Immunologic Research 1998; 17/3:313-327

The Intercellular Adhesion Molecule (ICAM) Family of Proteins

New Membersand NovelFunctions
ICOS Corporation, Bothell, WA.

Abstract
Macromolecular adhesive associations between cells are important for transmitting spatial and temporal information that is critical for immune system function. One such group of proteins, the intercellular adhesion molecules (ICAMs), has grown as newly identified members are revealed. In addition, the functions of the ICAMs, in general, have begun to be better understood, including intracellular signaling events. This information has led to the design of novel therapeutic agents that may prove effective in a variety of disease states.

Key Words
Intercellular adhesion molecules Signaling Therapeutics

Introduction
Intercellular adhesion molecules (ICAMs), are a family of related cell surface glycoproteins as defined by the criteria of polypeptide sequence homology, conserved [~2 integrin ligands, and, in most cases, genetic linkage. Each ICAM protein has extracellular C-type immunoglobulin-like domains ranging in number from two to nine making them members of the immunoglobulin superfamily (1). Despite their structural similarity and integrinbinding capability, these proteins are expressed on a wide range of cell types, with very distinct patterns of gene regulation and downstream effector behaviors. Recent data reveal

that ICAM engagement triggers outside-in signals that effect cellular responses. This article summarizes the current level of understanding regarding ICAM-triggered outside-in signaling events and recent developments in the area of newly characterized ICAM proteins. With the discovery of new members of this protein family, we propose that a common naming system be adopted for the currently known ICAM family members. These and any additional members should be given numerical designations based on the chronology of their discovery. Given the numerical assignments to ICAM- 1, ICAM-2, and ICAM-3, we believe it is appropriate to designate the

Dr. Joel S. Hayt]ick [COS Corp. 22021 20th Ave. SE Bothell, WA 98021 E-mail: jhayflick@icos.com

9 1998 Humana Press Inc. 0257-277X/98/ 17/3:313-327/$11.75

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Landsteiner-Weiner (LW) blood group antigen as ICAM-4 and telencephalin as ICAM-5.

ICAM-! (CD54)

ICAM-I is a 76-114-kDa surface glycoprotein that has five extracellular immunoglobulin-like domains (2,3). A motif within domains 1 and 3 of ICAM-1 has been shown to be critical for binding to the ~2 integrins lymphocyte function-associated antigen (LFA)- 1 (CD 1 I a/CD 18) and Mac- 1, respectively (CD1 lb/CDI8) (4,5). Integrin binding to ICAM- 1 is regulated by affinity and avidity changes. Through these interactions with LFA- 1 and Mac- 1, ICAM- 1 plays an important role in leukocyte-leukocyte, epithelialleukocyte, and endothelial-leukocyte contact. ICAM-2 (CD1021 As part of both the cellular and humoral immune respone during leukocyte-leukocyte The existence of ICAM-2, a second [32 contact, ICAM-1 serves as an accessory pro- integrin ligand, was inferred by the inability tein for antigen receptor activation on B and T of ICAM- 1-specific antibodies to completely cells (6) (see Recent Developments in Out- block the adhesion of leukocytes to endotheside-In Signaling and Cytoplasmic Functions lial monolayers (13). ICAM-2 is expressed of ICAM Proteins). Expressed basally on constitutively at low levels on resting lymphomany cell lineages ofmesynchymal origin, the cytes and monocytes and at higher levels on levels of surface ICAM-1 protein are mark- vascular endothelial cells, but is not upregulated edly upregulated at the transcriptional level in response to inflammatory stimuli as is by inflammatory stimuli like tumor necrosis ICAM-1 (14). Its two extracellular immunofactor (TNF), interferon yand lipopolysaccha- globulin-like domains are most closely related ride (LPS). The protein also plays a major role to the amino terminal domains of ICAM- l (15) during leukocyte transmigration from the vas- (see Table I for comparisons) and via binding cular compartment during immune surveil- of LFA-l, ICAM-2 plays an important role in lance. In this context, leukocytes are tethered endothelial-leukocyte interactions associated via specific carbohydrate moities to selectins, with inflammation (14). Evidence also exists as well as very late antigen-4 (VLA-4) bind- to suggest that Mac-1 is a ligand for ICAM-2 ing to VCAM- 1, both of which mediate leuko- in a manner similar to that previously demoncyte rolling on cytokine-activated endothelial strated for ICAM-1 (16). cells. Leukocytes bearing high-affinity LFA- 1 An outstanding issue yet to be resolved is or Mac-I then adhere firmly to the endothe- why endothelial cells express two different lium by binding to ICAM-I to initiate the ICAM proteins that have the same integrin process of transmigratory egress from the ligand, LFA-I. Could their relative sizes be a vasculature (7). Given the aforementioned clue to their roles in a multi-step process that roles of ICAM- 1, mice deficient in its expres- results in firm leukocyte attachment to actision through targeted disruption of the gene vated endothelium? The shape of the ICAM-1

have numerous inflammatory response abnormalities, including impaired neutrophil emigration, an inability to stimulate T cell proliferation in mixed allogeneic lymphocyte cultures, resistance to septic shock, and a reduced susceptibility to cerebral ischemiareperfusion injury (8-10). Interestingly, ICAM-I is utilized by several pathogens to infect host cells and evade immune detection. It was identified as the major cell-surface receptor for rhinovirus and Plasmodium falciparum (4,11). HIV particles have been found to incorporate ICAM-1 into the viral envelope during the budding process, resulting in increased infectivity that is ICAM1/LFA-I dependent (12).

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Table !, Human ICAM family Domains I and 2 (% amino acid identity)"

ICAM-1 ICAM-2 ICAM-3 ICAM-4 ICAM-5 34 38 30 25

34

77 38

26 27 27

64 42 64 28

36 30 33 30 31

36

aDomain 1, boldface; domain 2, italic.

extracellular region was found to be an extended rod of approx 16.6 nm (4). Each immunoglobulin domain therefore forms subregions of 3-3.5 nm. ICAM-2 should measure 6.5 nm putting its LFA-1 binding site much closer to the negatively charged plasma membrane. In a hypothetical stepwise model, LFA- 1 would engage ICAM-1 in its first domain at about 16 nm from the lipid surface. This binding could displace negatively charged surface molecules on the opposing cells and elicit bending of the ICAM bringing domain 3, at approx 9 nm from the surface, within proximity of Mac- 1 for binding to occur. As the cells are drawn together and if repulsion by the negatively charged glycocalyx on each cell's surface is reduced in this manner, then LFA-1 binding to ICAM-2 at about 6 nm from the surface could then occur resulting in firm attachment.

ICAM- (CDSO,ICAM-R)
ICAM-3 was sought by us and others based on data that ICAM-l-specific and ICAM-2specific antibody treatment partially blocked LFA- l-dependent homotypic T cell aggregation (14). Utilizing primer degeneracy and the a m i n o acid sequence conservation between ICAM-1, -2 and other C2-type immunoglobulin domains, a polymerase chain reaction product was generated and used to probe a cDNA library from the human myelomonocytic cell line, HL60. The deduced amino acid sequence revealed that ICAM-3,

like ICAM-1, was composed of five extracellular i m m u n o g l o b u l i n - l i k e domains (17-19). Domain-specific sequence homology with ICAM-1 ranged from 36-77%, with domain 2 being most highly conserved (see Table 1 for comparison of the first two domains) and the region of least homology being the cytoplasmic tail, which may be reflected by differences in outside-in signals triggered by engagement of ICAM-3 versus ICAM-1 (see Recent Developments in Outside-In Signaling and Cytoplasmic Functions of ICAM Proteins). The integrin-binding properties and expression pattern of ICAM-3 partially overlap with those of ICAM-1 and -2. ICAM-3 binds LFA- l, but apparently not Mac- 1 (20,21). The ICAM-3 binding site within LFA-1 has been localized to regions within the evolutionarily conserved I domain of the ~-chain (CDI l a) that overlaps with, but is distinct from, the site for ICAM- 1 binding. This observation is useful for those laboratories trying to develop ICAM-specific functional inhibitors (see Novel Areas of Therapeutic Development Involving ICAM Biology) (22,23). In addition to LFA- 1 binding, ICAM-3 has been identified as the prefered ligand for O~d~2, a recently discovered member of the [~2 integrin family (24). In humans, this integrin is expressed highly on certain tissue leukocytes, particularly macrophages, and on subsets of blood leukocytes (24). Given that the normal expres-

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sion pattern of ICAM-3 is high on all leukocytes, but lacking on endothelium, it is likely that it plays an important role in early events during leukocyte-leukocyte contact, for example B cell activation mediated by T cell help or T cell activation by antigen-presenting cells (APCs). Monoclonal antibodies (MAbs) to ICAM-3 have been shown to inhibit peripheral blood lymphocyte proliferation in response to phytohemagglutinin, allogeneic stimulator cells, and specific antigen (25,26). ICAM-3 is highly expressed on epidermal Langerhans cells where presentation of alloantigen or peptide antigen by these cells to CD4 + T cells may require an interaction between ICAM-3 and LFA- 1 (27-30). Thus, ICAM-3 has an important role in T cell stimulation driven by Langerhans cells. However, when a chronic inflammatory reaction in the skin occurs, T cell stimulation leads to pathological conditions such as psoriasis, so blockade of ICAM-3/LFA-1 binding may be an attractive therapeutic target (see Novel Areas of Therapeutic Development Involving ICAM Biology) (31). Unlike its close relatives, ICAM-3 is not expressed on normal or activated endothelium precluding its involvement in leukocyte-endothelial adhesion under normal circumstances. Interestingly, during neovascularization associated with tumor growth, ICAM-3 expression is induced on the vascular endothelium where its role is yet to be defined (32,33). Messenger RNA blot analysis demonstrated the presence of transcripts for ICAM-3 in leukocytes isolated from numerous primates species, dogs, and rabbits, but not in rodent cells (P Kilgannon, unpublished data). Consistent with these results, attempts to clone the rodent ICAM-3 homolog have been unsuccessful and it is likely that rodents do not have the genetic homolog of ICAM-3. Although this situation is unusual, it is not without precedent. Interleukin 8 (IL8) is another case wherein an

important immune cell mediator found in humans is absent from the rodent genome (34). Since the major integrin receptors for ICAM-3 are conserved in rodent species, we hypothesize that another protein performs the functions of ICAM-3, thus compensating for the absence of the gene.

ICAM-4 (ICAM-LW, LW Blood Group Glycoprotein)

The fourth member of the ICAM family, as determined by the chronology of its discovery as such, is a blood group antigen known as the Landsteiner Weiner (LW) glycoprotein (35). It was discovered at the same time as, and for many years confused with, the Rh blood group system, since both Rh and LW deficiencies are phenotypically linked (36). Genetically, however, these loci are distinct (37,38). This 42-kDa glycoprotein has been found only on erythrocytes where it is linked to the submembranous cytoskeleton. Recently, the deduced polypeptide sequence showed that ICAM-4 contains two extracellular immunoglobulin-like domains with significant homology to the first two domains of the other members of the family (see Table 1). Consistent with this homology, ICAM-4 has been shown to bind to LFA- 1 and to Mac- 1 in static adhesion assays (39). This interaction might be involved in erythrocyte turnover within the spleen, given the selective expression of ~d~2 on macrophages in splenic red pulp cords and ICAM-4 on erythrocytes (24).

ICAM-$ (Telencephalin)
Related to the other ICAM proteins previously described by polypeptide sequence conservation, ICAM-5 is the most recently identified and perhaps the most unique member of the ICAM family. The deduced protein sequence has nine i m m u n o g l o b u l i n - l i k e domains and is expressed in rats and humans

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only on telencephalic neurons where it is localized to somatic and dendritic membranes, hence its initial name telencephalin (40,41) (P Kilgannon, unpublished data). The five amino terminal immunoglobulin domains of ICAM-5 are homologous to the five domains of ICAM-1 and -3 whereas domains 6-8 appear to have arisen from exon duplication of domain 5. The ninth domain is least like those of the other ICAMs, resembling more closely the C-type immunoglobulin domains of other adhesion molecules expressed in the central nervous system such as chick axonin-1 and neurofascin. Domain 1 of ICAM-5 preserves the binding motif shown to be required for other ICAM/integrin interactions and recent results found that ICAM-5 bound to LFA-1 and Mac- 1 (41,42) (P Kilgannon, unpublished data). From these data, it is proposed that ICAM-5 is involved in neuronal-glial cell interactions under normal conditions or neuronal-leukocytic interactions in pathologic settings where lymphocyte influx into this "immunologically priviledged" site does not normally occur.

Similarities and Differences Within the ICAM Family

Structural, genetic, and functional similarities among the ICAM proteins serve to group these proteins together. It is likely that one of the ICAM genes is the primordial gene from which all others were derived. The genes for ICAM- 1, -3, -4, and -5 have all been mapped to human chromosome 19pl 3.2-p 13.3, whereas ICAM-2 resides on human c h r o m o s o m e 17q23-25 (37,43-46) (P Kilgannon, WM Gallatin, unpublished data). Since four of the five known ICAM genes in humans are clustered, this suggests that localized gene duplication events are responsible for generating diversity, a situation that is quite common for related genes within a given family, for instance, chemokines and their receptors (47-49).

As the basis for ICAM gene duplication, the individual immunoglobulin-like domains are each found within different exons reflecting what is the smallest functional unit that has been maintained by evolutionary conservation (1,50). As discussed previously, the ICAM proteins differ in the number of immunoglobulin-like domains that are linked together and whether these domains contain an integrin binding site. The importance of multiple immunoglobulin domains linked together means that several spatially distinct integrinbinding sites (IBSs) may exist within a single polypeptide, as is the case for ICAM-1. As such, ICAM-1/integrin-binding interactions may be occurring simultaneously to bring opposing cells more tightly together, as might be required for firm adhesion under shear stress or they could be timed to occur in a regulated manner as discussed previously. In such cases, the resulting outside-in signals delivered by the ICAM may differ. Yet the fact that the basic unit is replicated multiple times within a single polypeptide, regardless of the presence or absence of an IBS, indicates that its structure is critical to its function. Therefore, the form of the domain which makes up a stalk may project the IBSs away from the plasma membrane. Overall protein sequence identities between family members in the amino terminal domain ranges from 21-38% (see Table 1). However, a conserved pentapeptide sequence known to be critical for ICAM binding to ]32 integrins, denoted as the IBS, I(L)-D(E)-S(T)-P(X)L(X), originally identified in ICAM-1 by Staunton and colleagues (4,20), is found in all immunoglobulin-like domains of ICAM proteins that bind to integrins. Although the predicted secondary structure scaffold of the domains remains similar, the relatively low level of sequence conservation may affect binding affinity between different ICAM proteins and [~2 integrins. For example, LFA-1

