In vitro HIV1 infection of CD34+ progenitor-derived dendritic/Langerhans cells at different stages of their differentiation in the presence of GM-CSF/TNF alpha.
Langerhans cells (LC) are antigen-presenting cells which are found in areas at risk of inoculation by the human immunodeficiency virus (HIV). LC were shown to be sensitive to in vitro infection by HIV1. They could be generated in vitro by culturing CD34+ haematopoietic progenitors with GM-CSF+TNF alpha. In this study, we tested the sensitivity to HIV1 infection of in vitro generated LC throughout their differentiation and we investigated the effect of such an infection on in vitro differentiation. Phenotypic controls were performed using FACS analysis on day 6 for the presence of a CD1a+ cell population, and differentiation was assessed by transmission electron microscopy on day 13 for the presence of Birbeck granules. CD34+ cells were purified from cord blood mononuclear cells by magnetic separation. Cell suspensions were infected with either a T-lymphotropic, syncytium-inducing isolate (HXB2) or a macrophage-tropic, non-syncytium-inducing isolate (Ba-L). Viral particle release was measured by p24 antigen production in the culture supernatant. A high level of p24 production was noted on day 13 of postinfection only when infection was carried out with Ba-L isolate on cells generated after 6 days in culture with GM/CSF+TNF alpha. No infection of CD34+ progenitor cells was obtained either with Ba-L isolate or HXB2. The sensitivity of Langerhans cell/dendritic cell (LC/DC) precursors to NSI isolate (Ba-L) seemed to coincide with the early stage of differentiation (CD1a antigen appearance). The infection did not alter the differentiation of in vitro generated LC, which presented their specific ultrastructural marker of epidermal environment, i.e. Birbeck granules from day 15 of the culture as compared to control culture. These results highlight the HIV infectibility of a differentiated population of LC/DC generated in vitro from CD34+ progenitors.
INSERM U346, Peau humaine et immunité, Hôpital Edouard Herriot, Lyon, France.
A S
AS
Charbonnier
B
B
Verrier
C
C
Jacquet
C
C
Massacrier
M M
MM
Fiers
F
F
Mallet
C
C
Dezutter-Dambuyant
D
D
Schmitt
D018949:Antigens, CD1 / Q000096:biosynthesis; D018952:Antigens, CD34 / Q000096:biosynthesis; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000648:ultrastructure / Q000821:virology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D016655:HIV Core Protein p24 / Q000032:analysis; D015497:HIV-1; D006801:Humans; D007231:Infant, Newborn; D007801:Langerhans Cells / Q000648:ultrastructure / Q000821:virology*; D008854:Microscopy, Electron; D013234:Stem Cells / Q000821:virology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D018949:Antigens, CD1; D018952:Antigens, CD34; D016655:HIV Core Protein p24; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor