title: Reactivation from latency displays HIV particle budding at plasma membrane, accompanying CD44 upregulation and recruitment.
abstract: BACKGROUND
  It has been accepted that HIV buds from the cell surface in T lymphocytes, whereas in macrophages it buds into intracellular endosomes. Recent studies, on the other hand, suggest that HIV preferentially buds from the cell surface even in monocytic cells. However, most studies are based on observations in acutely infected cells and little is known about HIV budding concomitant with reactivation from latency. Such studies would provide a better understanding of a reservoir for HIV.
  
  
  RESULTS
  We observed HIV budding in latently infected T lymphocytic and monocytic cell lines following TNF-alpha stimulation and examined the upregulation of host factors that may be involved in particle production. Electron microscopy analysis revealed that reactivation of latently infected J1.1 cells (latently infected Jurkat cells with HIV-1) and U1 cells (latently infected U937 cells with HIV-1) displayed HIV particle budding predominantly at the plasma membrane, a morphology that is similar to particle budding in acutely infected Jurkat and U937 cells. When mRNA expression levels were quantified by qRT-PCR, we found that particle production from reactivated J1.1 and U1 cells was accompanied by CD44 upregulation. This upregulation was similarly observed when Jurkat and U937 cells were acutely infected with HIV-1 but not when just stimulated with TNF-alpha, suggesting that CD44 upregulation was linked with HIV production but not with cell stimulation. The molecules in endocytic pathways such as CD63 and HRS were also upregulated when U1 cells were reactivated and U937 cells were acutely infected with HIV-1. Confocal microscopy revealed that these upregulated host molecules were recruited to and accumulated at the sites where mature particles were formed at the plasma membrane.
  
  
  CONCLUSIONS
  Our study indicates that HIV particles are budded at the plasma membrane upon reactivation from latency, a morphology that is similar to particle budding in acute infection. Our data also suggest that HIV expression may lead to the upregulation of certain host cell molecules that are recruited to sites of particle assembly, possibly coordinating particle production.
authors: Kitasato University, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan. maris@lisci.kitasato-u.ac.jp
authors: Mari
authors: M
authors: Suyama
authors: Eriko
authors: E
authors: Daikoku
authors: Toshiyuki
authors: T
authors: Goto
authors: Kouichi
authors: K
authors: Sano
authors: Yuko
authors: Y
authors: Morikawa
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure / Q000821:virology*; D056827:Endosomal Sorting Complexes Required for Transport; D020869:Gene Expression Profiling; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis*; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D009000:Monocytes / Q000648:ultrastructure / Q000821:virology; D010750:Phosphoproteins / Q000096:biosynthesis; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis; D013601:T-Lymphocytes / Q000648:ultrastructure / Q000821:virology; D060149:Tetraspanin 30; D015854:Up-Regulation; D019065:Virus Assembly
chemical_list: D015703:Antigens, CD; C497483:CD44 protein, human; C554586:CD63 protein, human; D056827:Endosomal Sorting Complexes Required for Transport; D018960:Hyaluronan Receptors; D010750:Phosphoproteins; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; C096164:hepatocyte growth factor-regulated tyrosine kinase substrate