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binds to ICAM-1 with greater avidity than it does to 1CAM-3 and conversely, (~d[~2 binds to ICAM-3 preferentially over ICAM-I (24). Now that a crystal structure for immunoglobulin domains of ICAM-2 has been solved, the structures can be more readily compared and comparisons with other ICAMs may be insightful with respect to their relative integrin-binding preferences (51). Dramatic differences exist in ICAM expression patterns and the regulation of their genes by inflammatory cytokines. ICAM-1, -2, -3, and -4 expression is found on cells of hematopoietic lineages, however, none are expressed on all lineages. For instance, ICAM-4 is found only on erythrocytes whereas none of the others are. Disparate tissue expression patterns of these proteins is a feature that is the result of differential transcription factor expression or receptor/effector pathway regulation. Key differences also exist in whether the level of ICAM expression on leukocytes are regulated or not. For example, ICAM-1 is induced to high levels on leukocyte activation by cytokines such as TNF, whereas ICAM-3 is constitutively expressed at high levels. These differences regulate the timing with which ICAM/integrin engagement can occur and manifest themselves in the manner in which integrin engagement triggers outside-in signals and cellular behaviors that are ICAM specific.

Recent Developments in Outside-In Signaling and Cytoplasmic Functions of ICAM Proteins

A cross-species comparison of the amino acid sequences comprising the predicted cytoplasmic regions of the five known ICAM family members can be summarized in the following way. There is greater amino acid conservation amongst sequences of a particular ICAM protein from different species than there is between different ICAM proteins within a single species. For example, 71-93%

of residues are conserved in the cytoplasmic domains of ICAM-I from mouse, dog, and humans. For ICAM-3 from dog, cow, and humans, 68-90% of cytoplasmic domain residues are conserved. However, only 13% of residues are conserved between ICAM-1 and ICAM-3 from h u m a n s . I C A M - s p e c i f i c sequence conservation of the cytoplasmic regions implies that evolution has maintained them for specific functions involving specific cytoplasmic protein-protein interactions, which are probably essential for ICAM function. Mechanistically, studies of the ICAM proteins are not as far along as those of other plasma membrane receptors such as the epidermal growth factor or platelet-derived growth factor receptors, however some themes are beginning to emerge. Growth factor receptor dimerization following ligand binding is a mechanism known for receptor activation. Recent observations suggest that ICAM-I forms a noncovalent dimer at the cell surface and that this form exhibits higher avidity for LFA-1 binding than does the monomer form (52,53). Under the conditions that yielded ICAM- l dimers, ICAM-2 was found to remain monomeric although locally enriched plasma membrane regions of ICAM-2 (see below) would potentially exhibit greater avidity for LFA-1 binding despite its apparent monomeric form. The requirement for chemical crosslinkers during the experimental isolation and characterization of the ICAM- 1 dimer suggests that this is a labile form of the protein and that unidentified cofactors or posttranslational modifications such as phosphorylation may be required to stabilize the interaction in situ. Interestingly, ICAM-5 has been isolated from cell extracts as a tetrameric complex of high molecular weight and ICAM-3 has been found to form a high molecular weight complex that probably represents a homodimer (54) (J Hayflick, unpublished data). If multimeric ICAM molecules are important for function, then

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heteromeric complexes may form on those cells that express more than a single ICAM polypeptide. This type of combinatorial diversity may regulate integrin/ICAM binding interactions and the resulting intracellular events. Just as the dimeric form of ICAM- 1 increases binding affinity for LFA- 1, this complex may be the basic functional unit for transmitting ICAM-triggered signals into cells. Early signaling work using co-immobilized ICAM-I MAb and a suboptimal amount of anti-CD3 MAb to ligate surface proteins demonstrated that ICAM-l is a co-stimulatory molecule. More recently, ligation ofICAM- 1 was observed to trigger an accumulation of tyrosine phosphoproteins, including cdc2 in blood-derived leukocytes and the human T leukemic cell line, Molt-3 (55), and cortactin in immortalized rat brain endothelial cells (56). Additionally, ligation of ICAM-1 by antibody treatment of SV-40 large T antigen (Ag) transformed synovial cells induced activation of the AP- 1 transcription factor as measured by nuclear translocation and DNA binding (57). Upregulation of IL-113 message and chloramphenicol acetyl transferase (CAT) activity from promoter driven constructs that contain the IL- 1 [3 AP-1 binding site was also observed. Further evidence of the capacity for signaling mediated by ICAM- 1 ligation on blood leukocytes was found using MAb treatment of peripheral blood mononuclear cells (PBMC) in the presence of a suboptimal dose of N-formylmethionyl-leucyl-phenylalanine(FMLP) which together induced an oxidative burst response (58). These observations taken together, suggest a model for ICAM-I function in the development of cellular immune responses whereby early activation events such as cytokine binding to receptors or T cell antigen receptor binding to antigen/major histocompatibility complex (MHC) complexes triggers ICAM-1 surface expression along with other components of a signal transducing

complex. These complexes might include kinase/phosphatase activities since no intrinsic enzymatic activities have been demonstrated for any ICAM cytoplasmic region. Such assemblages may form stabilized ICAM-1 dimers and could bind to activated LFA-1 on opposing cells with higher avidity. This tight cell-cell contact could present other receptor/ligand pairs with an opportunity for interaction, perhaps ligating ICAM-1 into even larger molecular assemblages, eventually triggering ICAM-1-specific activation events. The other member of the ICAM family most frequently associated with signal transduction and activation events is ICAM-3. The fact that ICAM-3 was constitutively expressed at high levels on all cells of the hematopoietic lineages involved in cellular immune responses implied that it has a role during the initial phase of activation by maintaining cell-cell contact. Recent observations with peripheral blood T cells and T leukemic cell lines stimulated by co-immobilized ICAM-3 MAb and suboptimal amounts of CD3 MAb provided support for an early activation role of ICAM-3 (26,59,60). Some have observed that on peripheral blood leukocytes and T cell lines, ICAM-3 antibody treatment triggered tyrosine phosphorylation of specific proteins and fluxed intracellular calcium concentrations (55,61,62). Others however, ourselves included, have failed to see these responses. The reasons for these disparities may be caused by antibody epitope specificity or to the inherent differences in cell lines or activation states resulting from isolation procedures. ICAM-3-dependent activation of other surface proteins has also been reported. In particular, upregulation of [31 and [32 integrin function on PBMC blasts has been observed. However the biochemical signaling pathways leading to these effects are as yet unclear (63,64). ~ integrin upregulation by a ligand for ]32integrins suggests that an interesting circular activation pathway could exist.

