The Journal of Experimental Medicine

Lauvau, Gregoire; Pamer, Eric G.

doi: 10.1084/jem.193.10.F35pmid: 11369794

Hamad, Abdel Rahim A.; Srikrishnan, Ananth; Mirmonsef, Paria; Broeren, Chris P.M.; June, Carl H.; Pardoll, Drew; Schneck, Jonathan P.

doi: 10.1084/jem.193.10.1113pmid: 11369783

Lymphoproliferative diseases are characterized by massive accumulation of CD4 − CD8 − B220 + (double-negative DN) T cells in peripheral organs. Although evidence indicates these cells are derived from mature autoreactive α/β T cells, the significance of coreceptor downregulation is not known. In this study, we examined the role CD4 coreceptor plays in the survival of repeatedly stimulated T cells. CD4 +/+ and CD4 −/ − T cells from AND T cell receptor (TCR) transgenic mice exhibited similar phenotypes after antigenic stimulation, but the CD4 −/ − T cells survived in much larger numbers than the CD4 +/+ cells upon primary and secondary major histocompatibility complex (MHC)/peptide stimulation. Enhanced survival of CD4 −/ − T cells was due to decreased apoptosis rather than enhanced proliferation. Similarly, circumvention of the CD4/MHC interaction by using a surrogate TCR ligand that does not engage CD4 led to significant enhancement of CD4 +/+ cells than when stimulated with MHC/peptide. Finally, we generated DN B220 + T cells using an in vitro model system and showed they were more tolerant to chronic stimulation than CD4 +/+ cells. Together, these results indicate that coreceptor engagement controls expansion of normal T cells. In the absence of coreceptor, T cells survive chronic stimulation and express B220 as seen in autoimmune lymphoproliferative diseases. CD4 coreceptor double-negative T cell lymphoproliferation B220 apoptosis Footnotes C.H. June's present address is Abrameson Family Cancer Research Institute and Dept. of Molecular and Cellular Engineering, University of Pennsylvania, Philadelphia, PA 19104. Abbreviations used in this paper: AICD, activation-induced cell death; CFSE, 5- and 6-carboxyfluorescein diacetate succinimidyl ester; DN, double-negative; MCC, moth cytochrome c; tg, transgenic. Submitted: 13 October 2000 Revision requested 21 February 2001 Accepted: 9 April 2001

Forsyth, Christopher B.; Solovjov, Dmitry A.; Ugarova, Tatiana P.; Plow, Edward F.

doi: 10.1084/jem.193.10.1123pmid: 11369784

Leukocyte migration is the hallmark of inflammation, and integrin α M β 2 and its ligand fibrinogen (Fg) are key participants in this cellular response. Cells expressing wild-type or mutant α M β 2 and Fg or its derivatives have been used to dissect the molecular requirements for this receptor–ligand pair to mediate cell migration. The major conclusions are that (a) Fg, its D fragment, and its P1 and P2 α M β 2 recognition peptides support a chemotactic response; (b) when the I domain of α L was replaced with the I domain of α M , the chimeric receptor supported cell migration to Fg; however, the α M subunit, containing the I domain but lacking the β 2 subunit, supported migration poorly, thus, the α M I domain is necessary but not sufficient to support chemotaxis, and efficient migration requires the β 2 subunit and α M I domain; and (c) in addition to supporting cell migration, P2 enhanced α M β 2 -mediated chemotaxis to Fg and the P1 peptide. This activation was associated with exposure of the activation-dependent epitope recognized by monoclonal antibody 7E3 and was observed also with human neutrophils. Taken together, these data define specific molecular requirements for α M β 2 to mediate cell migration to Fg derivatives and assign a novel proinflammatory activity to the P2 peptide. adhesion molecules fibrinogen integrin CD11b/CD18 inflammation Footnotes Abbreviations used in this paper: Fg, fibrinogen; Fn, fibronectin; HPF, high power field; ICAM, intercellular adhesion molecule; MIDAS, metal ion–dependent adhesion site; NIF, neutrophil inhibitory factor; WT, wild-type. Submitted: 3 December 1999 Revision requested 22 March 2001 Accepted: 9 April 2001

