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    Unit 2 Virology

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    Uday Pratap Singh

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    © All Rights Reserved
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    Poxviruses oar emical properties mse INTRODUCTION largest viruses that infect verte- ss ie large enough 10 be seen-under the qoiwe a the sie TAS ope This group contains several viruses gt rs aly Posirdae that infect humans, sad oe past on genetic, antigenic segs eri tas been classified into two Sales. Family Poxviridae sobfily Chordopowirinas: sea te prsass of vertebrates. They ae clas- “dint six genera or SUDEFOUPS: jus: Mammalian poxvituses that tend to |e generalised infection with rash—varila, vac- cénia, cowpox, monkeypox, rabbitpox, buffalopox, | ipox, mousepOx t aoe ‘Viruses of ungulates that may occasion- + “ay infect humans—orf (contagious pustular derma- fis), paraveccinia (milker’s nodes, bovine papular siomgits) : ‘pansies: Viruses of goats and sheep—sheep-pox, ‘goatpox, lumpy skin disease fs ipox virus: Viruses of leporids (rabbits, hares, | ~squiels) myxoma and fibromas | Abposvirus: Viruses of birds—fowlpox, turkeypox, Pigeonpox, canarypox Supowirus: Swinepox | Subfamily Entomopoxviri the poxviruses of insects which do not infect Unclassified Poxviruses that have not been offically ass ‘genus include the virus of molluscum co" tanapox and the yaba monkey tumour. Human infections: Poxvirus diseases are char acterised by skin lesions which may be localised or generalised. The most important of these was smallpox caused by the variola virus. Other poxviruses which can infect humans are vaccinia, cowpox, monkeyPor: tanapox, molluscum contagiosum, paravaccinia and onl Buffalopox and camelpox may occasionally infect humans, causing lesions resembling vaccination. jgned to any ntagiosum, VARIOLA AND VACCINIA ‘The variola virus is the causative agent of smallpox. For thousands of years, smallpox raged as a scourge of humans, eausing death and disfigurement. The global cradication of smallpox, achieved after 10 years of concerted campaigns under the auspices of the WHO, has been a most impressive medical achievement. Naturally occurring smallpox came to an end in 1977. On 8 May, 1980, the WHO formally announced the global eradication of smallpox. Variola vi virus causing classical smallpox was called variola major and that causing alastrim variola minor. Variola major and minor were antigenically identical but they differed in certain biological char- acteristics. ee stable variants as the disease produced by each always bred true; alastrim did not lead to smallpox and vice versa. Vaccinia and the small pox vaccine: The vaccinia virus was used as the smallpox vaccine. Jenner origi- nally used the cowpox virus for vaccination against smallpox, but during the several years in which the original vaccine virus was maintained by arm-to-arm passage in humans, it underwent some permanent © scanned with OKEN Scanner Par VIROLOGY — ieroscope. The variola ViTUs was Figg, nstrated! microscopically by I iee7, me vinipoie developed a staining technique for the anc! demonstrated the clementary from smallpox lesion, the light adily differentiated from ges 50 that could {coupe and smallpon viases, THe MC sean that itis aati Sa nature as such, thas boon studied itis safer to work with rus ing wel as. 2 i ( the physieal and chemical properties binant saseines. The vaccinia ial es sat 25,000 foreign base Poxvirusesarestable and if p sug «an accommodate about 25.000 Fie en ible for months At room temperature, jg? cent fo ine ing these coding cold or when freze dred, they survive for years, The Sen inserted eR vies, HIN, rabies, are susceptible to ultraviolet light and other ira ‘ iN i mportant products such tions, They are resistant to 50% glycerol and 1% Phengy sly inpeeinia virus fs not but are readily inactivated by formalin and oxi FORE nice due (0 ts disinfectants. The virion consists essentially of py Be we protein and lipid. Though enveloped the virus ism ee j variola viruses are so similar in jnactivated by ether. The virion contains a lily at of enzymes, The entire multiplication ofthe virus ake in a parti i (Paschen bodies) nm The their properties tat they sidered together. pein seen place in the eytoplasm of the infected cel, Morphology ce «sian i Bick shaped. In vertical section, it con- Antigenic structure Al poxviruses share a common nucleoprotein (Np) antigen, By immunodiffusion, some twenty differen, ther side of the nucleoid is a lens-shaped structure antigens have been identified, These include the Lg Can he lateral Body Fg. 50.1). The virion measures antigen (a complex of two antigens, the heat lbis about 500 x 200 x 100 nm and so can be seen under |. and the heat stable S antigens), agglutinogen, ang hemagglutinin, which is