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... CCR5 coreceptors for transmission of virus from DCs to T cells. We next tested cells with a CCR5 coreceptor for M-tropic virus that was nonfunctional as a result of a 32-bp deletion in the CCR5 gene =-=(15)-=-. By preparing DCs and T blasts from these donors, we could analyze the role of HIV-1 viral entry into each cell type in DC-T cell cocultures. In a prior study, we found that infection of mature DCs w...

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...he DC pathway for enhancing viral replication. Immature DCs selectively replicate M-tropic HIV-1 and therefore can account for the “bottleneck” that selects for this type of virus during transmission =-=(27, 32)-=-. However, when maturation takes place, both macrophagetropic (M-tropic) and T-tropic virus can be transmitted by DCs to T cells. An important role for DCs during the course of HIV-1 infection is indi...

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...repancies could be due to several parameters, one of which is the state of DC maturation, as we now report. MATERIALS AND METHODS Cells. Dendritic cells were generated from the blood of normal donors =-=(2, 23)-=-. Peripheral blood mononuclear cells PBMC were isolated by sedimentation in Ficoll-Hypaque, and 5 � 10 7 PBMC were plated per 100-mm-diameter culture plastic dish in RPMI (Gibco) supplemented with 1% ...

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...mpairs their antigen-presenting cell function (10, 16), while other reports demonstrate that DCs are not productively infected but nevertheless promote viral replication upon interaction with T cells =-=(5, 20)-=-. These discrepancies could be due to several parameters, one of which is the state of DC maturation, as we now report. MATERIALS AND METHODS Cells. Dendritic cells were generated from the blood of no...

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...he DC pathway for enhancing viral replication. Immature DCs selectively replicate M-tropic HIV-1 and therefore can account for the “bottleneck” that selects for this type of virus during transmission =-=(27, 32)-=-. However, when maturation takes place, both macrophagetropic (M-tropic) and T-tropic virus can be transmitted by DCs to T cells. An important role for DCs during the course of HIV-1 infection is indi...

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...repancies could be due to several parameters, one of which is the state of DC maturation, as we now report. MATERIALS AND METHODS Cells. Dendritic cells were generated from the blood of normal donors =-=(2, 23)-=-. Peripheral blood mononuclear cells PBMC were isolated by sedimentation in Ficoll-Hypaque, and 5 � 10 7 PBMC were plated per 100-mm-diameter culture plastic dish in RPMI (Gibco) supplemented with 1% ...

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...HIV-1 infection during DC differentiation. The DCs develop from CD14 � blood monocytes when cultured with IL-4 and GM-CSF and then complete their differentiation and maturation in the presence of MCM =-=(2, 17, 23)-=- that includes products of inflammation like IL-1 and tumor necrosis factor alpha (21). The DCs generated do not divide; the cells differentiated only. Cells are in G0/G1 as determined by propidium io...

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...ction of monocytes, immature DCs (iDC), and mature DCs (mDC). (A) Early and late stage of reverse transcription. The different cell populations were pulsed with HIV-1 Ba-L (2 ng of p24 for 10 5 cells =-=[11]-=-). Cell lysates were prepared from 5 � 10 4 cells at the indicated times postinfection and reverse transcripts were amplified by PCR and compared to graded doses of ACH-2 cells. The left panel has T- ...

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...ultured with IL-4 and GM-CSF and then complete their differentiation and maturation in the presence of MCM (2, 17, 23) that includes products of inflammation like IL-1 and tumor necrosis factor alpha =-=(21)-=-. The DCs generated do not divide; the cells differentiated only. Cells are in G0/G1 as determined by propidium iodide and fluorescenceactivated cell sorting (FACS) (see Fig. 3C). Also, these cells do...

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...udy virus was scratched into skin organ cultures, and then the emigrating DCs were analyzed for capture of infectious HIV-1. It was found that only M-tropic virus was captured by DCs in the epidermis =-=(22)-=-. Our findings are entirely compatible and provide large numbers of these immature DCs for further study. Selective replication of M-tropic HIV-1 by DCs derived by CD34 � blood progenitors has also be...

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...nsive replication and leading to the death of CD4 � T cells (4–6, 19, 28). On the other hand, there are divergent findings with respect to the direct contribution of purified DCs to HIV-1 replication =-=(3, 12, 29)-=-. There is evidence that DCs replicate virus (14, 18, 26) and * Corresponding author. Mailing address: Laboratory of Cellular Physiology and Immunology, Rockefeller University, 1230 York Ave., New Yor...