Angeli, Véronique; Faveeuw, Christelle; Roye, Olivier; Fontaine, Josette; Teissier, Elisabeth; Capron, André; Wolowczuk, Isabelle; Capron, Monique; Trottein, François

doi: 10.1084/jem.193.10.1135pmid: 11369785

Epidermal Langerhans cells (LCs) play a key role in immune defense mechanisms and in numerous immunological disorders. In this report, we show that percutaneous infection of C57BL/6 mice with the helminth parasite Schistosoma mansoni leads to the activation of LCs but, surprisingly, to their retention in the epidermis. Moreover, using an experimental model of LC migration induced by tumor necrosis factor (TNF)-α, we show that parasites transiently impair the departure of LCs from the epidermis and their subsequent accumulation as dendritic cells in the draining lymph nodes. The inhibitory effect is mediated by soluble lipophilic factors released by the parasites and not by host-derived antiinflammatory cytokines, such as interleukin-10. We find that prostaglandin (PG)D 2 , but not the other major eicosanoids produced by the parasites, specifically impedes the TNF-α–triggered migration of LCs through the adenylate cyclase–coupled PGD 2 receptor (DP receptor). Moreover, the potent DP receptor antagonist BW A868C restores LC migration in infected mice. Finally, in a model of contact allergen-induced LC migration, we show that activation of the DP receptor not only inhibits LC emigration but also dramatically reduces the contact hypersensitivity responses after challenge. Taken together, we propose that the inhibition of LC migration could represent an additional stratagem for the schistosomes to escape the host immune system and that PGD 2 may play a key role in the control of cutaneous immune responses. dendritic cells migration Schistosoma eicosanoids cAMP Footnotes Abbreviations used in this paper: AC, adenylate cyclase; CHS, contact hypersensitivity; CCR, CC chemokine receptor; DC, dendritic cell; DP receptor, PGD 2 receptor; EIA, enzyme immunoassay; ES, excreted/secreted; HETE, 5-hydroxyeicosatetranoic acid; KC, keratinocyte; KO, knockout; LC, Langerhans cell; LT, leukotriene; RT, reverse transcription; SESP, schistosomula ES products; SLN, skin-draining LN; WT, wild-type. Submitted: 30 November 2000 Revision requested 15 March 2001 Accepted: 17 April 2001

Chan, Jason R.; Hyduk, Sharon J.; Cybulsky, Myron I.

doi: 10.1084/jem.193.10.1149pmid: 11369786

Chemoattractants and chemokines induce arrest of rolling monocytes during emigration from blood into tissues. In this study, we demonstrated that α4 integrin affinity for vascular cell adhesion molecule (VCAM)-1 was upregulated rapidly and transiently by chemoattractants and stromal cell–derived factor (SDF)-1α and mediated monocyte arrest. α4 integrin affinity changes were detected and blocked using soluble VCAM-1/Fc (sVCAM-1/Fc). In a flow cytometry assay, markedly increased sVCAM-1/Fc binding to human blood monocytes or U937 cells transfected with formyl peptide (FP) receptor was detected 30 s after FP or SDF-1α treatment and declined after 2 min. In a parallel plate flow chamber assay, FP, C5a, platelet-activating factor, or SDF-1α coimmobilized with VCAM-1 induced leukocyte arrest, which was blocked by inclusion of sVCAM-1/Fc but not soluble nonimmune immunoglobulin G in the assay buffer. stromal cell–derived factor 1α chemokines formyl peptide very late antigen 4 inflammation Footnotes Abbreviations used in this paper: CXCR, CXC chemokine receptor; cytD, cytochalasin D; FBS, fetal bovine serum; FP, formyl peptide; ICAM, intercellular adhesion molecule; MCP, monocyte chemotactic protein; PAF, platelet-activating factor; PTx, pertussis toxin; RANTES, regulated upon activation, normal T cell expressed and secreted; SDF, stromal cell–derived factor; VCAM, vascular cell adhesion molecule. Submitted: 22 November 2000 Revision requested 30 March 2001 Accepted: 16 April 2001

Yen, Jeng-Hsien; Moore, Brenda E.; Nakajima, Takako; Scholl, Dirk; Schaid, Daniel J.; Weyand, Cornelia M.; Goronzy, Jörg J.

