Virology VN Virus: obligate intracellular parasite; depends on host cells for everything; uses host machinery/resources to make prots

Structure of Virion (virus particle) - Enveloped or Naked envelope surrounds nucleocapsid of some viruses; composed of viral specific prots & lipids; may have spikes that act as cell surface receptors (interact w/ host cells) i.e. hemagglutinin spikes on influenza virus (allows virus to attach to respiratory tract lining) naked more able to withstand harsh environment (less complex structure --> less susceptible to degradation) - protein capsid or repeating protein subunits surrounds nucleic acids = nucleocapsid - nucleic acid core (either ss/ds RNA/DNA) encodes structural proteins and enzymes --> replication & lifecycle Ex. Influenza Virus: envelope w/ spikes essential for survival - Hemagglutinin - spikes that help virus fuse w/ host cell membrane - Neuraminidase - enz that allows virus to be released from infected cell (and spread to other cells) - matrix protein makes viral capside surrounding RNA core Transmission: m/c aerosols (also contaminated food/water, fecal-oral route, on inanimate objects (fomites), sexual contact, contaminated blood --> organ transplant, insects, IV drug users) - depends on source of infxn (diff hosts), environment (can virus withstand temp, pH, etc?) - enveloped virus more fragile --> more complicated strxr susceptible to environmental stress - naked virus more stable and less complex; less susceptible to environmental stress - Horizontal: person-to-person mode of transmission - Vertical: mother to fetus (through placenta, birth canal, or in germ cell): herpes, HBV (hep B), VZV (varicella zoster) Viral Life-Cycle - enters host cell, uncoats --> releases nucleic acid core, viral prots synthesized, new viruses form - can cause host cell lysis; can cause neoplasm (insert their nucleic acids into host DNA --> mutation); can cause attack from T cells (viral Ags shed intracellularly are brought to surface, allowing recognition by immune system)

Viral Invasion: m/c is inhalation of infected respiratory droplets - entry via breaks in barriers (skin, mucous membranes, epithelia) Viral Incubation: some common virus incubation periods - Influenza: 1-2days, Hep B: 50-150days, Papilloma: 50-150days, HIV: 1-10years Viral Infection: amt of spread / length of infxn depend on virus - acquisition (prodrome) --> primary infxn replication --> spreading (local, neuronal, blood, lymph) --> secondary site replication --> target (specific matching w/ prots on viral surface = tropism --> drawn to compatible cells) Pathogenesis - How do viruses cause disease? - viruses collide and attach to host cells via cell surface receptors (GP spikes and viral surface prots) - virus endocytosed into host cell / fuse w/ plasma membrane (esp. enveloped viruses) - virus uncoats and starts replicating in human host cell viral genome replication and viral prot synthesis (transcription/translation) DNA-->mRNA-->ribosome-->prot - viruses use our enzs/resources to replicate and make new prots - viral Ags can be brought to host cell surface to flag immune system - after replication & synth of viral proteins, accumulation of these products causes host cell lysis --> releases virions to infect other cells (spread); enveloped viruses are released from infected cells by budding (can use part of host cell
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membrane in their envelope) Viral Nucleic Acids - DNA viruses (ss or ds) ssDNA: parvovirus, dsDNA: smallpox, herpes, papovavirus - RNA viruses (ss or ds) ssRNA: orthomyxovirus (influenza), rhinovirus, coronavirus retroviruses: ssRNA--reverse transcriptase --> dsDNA (ex. HIV) Pathobiology of viruses (mechs of cell/tissue injury) - Sgs & Sxs caused by virus --> cell death AND host immune response to virus - after virus enters, attaches, invades, and infects host cells, it may spread via lymph/blood (viremia) to other organs (secondary sites), CNS, lymph tissue - mobile virus ex: Herpes & Varicella zoster (travel along nerves) Stages of Viral Infection - entry --> initiation of infxn at primary site (skin, mucous membrane, resp tract, GI tract, etc) - incubation --> no symptoms, but virus actively replicating & may spread to secondary site (length varies w/ virus) - prodrome --> nonspecific flu-like Sxs that preced organ-specific Sxs - virus-specific disease signs appear shortly after prodrome; nature and severity depends on type of virus, which organ(s) are affected, and how rigorous the immune response is Potential Outcomes to Viral Infxn - acute infection: rapidly controlled by IS; self-limited (days-->weeks) ex. common cold, rotovirus, influenza - inapparent