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To undertake a description of how intracellular events are triggered by ICAM-3, a functional map of the cytoplasmic region of the protein was developed using an ICAM-3deficient Jurkat cell line and gene transfer techniques. Human ICAM-3 cytoplasmic tail truncations and single amino acid mutants were tested with respect to CD3/ICAM-3dependent cytokine secretion, homotypic aggregation, and cellular spreading (65). The majority of amino acids in the cytoplasmic region were required for CD3 co-stimulation. Deletion of the membrane distal or proximal thirds of the cytoplasmic region significantly affected aggregation and spreading responses. Point mutation of serine 489 or serine 515 to alanine (located in the membrane proximal or distal regions, respectively) also blocked ICAM-3 function. Phosphorylation of serine 489 in Jurkat cells was enhanced by CD3 crosslinking or by directly stimulating protein kinase C (PKC) isoforms with phorbol ester. Furthermore, serine 489 was phosphorylated specifically by purified isoforms of PKC suggesting that this enzyme may be a significant contributor to ICAM-3-mediated signal transduction events. Efficient activation of T cells is thought to require signaling at or greater than a threshold level from both antigen receptor complexes and numerous other surface proteins for which ligands exist on the APC surface within a specialized contact zone between these cells. At this adhesive interface, ICAM-3 probably performs important functions in support of and to amplify signaling events. For naive T cells (CD45RA+), the signaling threshold required for activation has been observed to be greater than it is for memory T cells (CD45RO +) (66). This may be the result of increased levels in surface expression of activation-promoting proteins on memory vs naive T cells or decreased levels of more negatively charged proteins, such as the larger isoforms of CD45,

providing less repulsion between cells. One means for naive T cells to achieve productive activation is for them to form greater cell-cell contact zones with an APC allowing for surveillance of cognate antigenic peptides or to increase the duration of the contact. Interestingly, ICAM-3, but not ICAM- 1, triggered the induction of T cell spreading. This lends support to the notion that one of the functions ICAM-3 is to increase the contact area between cells, for example, during the early events attendant T cell/APC engagement. Thus, high constitutive expression levels of T cell ICAM-3 complexed with LFA-1 may allow productive activation to occur by increasing the probability of TCR/Ag interactions or maintaining that interaction in case of naive T cell contact or low antigen affinity. In light of the previous discussions, a model of the relationship between ICAM-3 function and activation can be proposed. At initial contact between an APC and a T cell, surface adhesive proteins such as ICAM-3/LFA- 1 interact. Under conditions of high-affinity integrin binding, which would be the case if the APC is activated, these cell-cell contacts would be stablilized. T cell receptor (TCR) engagement with cognate antigen/MHC complexes leads to kinase activation and phosphorylation of functionally essential cytoplasmic residues of ICAM-3 (i.e., serine 489). Dimers of ICAM-3 form or are stabilized, possibly as a result of phosphorylation and recruitment of cytosolic factors, adding stability to the cellular contact zone and the program for T cell activation and clonal proliferation ensues. Several objectives are achieved by engagement of ICAM-3 in this context. The number of antigen-receptor interactions are increased in the enlarged contact zone. The contact is stabilized allowing the signaling threshold to be met in the case of a naive T cell. Also, it brings the contact area between the two cells into tight juxtaposition allowing other receptor pairs (i.e., CD40-CD40L) with

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an opportunity to interact also contributing to attainment of the activation threshold. With clonal proliferation comes the need for dissociation of the T cell from the APC, and this could be acheived by de-phosphorylation of ICAM-3, which would lead to loss of surface dimers, lower LFA-1 binding avidity, and destabilized adhesive interactions. Re-organization of the actin cytoskeleton in T cells is necessary for the development of antigen-induced activation, which includes recruitment of important cytoskeleton-associated proteins to the cortical actin network and morphological changes accompanying cellcell contact (67). Mechanistically, the polymerization of actin has been hypothesized as the driving force for protrusion of the plasma membrane leading edge during cell spreading as occurs when a T cell engulfs an APC or it migrates during immune surveillance within T cell zones of secondary lymphoid organs (68). Since ICAM-3 ligation triggers cell spreading, changes in cellular f-actin content would be expected to occur. Jurkat cells treated with ICAM-3 MAb responded with a slow and prolonged increase in cellular f-actin and a decrease in g-actin. Also, ICAM-3-triggered spread cells exhibited fluorescent staining for f-actin at the edges and in a punctate distribution, which colocalized with ICAM-3 (J Hayflick, unpublished data). In contrast, anti-CD3 MAb elicited a rapid, but transient increase in total cellular f-actin which was not colocalized in spread cells (69-71) (J Hayflick, unpublished data). Interestingly, in human lymphocytes, PKC activity triggered by TCR engagement has been implicated in regulating cell shape and motility by modulating the cellular content of f-actin and actin-binding protein function (72-74). An additional consequence of re-organization of the cytoskeleton is its effect on gene expression by activation of transcription factors of the NF-kB family (75). ICAM-3-triggered

cytoskeletal reorganization and gene activation is not limited to T cells, since immobilized MAb engagement of ICAM-3 on PBMC and the monocytic cell line, THP-1 triggered cellular spreading and a dose-dependent increase in chemokine gene expression and secretion (75a). The consequences of ICAM-1 and -3 triggering on cellular behavior are most likely mediated by direct interactions with plasma membrane-associated cytoskeletal components. ICAM- 1 has been observed in association with microfilament-rich, detergent-extracted fibroblasts (76). Changes in ICAM-3 surface localization by antibody-induced capping and coincident increase in resistance to detergent extraction have been found (S Rosenman, M Gallatin, unpublished data). Anti-ICAM-3 MAb treatment led to cap formation and increased detergent resistance in an epitopedependent manner indicating that linkage of ICAM-3 to the cytoskeleton is a regulated event perhaps mediated by integrin binding or ICAM-3 dimerization. Direct cytosolic protein-protein interactions are presumably the underlying mechanism for these findings, and several such binding interactions with the ICAMs have already been reported. Among the cytoskeleton-associated proteins that have been observed by in vitro binding to ICAM-1 cytoplasmic domain are actin, (x-actinin, ~-tubulin, and glyceraldehyde-3phosphate dehydrogenase, a protein with microtubule-bundling activity (76-78). These interactions could link ICAM-1 with the cytoskeleton and perhaps participate in regulating cytoskeletal organization. ICAM-2 has also been observed to bind directly to ~-actinin in vitro (79). In fact, the binding sites on both ICAM-1 and -2 for oc-actinin share features in common with other o~-actinin binding proteins that span the plasma membrane, namely a preponderance of basic and hydrophobic amino acids.

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Changes in cell-surface protein localization are commonly observed during or resulting from cellular activation or differentiation state, which are likely to be mediated by direct interactions between the transmembrane proteins themselves and the peripheral actin cytoskeleton. As noted previously, evidence for direct cytoplasmic domain interactions with intracellular proteins has been found for the cytoplasmic domains of ICAM-1 and -2. In addition, high-level expression of certain cytoskeleton-associated proteins has been found to profoundly affect surface ICAM localization. For example, when ezrin was overexpressed in a natural killer (NK) target cell variant that was resistant to cell killing, it localized predominantly to uropodial protrusions (80). Coincident re-distribution of ICAM-2 into the same structures was also seen and this contributed to the cells acquiring sensitivity to killing by NK cells. This cytotoxicity was also inhibited by ~32 integrin MAb treatment. These data suggest that NK cytolytic activity is dependent on re-localization of ICAM-2 and P2 integrin function. Ezrin, a member of the ezrin-radixin-moesin (ERM) family of cytoskeletal-membrane protein linkers, is thought to contain cryptic f-actin binding sites that are exposed during microvillus or uropod formation (81). Ezrin binding to microfilaments might then laterally redistribute microfilament bound c~-actinin/ ICAM-2 complexes. Cortical flow of actin filaments and their associated proteins (e.g., ezrin and c~-actinin) has been postulated as the basis for generating highly enriched membrane subdomains like those found in uropod structures (82). Such enriched domains could also lead to ICAM multimers that would foster high avidity integrin-binding interactions which has been reported for dimers of ICAM-1. Localization to uropods is a feature of ICAM-2 that is shared by other ICAMs. ICAM-1 has also been found to localize to

uropod structures in JY B cells when seeded onto purified LFA-1 bilayers (83). Uropod protrusions on activated peripheral blood T lymphoblasts and Jurkat T cells have been found to contain ICAM-3 (63,84,85). Interestingly, ICAM-3 redistribution on T cells was found to occur when these cells were seeded onto monolayers of TNF-activated endothelial cells, as well as onto purified ICAM-1 or vascular cell adhesion molecule- 1 (VCAM- 1), in the presence of the chemokines RANTES and monocyte chemotactic protein- 1 (MCP- 1), suggesting that multiple signaling pathways are involved in the membrane distribution changes. As proposed for ICAM-2, a highly concentrated membrane domain of ICAM-1 or -3, probably as a dimer, would be expected to exhibit greater binding avidity for P2 integrins. The occurance of simultaneous localization of different ICAMs to the uropod or whether a temporal sequence of one ICAM followed by another into such structures will require further studies.

