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The Journal of Immunology, 1998, 161: 3087-3095.
Copyright © 1998 by The American Association of Immunologists

SFA-1/PETA-3 (CD151), a Member of the Transmembrane 4 Superfamily, Associates Preferentially with {alpha}5ß1 Integrin and Regulates Adhesion of Human T Cell Leukemia Virus Type 1-Infected T Cells to Fibronectin1

Hitoshi Hasegawa2, Tetsuhiko Nomura, Kyoko Kishimoto, Kohsuke Yanagisawa and Shigeru Fujita

First Department of Internal Medicine, Ehime University School of Medicine, Shigenobu, Ehime, Japan


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
In this study we have analyzed the adhesion molecules associated with and the biologic function of SFA-1/PETA-3 (CD151) in human T cell leukemia virus type 1 (HTLV-1)-infected T cells and in freshly isolated adult T cell leukemia (ATL) cells using an anti-CD151 mAb. The anti-CD151 mAb coprecipitated {alpha}5ß1 integrin from HTLV-1-infected T cells. Conversely, an anti-{alpha}5 integrin mAb coprecipitated CD151. The anti-CD151 mAb inhibited the adhesion of HTLV-1-infected T cells to fibronectin but did not have any effect on their adhesion to laminin, collagen type I, or collagen type IV. Moreover, antisense CD151 oligonucleotide-treated HTLV-1-infected T cells showed significant inhibition of adhesion to fibronectin. These findings showed that the CD151 molecule was associated with the {alpha}5ß1 integrin molecule and that it enhanced {alpha}5ß1 integrin-mediated adhesion to fibronectin. In addition, the expression levels of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells from lymph nodes of lymphoma-type ATL patients were significantly higher than those on circulating ATL cells from leukemia-type ATL patients. This suggests that the increased expression of these integrins may contribute to lymphoma formation through the adhesion of ATL cells to the extracellular matrix and dendritic cells, rather than contributing to transmigration.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Human T cell leukemia virus type 1 (HTLV-1)3 is an exogenous human retrovirus closely linked with adult T cell leukemia (ATL). HTLV-1 has also been reported to be associated with myelopathy, alveolitis, arthropathy, Sjögren syndrome, and uvenitis, which may result from immunologic alterations induced by HTLV-1 infection (1, 2, 3, 4, 5). The HTLV-1 genome encodes a 40-kDa protein, Tax, that functions as a transcriptional trans-activator of viral and cellular gene expression. T cell proliferation and immunologic alterations observed during HTLV-1 infection appear to be due to the effect of Tax on viral and cellular gene expression. Tax stimulates the expression of various cellular genes, including IL-2, IL-2R{alpha}, granulocyte-macrophage CSF, TNF-ß, TGF-ß, c-fos, c-jun, Krox-20, and Krox-24, and vimentin and suppresses the expression of the genes, ß-polymerase, p53, NF-1, and lck (5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17). However, the mechanism of HTLV-1-induced disease still remains to be elucidated.

Previously, to examine the changes in CD4+ T cells after HTLV-1 infection, we have cloned SFA-1 by differential hybridization of a cDNA library, using probes obtained from an HTLV-1-infected T cells as well as probes obtained from normal CD4+ T cells and the MOLT-4 cell line (18). SFA-1 and PETA-3 were assigned CD151 at the Sixth Human Leukocyte Differentiation Antigen Workshop. Human SFA-1 (CD151) was found to be up-regulated upon transformation by HTLV-1 and is trans-activated by Tax. The mRNA of the human SFA-1 (CD151) gene is comprised of approximately 1.6 kb and encodes a protein of 253 amino acids. SFA-1 is a member of the transmembrane 4 superfamily (TM4SF). The human SFA-1/PETA-3 (CD151) gene is a single gene located on chromosome 11p15.5 (19). Moreover, CD151 is conserved between human and mouse (20). PETA-3 was originally identified as a human platelet surface glycoprotein, and it has been reported to regulate platelet aggregation and mediator release (21, 22, 23).

