title: Perforin is not co-expressed with granzyme A within cytotoxic granules in CD8 T lymphocytes present in lymphoid tissue during chronic HIV infection.
abstract: BACKGROUND
  Residual HIV-1-infected cells are poorly eliminated from lymphoid tissue (LT) reservoirs by effector cytotoxic T lymphocytes (eCTL) despite antiretroviral therapy. Perforin and granzyme A (grA) constitute major effector molecules within eCTL granules that induce apoptosis and lysis of virally infected cells.
  
  
  OBJECTIVE
  Expression of perforin and grA was studied at the single cell level in LT and blood from 16 patients infected with HIV-1 (stage A1-C) who were not taking antiretroviral therapy.
  
  
  METHODS
  Immunohistochemical analysis by in situ imaging of cells from blood and LT.
  
  
  RESULTS
  Quantitative in situ imaging showed that perforin-expressing CD8 T cells comprised 0.3-1.5% of total cells within the LT from recent HIV-1 seroconverters, while grA was found in 2.1-7.2% of total cells. However, despite high-level grA upregulation (1.5-4.5% of total cells) compared with that in non-infected individuals (0.4-0.9%), perforin expression remained low (< 0.1% of total cells) (P < 0.02) in LT from patients with chronic HIV-1 infection (stage A2-C). This contrasted with findings in peripheral blood mononuclear cells (PBMC) from the same HIV-1 infected cohort where perforin was detected in 13-31% of all PBMC, which was 10- to 100-fold higher than in lymphoid tissue (P < 0.001); grA was found in 14-32% of total PBMC. Two-colour staining showed that granular expression of perforin and grA was restricted to CD8 T cells in over 90% of total cells in both LT and blood.
  
  
  CONCLUSIONS
  These findings indicate that cytotoxic perforin expression is impaired at local sites of HIV replication within lymphoid tissue. Since perforin is required together with grA for granule-mediated cytolysis, the low perforin expression in the LT may limit the ability of eCTL to eliminate HIV-1 infected cells in lymphoid tissue.
authors.affiliation: Department of Infectious Diseases, Karolinska Institutet, Huddinge University Hospital, Stockholm, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Andersson
authors.forename: H
authors.initials: H
authors.lastname: Behbahani
authors.forename: J
authors.initials: J
authors.lastname: Lieberman
authors.forename: E
authors.initials: E
authors.lastname: Connick
authors.forename: A
authors.initials: A
authors.lastname: Landay
authors.forename: B
authors.initials: B
authors.lastname: Patterson
authors.forename: A
authors.initials: A
authors.lastname: Sönnerborg
authors.forename: K
authors.initials: K
authors.lastname: Loré
authors.forename: S
authors.initials: S
authors.lastname: Uccini
authors.forename: T E
authors.initials: TE
authors.lastname: Fehniger
mesh_terms: D018791:CD4 Lymphocyte Count; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism*; D053804:Granzymes; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000502:physiology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000276:immunology; D008221:Lymphoid Tissue / Q000276:immunology; D008562:Membrane Glycoproteins / Q000378:metabolism*; D014066:Palatine Tonsil / Q000276:immunology; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012367:RNA, Viral / Q000097:blood; D012697:Serine Endopeptidases / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D012367:RNA, Viral; D054353:Perforin; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human