Novel Areas of Therapeutic Development Involving ICAM Biology

The notion that therapeutic intervention during chronic and acute immune responses at the level of ICAM/integrin interactions has gained much favor and is being pursued vigorously by the pharmaceutical industry. The following strategies are being explored: 1. Physical blockage of extracellular binding domain interactions; 2. Physical blockage of intracellular interactions; and 3. Interruption of protein expression. The idea that physically blocking an ICAM/ integrin interaction by competitive antibody binding has led to the development of numerous therapeutic candidates that have entered clinical trials for a variety of indications. An alternative approach stemmed from the obser-

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vations of naturally occuring soluble forms of several ICAM proteins (86-88). Interestingly, serum levels of both soluble ICAM-1 and -3 fluctuate, appearing to correlate with distinct pathologies such as neutropenic pneumonia, multiple sclerosis, and asthma, leaving available the possibility that their detection may be of diagnostic value (89-91). Preclinically, administration of soluble ICAM-1 has been found to be effective in prevention of rhinoviral infection in chimpanzees (92). The role of soluble ICAM proteins in vivo is unclear, but they may serve to downregulate integrin binding and therefore cell-cell contacts after an immune response by specifically shedding the integrin-binding domain or to release an antagonistic form that could compete for integrin binding. Lower integrin-binding affinity of monomeric ICAM-1 compared to the dimer form suggests that the soluble protein would have to be in a dimer to effectively compete. These observations have given rise to the idea that antibody Fc/ICAM fusion proteins which possess two respective integrinbinding sites might prove to be a more effective soluble antagonist. An extension of this is the proposal that peptides or similar space filling, low molecular weight entities, might competively inhibit binding thereby proving to be efficacious. From observations reviewed in the section entitled Recent Developments in Outside-In Signaling and Cytoplasmic Functions of ICAM Proteins regarding the roles of the ICAMs in intracellular events such as direct protein interactions or generation of second messenger effector pathways, it is plausible that inhibiting either of these may prove to be beneficial. Similar to the extracellular binding interactions, targeted drug screens against ICAM-1/o~-actinin interaction, for example, could serve the purpose of developing drug leads that interrupt cytoskeletal association. Interruption of such associations in endothelial cells, for instance,

may be effective in reducing lymphocyte infiltration in disease settings such as rheumatoid arthritis or Crohns disease. An intriguing alternative approach where the drug targets are kinase/phosphatase effector molecules that are triggered on cellular activation leading to posttranslational modification of ICAM proteins or that are activated as a result of ICAM ligation. Catalytic site inhibitors of these activities may be relatively nonspecific however, unless their targets are sufficiently limited in their distribution. Accessability to the internal milieu is a further hurdle to be overcome, since these targets reside within the cell, therefore small molecule inhibitors with favorable lipid transport qualities must be sought. Finally, a strategy based on reducing synthesis of specific ICAM proteins or important cytoplasmic components, such as second messenger effector proteins, utilizing mainly antisense technology have been reported (93-96). Important factors impinging on the effectivness of this approach include the turnover rate of the target protein itself, lysosomal degradation ofantisense nucleic acids, and targeting to the messenger RNA. This strategy has also yielded drug candidates that are presently in clinical trials. In the future, the mechanisms of how ICAM-triggered intracellular events are regulated by integrin engagement should become clarified. Furthermore, the relationship between ICAM/cytoskeleton interactions and ICAM/ integrin avidity changes may lead to the elucidation of novel therapeutic targets.

Acknowledgments
The authors thank their colleagues for allowing them to include unpublished data and apologize to those whose work we did not include because of space limitations. We also thank Louise Band and Alice Dersham for critical reading and help in preparing the manuscript, respectively.

