title: HIV integration and the establishment of latency in CCL19-treated resting CD4(+) T cells require activation of NF-κB.
abstract: Eradication of HIV cannot be achieved with combination antiretroviral therapy (cART) because of the persistence of long-lived latently infected resting memory CD4(+) T cells. We previously reported that HIV latency could be established in resting CD4(+) T cells in the presence of the chemokine CCL19. To define how CCL19 facilitated the establishment of latent HIV infection, the role of chemokine receptor signalling was explored.
  
  
  
  In resting CD4(+) T cells, CCL19 induced phosphorylation of RAC-alpha serine/threonine-protein kinase (Akt), nuclear factor kappa B (NF-κB), extracellular-signal-regulated kinase (ERK) and p38. Inhibition of the phosphoinositol-3-kinase (PI3K) and Ras/Raf/Mitogen-activated protein kinase/ERK kinase (MEK)/ERK signalling pathways inhibited HIV integration, without significant reduction in HIV nuclear entry (measured by Alu-LTR and 2-LTR circle qPCR respectively). Inhibiting activation of MEK1/ERK1/2, c-Jun N-terminal kinase (JNK), activating protein-1 (AP-1) and NF-κB, but not p38, also inhibited HIV integration. We also show that HIV integrases interact with Pin1 in CCL19-treated CD4(+) T cells and inhibition of JNK markedly reduced this interaction, suggesting that CCL19 treatment provided sufficient signals to protect HIV integrase from degradation via the proteasome pathway. Infection of CCL19-treated resting CD4(+) T cells with mutant strains of HIV, lacking NF-κB binding sites in the HIV long terminal repeat (LTR) compared to infection with wild type virus, led to a significant reduction in integration by up to 40-fold (range 1-115.4, p = 0.03). This was in contrast to only a modest reduction of 5-fold (range 1.7-11, p > 0.05) in fully activated CD4(+) T cells infected with the same mutants. Finally, we demonstrated significant differences in integration sites following HIV infection of unactivated, CCL19-treated, and fully activated CD4(+) T cells.
  
  
  
  HIV integration in CCL19-treated resting CD4(+) T cells depends on NF-κB signalling and increases the stability of HIV integrase, which allow subsequent integration and establishment of latency. These findings have implications for strategies needed to prevent the establishment, and potentially reverse, latent infection.
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Suha
authors.initials: S
authors.lastname: Saleh
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Hao K
authors.initials: HK
authors.lastname: Lu
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Evans
authors.affiliation: Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: David
authors.initials: D
authors.lastname: Harisson
authors.affiliation: Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: Jingling
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: Anthony
authors.initials: A
authors.lastname: Jaworowski
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Georgina
authors.initials: G
authors.lastname: Sallmann
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Karey Y
authors.initials: KY
authors.lastname: Cheong
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.
authors.forename: Talia M
authors.initials: TM
authors.lastname: Mota
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Surekha
authors.initials: S
authors.lastname: Tennakoon
authors.affiliation: Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Angelovich
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.
authors.forename: Jenny
authors.initials: J
authors.lastname: Anderson
authors.affiliation: Centre for Virus Research, Westmead Millennium Institute, University of Sydney, Sydney, Australia.
authors.forename: Andrew
authors.initials: A
authors.lastname: Harman
authors.affiliation: Centre for Virus Research, Westmead Millennium Institute, University of Sydney, Sydney, Australia.
authors.forename: Anthony
authors.initials: A
authors.lastname: Cunningham
authors.affiliation: Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: Lachlan
authors.initials: L
authors.lastname: Gray
authors.affiliation: Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.|Department of Microbiology, Monash University, Melbourne, Australia.
authors.forename: Melissa
authors.initials: M
authors.lastname: Churchill
authors.affiliation: Department of Infectious Diseases, School of Medicine, Deakin University, Melbourne, Australia.
authors.forename: Johnson
authors.initials: J
authors.lastname: Mak
authors.affiliation: Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.|Department of Microbiology, Monash University, Melbourne, Australia.
authors.forename: Heidi
authors.initials: H
authors.lastname: Drummer
authors.affiliation: Division of Hematology-Oncology, Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.|University California Los Angeles (UCLA) AIDS Institute, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
authors.forename: Dimitrios N
authors.initials: DN
authors.lastname: Vatakis
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.
authors.forename: Sharon R
authors.initials: SR
authors.lastname: Lewin
authors.affiliation: Doherty Institute for Infection and Immunity, The University of Melbourne, Melbourne, Australia. paul.cameron@unimelb.edu.au.|Department of Infectious Diseases, Alfred Health, Monash University, Melbourne, Australia. paul.cameron@unimelb.edu.au.|Centre for Biomedical Research, Burnet Institute, Melbourne, Australia. paul.cameron@unimelb.edu.au.
authors.forename: Paul U
authors.identifier: 0000-0002-1906-6945
authors.initials: PU
authors.lastname: Cameron
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000502:physiology / Q000821:virology*; D054415:Chemokine CCL19 / Q000494:pharmacology*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D019427:HIV Integrase / Q000235:genetics; D015497:HIV-1 / Q000201:enzymology / Q000502:physiology; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D054388:Receptors, CCR / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D016662:Virus Integration / Q000187:drug effects; D017735:Virus Latency / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: C515460:CCL19 protein, human; D054415:Chemokine CCL19; D016328:NF-kappa B; D054388:Receptors, CCR; D019427:HIV Integrase
keywords: CD4+ T cells; Chemokine signalling; HIV latency; Integration; NF-κB