVMTLSHVDRMNLRTWTMAIACCLLSFVNIVVFSVAAHFPGIGFPCYY PRIIDFDNMNLTMYNAIHHLTPQLFLDPVQLIVYVIFTELIFFCVLSYYIVCWVQIYFRSEHGTQVNQST RDINFMGDSATCFTFVLTMDTFQIFLLSLSFRLPSMVAFSKCMYFMCLTAFVVTLVTHYESRERSAFALS KIHPKLQGTIRYRTAVVNLTQLILGFATMVLAMSLALGFGNSFFVKTAHVVFGAMVAFAIVACVYFSIIE SVLSRYMKVQFGYHIGTILGVCGAMYPIIRYEALNASSYARDINIGITVLLLLCVAFSVIRTVRFLLRRN KRYRALALDNEEIRALRSDAE Protective antigen gN Murine cytomegalovirus (strain Smith) 190358890 CDD:281543 10367 ? Envelope glycoprotein N 7.5 15087.85 202 Envelope glycoprotein N. /FTId=PRO_0000116221. >sp|Q69150.2|GN_MUHVS RecName: Full=Envelope glycoprotein N MACGKTESGDDSGRFGRTGAGMFGFIMPGFVGIFRLSFFLLLSFAMASGSSSPASVPVSVAASVPDTTVN KIVISDGSEAHNINEFYDVKCHSHFYGLSVSSFASIWMMVNAIVFICAFGVFMRHWCYKAFTSDTAKGY Protective antigen IE1 Murine cytomegalovirus 138479 10367 ? Immediate-early protein 1 4.46 65528.13 731 IE1; Immediate-early phosphoprotein PP89 >sp|P11210.1|VIE1_MUHVS RecName: Full=Immediate-early protein 1; Short=IE1; AltName: Full=Immediate-early phosphoprotein PP89 MEPAAPSCNMIMIADQASVNAHGRHLDENRVYPSDKVPAHVANKILESGTETVRCDLTLEDMLGDYEYDD PTEEEKILMDRIADHVGNDNSDMAIKHAAVRSVLLSCKIAHLMIKQNYQSAINSATNILCQLANDIFERI ERQRKMIYGCFRSEFDNVQLGRLMYDMYPHFMPTNLGPSEKRVWMSYVGEAIVAATNIDHALDERAAWAK TDCSLPGEFKPELCVLVGAIRRLHDPPCYTKPFLDAKSQLAVWQQMKAIESESVSTHVVVVEALKLRENL AKAVQETIAYERHQYHRVCQMMCNNMKDHLETTCMLARGRTLATLADLRSTRYNLALFLLSEMHIFDSFT MPRIRGAMKQARCMSYVERTISLAKFRELADRVHNRSAPSPQGVIEEQQQAGEEEQQQQQEIEYDPEMPP LEREEEQEDEQVEEEPPADEEEGGAVGGVTQEEPAGEATEEAEEDESQPGPSDNQVVPESSETPTPAEDE ETQSADEGESQELEGSQQLILSRPAAPLTDSETDSDSEDDDEVTRIPVGFSLMTSPVLQPTTRSATAAAS SGTAPRPALKRQYAMVHTRSKSSENQQQPKKKSKK Protective antigen González et al., 1996 journal González Armas JC, Morello CS, Cranmer LD, Spector DH 1996 70 11 7921-7928 Journal of virology Krmpotic et al., 2003 journal Krmpotic A, Bubic I, Polic B, Lucin P, Jonjic S 2003 5 13 1263-1277 Microbes and infection / Institut Pasteur Wang et al., 2013 journal Wang H, Yao Y, Huang C, Chen Q, Chen J, Chen Z 2013 Virologica Sinica Wang et al., 2015 journal Wang H, Huang C, Dong J, Yao Y, Xie Z, Liu X, Zhang W, Fang F, Chen Z 2015 10 3 e0119964 PloS one