HUMAN IMMUNODEFICIENCY VIRUS (HIV) Family: Retroviridae Genus: Lentivirus (known to be slow viruses, long term chronic infections)

HIV: virus itself AIDS: condition that results from the infection; specific, advanced stage of the disease HIV is a blood-borne virus Transmission from a positive carrier via saliva or blood Both a medical and social problem Sub-Saharan Africa has the most number of cases; 22.5M cases High Risk Groups: Commercial Sex Workers, Men to Men Sex High risk group: 15-24 y/o Low but slow: spread of virus Affects more male than female: because of male to male sex Important Properties of Lentivirus (Nononcogenic Retrovirus): Cylindrical nucleocapsid in mature virion RNA genome more complex than that of transforming retroviruses Icosahedral symmetry and vaseshaped protein core Virion: spherical, 80-100 nm in diameter with cylindrical core Genome: +ssRNA, linear, 9-10 kb, diploid; genome more complex than that of oncogenic retrovirus, contains up to 6 additional replication genes b/c of positive polarity, first step after entering host cell is translation, not transcription, capable of initiating translation Proteins: Envelope protein undergoes antigenic variation; reverse transcriptase enzyme contained inside virions, protease required for production of infections virus Envelope: Present

Envelope proteins consist of the glycoprotein 120 (gp120) and gp41. GP 41 is the transmembrane protein which serves as a stem that links gp120 to the viral envelope - This glycoprotein complex allows attachment and fusion to target cell Capsid: conical in shape viral protein p24 spread over capsid Diploid genome: carries two copies instead of one Replication: Reverse transcriptase makes DNA copy from genomic RNA; provirus DNA is template for viral RNA. Genetic variability is common Maturation: Particles bud from plasma membrane Outstanding characteristics: Members are non-oncogenic and may be cytocydal Infect cells of immune system Proviruses remain permanently associated with cells Viral expression is restricted in some cells in vivo Cause slowly progressive, chronic disease Replication is usually speciesspecific Group includes the causative agents of AIDS Structure of HIV: Carries 3 types of enzymes: a. Reverse transcriptase- RNA dependent DNA polymerase b. Integrase- responsible for integrating c. Protease- cleaves products of translation to specific products Genes required for replicating retrovirus: Gag, pro, pol and env

*Env gene- which codes for viral envelope proteins - product is SU/ gp120 SU/ gp120- contains binding domains responsible for illness attachment to the CD4 molecule *Co-receptor- determines lymphocyte and macrophage tropisms and caries the major antigenic determinants that elicit neutralizing antibodies *HIV glycoprotein has 5 variable (V) regions that diverge among isolates, with V3 region important in neutralization *TM (gp41) env product contains both a transmembrane domain that anchors the glycoprotein in the viral envelope and fusion domain that facilitates viral penetration into target cells HIV membrane proteins facilitate viral entry into cells, interact with antibodies and are major vaccine candidates Mechanism of CKR5 co- receptor function: 1. Attachment -gp120 binds to CD4 receptors in Macrophages and helper T cells 2. Binding to second receptor -gp120 binds to CKR5 which partially exposes the transmembrane protein gp 41 which contains fusion proteins 3. Fusion -fusion peptides from gp41 is then exposed allowing viral and cell membrane fusion *entry: fusion *exit: budding Other important points: - expression of coreptors can indicate the stage of infection

- CCR5 indicates acute infection -CXCR4 indicates advanced infection -during advanced infection there are more virus particles now compared to antibodies, the virus is easier to isolate Upon entry of DNA into host cell: A) reverse transcriptase vital, 3 functions: 1. synthesizes complementary DNA (cDNA) using the ssRNA genome of the virus 2. also contains ribonuclease activity which degrades the viral RNA after synthesizing the cDNA 3. has DNA-dependent polymerase activity which uses the cDNA as the complementary strand to create the sense strand cDNA=antisense strand B) the dsDNA is then integrated into the host genome in the nucleus via the integrase protein C) The virus may then lie dormant for years known as the laten period until initiation of transcription of viral mRNA is produced D) viral proteins are transcribed from the Env, Gag, and Pol genes and the mRNA translated into the respective proteins E) Packaging of viral proteins occurs in the cytoplasm F) release occurs through budding Six additional replicating genes:

a. Tat protein- functions in transactivation whereby a viral gene product is involved in transcriptional activation of other viral genes b. Rev protein- required for expression of viral structural protein - Facilitates the export of unspliced viral transcripts from nucleus

*Structural proteins are translated from unspliced mRNAs during the late phase of viral replication c. Nef protein- increases viral infection; facilitates activation of resting T cells and down regulates expression of CD4 and MHC class I d. Vpr protein- increases transport of viral preintegration complex into the nucleus and also arrests cells in G2 phase of the cell e. Vpu protein- promotes CD4 degredation f. Vif- responsible for infectivity **** I. LTR- binding sites for host transcription factors Gag- capsid protein, matrix protein, nucleocapsid protein Pol- codes for reverse transcriptase, protease, integrase and ribonuclease Env- encodes for gp160 which is cleaved by protease to form gp120 and gp41 Vif- for maturation of HIV virion. Promotes integrity of cell- free virus Vpr- transcriptional activator Classification of HIV: HIV-2 -limited to Africa