The TM4SF is a family of membrane proteins that are characterized by the presence of four highly conserved transmembrane domains (reviewed by Refs. 24–26). This family currently has 19 members that are found in species from Schistosoma to human: CD9, CD37, CD53, CD63/ME491, CD81/TAPA-1, CD82/C33/R2/KAI1, CO-029, A15, lbl, CD151/SFA-1/PETA-3, SAS, sm23, sj23, il-TMP, L6, peripherin, Rom-1, uroplakin Ia, and uroplakin Ib. TM4SF members play roles in signal transduction pathways and regulate cell activation, development, proliferation, motility, and adhesion of a number of cell types. In addition, TM4SF members can form noncovalent associations with each other and with other molecules, such as those involved in signal transduction and adhesion. In this report, we describe the adhesion molecules associated with CD151 and its biologic function in HTLV-1-infected T cells and freshly isolated ATL cells.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Cells

Human T cell lines, MOLT-4 and Jurkat; a human erythroleukemia cell line, K562; two human myelomonocytoid cell lines, HL60 and U937; and NIH-3T3 cells were obtained from the American Type Culture Collection (Rockville, MD). A human glioblastoma cell line, A172, and a human renal carcinoma cell line, Caki-1, were obtained from the Japanese Cancer Research Resources Bank (Tokyo, Japan). These cell lines were maintained in RPMI 1640 or DMEM supplemented with 10% heat-inactivated FCS (Life Technologies, Gaithersburg, MD). Three HTLV-1-infected T cell lines, MT-2, MT-4, and HUT 102, were maintained in RPMI 1640 medium supplemented with 10% FCS. The HTLV-1-infected T cell line, SF-HT (27), was maintained in RPMI 1640 medium supplemented with 10% FCS and 100 U/ml human rIL-2 (Takeda Chemical Industries, Osaka, Japan). Monocytes were enriched as described previously (28). Normal T and B cells were separated using the sheep erythrocyte-rosetting method (29).

ATL patients

Mononuclear cells were isolated from peripheral blood samples from 14 patients with leukemia-type and from the lymph nodes of 10 patients with lymphoma-type by a Ficoll-Conray density gradient centrifugation. Control PBMCs were obtained from 10 normal healthy volunteers. The control lymph nodes samples were prepared from the reactive lymph nodes of eight HTLV-1-seronegative individuals who had undergone abdominal surgery. ATL was diagnosed according to the following clinical criteria: serum Abs against HTLV-1-associated Ags; morphologic characteristics showing highly convoluted nuclei; phenotypic analysis of ATL cells with anti-CD2, anti-CD4, and anti-CD25 mAbs; and monoclonal integration of the HTLV-1 proviral genome in the cells. Using Shimoyama’s criteria (30), 11 of 14 leukemia-type ATL patients were diagnosed with acute ATL, and 3 had chronic or smoldering ATL. Six of the leukemia-type ATL patients had involvement of the skin, gut, or lymphoid organs such as the lymph nodes, liver, or spleen. The infiltration of ATL cells into the skin, gut, or lymph node was confirmed histochemically using biopsy samples or following autopsy. Of the eight patients with noninfiltrating ATL, five had acute ATL, and three had chronic or smoldering ATL.

Highly purified CD4+ T cells were enriched by the negative immunoselection from mononuclear cells by using a multiple mAb mixture and immunomagnetic beads, as described by Ishikawa et al. (31). Briefly, the mononuclear cells prepared from peripheral blood and lymph nodes of ATL patients and control samples were incubated for 30 min at 4°C with a mixture of mAbs against CD8, CD11b, CD14, CD16, and CD20. After washing, Dynabeads (Japan Dynal, Tokyo, Japan) were added and incubated for 1 h at 4°C, which were then removed with a Dynal magnetic particle concentrator. In all samples from ATL patients, the proportion of ATL cells (CD4+ and CD25+ cells) in the negatively selected cells was >90%. ATL cells from all samples expressed CD45RO. The CD4+CD45RO+ T cells from control samples were purified by the same negative immunoselection technique using additional mAbs against HLA-DR and CD45RA. The percentage of CD4+CD45RO+ T cells was approximately 90%.

Antibodies

Mouse mAbs to CD8 (OKT8), CD11b (OKM1), and HLA-DR (OKIa1) were obtained from the American Type Culture Collection (Rockville, MD). Mouse mAbs, anti-CD14 (322A-1), anti-CD20 (B1), anti-CD45RA (2H4), FITC-conjugated anti-CD4 (OKT4), phycoerythrin (PE)-conjugated anti-CD25 (B1.49.9), and PE-anti-CD45RO (UCHL1) were purchased from Coulter (Hialeah, FL). Mouse mAbs to CD16 (3G8), integrin {alpha}2 (PIE6), {alpha}3 (PIB5), and {alpha}v (VNR147) were purchased from Chemicon International. Mouse mAbs to integrin {alpha}4 (HP2/1), {alpha}5 (SAM1), {alpha}6 (GoH3), and ß1 (K20) were purchased from Immunotech (Marseille, France).