doi: 10.1084/jem.193.10.1159pmid: 11369787

Rheumatoid arthritis (RA) is a heterogeneous syndrome of which a subset of patients develops vascular inflammation. The genetic determinants that confer risk for rheumatoid vasculitis are not known, but patients with vascular complications are known to have an expansion of CD4 + CD28 null T cells, a cell population potentially involved in endothelial damage. CD4 + CD28 null T cell clones isolated from RA patients with vasculitis were found to express killer cell immunoglobulin–like receptors (KIRs) with the stimulatory KIR2DS2 often present in the absence of opposing inhibitory receptors with related specificities. To test the hypothesis that the KIR2DS2 gene is involved in the development of vasculitis, association studies were performed. The KIR2DS2 gene was significantly enriched among patients with rheumatoid vasculitis compared with normal individuals (odds ratio 5.56, P = 0.001) and patients with RA but no vasculitis (odds ratio 7.96, P = 0.001). Also, the distribution of human histocompatibility leukocyte antigen (HLA)-C, the putative ligand for KIRs, was significantly different in patients with rheumatoid vasculitis in comparison with the control populations. These data suggest that HLA class I–recognizing receptors and HLA class I genes are genetic risk determinants that modulate the pattern of RA expression. Specifically, KIR2DS2 in conjunction with the appropriate HLA-C ligand may have a role in vascular damage by regulating CD4 + CD28 null T cells. killer cell immunoglobulin–like receptor T cell genetic susceptibility rheumatoid arthritis vasculitis Footnotes Abbreviations used in this paper: KIR, killer cell Ig–like receptor; RA, rheumatoid arthritis; RT, reverse transcription. Submitted: 20 April 2000 Revision requested 20 March 2001 Accepted: 26 March 2001

Dingjan, Gemma M.; Middendorp, Sabine; Dahlenborg, Katarina; Maas, Alex; Grosveld, Frank; Hendriks, Rudolf W.

doi: 10.1084/jem.193.10.1169pmid: 11369788

Bruton's tyrosine kinase (Btk) is a nonreceptor tyrosine kinase involved in precursor B (pre-B) cell receptor signaling. Here we demonstrate that Btk-deficient mice have an ∼50% reduction in the frequency of immunoglobulin (Ig) λ light chain expression, already at the immature B cell stage in the bone marrow. Conversely, transgenic mice expressing the activated mutant Btk E41K showed increased λ usage. As the κ/λ ratio is dependent on (a) the level and kinetics of κ and λ locus activation, (b) the life span of pre-B cells, and (c) the extent of receptor editing, we analyzed the role of Btk in these processes. Enforced expression of the Bcl-2 apoptosis inhibitor did not alter the Btk dependence of λ usage. Crossing 3-83μδ autoantibody transgenic mice into Btk-deficient mice showed that Btk is not essential for receptor editing. Also, Btk-deficient surface Ig + B cells that were generated in vitro in interleukin 7-driven bone marrow cultures manifested reduced λ usage. An intrinsic defect in λ locus recombination was further supported by the finding in Btk-deficient mice of reduced λ usage in the fraction of pre-B cells that express light chains in their cytoplasm. These results implicate Btk in the regulation of the activation of the λ locus for V(D)J recombination in pre-B cells. Btk B lymphocytes Ig L chain receptor editing V(D)J rearrangements Footnotes Abbreviations used in this paper: BCR, B cell receptor; Btk, Bruton's tyrosine kinase; PH, pleckstrin homology; WT, wild-type; Xid, X-linked immunodeficiency; XLA, X-linked agammaglobulinemia. Submitted: 26 January 2001 Revision requested 10 April 2001 Accepted: 13 April 2001

Wong, Phillip; Barton, Gregory M.; Forbush, Katherine A.; Rudensky, Alexander Y.

doi: 10.1084/jem.193.10.1179pmid: 11369789

Intrathymic self-peptide–major histocompatibility complex class II (MHC) molecules shape the T cell repertoire through positive and negative selection of immature CD4 + CD8 + thymocytes. By analyzing the development of MHC class II–restricted T cell receptor (TCR) transgenic T cells under conditions in which the endogenous peptide repertoire is altered, we show that self-peptide–MHC complexes are also involved in setting T cell activation thresholds. This occurs through changes in the expression level of molecules on thymocytes that influence the sensitivity of TCR signaling. Our results suggest that the endogenous peptide repertoire modulates T cell responsiveness in the thymus in order to enforce tolerance to self-antigens. T cell antigen receptors thymus tolerance transgenic mice CD5 Footnotes Abbreviations used in this paper: APC, allophycocyanin; β2m, β2-microglobulin; CLIP, class II–associated Ii peptide; DP, double positive; Ii, invariant chain; PerCP, peridinin chlorophyll protein; RAG, recombination activating gene; SP, single positive. Submitted: 6 December 2000 Revision requested 5 March 2001 Accepted: 16 April 2001