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...ium fluxes in immature DCs (9). The behavior of immature DCs was strikingly different from that of mature DCs, which did not support the formation of LTR/gag sequences and produced little or no RTase =-=(5, 11, 20, 30)-=- (Fig. 1). Mature DCs efficiently transmit HIV-1 to T cells. To pursue the capacity of immature and mature DCs to disseminate HIV1, each type of DC was pulsed with virus and washed, and 24 h later, T ...

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...d by HIV-1 produced by immature DCs after 11 days of infection. 6 days in GM-CSF and IL-4, they acquired the characteristics of immature DCs (HLA-DR � , CD14 low , CD83 low , p55 low , and CD25 low ) =-=(9)-=-. At this stage of maturation, HIV-1 entered and replicated extensively, forming large numbers of full-length, LTR/gag-containing sequences (Fig. 1A, right) and releasing RTase (Fig. 1B). Moreover, th...

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...lls (4–6, 19, 28). On the other hand, there are divergent findings with respect to the direct contribution of purified DCs to HIV-1 replication (3, 12, 29). There is evidence that DCs replicate virus =-=(14, 18, 26)-=- and * Corresponding author. Mailing address: Laboratory of Cellular Physiology and Immunology, Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7986. Fax: (212) 327-8875. ...

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...ls Efficiently Transmit both M- and T-Tropic Virus to T Cells REFERENCES CONTENT ALERTS Angela Granelli-Piperno, Elena Delgado, Victoria Finkel, William Paxton and Ralph M. Steinman J. Virol. 1998, 72=-=(4)-=-:2733. Updated information and services can be found at: http://jvi.asm.org/content/72/4/2733 These include: This article cites 25 articles, 11 of which can be accessed free at: http://jvi.asm.org/con...

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...gs are entirely compatible and provide large numbers of these immature DCs for further study. Selective replication of M-tropic HIV-1 by DCs derived by CD34 � blood progenitors has also been reported =-=(7)-=-. When DCs are derived from the CD34 � progenitors, there is extensive cell replication. However, DCs that develop from monocytes with cytokines do not proliferate (Fig. 3C). It is known that there is...

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...rast to immature DCs, do not support HIV-1 replication (Fig. 1). Also monocytes only migrate to macrophage-rich regions of lymphoid tissues (24), whereas DCs migrate to the T-cell areas of the organs =-=(1, 13)-=-. A second role for DCs is to transfer HIV-1 during their normal interactions with T cells. We find that when DCs mature, they lose the capacity to replicate HIV-1 but can nonetheless transmit both M-...

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...nsive replication and leading to the death of CD4 � T cells (4–6, 19, 28). On the other hand, there are divergent findings with respect to the direct contribution of purified DCs to HIV-1 replication =-=(3, 12, 29)-=-. There is evidence that DCs replicate virus (14, 18, 26) and * Corresponding author. Mailing address: Laboratory of Cellular Physiology and Immunology, Rockefeller University, 1230 York Ave., New Yor...

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...lls (4–6, 19, 28). On the other hand, there are divergent findings with respect to the direct contribution of purified DCs to HIV-1 replication (3, 12, 29). There is evidence that DCs replicate virus =-=(14, 18, 26)-=- and * Corresponding author. Mailing address: Laboratory of Cellular Physiology and Immunology, Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7986. Fax: (212) 327-8875. ...

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...le for HIV-1 transmission (27, 32). Blood monocytes, in contrast to immature DCs, do not support HIV-1 replication (Fig. 1). Also monocytes only migrate to macrophage-rich regions of lymphoid tissues =-=(24)-=-, whereas DCs migrate to the T-cell areas of the organs (1, 13). A second role for DCs is to transfer HIV-1 during their normal interactions with T cells. We find that when DCs mature, they lose the c...

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...nsive replication and leading to the death of CD4 � T cells (4–6, 19, 28). On the other hand, there are divergent findings with respect to the direct contribution of purified DCs to HIV-1 replication =-=(3, 12, 29)-=-. There is evidence that DCs replicate virus (14, 18, 26) and * Corresponding author. Mailing address: Laboratory of Cellular Physiology and Immunology, Rockefeller University, 1230 York Ave., New Yor...

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...mpairs their antigen-presenting cell function (10, 16), while other reports demonstrate that DCs are not productively infected but nevertheless promote viral replication upon interaction with T cells =-=(5, 20)-=-. These discrepancies could be due to several parameters, one of which is the state of DC maturation, as we now report. MATERIALS AND METHODS Cells. Dendritic cells were generated from the blood of no...

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...kefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-7986. Fax: (212) 327-8875. E-mail: piperno@rockvax.rockefeller.edu. that HIV-1 impairs their antigen-presenting cell function =-=(10, 16)-=-, while other reports demonstrate that DCs are not productively infected but nevertheless promote viral replication upon interaction with T cells (5, 20). These discrepancies could be due to several p...