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References
1 Williams AF: The immunoglobulin superfamily takes shape (news). Nature (Lond) 1984;308 (5954):12-13. Staunton DE, Marlin SD, Stratowa C, Dustin ML, Springer TA: Primary structure of ICAM-I demonstrates interaction between members of the immunoglobulin and integrin supergene families. Cell 1988;52(6):925-933. 3 Simmons D, Makgoba MW, Seed B: ICAM, an adhesion ligand of LFA- 1, is homologous to the neural cell adhesion molecule NCAM. Nature (Lond) 1988;331(6157): 624-627. 4 Staunton DE, Dustin ML, Erickson HP, Springer TA: The arrangement of the immunoglobulinlike domains of ICAM-I and the binding sites for LFA- 1and rhinovirus (published errata appear in Cell Jun 15 190;61(2):1157 and Sep201991 ;66(6):following1311). Cell 1990;61(2):243-254. 5 Vonderheide RH, Tedder TF, Springer TA, Staunton DE: Residues within a conserved amino acid motif of domains 1 and 4 of VCAM-1 are required for binding to VLA-4. J Cell Bio11994;125(1): 215-222. 6 Altmann DM, Hogg N, Trowsdale J, Wilkinson D: Cotransfection of ICAM-I and HLA-DR reconstitutes human antigen-presenting cell function in mouse L cells. Nature (Lond) 1989;338(6215): 512-514. 7 Springer TA: Adhesion receptors of the immune system. Nature (Lond) 1990;346(6283):425-434. 8 Sligh JE, Jr., Ballantyne CM, Rich SS, Hawkins HK, Smith CW, Bradley A, Beaudet AL: Inflammatory and immune responses are impaired in mice deficient in intercellular adhesion molecule 1. Proc Natl Acad Sci USA 1993; 90(I 8):8529-8533. 9 Xu H, Gonzalo JA, St Pierre Y, Williams IR, Kupper TS, Cotran RS, Springer TA, GutierrezRamos JC: Leukocytosis and resistance to septic shock in intercellular adhesion molecule ldeficient mice. J Exp Meal 1994; 180( I):95-109. 10 Soriano SG, Lipton SA, Wang YF, Xiao M, Springer TA, GutierrezRamos JC, Hickey PR: Intercellular adhesion molecule-ldeficient mice are less susceptible to cerebral ischemia-reperfusion injury. Ann Neurol 1996;39(5): 618-624. 11 Berendt AR, Simmons DL, Tansey J, Newbold CI, Marsh K: Intercellular adhesion molecule- 1 is an endothelial cell adhesion receptor for Plasmodium falciparum. Nature (Lond) 1989;341(6237):57-59. 12 Fortin JF, Cantin R, Lamontagne G, Tremblay M: Host-derived ICAM-1 glycoproteins incorporated on human immunodeficiency virus type 1 are biologically active and enhance viral infectivity. J Virol 1997;71(5):3588-3596. 13 Dustin ML, Springer TA: Lymphocyte function-associated antigen-I (LFA-I) interaction with intercellular adhesion molecule- 1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to cultured endothelial cells. J Cell Biol 1988;107(1): 321-331. 14 de Fougerolles AR, Stacker SA, Schwarting R, Springer TA: Characterization of ICAM-2 and evidence for a third counter-receptor forLFA-l.J Exp Med 1991;174(1): 253-267. 15 Staunton DE, Dustin ML, Springer TA: Functional cloning of ICAM-2, a cell adhesion ligand for LFA-I homologous to ICAM-1. Nature (Lond) 1989;339(6219): 61-64. 16 Xie J, Li R, Kotovuori P, VermotDesroches C, Wijdenes J, Arnaout MA, Nortamo P, Gahmberg CG: Intercellular adhesion molecule-2 (CDI02) binds to the leukocyte integrin CDI Ib/CD18 through the A domain. J Immunol 1995;155 (7):3619-3628. 17 Vazeux R, Hoffman PA, Tomita JK, Dickinson ES, Jasman RL, St. John T, Gallatin WM: Cloning and characterization of a new intercellular adhesion molecule ICAMR. Nature (Lond) 1992;360: 485-488. 18 Fawcett J, Holness CLL, Needham LA, Turley H, Gatter KC, Mason DY, Simmons DL: Molecular cloning of ICAM-3, a third ligand for LFA-I, constitutively expressed on resting leukocytes. Nature (Lond) 1992;360:481-484. de Fougerolles AR, Klickstein LB, Springer TA: Cloning and expression of intercellular adhesion molecule 3 reveals strong homology to other immunoglobulin family counter-receptors for lymphocyte function-associated antigen 1. J Exp Med 1993;177(4): 1187-1192. Sadhu C, Lipsky B, Erickson HP, Hayflick J, Dick KO, Gallatin WM, Staunton DE: LFA-I binding site in ICAM-3 contains a conserved motif and non-contiguous amino acids. Cell Adhes Comrnun 1994;2:429440. de Fougerolles AR, Diamond MS, Springer TA: Heterogenous glycosylation of ICAM-3 and lack of interactionwith Mac-I and p 150,95. EurJ Immunol1995;25(4):1008-1012. van Kooyk Y, Binnerts ME, Edwards CP, Champe M, Berman PW, Figdor CG, Bodary SC: Critical amino acids in the lymphocyte function-associated antigen-1 I domain mediate intercellular adhesion molecule 3 binding and immune function. J Exp Med 1996;183(3):1247-1252. Klickstein LB, York MR, Fougerolles AR, Springer TA: Localization of the binding site on intercellular adhesion molecule-3 (ICAM-3)for lymphocytefunctionassociated antigen 1 (LFA-I). J Biol Chem 1996;271(39):23,92023,927. Van der Vieren M, Le Trong H, Wood CL, Moore PF, St. John T, Staunton DE, Gallatin WM: A novel leukointegrin, alpha-d beta2, binds preferentially to ICAM-3. Immunity 1995;3:683-690. Vilella R, Mila J, Lozano F, A1berola-Ila ,I, Places L, Vires J: Involvement of the CDw50 molecule in allorecognition. Tissue Antigens 1990;36(5):203-210. de Fougerolles AR, Qin X, Springer TA: Characterization of the function of intercellular adhesion molecule (ICAM)-3 and comparison with ICAM-I and ICAM-2 in

19

20

21

22

23

24

25

26

324

Hayflick, Kilgannon, and Gallatin

27

28

29

30

31

32

33

34

35

immune responses. J Exp Med 1994;179(2):619-629. Acevedo A, de! Pozo MA, Arroyo AG, Sanchez-Mateos P, Gonzalez-Amaro R, Sanchez-Madrid F: Distribution of ICAM-3-bearing cells in normal human tissues. Expression of a novel counterreceptor for LFA-1 in epidermal Langerhans cells. Am J Pathol 1993;143(3):774-783. Staquet MJ, Peguet J, Jacquet C, Dezutter-Dambuyant C, Schmitt D: Expression of ICAM-3 on human epidermal dendritic cells. Immunobiology 1995;192(3-4): 249-261. Teunissen MB, Koomen CW, Bos JD: Intercellular adhesion molecule-3 (CD50) on human epidermalLangerhanscellsparticipates in T cell activation. J Invest Dermatol 1995;104(6):995-998. Griffiths CE, Railan D, Gallatin WM, Cooper KD: The ICAM-3/ LFA-I interaction is critical for epidermal Langerhans cell alloantigen presentation to CD4+ T cells. Br J Dermatol 1995;133(6): 823-829. Christophers E: The immunopathology of psoriasis. Int Arch Allergy Immunol 1996;110(3): 199-206. Doussis-Anagnostopoulou I, Kaklamanis L, Cordell J, Jones M, Turley H, Pulford K, Simmons D, Mason D, Garter K: ICAM-3 expression on endothelium in lymphoid malignancy. Am J Pathol 1993;143(4):1040-1043. Patey N, Vazeux R, Canioni D, Potter T, Gallatin WM, Brousse N: Intercellular adhesion molecule-3 on endothelial cells. Expression in tumors but not in inflammatory responses. Am J Pathol 1996;148 (2):465-472. YoshimuraT, JohnsonDG: cDNA cloning and expression of guinea pig neutrophil attractant protein-1 (Nap-l). Nap-I is highly conserved in guinea pig. J Immunol 1993;151(11):6225-6236. Bailly P, Hermand P, Callebaut I, Sonneborn HH, Khamlichi S, Mornon JP, Cartron JP: The LW blood group glycoprotein is homologous to intercellular adhesion molecules. Proc Natl Acad Sci USA 1994;91(12):5306-5310.

36 Levine P, Stetson RE: An unusual case of intragroup agglutination. JAMA 1939;113:126,127. 37 Sistonen P: Linkage of the LW blood group locus with the complement C3 and Lutheran blood group loci. Ann Hum Genet 1984;48(Pt 3):239-242. 38 Marsh WL, Chaganti RS, Gardner FH, Mayer K, Nowell PC, German J: Mapping human autosomes: evidence supporting assignment of rhesus to the short arm of chromosome no. 1. Science (Wash DC) 1974;183(128):966-968. 39 Bailly P, Tontti E, Hermand P, Cartron JP, Gahmberg CG: The red cell LW blood group protein is an intercellular adhesion molecule which binds to CD11/CD18 leukocyte integrins. Eur J Immunol 1995;25(12):3316-3320. 40 Yoshihara Y, Oka S, Nemoto Y, Watanabe Y, Nagata S, Kagamiyama H, Mori K: An ICAM-related neuronal glycoprotein, telencephalin, with brain segmentspecific expression. Neuron 1994; 12(3):541-553. 41 Mizuno T, Yoshihara Y, Inazawa J, Kagamiyama H, Mori K: cDNA cloning and chromosomal localization of the human telencephalin and its distinctive interaction with lymphocyte function-associated antigen-1. J Biol Chem 1997;272(2): 1156-1 t63. 42 Tian L, Yoshihara Y, Mizuno T, Mori K, Gahmberg CG: The neuronal glycoprotein telencephalin is a cellular ligand for the CD1 la/ CD18 leukocyte integrin. J Immunol 1997;158(2):928-936. 43 Lewis M, Kaita H, Coghlan G, Philipps S, Belcher E, McAlpine PJ, Coopland GR, Woods RA: The chromosome 19 linkage group LDLR, C3, LW, APOC2, LU, SE in man. Ann Hum Genet 1988;52 (Pt 2):137-144. 44 Sansom D, Borrow J, Solomon E, Trowsdale J: The human ICAM2 gene maps to 17q23-25. Genomics 1991;11 (2):462-464. 45 Trask B, Fertitta A, Christensen M, Youngblom J, Bergmann A, Copeland A, de Jong P, Mohrenweiser H, Olsen A, Carrano A, et al.: Fluorescence in situ hybridization mapping of human chromosome 19: cytogenetic band location of