Cell transfection

A recombinant plasmid, pL2neoSR{alpha}IIISFA-1, was constructed by inserting the 2.5-kb SalI fragment of pCDSR{alpha}SFA-1 into the SalI site of the pL2neoSR{alpha}III vector (32). The expression of the SFA-1 cDNA is regulated by the SR{alpha} promoter. No transcription of the SFA-1 (CD151) gene was observed in NIH-3T3 cells (20). To transfect NIH-3T3 cells, cells at about 50% confluence in a 10-cm dish were rinsed twice with Opti-MEM (Life Technologies) and were transfected with 10 µg of pL2neoSR{alpha}IIISFA-1 using lipofectin (Life Technologies) and following the instructions of the manufacturer. After incubation at 37°C for 6 h, the transfection mixture was removed. The cells were allowed to grow in fresh RPMI 1640 medium supplemented with 10% FCS for 24 h and then were added to G418 (Life Technologies) at the concentration of 400 µg/ml. After 2 wk, several stable transformants were isolated and examined for the expression of SFA-1 by Northern blot analysis as described previously (33). Of these transformants, the clone with highest expression of SFA-1 is designated NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells.

Preparation of mAbs

mAbs were produced by hybridoma technology as described previously (34). In brief, hybridomas were produced through the fusion of P3X63Ag8.653 cells with spleen cells from BALB/c mice immunized against the NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells. The hybridoma culture supernatants that bound to NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells but not to NIH-3T3/pL2neoSR{alpha}III were screened. After screening, selected colonies were cloned twice or more by the limiting dilution method. After cloning, the Ab-producing hybridomas were inoculated into BALB/c mice treated previously with pristane (Aldrich, Milwaukee, WI). Ascitic fluid containing Ab was obtained after about 2 wk. The mAbs from the ascitic fluids were purified by affinity chromatography on a DEAE column.

Immunoprecipitation

Cell surface proteins were labeled with 125I using carrier-free Na 125I and lactoperoxidase (ICN, Costa Mesa, CA) following the instructions of the manufacturer. Labeled cells were lysed in either Nonidet P-40 lysis buffer (10 mM Tris-HCl (pH 8.0), 0.15 M NaCl, 3 mM MgCl2, 2 mM PMSF, 0.5% Nonidet P-40), or CHAPS lysis buffer (10 mM Tris-HCl (pH 8.0), 0.15 M NaCl, 2 mM PMSF, 1 µg/ml antipain, 1 µg/ml pepstatin, 1 µg/ml leupeptin, 1 µg/ml chymostatin, 10 mM iodoacetamine, and 1% CHAPS; all reagents were purchased from Sigma, St. Louis, MO). The lysates were centrifuged, and the supernatants were precleared overnight at 4°C with protein G-Sepharose (Pharmacia, Piscataway, NJ) precoated with normal rabbit serum. The lysates were then incubated with protein G-Sepharose precoated with each mAb. After washing extensively with lysis buffer, samples were boiled for 2 min in Laemmli’s electrophoresis sample buffer and fractionated by SDS-PAGE. Radioactive bands were detected by autoradiography.

In vitro adhesion assay

Adhesion assays were performed as described by Sonnenberg et al. (35). The 96-well plates were coated overnight at 4°C with fibronectin, collagen type I, collagen type IV, or laminin at 100 µg/ml (all obtained from Sigma). Plates were washed three times with PBS and blocked with 1% BSA in PBS for at least 1 h at room temperature. The cell suspension was diluted to contain 1 x 104 cells/ml in serum-free DMEM and incubated with SFA1.4F11 mAb at the saturating concentration of 100 µg/ml for 30 min at room temperature, and aliquots of 100 µl were added to each well. After incubation for 1 h, the wells were washed, and the number of adherent cells was estimated using spectrophotometric absorbance following uptake of 3-(4, 5)-dimethylthiazol-2-yl-2,5 diphenyltetrazolium bromide (Sigma) by live cells (36). All experiments were conducted in triplicate at 37°C in a CO2 incubator.

