The Journal of Experimental Medicine

Abi-Rached, Laurent; Parham, Peter

doi: 10.1084/jem.20042558pmid: 15837816

Expression of killer cell Ig-like receptors (KIRs) diversifies human natural killer cell populations and T cell subpopulations. Whereas the major histocompatibility complex class I binding functions of inhibitory KIR are known, specificities for the activating receptors have resisted analysis. To understand better activating KIR and their relationship to inhibitory KIR, we took the approach of reconstructing their natural history and that of Ly49, the analogous system in rodents. A general principle is that inhibitory receptors are ancestral, the activating receptors having evolved from them by mutation. This evolutionary process of functional switch occurs independently in different species to yield activating KIR and Ly49 genes with similar signaling domains. Selecting such convergent evolution were the signaling adaptors, which are older and more conserved than any KIR or Ly49. After functional shift, further activating receptors form through recombination and gene duplication. Activating receptors are short lived and evolved recurrently, showing they are subject to conflicting selections, consistent with activating KIR's association with resistance to infection, reproductive success, and susceptibility to autoimmunity. Our analysis suggests a two-stage model in which activating KIR or Ly49 are initially subject to positive selection that rapidly increases their frequency, followed by negative selection that decreases their frequency and leads eventually to loss. Footnotes Abbreviations used: CYT, cytoplasmic tail; ITIM, inhibitory tyrosine-containing immunomotif; KIR, killer cell Ig-like receptor; LTK, long-tailed KIR; MYA, million yr ago; NJ, neighbor joining; STK, short-tailed KIR; TM, transmembrane domain. Submitted: 16 December 2004 Accepted: 10 March 2005

Van Epps, Heather L.

doi: 10.1084/jem2018iti1pmid: N/A

Katayama, Yoshio; Hidalgo, Andrés; Chang, Jungshan; Peired, Anna; Frenette, Paul S.

doi: 10.1084/jem.20042014pmid: 15824084

The selectin family of adhesion molecules and their glycoconjugated ligands are essential for blood polymorphonuclear neutrophil (PMN) extravasation into inflammatory and infectious sites. However, E-selectin ligands on PMNs are not well characterized. We show here that CD44 immunopurified from G-CSF–differentiated 32D cells or from peripheral blood PMNs binds specifically to E-selectin. In contrast, CD44 extracted from bone marrow stromal or brain endothelial cell lines does not interact with E-selectin, suggesting cell-specific posttranslational modifications of CD44. PMN-derived CD44 binding activity is mediated by sialylated, α(1,3) fucosylated, N-linked glycans. CD44 enables slow leukocyte rolling on E-selectin expressed on inflamed endothelium in vivo and cooperates with P-selectin glycoprotein ligand–1 to recruit neutrophils into thioglycollate-induced peritonitis and staphylococcal enterotoxin A–injected skin pouch. CD44 extracted from human PMNs also binds to E-selectin. Moreover, we demonstrate that CD44 is hypofucosylated in PMNs from a patient with leukocyte adhesion deficiency type II, suggesting that it contributes to the syndrome. These findings thus suggest broader roles for CD44 in the innate immune response and uncover a potential new target for diseases in which selectins play a prominent role. Footnotes Abbreviations used: DKO, double knockout; ESL, E-selectin ligand; FucT, fucosyltransferase; HA, hyaluronic acid; LADII, leukocyte adhesion deficiency type II; OSGE, O -sialoglycoprotein endopeptidase; PB, peripheral blood; PMN, polymorphonuclear neutrophil; PSGL1; P-selectin glycoprotein ligand-1; SEA, staphylococcal enterotoxin A. Y. Katayama, A. Hidalgo, and J. Chang contributed equally to this work. Y. Katayama's present address is Department of Hematology, Oncology and Respiratory Medicine, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Submitted: 29 September 2004 Accepted: 28 February 2005

Jameson, Julie M.; Cauvi, Gabrielle; Sharp, Leslie L.; Witherden, Deborah A.; Havran, Wendy L.