46

47

48

49

50

51

52

53

54

55

540 cosmids and 70 genes or DNA markers. Genomics 1993;15(1): 133-145. Bossy D, Mattei MG, Simmons DL: The human intercellular adhesion molecule 3 (ICAM3) gene is located in the 19p13.2-p13.3 region, close to the ICAM1 gene. Genomics 1994;23(3):712,713. Naruse K, Ueno M, Staoh T, Nomiyama H, Tel H, Takeda M, Ledbetter DH, Coillie EV, Opdenakker G, Gunge N, Sakaki Y: A YAC contig of the human CC chemokine genes clustered on chromosome 17ql 1.2. Genomics 1996;34: 236-240. Alvarez V, Coto E, Setien F, Lopez-Larrea C: A physical map of two clusters containing the genes for six proinflammatory receptors. Immunogenetics 1994; 40(2):100-103. Raport CJ, Schweickart VL, Chantry D, Eddy RL, Jr., Shows TB, Godiska R, Gray PW: New members of the chemokine receptor gene family. J Leukoc Biol 1996;59(1):18-23. Jensenius JC, Williams AF: The T lymphocyte antigen receptor-paradigm lost. Nature (Lond) 1982;300(5893):583-588. Casasnovas JM, Springer TA, Liu JH, Harrison SC, Wang JH: Crystal structure of ICAM-2 reveals a distinctive integrin recognition surface. Nature (Lond) 1997;387 (6630):312-315. Reilly PL, Woska JR, Jr., Jeanfavre DD, McNally E, Rothlein R, Bormann BJ: The native structure of intercellular adhesion molecule-1 (ICAM-1) is a dimer. Correlation with binding to LFA-1. J lmmunol 1995;155(2):529-532. Miller J, Knorr R, Ferrone M, Houdei R, Carton CP, Dustin ML: Intercellular adhesion molecule- 1 dimerization and its consequences for adhesion mediated by lymphocyte function associated-l. J Exp Med 1995;182(5):1231-1241. Oka S, Mori K, Watanabe Y: Mammalian telencephalic neurons expressa segment-specific membrane glycoprotein, telencephalin.Neuroscience 1990;35(1):93-103. Chirathaworn C, Tibbetts SA, Chan MA, Benedict SH: Crosslinking of ICAM-1 on T cells

The ICAM Protein Family

325

56

57

58

59

60

61

62

induces transient tyrosine phosphorylation and inactivation of cdc2 kinase. J Immunol 1995; 155(12):5479-5482. Durieu-Trautmann O, Chaverot N, Cazaubon S, Strosberg AD, Couraud PO: Intercellular adhesion molecule 1 activation induces tyrosine phosphorylation of the cytoskeleton-associated protein cortactin in brain microvessel endothelial cells. J Biol Chem 1994;269(17): 12,536-12,540. Koyama Y, Tanaka Y, Saito K, Abe M, Nakatsuka K, Morimoto 1, Auron PE, Eto S: Cross-linking of intercellular adhesion molecule 1 (CD54) induces AP-I activation and IL-l-beta transcription. J Immunol 1996;157:5097-5103. Rothlein R, Kishimoto TK, Mainolfi E: Cross-linking of ICAM-I induces co-signaling of an oxidative burst from mononuclear leukocytes. J Immunol 1994;152 (5):2488-2495. Hernandez-Caselles T, Rubio G, Campanero MR, del Pozo MA, Muro M, Sanchez-MadfidF, Aparicio P: ICAM-3, the third LFA-1 counterreceptor, is a co-stimulatory molecule for both resting and activated T lymphocytes. Eur J Immunol 1993;23(11): 2799-2806. del Pozo MA, Campanero MR, Sanchez-Mateos P, Arroyo AG, Pulido R, Munoz C, HernandezCaselles T, Aparicio P, SanchezMadrid F: Role of ICAM-3 in intercellular adhesion and activation ofT lymphocytes. Cell Adhes Commun 1994;2(3):211-218. Arroyo AG, Campanero MR, Sanchez-Mateos P, Zapata JM, Ursa MA, del Pozo MA, SanchezMadrid F: Induction of tyrosine phosphorylation during ICAM-3 and LFA-l-mediated intercellular adhesion, and its regulation by the CD45 tyrosine phosphatase. J Cell Biol 1994;126(5):1277-1286. Juan M, Vinas O, Pino-Otin MR, Places L, Martinez-Caceres E, Barcelo JJ, Miralles A, Vilella R, de la Fuente MA, Vires J, et al.: CD50 (intercellular adhesion molecule 3) stimulation induces calcium mobilization and tyrosine phosphorylation through p59fyn

63

64

65

66

67

68

69

70

and p561ck in Jurkat T cell line. J Exp Med 1994;179(6):17471756. Campanero MR, Sanchez-Mateos P, del Pozo MA, Sanchez-Madrid F: ICAM-3 regulates lymphocyte morphology and integrin-mediated T cell interaction with endothelial cell and extracellular matrix ligands. J Cell Biol 1994; 127(3):867-878. Cid MC, Esparza J, Juan M, Miralles A, Ordi J, Vilella R, UrbanoMarquez A, Gaya A, Vives J, Yague J: Signaling through CD50 (ICAM-3) stimulates Y lymphocyte binding to human umbilical vein endothelial cells and extracellular matrix proteins via an increase in beta 1 and beta 2 integrin function. Eur J Immunol 1994;24(6):1377-1382. Hayflick JS, Stine J, Fox R, Hoekstra D, Gallatin WM: Functional mapping of the cytoplasmic region of intercellular adhesion molecule 3 reveals important roles for serine residues. J Biol Chem 1997;272(35):22,207-22,214. Croft M, Bradley LM, Swain SL: Naive versus memory CD4 Y cell response to antigen. Memory cells are less dependent on accessory cell costimulation and can respond to many antigen-presenting cell types including resting B cells. J lmmuno11994;152(6):2675-2685. Valitutti S, Dessing M, Aktories K, Gallati H, Lanzavecchia A: Sustained signaling leading to T cell activation results from prolonged Y cell receptor occupancy. Role ofT cell actin cytoskeleton. J Exp Med 1995;181(2):577-584. Theriot JA, Mitchison TJ: Actin microfilament dynamics in locomoting cells (see comments). Nature (Lond) 1991;352(6331): 126-131. DeBell KE, Conti A, Alava MA, Hoffman T, Bonvini E: Microfilament assembly modulates phospholipase C-mediated signal transduction by the TCR/CD3 in murine T helper lymphocytes. J Immunol 1992;149(7):2271-2280. Parsey MV, Lewis GK: Actin polymerization and pseudopod reorganization accompany antiCD3-induced growth arrest in