responsible for the agglut. nation of erythrocytes of those fowls, which are aso agglutinated non-specifically by tissue lipids, sists of 2 double-layered membrane which surrounds a biconcave “aucleoid’ containing the DNA core. On Cultivation and host range The variola and vaccinia viruses can be differentiated by their growth characteristics and host range, Chick embryo: Both viruses grow on the CAM of 11-13-day-old chick embryo, producing pocks in 48-72 hours. Variola pocks are small, shiny, whit, convex, non-necrotic, non-hemorthagic lesions. Vaccinia pocks are larger, irregular, flat, greyi necrotic lesions, some of which are hemorthagc (Fig. 50.2). The viruses may also be differentiated by their ‘ceiling temperatures’, the highest temperature above which pocks are not produced. The ceiling tem peratures are 41°C for vaccinia, 38°C for variola major and 37.5°C for variola minor. Tissue culture: Variola and vaccinia viruses can be grown in tissue cultures of monkey kidney, HeLa and : chick embryo cells. Cytopathic effects are produced by Fig. 50.1 The structure of vaccinia virus; top: surface vaccinia in 24~48 hours and more slowly by vatiols. structure; bottom: section view Eosinophilic inclusion bodies—Guarnieri bodies—a" Section of virus 4 © scanned with OKEN Scanner pots ‘Vaccinia virus a pocks on CAM vaccinia po* jot a ‘variola, produces plaques in a os Ot not ah ue cultures. i be i viru can infect a wide range 1 Tally, Monkeys, calves, sheep tags CPT fected by scarification, leading ois om bee variola virus produces similar 1 rks Scarification of rabbit cornea ely in monkey keratitis, and sections ofthe ‘ ical Guarnieri bodies. Intranasal and ne in the monkey produces atte attack of smallpox with generalised aft ei SMALLPOX dicated. The lst natural case of sero eal ‘was Saiban Bibi, a Bangladeshi ‘ wth smallpox on the Karimgan) railway sot io Mey, 1975. The last case of Merce, Somalia, in October ing generations are unlikely to witness se nore altogether. Arie account of smal hefore being presented ; Sealp nas an exclusively human infection, ‘eho rina reservoir, There were no carriers as ‘vis was eliminated completely from the patient fencer, The source of infection was a patient i te ely phase of the disease, though infectiv- is tended from the appearance of buccal mucosal bins (enanthems) to the disappearance of all the sn sin (xanthems). Infection usually ocourred céyincse contacts, Virus entered the body by inha- Aer nt muliplcaton inthe local lymphoid Ss, he vius reached the reticuloendothelil cells, ‘Hee further multiplication took place, leading to severe viremia with seeding of the muvor heralding: the clini W disease, ‘The incubation period was around 12 days, The single crop of centrifugal exanthems passed through macular, papular, vesicular and pustular stages, belote scabbing and healing by sear formation in 2-4 weeks. The exanthems varied in severity from the hemorthagic, fat, ordinary or modified form, in descending order, Smallpox could be diagnosed in the laboratory by detection of virus antigen or by isolation of the virus from the blood in the early phase (in severe cases only) or from the eruptive lesion (in all eases). On account of the distinctive morphology of the virion, rapid diagnosis was possible by electron microscopy, where the facility was available ‘Two factors which contributed to the success of the small pox eradication programme were the use of freeze dried vaccine (in place of the unreliable liquid vaccine used earlier) and the technique of vaccination by multiple puncture with the bifurcated needle, which ‘was simple, effective and economical. ‘Though natural smallpox ceased in 1977, a small outbreak of variola major occurred in August 1978, in Birmingham, following the accidental spread of the virus from the virus laboratory in the medical school. It ‘was promptly identified and controlled but the incident showed the hazard of keeping variola stocks in labo- ratories. Following a directive by the WHO, all such laboratory stocks of the virus have been destroyed. The last stocks of smallpox virus were held under high secu- rity in the Centers for Disease Control and Prevention, Atlanta, Georgia (USA) and the Centre for Research on Virology and Biotechnology, Koltsova (Russia). ‘They were to have been destroyed by June 30, 1999. As a measure of protection against the remote danger of smallpox re-emerging or posing a bioterrorism threat, large stocks of smallpox vaccine are maintained by the WHO for rapid deployment, if needed. OTHER POXVIRUS DISEASES With the elimination of smallpox, it has become impor- tant to identify and characterise other orthopoxviruses which can infect human beings and cause disease resembling smallpox (Table 50.1). Monkeypox: This virus was first isolated in 1958 from an outbreak of pox disease in a captive monkey colony in Copenhagen. Monkeys are only incidental, © scanned with OKEN Scanner uses a Table S0.1._ Comparison of properties of some ontop _— pie ‘ BEE ‘wile Mowheyra DE One Toe Wa unas monte CUE, age felis Isolates foo area uke Hemorrhage Smal Whig al pi Arctson cal small wite fe = Cesta termperatre : . 