doi: 10.1084/jem.20042057pmid: 15837812

Nonhealing wounds are a major complication of diseases such as diabetes and rheumatoid arthritis. For efficient tissue repair, inflammatory cells must infiltrate into the damaged tissue to orchestrate wound closure. Hyaluronan is involved in the inflammation associated with wound repair and binds the surface of leukocytes infiltrating damaged sites. Skin γδ T cells play specialized roles in keratinocyte proliferation during wound repair. Here, we show that γδ T cells are required for hyaluronan deposition in the extracellular matrix (ECM) and subsequent macrophage infiltration into wound sites. We describe a novel mechanism of control in which γδ T cell–derived keratinocyte growth factors induce epithelial cell production of hyaluronan. In turn, hyaluronan recruits macrophages to the site of damage. These results demonstrate a novel function for skin γδ T cells in inflammation and provide a new perspective on T cell regulation of ECM molecules. Footnotes The online version of this article contains supplemental material. Abbreviations used: DETC, dendritic epidermal T cells; IEL, intraepithelial lymphocyte; ECM, extracellular matrix; GAG, glycosaminoglycan; HAS, hyaluronan synthases; FGF-7, keratinocyte growth factor-1; FGF-10, keratinocyte growth factor-2. Submitted: 6 October 2004 Accepted: 7 March 2005

Van Epps, Heather L.

doi: 10.1084/jem2018iti3pmid: N/A

Bende, Richard J.; Aarts, Wilhelmina M.; Riedl, Robert G.; de Jong, Daphne; Pals, Steven T.; van Noesel, Carel J.M.

doi: 10.1084/jem.20050068pmid: 15837810

We analyzed the structure of antigen receptors of a comprehensive panel of mature B non-Hodgkin's lymphomas (B-NHLs) by comparing, at the amino acid level, their immunoglobulin (Ig)V H -CDR3s with CDR3 sequences present in GenBank. Follicular lymphomas, diffuse large B cell lymphomas, Burkitt's lymphomas, and myelomas expressed a CDR3 repertoire comparable to that of normal B cells. Mantle cell lymphomas and B cell chronic lymphocytic leukemias (B-CLLs) expressed clearly restricted albeit different CDR3 repertoires. Lymphomas of mucosa-associated lymphoid tissues (MALTs) were unique as 8 out of 45 (18%) of gastric- and 13 out of 32 (41%) of salivary gland-MALT lymphomas expressed B cell antigen receptors with strong CDR3 homology to rheumatoid factors (RFs). Of note, the RF-CDR3 homology without exception included N-region–encoded residues in the hypermutated IgV H genes, indicating that they were stringently selected for reactivity with auto-IgG. By in vitro binding studies with 10 MALT lymphoma–derived antibodies, we showed that seven of these cases, of which four with RF-CDR3 homology, indeed possessed strong RF reactivity. Of one MALT lymphoma, functional proof for selection of subclones with high RF affinity was obtained. Interestingly, RF-CDR3 homology and t(11;18) appeared to be mutually exclusive features and RF-CDR3 homology was not encountered in any of the 19 pulmonary MALT lymphomas studied. Footnotes Abbreviations used: Ag, antigen; B-CLL, B cell chronic lymphocytic leukemia; B-NHL, B non-Hodgkin's lymphoma; BCR, B cell antigen receptor; BL, Burkitt's lymphoma; D, diversity; DLBCL, diffuse large B cell lymphoma; FL, follicular lymphoma; FR, framework region; GC, germinal center; HCV, hepatitis C virus; HID, healthy immunized donor; ICV, intraclonal sequence variation; J, joining; LIDA lymphoma- idiotype-derived Ab; MALT, mucosa-associated lymphoid tissue; MCL, mantle cell lymphoma; MZBCL, marginal zone B cell lymphoma; RF, rheumatoid factor; V, variable. Submitted: 7 January 2005 Accepted: 3 March 2005

Van Epps, Heather L.

doi: 10.1084/jem2018iti5pmid: N/A

Pop, Shannon M.; Wong, Carmen P.; Culton, Donna A.; Clarke, Stephen H.; Tisch, Roland