71

72

73

74

75

75a

76

77

78

Jurkat T cells. J Immunol 1993; 151(4):1881-1893. Phatak PD, Packman CH: Engagement of the T cell antigen receptor by anti-CD3 monoclonal antibody causes a rapid increase in lymphocyte F-actin. J Cell Physio11994; 159(2):365-370. Phatak PD, Packman CH, Lichtman MA: Protein kinase C modulates actin conformation in human T lymphocytes. J Immunol 1988;141(9):2929-2934. Brock MA, Chrest F: Differential regulation of actin polymerization following activation of resting T lymphocytes from young and aged mice. J Cell Physiol 1993;157(2): 36%378. Trachsel S, Keller HU: Selective responses (actin polymerization, shape changes, locomotion, pinocytosis) to the PKC inhibitor Ro 31-8220 suggest that PKC discriminately regulates functions of human blood lymphocytes. J Leukoc Bio11995 ;57(4):587-591. Rosette C, Karin M: Cytoskeletal control of gene expression: depolymerization of microtubules activates NF-kappa B. J Cell Biol 1995;128(6):1111-1119. Kessel JM, Hayflick J, Weyrich AS, Hoffman PA, Gallatin M, Mclntyre TM, Prescott SM, Zimmerman GA: Engagement of intercellular adhesion molecule-3 (ICAM-3) induces chemokine secretion and spreading by myeloid leukocytes. J Immunol 1998; in press. Vogetseder W, Dierich MP: Intercellular adhesion molecule-i (ICAM-I, CD 54) is associated with actin-filaments. Immunobiology 1991 ;182(2): 143-151. Carpen O, Pallai P, Staunton DE, Springer TA: Association of intercellular adhesion molecule-1 (ICAM-1) with actin-containing cytoskeleton and alpha-actinin. J Cell Bio11992;118(5): 1223,1234. Federici C, Camoin L, Hattab M, Strosberg AD, Couraud PO: Association of the cytoplasmic domain of intercellular-adhesionmoleculel with glyceraldehyde-3-phosphate dehydrogenase and beta-tubulin. Euro J Biochem 1996;238 (1):173-180.

326

Hayflick, Kilgannon, and Gallatin

79 Heiska L, Kantor C, Parr T, Critchley DR, Vilja P, Gahmberg CG, Carpen O: Binding of the cytoplasmic domain of intercellular adhesion molecule-2 (ICAM-2) to alpha-actinin. J Biol Chem 1996; 271 (42):26,214-26,219. 80 Helander TS, Carpen O, Turunen O, Kovanen PE, Vaheri A, Timonen T: ICAM-2 redistributed by ezrin as a target for killer cells. Nature (Lond) 1996;382(6588): 265-268. 81 Jockusch BM, Rudiger M: Crosstalk between cell adhesion molecules: viniculin as a paradigm for reguation by conformation. Cell Biology 1996;6:311-315. 82 Bray D, White JG: Cortical flow in animal cells. Science (Wash DC) 1988;239(4842):883-888. 83 Dustin ML, Carpen O, Springer TA: Regulation of locomotion and cell-cell contact area by the LFA- I and ICAM-I adhesion receptors. J Immunol 1992;148(9):2654-2663. 84 Campanero MR, del Pozo MA, Arroyo AG, Sanchez-Mateos P, Hernandez-Caselles T, Craig A, Pulido R, Sanchez-Madrid F: ICAM-3 interacts with LFA- 1 and regulates the LFA- I/ICAM- 1 cell adhesion pathway. J Cell Biol 1993;123(4):1007-1016. 85 del Pozo MA, Sanchez-Mateos P, Nieto M, Sanchez-Madrid F: Chemokines regulate cellular polarization and adhesion receptor redistribution during lymphocyte interaction with endothelium and extracellular matrix. Involvement of cAMP signaling pathway. J Cell Biol 1995;131(2):495-508. 86 del Pozo MA, Pulido R, Munoz C, AIvarez V, Humbria A, Campanero MR, Sanchez-Madrid F:

87

88

89

90

91

Regulation of ICAM-3 (CD50) membrane expression on human neutrophils through a proteolytic shedding mechanism. Eur J Immunol 1994;24(11):2586-2594. Pino-Otin MR, Vinas O, de la Fuente MA, Juan M, Font J, Torradeflot M, Pallares L, Lozano F, Alberola-IlaJ, Martorell J, Yague J, Vives J, Gaya A: Existence of a soluble form of CD50 (intercellular adhesion molecule-3) produced upon human lymphocyte activation. J Immunol 1995; 154:3015-3024. Martin S, Rieckmann P, Melchers I, Wagner R, Bertrams J, Voskuyl AE, Roep BO, Zielasek J, Heidenthal E, Weichselbraun I, et a: Circulating forms of ICAM-3 (cICAM-3). Elevated levels in autoimmune diseases and lack of association with cICAM-1. J Immunol 1995;154(4): 1951-1955. Sudhoff T, Wehmeier A, Arning M, Bauser U, Schlomer P, Aul C, Schneider W: Increases of sICAMI during neutropenic pneumonia in leukemic patients. Leukemia 1997; 11(3):346-351. Rieckmann P, Altenhofen B, Riegel A, Baudewig J, Felgenhauer K: Soluble adhesion molecules (sVCAM-1 and sICAM-1) in cerebrospinal fluid and serum correlate with MRI activity in multiple sclerosis. Ann Neurol 1997;41 (3):326-333. Gonokami Y, Konno S, Kurokawa M, Kawazu K, Ueno K, Tomita K, Ike M, Nyui M, Adachi M: Circulating intracellular adhesion molecule-I concentrations following bronchial provocation in atopic asthma, lnt Arch Allergy Immunol 1997;112(4):386-391.

92 Huguenel ED, Cohn D, Dockum DP, Greve JM, Fournel MA, Hammond L, Irwin R, Mahoney J, McClelland A, Muchmore E, Ohlin AC, Scuderi P: Prevention of rhinovirus infection in chimpanzees by soluble intercellular adhesion molecule-1. Am J Respir Crit Care Med 1997;155(4):1206-1210. 93 Stepkowski SM, Yu Y, Condon TP, Bennett CF: Blocking of heart allograft rejection by intercellular adhesion molecule-I antisense oligonucleotides alone or in combination with other immunosuppressive modalities (published erratum appears in J Immunol 1995 Feb 1:154(3): 1521). J Immunol 1994;153(11): 5336-5346. 94 Stepkowski SM, Tu Y, Condon TP, Bennett CF: Induction of transplantation tolerance by treatment with ICAM-I antisense oligonucleotides and anti-LFA-1 monoclonal antibodies. Transplant Proc 1995;27(1):113. 95 Katz SM, Browne B, Phan T, Wang ME, Bennett CF, Stepkowski SM, Kahan BD: Efficacy ofICAM-1 antisense oligonucleotide in pancreatic islet transplantation. Transplant Proc 1995;27 (6):3214. 96 Kumasaka T, Quinlan WM, Doyle NA, Condon TP, Sligh J, Takei F, Beaudet A, Bennett CF, Doerschuk CM: Role of the intercellular adhesion molecule-I (ICAM-1) in endotoxin-induced pneumonia evaluated using ICAM-1 antisense oligonucleotides, anti-ICAM-1 monoclonal antibodies, and ICAM-I mutant mice. J Clin Invest 1996;97(10): 2362-2369.

The ICAM Protein Family

327