3 en ca (°C) gras : : Growthon cs sort rabbit kin : : : Trymicine hinase : : : ~ High Low s baby ae low High High & Ree ec - ‘ ; : Spectetor {variola : : 2 : a Character of Character of Characer of charcterof—? pee oe monkeypox vaccinia comp petern The first human case was rodents being the hosts. n cas man infection is com- reported in 1970 from Zaire. Hur ron in Central and West Africa, wit a fatality rate of 5-10 per cent. The outbreak in America occurred in 2005, in Wisconsin, USA, affecting 11 local persons ‘and many prairie dogs. The source of infection is said to be an imported African rodent, which had infected local human contacts and prairie dogs. “The cases clinically resembled smallpox. However, person-to-person transmission appears to be rare. Serological studies have shown evidence of widespread natural infection in monkeys in Africa. The virus can be distinguished from variola. Buffalopox was identified in cattle in India in 1934 and was considered as an outbreak of vaccinia. Epizootics had occurred in buffaloes and lesions had been observed on the hands of persons in contact with infected animals. Two decades after the eradic of smallpox and cessation of vaccination, buffalopox still occurs, proving it to be distinct from variola and vaccinia, Though it resembles them closely, itis pos sible to distinguish between them in the laboratory. ‘The smallpox vaccine does not seem to protect persons against occupational buffalopox. Cowpox and milker’s nodes: Both these infections are obtained from cows. Cowpox lesions are seen on the udder and teats of cows and may be transmitted to hhumans during milking. The lesions in humans usually appear on the hands or fingers and resemble primary vaccinia. The disease is associated with some fever and constitutional symptoms, The cowpox virus resembles variola and vaccinia antigenically but can be differen. ted by the hemorrhagic lesions it produces on CaN ‘and rabbit skin. Restriction endonuclease maps of vac. Ginia and cowpox genomes show distinct differences, ‘Cowpox infection has been observed only in Britain and Europe. There have been outbreaks of fatal cow. pox infection in wild animals kept in zoos, inclu Cheetahs and elephants. Natural infection has been observed in domestic cats. It has been suggested that the primary host of cowpox may not be cows but more likely wild rodents or cats. Milker’s node (paravaccinia) is a trivial occupa. tional disease that humans contract by milking infected cows. The lesions are small ulcerating nodules. The virus is unrelated to cowpox and does not grow in eggs. It can be grown in bovine kidney cultures. It resembles the orf virus morphologically. Orf (Contagious pustular dermatitis): Orf is « disease of sheep and goats transmitted to humans by contact. In humans, the disease occurs as a single papulovesicular lesion with a central ulcer, usually on the hand, forearm or face. The virus is unrelated tothe variola-vaccinia group and resembles the paravaccinia virus morphologically, ‘Tanapox: This virus was isolated from epidemics ofa febrile illness along the Tana river in Kenya in 1957-19. The patients had a single pock-like lesion on the upPet part of the body. The virus is antigenically unrelated © other poxviruses and does not grow in eggs. It canbe grown in human and monkey tissue cultures. Monke}s are the only animals susceptible. The virus is 10 4 © scanned with OKEN Scanner Poxviruses 465 active in Africa, particularly in Zaire. A similar virus tas been isolated from outbreaks of disease in primate colonies in America. Molluscum contagiosum: This disease, seen usually inchildren and young adults, is characterised by pink or aly white wart-like nodules on the skin. Sections of the lesions show large (20-30 um) eosinophilic hyaline inclusion bodies which displace the nuclei to the margin, These molluscum, bodies are composed of large num- bers of virus particles, embedded in a protein matrix. Humans are the only susceptible hosts. The virus cannot be grown in eggs, tissue cultures or animals. The incidence of molluscum contagiosum as a sexu- ally transmitted disease in young adults is increasing. Whenit occursin the genital area, it may become inflamed and ulcerated and may simulate HSV infections. Yabapox: This is a monkey tumour virus which is related to poxviruses, RECAP * The ponvirticae sen 1 © scanned with OKEN Scanner RUS (CHICKENPOX) athogenicity Laboratory diagnosis Prophylaxis and treatment HINGLES, ZONA) Pathogenicity 2boratory diagnosis CYTOMEGALOVIRUSES Clinical features Epidemiology Laboratory diagnosis Prevention and trestment EPSTEIN-BARR VIRUS