doi: 10.1084/jem.20042398pmid: 15837817

Natural CD4 + CD25 + regulatory T (CD4 + CD25 + T reg) cells play a key role in the immunoregulation of autoimmunity. However, little is known about the interactions between CD4 + CD25 + T reg cells and autoreactive T cells. This is due, in part, to the difficulty of using cell surface markers to identify CD4 + CD25 + T reg cells accurately. Using a novel real-time PCR assay, mRNA copy number of FoxP3, TGFβ1, and interleukin (IL)-10 was measured in single cells to characterize and quantify CD4 + CD25 + T reg cells in the nonobese diabetic (NOD) mouse, a murine model for type 1 diabetes (T1D). The suppressor function of CD4 + CD25 + CD62L hi T cells, mediated by TGFβ, declined in an age-dependent manner. This loss of function coincided with a temporal decrease in the percentage of FoxP3 and TGFβ1 coexpressing T cells within pancreatic lymph node and islet infiltrating CD4 + CD25 + CD62L hi T cells, and was detected in female NOD mice but not in NOD male mice, or NOR or C57BL/6 female mice. These results demonstrate that the majority of FoxP3-positive CD4 + CD25 + T reg cells in NOD mice express TGFβ1 but not IL-10, and that a defect in the maintenance and/or expansion of this pool of immunoregulatory effectors is associated with the progression of T1D. Footnotes Abbreviations used: Ab, antibody; CD62L hi T cells, CD4 + CD25 + CD62L hi T cells; CD62L lo T cells, CD4 + CD25 + CD62L lo T cells; CTLA-4, cytotoxic T lymphocyte–associated antigen 4; GITR, glucocorticoid-induced TNF receptor gene; NOD, nonobese diabetic; PLN, pancreatic LN; rh, recombinant human; T reg, regulatory T; T1D, type 1 diabetes. Submitted: 24 November 2004 Accepted: 11 March 2005

van Gisbergen, Klaas P.J.M.; Sanchez-Hernandez, Marta; Geijtenbeek, Teunis B.H.; van Kooyk, Yvette

doi: 10.1084/jem.20041276pmid: 15837813

Neutrophils are key players of the innate immune system that provide a first line of defense against invading pathogens. However, it is unknown whether neutrophils can interact with dendritic cells (DCs) to modulate adaptive immune responses. We demonstrate that neutrophils strongly cluster with immature DCs and that activated, not resting, neutrophils induce maturation of DCs that enables these DCs to trigger strong T cell proliferation and T helper type 1 polarization of T cells. This neutrophil–DC interaction is driven by the binding of the DC-specific, C-type lectin DC-SIGN to the β 2 -integrin Mac-1. Strikingly, DC-SIGN only interacts with Mac-1 from neutrophils, but not from other leukocytes, mainly because of specific Lewis x carbohydrates that are present on the α M chain of Mac-1 from neutrophils. Furthermore, we show that besides the formation of cellular contact, the tumor necrosis factor-α produced by activated neutrophils is essential for inducing DC maturation. Our data demonstrate that DC-SIGN and Mac-1 define a molecular pathway to establish cellular adhesion between DCs and neutrophils, thereby providing a novel cellular link between innate and adaptive immunity. Footnotes Submitted: 25 June 2004 Accepted: 4 March 2005

Bronte, Vincenzo; Kasic, Tihana; Gri, Giorgia; Gallana, Keti; Borsellino, Giovanna; Marigo, Ilaria; Battistini, Luca; Iafrate, Massimo; Prayer-Galetti, Tommaso; Pagano, Francesco; Viola, Antonella

doi: 10.1084/jem.20042028pmid: 15824085

Immunotherapy may provide valid alternative therapy for patients with hormone-refractory metastatic prostate cancer. However, if the tumor environment exerts a suppressive action on antigen-specific tumor-infiltrating lymphocytes (TIL), immunotherapy will achieve little, if any, success. In this study, we analyzed the modulation of TIL responses by the tumor environment using collagen gel matrix–supported organ cultures of human prostate carcinomas. Our results indicate that human prostatic adenocarcinomas are infiltrated by terminally differentiated cytotoxic T lymphocytes that are, however, in an unresponsive status. We demonstrate the presence of high levels of nitrotyrosines in prostatic TIL, suggesting a local production of peroxynitrites. By inhibiting the activity of arginase and nitric oxide synthase, key enzymes of L-arginine metabolism that are highly expressed in malignant but not in normal prostates, reduced tyrosine nitration and restoration of TIL responsiveness to tumor were achieved. The metabolic control exerted by the tumor on TIL function was confirmed in a transgenic mouse prostate model, which exhibits similarities with human prostate cancer. These results identify a novel and dominant mechanism by which cancers induce immunosuppression in situ and suggest novel strategies for tumor immunotherapy. Footnotes Abbreviations used: ARG, arginase; L-Arg, L-arginine; L-NMMA, N -monomethyl- L-arginine; NO, nitric oxide; NOHA, N -hydroxy-L-arginine; NOS, nitric oxide synthase; PCa, prostate carcinoma; TIA-1, T cell intracellular antigen 1; TIL, tumor-infiltrating lymphocytes; TRAMP, transgenic adenocarcinoma mouse prostate. Submitted: 30 September 2004 Accepted: 2 March 2005