Probst-Kepper, Michael; Stroobant, Vincent; Kridel, Robert; Gaugler, Béatrice; Landry, Claire; Brasseur, Francis; Cosyns, Jean-Pierre; Weynand, Birgit; Boon, Thierry; Van den Eynde, Benoit J.

doi: 10.1084/jem.193.10.1189pmid: 11369790

We show that cytotoxic T lymphocytes (CTLs) infiltrating a kidney tumor recognize a peptide encoded by an alternative open reading frame (ORF) of the macrophage colony-stimulating factor (M-CSF) gene. Remarkably, this alternative ORF, which is translated in many tumors concurrently with the major ORF, is also translated in some tissues that do not produce M-CSF, such as liver and kidney. Such a dissociation of the translation of two overlapping ORFs from the same gene is unexpected. The antigenic peptide encoded by the alternative ORF is presented by human histocompatibility leukocyte antigen (HLA)-B*3501 and has a length of 14 residues. Peptide elution indicated that tumor cells naturally present this 14 mer, which is the longest peptide known to be recognized by CTLs. Binding studies of peptide analogues suggest that it binds by its two extremities and bulges out of the HLA groove to compensate for its length. renal cell carcinoma HLA-B35 translation peptide binding natural peptide Footnotes Abbreviations used in this paper: FL, fluorescein; ORF, open reading frame; RCC, renal cell carcinoma; TIL, tumor-infiltrating lymphocyte. B. Gaugler's present address is Laboratoire d'Immunologie des Tumeurs, Institut Paoli-Calmettes, Marseille 13009, France. Submitted: 27 March 2001 Accepted: 18 April 2001

Drake, Penelope M.; Gunn, Michael D.; Charo, Israel F.; Tsou, Chia-Lin; Zhou, Yan; Huang, Ling; Fisher, Susan J.

doi: 10.1084/jem.193.10.1199pmid: 11369791

During human pregnancy, the specialized epithelial cells of the placenta (cytotrophoblasts) come into direct contact with immune cells in several locations. In the fetal compartment of the placenta, cytotrophoblast stem cells lie adjacent to macrophages (Hofbauer cells) that reside within the chorionic villus stroma. At sites of placental attachment to the mother, invasive cytotrophoblasts encounter specialized maternal natural killer (NK) cells (CD56 bright ), macrophages, and T cells that accumulate within the uterine wall during pregnancy. Here we tested the hypothesis that fetal cytotrophoblasts can direct the migration of these maternal immune cells. First, we assayed the chemotactic activity of cytotrophoblast conditioned medium samples, using human peripheral blood mononuclear cells as targets. The placental samples preferentially attracted NK cells (both CD56 dim and CD56 bright ), monocytes, and T cells, suggesting that our hypothesis was correct. A screen to identify chemokine activity through the induction of a Ca 2 + flux in cells transfected with individual chemokine receptors suggested that cytotrophoblasts secreted monocyte inflammatory protein (MIP)-1α. This was confirmed by localizing the corresponding mRNA and protein, both in vitro and in vivo. MIP-1α protein in conditioned medium was further characterized by immunoblotting and enzyme-linked immunosorbent assay. Immunodepletion of MIP-1α from cytotrophoblast conditioned medium showed that this chemokine was responsible for a significant portion of the induced monocyte and CD56 bright NK cell chemotax-is. These data suggest the specific conclusion that cytotrophoblasts can attract monocytes and CD56 bright NK cells by producing MIP-1α and the more general hypothesis that these cells may organize and act on leukocytes at the maternal–fetal interface. chemokine pregnancy leukocyte immune tolerance trophoblast Footnotes Abbreviations used in this paper: CM, conditioned medium; DGLs, decidual granulated leukocytes; MIP-1α, monocyte inflammatory protein 1α; SFM, serum-free medium. Submitted: 4 December 2000 Revision requested 9 April 2001 Accepted: 17 April 2001