infection (covert): no resulting disease; ex. vaccination - persistent infection: IS and virus battle (ex. Hep B&C) latent infection (ex. Varicella zoster - chickenpox; dormant virus can return as shingles) chronic infection (ex. HepB - acquired, spreads to bld & liver) - IS can completely clear, or virus can remain for years and shed a low level slow infection (ex. mad cow disease) infxn starts very low grade, then develops into disease Immune Response to Viral Infxn - how virus gets in and activates IS: skin barrier / mucous membranes are transgressed, direct infxn of blood, organ transplant, inhaled into respiratory duct, materal-fetal transmission, fecal-oral ingestion, etc - innate response include interferons (1st active defense from macrophages/fibroblasts) which are produced by cells in response to viral infxn (w/in hours) and keeps viral load in check by limiting viral prot synth in other infected cells --> flu-like symptoms stimulate other lymphokines (TNF, etc), activate NK cells, stimulate macrophages - later viral Ag-specific acquired response kicks in (primary and secondary) cell-mediated immunity plays a central role in viral infxn; T-cells very important vaccination w/ virus or viral Ags provokes an Ab response specific for that virus --> affords memory cells for longlasting immune response Types of Infxn: - failed infxn (abortive infxn) - virus doesnt multiply and disappears - lytic infxn - cause cell death & tissue damage, then resolve; cause acute disease (virus can make prots that have a cytopathologic effect - CPE; characteristic to certain virus); viruses interrupt host DNA/prot prodxn - persistent infxn - active infxn; may not cause cell death but lives in cells and may be chronic, latent, recurrent, or transforming infxn chronic - productive infxn where virus is being produced slowly w/out killing host cell (ex. Hep B); can be contagious during this time latent - dormant, non-infectious, non-productive but some viral molecules are produced in host cells; virus may be reactivated under stress and other stimuli such as immunosuppression (ex. HSV - herpes simplex virus) - oncogenic viruses - persistent viral infxns that may transform infected host cells -> malignant cells (common in viruses that integrate their own genome into host DNA); ex. EBV (epstein barr virus - B cell lymphoma, activate oncogene) Clinical Virology - Systemic Viruses: cytomegalovirus, human immunodef. virus (HIV), epstein barr virus (infectious mononucleosis) - Oncogenic Viruses: human papilloma virus, epstein barr virus, hepatitis B and C Specific Viruses:
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HERPES TYPE VIRUSES Herpes Simplex - type of human herpes virus (includes: HSV I & II, Epstein Barr Virus (EBV), Varicella Zoster Virus (VZV), and Cytomegalovirus (CMV)) - all dsDNA viruses, large & enveloped - HSV I & II transmitted via direct contact w/ infected saliva, sexual, and lesion fluid; causes lytic infxn of fibroblasts, epithelial cells, and latent infxn of innervating neurons - HSV I usually ABOVE the waist; causes 1o oral gingivostomatitis, recurrent herpes labialis, pharyngitis, and keratitis (corneal infxn) - HSV II usually BELOW the waist; causes genital herpes but can cause oral herpes (15%) - Sg/Sx: primary infxn: contact, 5day incubation, prodrome; may be asymptomatic, but when symptomatic are worse than recurrent episodes; recurrences cause painful itching vesicles and ulcerations on penis/vagina/cervix recurrent infxns (from reactivation of latent herpes living inside the nerve root) are less severe than 1o infxn and involve lips and perioral regions - clusters of vesicles --> ulcers --> scab (painful) - lab Dx: used to do Tzanck Smear; now do PCR for HSV DNA; can also do serological IgM/IgG to HSV Varicella Zoster Virus - another type of human herpes virus; vaccines available for both types - chickenpox (primary VZV infxn - worse in adults) and herpes zoster or shingles (latent, recurrent infxns) - pathogenesis - spread via resp. droplets and contact w/ skin vesicles (very contagious) - incubation & infxn in local nodes and upper respiratory tract ~13 days - primary viremia spreads --> liver, spleen --> skin (vesiculous rash on erythematous base: dew drop on rose petal --> burst and crust over); fever and characteristic skin lesion - contagious period: until all vesicles crusted over - herpes zoster = shingles = reactivation of latent VZV infxn --> dermatomal lesions, more neuropathic pain - lab Dx: PCR analysis, serum Ab levels Smallpox Virus - largest, most complex viruses; pox virus (herpes type virus); many diff types of smallpox - exposure --> incubation 1-2wks (not contagious) --> prodrome 2-4days --> rash 3-4days (small macules/papules on mouth) --> sores lyse and spread more virus --> rash on face --> arms/legs/hands/fee --> pustular lesions day 4-5 w/ central umbilication --> scab over, fall off, form scars (not contagious) - prodrome: high fever, malaise, body aches, muscle pain Papovaviruses (dsDNA, non-enveloped) - Human Papilloma Virus causes warts; herpes type virus - long incubation: 50-100days; persistent, latent - epidemiology & pathogenesis: transmission is direct contact w/ infected skin or mucous membr, fomites, verticle transmission (mom-->fetus) infects and replicates in squamous cells of skin and mucous membrs; proliferation --> warts HPV16 & 18 are oncogenic (inactivate cell growth suppressor genes); cause cervical dysplasia --> cervical cancer - clincial manifestations: common warts, plantar warts, anogenital wars (condylomata acuminata), and cervical intraepithelial dysplasia --> cancer (cervical neoplasia) depend on subtypes (ex. genital warts = HPV 6,11; common warts = HPV 2,4) - lab Dx: clincial; pap smear w/ HPV-DNA probe; PCR Cytomegalovirus (CMV) - herpes type virus; persistent, latent infxn - systemic infxn usually asymptomatic (but immunocompromised pts at risk for severe disease - pneumonia, retinitis, colitis, esophagitis) - transmission: all body secretions, blood, organ transplants - some infxns cause heterophile-negative infectious mono-like syndrome (milder than w/ EBV) - neonates may acquire CMV transplacentally or during birth; severe TORCHS infxn (group of infectious diseases that cause congenital abnormalities); can have severe congenital abnormalities - lab Dx: serology (IgM/IgG to CMV titers), DNA probes, PCR of infected fluid, cell culture (owls-eye inclusion body) - potential outcomes (determined by pt immune system) - asymptomatic carrier, mild mono-like illness (heterophile/EBV
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neg mono), cytomegalic inclusion disease (congenital infxn in neonates -> abnormalities), multisite symptomatic disease (AIDS, immunocompromised pts) PICORNAVIRUSES: small, stable, naked RNA viruses Rhinovirus - causes common cold (clinical Dx); upper resp. infection (sore throat, cough), resolves w/in a week - transmitted via aerosol (resp. droplets) Enteroviruses begins in mouth (fecal-oral), replicates in oropharynx & GI, spreads to blood, --> target tissues (viremia can cause secondary sxs) - transmitted fecal-oral, impervious to stomach acid, and include: polio virus - vaccines to prevent; gets into GI --> blood --> CNS (brain & spinal cord) --> paralysis coxsackie viruses - herpangina (oral rash & lesions) w/ vesicles and ulcers on the soft palate & uvula; fever, HFMD w/ vesicles on tongue, hands, and feet; lasts a few days and resolves w/out issue (usually in babies < 3yo) coxsackie A: mild coxsackie B: can effect pleura & pericardial lining --> autoimmune in pancreas (DM) echo virus - Dx: most clinical w/out lab PARAMYXOVIRUSES: enveloped, ssRNA viruses dont see much MMR b/c of vaccination; transmitted via resp droplets & initiate infxn in resp tract acute; no long-lasting effects, cell-mediated immunity (CMI) causes many Sxs and then resolves the infxn Measles - measles is on of the childhood viral exanthems and causes CCC&P (= cough, coryza, conjunctivitis, pharyngitis) and high fever (prodrome), followed by Koplik spots and extensive maculopapular rash (usually starts on lower face and spreads to trunk) - vaccinations in MMR 1&2 in childhood - lab Dx Measles: Ab assay = MV sandwich: IgM rxn - blood test that measures measles specific IgM Mumps - mumps is another childhood disease causing parotitis (incubates/multiplies in parotid glands and then spreads) --> testes, ovaries, NS infxn, and is rare in developed countries (b/c of vaccination) - lab Dx Mumps: Parainfluenza causes croup in children esp. < 5yo; NO vaccine - upper resp tract disease (infects resp. epithelium) - cough, low-grade fever, stridor - usually happens in autumn (seasonal); no cure - lab Dx: immunofluorescent testing; also clinical Dx w/ stridor Respiratory Syncytial Virus (RSV) causes bronchiolitis in babies < 2yo; NO vaccine - direct invasion of respiratory epithelium - lab Dx: nasopharyngeal washings or swabs w/ RSV immunoassay ORTHOMYXOVIRUSES: influenza viruses (acute virus infecting lower resp. tract); usually self-limiting lab detection: rapid detection from nasopharyngeal swab --> Dx < 30min 70%Sn, 90%Sp also immunoassay test, viral cell culture (rare in clinical) - uncomplicated course: short incubation --> flu Sxs --> CMI, interferons --> healing; b/c immune system busy fighting flu, can have secondary bacterial infxn that complicates disease ADENOVIRUSES: dsDNA, cause common cold - causes resp tract infxn, pharyngoconjunctival fever (conjunctivitis, pharyngitis, fever), gastroenteritis - transm: fecal-oral or resp contact, fingers/fomites, poorly chlorinated pools - lab Dx: ELISA, fluorescent Ab tests, PCR & DNA probes ROTAVIRUS (reovirus family): dsRNA, non-enveloped, responsible for acute infantile gastritis - short incubation, short-lived Sxs (a few days) - transm: fecal material (esp. daycare setting); resilient, fecal-oral - vaccine available for infants to prevent - Sx: fever, vomitting, diarrhea; may cause dehydration (decreased abs of water and ions)
