HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Nimura, F. Articles by Tanaka, Y. Search for Related Content PubMed PubMed Citation Articles by Nimura, F. Articles by Tanaka, Y.

---

ORIGINAL RESEARCH ARTICLE

Cross-Linking Cell Surface Chemokine Receptors Leads to Isolation, Activation, and Differentiation of Monocytes into Potent Dendritic Cells

Fumikazu Nimura^*,, Li Feng Zhang^*, Kazu Okuma^*, Reiko Tanaka^*, Hajime Sunakawa, Naoki Yamamoto and Yuetsu Tanaka^*,1

^* Department of Immunology and Department of Oral and Maxillofacial Functional Rehabilitation, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan; and AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan

To whom requests for reprints should be addressed at ¹ Department of Immunology, Graduate School of Medicine, University of the Ryukyus, Uehara 207, Nishihara-cho, Nakagami-gun, Okinawa 903-0215, Japan. E-mail: yuetsu{at}s4.dion.ne.jp

Monocytes express on the cell surface several kinds of chemokine receptors that facilitate chemotaxis followed by differentiation in target tissues. In the present study, we found that a large number of monocytes from peripheral blood mononuclear cells (PBMCs) tightly adhered to plastic cell culture plates precoated with a monoclonal antibody (mAb, clone T312) specific for human CCR5 but not an isotype control after overnight incubation. Soluble T312 did not induce such adhesion, indicating that cross-linking of CCR5 is required for the enhanced adhesion of monocytes. The adhesion was blocked by a PI3-K inhibitor and an anti-CD18 blocking mAb. Following the cross-linking of CCR5, monocytes synthesized high levels of M-CSF, RANTES, MIP-1, and MIP-1ß associated with a readily detectable downmodulation of CD14, CD4, CCR5, and CXCR4 expression. The T312-enriched monocytes differentiated into dendritic cells (DCs) in the presence of interleukin-4 alone. After maturation with ß-interferon, the T312-induced DCs stimulated proliferation of allogeneic naïve CD4⁺ T cells accompanied by the synthesis of high levels of -interferon in vitro. Furthermore, the T312-induced DCs were capable of stimulating antigen-specific human T- and B-cell immune responses in our hu-PBL-SCID mouse system. Finally, screening of other anti-chemokine receptor mAbs showed that select clones of mAbs against CXCR4 and CCR3 were also capable of facilitating enrichment of monocytes similar to T312. These results show that cross-linking of chemokine receptors on monocytes by appropriate mAbs leads to activation and differentiation of monocytes and that the method described herein provides an alternate simple strategy for adherence-based isolation of monocytes and generation of functional DCs.

Key Words: dendritic cell • monocyte • chemokine receptor • human immunodeficiency virus (HIV)

This article has been cited by other articles:

				L. F. Zhang, K. Okuma, R. Tanaka, A. Kodama, K. Kondo, A. A. Ansari, and Y. Tanaka Generation of Mature Dendritic Cells with Unique Phenotype and Function by In Vitro Short-Term Culture of Human Monocytes in the Presence of Interleukin-4 and Interferon-{beta} Experimental Biology and Medicine, June 1, 2008; 233(6): 721 - 731. [Abstract] [Full Text] [PDF]