K- Congo, Cameron CRFs- A/E or A/G *in Phils B and A *E subtype is a modification of subtype A; now in CRFs (circulating recombinant form) HIV-1 genome Viral proteins are synthesized as precursor polyproteins (gag-pol (Pr160), gag (Pr--) and ENv (160)), which are enzymatically processed to yield virion proteins Gag-pol and Gag are cleaved by the viral protease PR to produce the indicated small proteins Env is cleaned by cellular PR, producing SU gp120 and TM gp41 *HIV-2 and SIV lack the vpu gene but contain vpx gene HIV-2 Restricted to west Africa No HIV-2 pandemic Less pathogenic than HIV-1 There is no evidence that HIV-2 infected person may be at decreased risk for acquiring HIV-1

Origin of AIDS HIV in humans originated from cross-species infections by simian viruses in rural Africa, probably due to direct human contact with infected primate blood. Mode of Transmission

HIV-1 M N O

A B C D F G H J K CRFs A- West and Central Africa B- Europe, America, Japan, Australia C- S and E Africa, India, Nepal D- East and Central Africa F-Central Africa, S America, E Europe G- W and E Africa, C Europe H- Central Africa J- Central America

1. Direct injection into the bloodstream Through sexual contact (mostly common to heterosexual) Injecting drug use Blood transfusion Needle stick injuries 2. Across or through breaks in mucosal surfaces 3. From mother to infant In utero

During birth Via breast milk

Biologic Risk factors for sex route High viral load Being the receptive partner Young female Uncircumcised male Damage to genital skin/ mucous membrane Having a sexually transmitted infection Transmission myths: blood sucking mosquito holding hands kissing toilet seats swimming pools *except if you have ulcerations Mechanism: Because HIV targets specifically helper T cells the immune system goes to whack other cells of the immune system cant function since CD4 T cells coordinate the immune system like a symphony opportunistic organisms, infections that usually do no harm (fungal infections, minor orgs, etc) produce destructive effects bc of the extremely suppressed immune system death ensues Primary infection = flu like symptoms AIDS symptoms: - Kaposis sarcoma - CMV retinitis - Oral candidiasis - PCP - Cryptococcus infection Dental infections: -cervical caries - candidiasis - oral hairy leukoplasia - PPV warts - HSV

- During acute infection (1-2 weeks) the p24 antigen is high and there is still little presence of antibodies - Long intermediate period: decline in p24, rise in antibodies (anti-HIV gp120) -When initiated, virus replication occurs super rapidlyimmune system tries to mount and build up more T cells macrophages etc but all this does is create more hosts for the virusimmune system cant catch up anymore and its stores are being depleteddeath Disinfection and Inactivation HIV completely inactivated ( 105 units of infection) by treatment for 10 mins at room temperature with any of the following: 10% household bleach 50 5 ethanol 35% isopropanol 1% Nonidet P40% 0.5% Lysol 0.5% paraformaldehyde 0.3% hydrogen peroxide Inactivated by extreme pH (1.0 and 13.0) Heat Readily inactivated in liquids or 10% serum by heating at 56 0C for 10 mins Lyophilized blood products would need to be heated at 68 0C for 72 hours Disinfectants Many detergents (NP40 but not Tween 20) and high and low pH solutions rapidly inactivate the virus Glutaraldehyde and hypochlorite within several mins in low concentration Isopropyl alcohol or ethanol (40%-70%) H2O2 phenolics and paraformaldehyde

 Rapidly inactivate HIV in suspension Less effective where HUV has dried on surfaces or in the presence of large amounts of protein Drying Redusec viral infectivity by more than ten fold HIV dired in salt solution on stainless steel surfaces may still be detected after a week Dried serum or blood is unlikely to be a significant infection risk Laboratory Diagnosis of HIV Screening assays -very sensitive but not specific ELISA -most common is western blot -used for confirmatory test Particle agglutination Test **** Viral markers -cell culture -p24 antigen test -PCR -viral load Markers for immune function -flow cytometry (CD4/CD8 counts)

Protease Inhibitors -mimic a peptide that can bind to the active site of the viral protease -Saquinavir, Ritonavir, Indinavir HAART -Highly active anti-retroviral therapy -considered as the standard in HIV therapy; monotherapy is no longer recommended Reasons for discontinuation of drug therapies: Toxicity Drug interactions Developmental drug resistance Latently infected cells

Why is it difficult for the immune system to fight off HIV infection? HIV attacks the immune system itself, weakening its ability to fight back HIV replicates in large quantities that are more than the compromised immune system can handle HIV has the ebility to mutate (change itself) very quickly, making it more difficult for the body to fight the infection. ABSTINENCE best preventive measure

Types of Drugs Nucleoside Inhibitors -competitive inhibitors of reverse transcriptase AZT, DDI, DDC, d4T, 3TC, Abacavir, Succinate, Previon Non-nucleoside -reverse transriptase inhibitors but bind to site other than the active site of the enzyme -Nevirapine, Atervidine, Detavirdine, Efavirenz