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- lab Dx: not necessary, but enz immunoassay to detect rotavirus in stool sample is available NORWALK AGENT: causes self-limited, acute (24-48hrs) gastroenteritis (vomitting, diarrhea) - transm.: fecal oral (cruise ships, hotels, schools, restaurants; large close quarters w/ poor hygiene) - lab Dx: generally not necessary, but ELISA used to detect Abs against Norwalk Agent HEPATITIS VIRUSES will go over more in Clin Med II types - at least 6 types (we will discuss A, B, C); infect hepatocytes and cause liver damage, jaundice, ^ liver enzymes Hepatitis A (HAV): ssRNA causing self-limited, acute hepatitis (diff from B&C, which are chronic) - transm: fecal-oral (contaminated food/water); (other hep viruses diff...) - vaccination now available for Hep A - incubation 15-50days; ingestion --> bloodstream via oral/intestinal lining --> liver (replication) - immune system attacks controls infxn - clinical manifestation: history of travel to endemic area; recent outbreak & incubation period w/ prodrome of nausea, fatigue, loss of appetite, and RUQ pain; 4-6 days later ICTERIC PHASE (jaundice, hepatomegaly, liver tenderness = big hint for Dx) - lab Dx: elevated liver enzymes ALT>AST (drug-induced --> AST is higher); anti-HAV IgM (Ab against HAV) Hepatitis B (HBV): dsDNA, small, enveloped; chronic hepatitis - causes hepatocellular carcinoma, cirrhosis - VERY diff from hep A; even diff family - Hepadnavirus family - transm: blood (IV drugs, transfusions), sexual contact, vertical - viremia spreads virus --> hepatocytes of liver, where it replicates/sheds (induces Ab response, may incubate 45-160days) - shedding --> expression of Ag on cell membr --> initiates T cells (CMI) - if immune system cant control infxn -> chronic hepatitis (contagious) - virus can integrate its DNA in hepatocytes and slowly replicate --> hepatic carcinoma, cirrhosis - Sxs: incubation (long) --> acute disease (prodrome - malaise, anorrexia, nausea, RUQ pain) --> icteric phase (jaundice, dark urine, light stool, itching) --> convalescence - outcomes: 90% --> resolution (not chronic) 1% --> fulminant hepatitis --> liver failure, possibly death (virulent infxn) 9% --> chronic carriers --> 10% of these have chronic active hepatitis --> cirrhosis, hepatic carcinoma (fatal) - lab Dx: clinical syndrome + elevated LFTs to Dx recent infxn --> serology (anti-HB core IgM and HB surface Ag) in chronic pts --> viral load (PCR) quanitfies # of viral copies of DNA in blood Hepatitis C (HCV): RNA, enveloped; chronic hepatitis - infects hepatocytes (receptor-mediated uptake into hepatocyte) - also diff from other hep viruses - Flaviviridae family - transm: infected blood (sometimes through sexual contact); 170million HCV carriers worldwide - pathogenesis: via bloodstream (viremia) --> Ab response --> virus travels to liver --> infects hepatocytes and replicates w/out killing the host cell but provokes continued immune response and tissue damage - persistent, chronic infection leads to increased risk of hepatocellular carcinoma and cirrhosis - outcomes: 15% --> resolution (not chronic): more difficult for the body to eliminate HepC than HBV 85%--> persistent infxn --> 30-35% of these have chronic hepatitis (actively replicating, but not killing host cells) --> 20% of those -->cirrhosis, 4%-->carcinoma, 6%-->liver failure - lab Dx: often discovered during screening or as part of workup for elevated LFTs serologic: Ig to HCV (Anti-HCV) by ELISA viral load (via PCR, RT PCR to quantify # of copies of viral nucleic acid in blood Labs to Dx Viral Infxns - cytologic examination - electron microscopy - virus isolation and growth - detection of viral proteins (Ags, enzs) - detection of viral genomes (viral load) - serology - links to review procedures of Dx viral infxns (animations!)
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 http://student.ccbcmd.edu/courses/bio141/labmanua/lab18/wb_flash.html http://student.ccbcmd.edu/courses/bio141/labmanua/lab18/eia_flash.html http://student.ccbcmd.edu/courses/bio141/labmanua/lab17/lab17.html