Epidemiology Pathogenicity CTIOUS MONONUCLEOSIS (GLANDULAR FEVER) Laboratory diagnosis - HUMAN HERPES VIRUS TYPES 6, 7, 8 HERPESVIRUS SIMIAE: B VIRUS -ANTTRODUCTION ‘The herpesvirus family contains over a hundred, spe- cies of enveloped DNA viruses thin affect humans and animals. They are characterised’ by their ability to" establish latent infections, enabling the virus to persist in infected hosts and to undergo pe, odie renctivation. Morphology ‘The herpesvirus capsid is icosahedral, composed of capsomers, aigLencosing the core containing “nso doubles anded DNA genom The nicl ae Sid is surrounded by the lipid envelope derived from, the modified host cell nuclear membrane through which the naked virions bud during replication, carties surface spikes, about 8 nm long (Fig. 51,1), Between the envelope and the capsid is an amorphous structure called the tegument, containing several pro. teins, The enveloped virion measures about 200 nm and the naked virion about 100 nm in diameter. Replication . Herpesviruses replicate in the host cell nucleus. They form Cowdry type A infranuclear (Lipschuta) inclusion Codie Resistance ike other enveloped viruses, herpesviruses are sus: ceptible to fat solvent Icohol, ether, chloroform and bile salts, They are heat labile and must Be Stored Capsid DNA core Fig. 51.1 Herpes simplex virus © scanned with OKEN Scanner ee evr heron eens se 2 Rear pepe PS es Pe ae Perit eS hoor Varcea sorter vias Epstein-dar virus Gyromegsiovius| Human 8 celliymphotropie somanarpssis 76 Rivas sors 9007 ara erpsvius Pe 8 a meson eres nse rime 6, and 8 (Table 51}. foc aPet ly Herpesviridae is divided into three ee con begs phe nd gente I nay vais eae SE veal maganla te ae wey nfcion memory engin er yep etopathic d etn tna rowed fon cll for empl, J Tape roy aril ots virus Se whch repeats sony” (224 he aro hot rage, Bro BO 8 ‘lati a tendency 10 enue eltgonen sete cls (yomera) snd ten ion Sana ims and ober ongans, In ele, he cpm ect slow andthe vies renin call associated, for exemple, cytomegalovirus. 7 py eGonnahereninses, whch bave 2 nar Yost ge repeat nimple a are pct foetier Bor T hmphocyes and frequent couse Tact inn in phot sue, for example te Epler rr Teresi family as no commen group a ‘en and the different species do not show any Git anigeni crossreacion, except betwen herpes simplex types 1 and 2. classifi rama ost Bernese eraser nnnert vez Herpes simplex vine Woe t a sipha— Gyoite iewrons Kidneys, other it wee - po lee Clinical cate 1A 25 year-old woman presented wth emplaines of gertal blisters forthe previous seven days ond eve hendacheandvoritingf: the previous ‘a days On examination. she was found t have neck i Fei ond' mbar puncte was pertormed for CSF i tamination The CSF was clea with cell count of 100 EBlutnn?nphoeytes were predominant. CSF glucose thes normal end the proteins sight elevated. The CSE Was inoculate inte Vero cel nes nthe laboratory and the Pet test was postive for herpesvirus type 2 DNA The ptient was started on treatment with goncyelowi {ipportive estmantwas als provided and the patient {esponded. Se was advised to recount this history to her doctor at any future pregnancy. ‘The herpes simplex virus (HSV) occuts naturally only in humans, but i ean produce experimental infection in many laboratory animals, The virus is of two types._ HSV type 1 ¢human herpes virus type 1 or HV ‘ap. & ay led fo ein in and. sround| ihe mouth andi transmitted by direct contactor drop- let spread from cases or carries. HSV type 2 (HAV type 2) is responsible for the majority of genital herpes infections and is commonly transmitted venereal. Intracerebral inoculation in rabbits and mice leads to encephalitis, and corneal scarification produces kera- toconjunctvits in rabbit. “The virus grows in a variety of primary and con- tinuous cell cultures (monkey or rabbit kidney, human ‘amnion, HeLa Fig. 512s) producing cytopathic changes, well-defined foci with heaped up cells and © scanned with OKEN Scanner Pes VROOUY showinyt inle_antibordy. Primary jf, usally acquire ‘lion in ealyclildlood, betyeun 1 five yeas of ae Humans are the only agg ‘inthe soureéoF infection are, sala, skin og MS regpirator ions, Asympter ony the more important souree of infeetion, e ion wit + espe genital infection with type 2 strains, aly Route of infection: Transmiss Y wach and may be venereal in virus enters throu, i > : o yembranes aie! multiplies spread. The virus enters cutaneous “ 7 intra -axonally (o the ganglia wherein sates Centrifugal migration OTTRE IFAS can ee Hom the ganglia to the skin and mucosa TOeAN Vg VE neous and mucos e Wis To in the ganglia, particularly of the tigeming! yet D Sh ptype ‘al Sacral (HSV wpe 2) ners to be reat ) kf ated perodeal in someind duals causing recutten, ay sidney S26 ole fou xan genial sions. p ‘ 3) Vervet monkey kidney'eells¢,,,4,

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