title: Quantification of HIV-1-specific T-cell responses at the mucosal cervicovaginal surface.
abstract: OBJECTIVE
  To characterize HIV-1 specific cellular immune responses at mucosal surfaces using a rapid, sensitive enzyme-linked immuno-spot (ELISPOT) technique.
  
  
  METHODS
  Cervicovaginal mononuclear cells obtained from cytobrush and cervicovaginal lavage were assessed for production of interferon-gamma (IFN-gamma) in response to stimulation by HIV-1 antigens. HIV-1 specific responses were compared in a cross-sectional study of two HIV-1-positive patient groups: women not currently on antiretroviral therapy with peripheral CD4 cell counts > 250 x 10(6)/l (n = 12); and women on highly active antiretroviral therapy (HAART) (n = 9).
  
  
  METHODS
  Mononuclear cells from peripheral blood or cervicovaginal specimens were assessed in an ELISPOT assay for responses to HIV-1 antigens expressed by recombinant vaccinia viruses. This assay detects primarily CD8 T cells and shows good correlation with MHC class I tetramer staining of cytotoxic T lymphocytes.
  
  
  RESULTS
  HIV-1 specific IFN-gamma spot-forming cells were detected in cervicovaginal samples of one out of nine women (11%) on HAART and five out of 12 women (42%) not currently on HAART. In peripheral blood mononuclear cells, HIV-1 specific IFN-gamma spot-forming cells were significantly more numerous in women not currently on HAART than in women on HAART (P = 0.009). In most cases, antigens recognized by mucosal T cells were also recognized by PBMC; however, there were exceptions.
  
  
  CONCLUSIONS
  HIV-1-specific antigen-reactive T cells may be detected in routine, noninvasive gynecological specimens. The results suggest that cervicovaginal HIV-1-specific T cells may be less numerous in individuals on HAART than in those not on HAART, as shown previously for HIV-1-specific cytotoxic T lymphocytes in the peripheral blood.
authors.affiliation: Aaron Diamond AIDS Research Center, The Rockefeller University, New York, NY 10016, USA.
authors.forename: B L
authors.initials: BL
authors.lastname: Shacklett
authors.forename: S
authors.initials: S
authors.lastname: Cu-Uvin
authors.forename: T J
authors.initials: TJ
authors.lastname: Beadle
authors.forename: C A
authors.initials: CA
authors.lastname: Pace
authors.forename: N M
authors.initials: NM
authors.lastname: Fast
authors.forename: S M
authors.initials: SM
authors.lastname: Donahue
authors.forename: A M
authors.initials: AM
authors.lastname: Caliendo
authors.forename: T P
authors.initials: TP
authors.lastname: Flanigan
authors.forename: C C
authors.initials: CC
authors.lastname: Carpenter
authors.forename: D F
authors.initials: DF
authors.lastname: Nixon
mesh_terms: D023241:Antiretroviral Therapy, Highly Active; D018791:CD4 Lymphocyte Count; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002584:Cervix Uteri / Q000166:cytology / Q000276:immunology*; D003430:Cross-Sectional Studies; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D015658:HIV Infections / Q000188:drug therapy / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D018928:Immunity, Mucosal; D007371:Interferon-gamma / Q000096:biosynthesis; D009092:Mucous Membrane / Q000166:cytology / Q000276:immunology; D014621:Vagina / Q000166:cytology / Q000276:immunology*
chemical_list: D007371:Interferon-gamma