HIV susceptibility and CCR5 Anna-Lisa Doebley Final Presentation May 6th 2014

The HIV/AIDS epidemic 35.3 Million Total

The only person who has been cured of HIV: Timothy Brown was infected with HIV in 1995. Controlled the disease with antiretroviral therapy. Developed Leukemia many years later. Treated with a bone marrow transplant from an HIV resistant donor. He no longer has any detectable HIV in his immune system. The success of this treatment has inspired research which may someday cure AIDS. How?

What is the HIV virus? Blood Borne Pathogen Infects Immune cells Causes AIDS if not treated

CCR5 is a gene associated with AIDS resistance Human 7 tm_1 352aa 352aa Cellular component: Biological process: Molecular function: Plasma membrane Cell communication Chemokine binding Chemokine

CCR5 is well conserved across vertebrates Human 7 tm_1 352aa Chimpanzee 7 tm_1 352aa 99% Orangutan 7 tm_1 352aa 99% Gorilla 7 tm_1 352aa 99% Mouse 7 tm_1 354aa 82% Chemokine receptor Natural function not well known Chicken 7 tm_1 355aa 62% Zebrafish 7 tm_1 358aa 42%

What is the function of CCR5 in HIV infection? HIV Virus CCR5 Host Cell

HIV binds CCR5 HIV Virus CCR5 Host Cell

HIV enters the cell Host Cell

CCR5 alleles can block HIV infection 7 tm_1 Human Wild Type 352aa CCR5 Δ32 allele: 32 bp deletion results in a frame shift and stop codon 7 tm_1 Human Δ32 215aa (31 after frame shift)

CCR5 alleles can block HIV infection WT CCR5 HIV CCR5 Δ32aaa a HIV

SIVs do not cause AIDS… Why? Where did HIV come from? SIV HIV HIV evolved from the family of Simian Immunodeficiency Viruses which infect non-human primates Transmitted from Chimpanzees to humans on several separate occasions SIVs do not cause AIDS… Why?

HIV does not cause AIDS in chimpanzees WT CCR5 SIV WT CCR5 HIV

Why don’t SIVs cause AIDS? Goal: understand how non-human primates are able to combat SIV/HIV infection Hypothesis: sequence similarities between human AIDS-resistant CCR5 alleles and primate CCR5 alleles may help explain AIDS resistance

Aim 1: Determine which regions of CCR5 are conserved in non-human primates 7 tm_1 352aa Chimpanzee 7 tm_1 352aa 99% Orangutan 7 tm_1 352aa 99% Chemokine receptor Natural function not well known Gorilla 7 tm_1 352aa 99%

Approach: Determine which regions of CCR5 are conserved in non-human primates VandeWoude and Apetrei, 2006 34 species of primates infected with SIVs (3 progress to AIDS) 22 sequences available Chemokine receptor Natural function not well known

CCR5 amino acid substitutions in AIDS resistant Primates 7 tm_1 Each blue bar represents an amino acid that is different between different species of primates

Aim 2: determine if primate CCR5 alleles are similar to AIDS-resistant human CCR5 alleles. Carrington et al. 1999 9 CCR5 sequences (3 believed to be AIDS resistant) **deletion alleles not included Combine data with Primate sequences

AIDS susceptible allele AIDS resistant allele

CCR5 amino acid substitutions 7 tm_1 Resistant Susceptible What is in these regions?

Aim 3: Determine if particular protein domains are found in the non-conserved regions of CCR5

CCR5 amino acid substitutions affect areas outside of transmembrane domains 7 tm_1 Transmembrane domains

Future directions Sequence the 5’ and 3’ UTRs of primate CCR5 genes Study the sequences of CCR5 genes in other species that are affected by immunodeficiency viruses

Sources Images: Info: Human: http://genetics.wisc.edu/Ikeda.htm Slide 2: Map: http://www.who.int/hiv/data/global_data/en/ Total statistic: http://www.who.int/gho/hiv/epidemic/hiv_001.jpg Slide 3: http://www.redpepper.co.ug/hivaids-patients-hit-hard-by-health-workers-strike/ Slide 4: http://en.wikipedia.org/wiki/Signs_and_symptoms_of_HIV/AIDS Slide 5: http://a.abcnews.com/images/Health/wabc_berlin_patient_120724_wg.jpg Slide 6: http://siklusair.com/what-does-hiv-virus-look-like Slide 7: Plasma membrane: http://www.scs.illinois.edu/~mlkraft/images/cell%20membrane%20smaller.jpg Communication: http://openclipart.org/image/800px/svg_to_png/183631/communication.png Chemokine: Modified image from Akihiro Ikeda Slides 9 to 11: Modified image from: Image: http://www.niaid.nih.gov/SiteCollectionImages/topics/hivaids/hivReplicationCycle.gif Slide 13: Sick Person: http://1.bp.blogspot.com/-aZDdz8HAmWU/TlWCfOxg3kI/AAAAAAAAAD0/Pr3HpiZ5yV4/s1600/sick- person-300x286.jpg Healthy Person: http://classroomclipart.com/images/gallery/Clipart/Sports/Jogging_Clipart/ jogging_cartoon_03.jpg Slide 15: Chimpanzee: http://images.nationalgeographic.com/wpf/media-live/photos/000/007/cache/young- chimp_763_600x450.jpg Human: http://genetics.wisc.edu/Ikeda.htm “SIV” virus: http://aidsthekiller.files.wordpress.com/2010/04/hiv-virus-picture.png Slide 16: Monkey: http://bestclipartblog.com/clipart-pics/monkey-clipart-6.jpg Info: Naif HM. 2013. Pathogenesis of HIV Infection. Infect Dis Rep 5:e6. de Groot NG, Bontrop RE. 2013. The HIV-1 pandemic: does the selective sweep in chimpanzees mirror humankind's future? Retrovirology 10:53. Wooding S, Stone AC, Dunn DM, Mummidi S, Jorde LB, Weiss RK, Ahuja S, Bamshad MJ. 2005. Contrasting effects of natural selection on human and chimpanzee CC chemokine receptor 5. Am J Hum Genet 76:291-301. Carrington M, Dean M, Martin MP, O'Brien SJ. 1999. Genetics of HIV-1 infection: chemokine receptor CCR5 polymorphism and its consequences. Hum Mol Genet 8:1939-1945. VandeWoude S, Apetrei C. 2006. Going wild: lessons from naturally occurring T-lymphotropic lentiviruses. Clin Microbiol Rev 19:728-762

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