Antisense CD151 oligonucleotides

Oligonucleotides (32 mer) corresponding to the antisense sequence flanking the translation initiation region of the mRNA for human CD151 were synthesized using phosphorothioate linkages because of their demonstrated resistance to nucleases. The sequences of the oligonucleotides were 5'-GGCAAACGGTGCCACATGTTGTCTTCTTCTCG-3' (antisense) and 5'-CGGATAGGCTCCGAGAAGATCTGTACATGTGG-3' (nonsense). After the cells had been treated for 24 h with 5 µM nonsense or antisense oligonucleotide, in vitro adhesion assays were performed.

Flow cytometric analysis and immunoblotting

Cells growing in monolayers were detached from the culture flasks by incubation at 37°C for 10 min with PBS containing 0.05% trypsin and 1 mM EDTA. The cells (2 x 105) were washed once with 3% FCS-PBS and incubated with mAb, SFA1.4F11, or IgG1 mouse mAb (negative control) at the saturating concentration (10 µg/ml) on ice for 30 min. After washing with 3% FCS-PBS, the cells were stained with FITC-conjugated goat anti-mouse IgG (Cappel, Westchester, PA) on ice for 30 min. After washing, the cells were resuspended in 3% FCS-PBS, and the fluorescence intensity was analyzed on using a profile flow cytometer (EPICS, Coulter).

Flow cytometric analysis of freshly isolated ATL cells and control CD4+CD45RO+ T cells was conducted as described by Ishikawa et al. (31). Briefly, cells (2 x 105) were incubated for 15 min at 4°C with human serum globulin (10 µg/ml; Green Cross, Osaka, Japan) to block Fc binding sites and then further incubated with FITC-anti-CD4 mAb for 30 min at 4°C. After washing, the cells were further incubated with PE-conjugated SFA1.4F11, PE-anti-{alpha}4 integrin mAb (HP2/1), PE-anti-{alpha}5 integrin mAb (SAM1), or PE-conjugated nonbinding control IgG1 mouse mAb at a saturating concentration (10 µg/ml) for 30 min at 4°C. After washing, the two-color immunostaining of cells was analyzed. The mean fluorescence intensity (MFI) values for staining of CD151, {alpha}4 integrin, and {alpha}5 integrin were corrected by subtracting MFI obtained with the nonbinding control IgG1 mouse mAb.

A recombinant plasmid, pQE30SFA-1, was constructed by inserting the 0.8-kb SacI fragment of pCDSR{alpha}SFA-1 into the SacI site of the pQE30 vector. Histidine-tagged SFA-1 protein was purified using QIAexpress system (Qiagen, Chatsworth, CA). Immunoblotting was performed by using mouse serum immunized against the purified histidine-tagged SFA-1 protein as described previously (34).


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Isolation of mAb against CD151

For use as a immunogen, we transfected the recombinant plasmid pL2neoSR{alpha}IIISFA-1 into NIH-3T3 cells. Nontransfected NIH-3T3 cells did not express any detectable levels of mRNA for CD151. After 2 wk, several stable transformants were isolated. Among these transformants, one stable neomycin-resistant transformant, designated NIH-3T3/pL2neoSR{alpha}IIISFA-1, which has highest expression of CD151 mRNA, was used as an immunogen. After injecting the NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells i.p. twice, hybridomas were produced through the fusion of P3X63Ag8.653 cells with spleen cells from immunized BALB/c mice. The anti-CD151 mAb, designated SFA1.4F11, which bound to NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells, but not to NIH-3T3/pL2neoSR{alpha}III cells, was obtained (Fig. 1GoA). SFA1.4F11 was an IgG1 Ig with {kappa} light chain. We confirmed that this Ab reacted to CD151 by immunoprecipitation and immunoblotting using histidine-tagged SFA-1 protein (Fig. 1Go, B and C).



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  		FIGURE 1. Reactivity of SFA1.4F11 mAb to CD151-transfected NIH-3T3 cells. A, Flow cytometric analysis of the expression of SFA1.4F11 mAb on CD151-transfected NIH-3T3 cells. CD151-transfected (NIH3T3/pL2neoSR{alpha}IIISFA-1; solid histogram) or control (NIH-3T3/pL2neoSR{alpha}III; clear histogram) NIH-3T3 cells were incubated with SFA1.4F11 mAb and stained with FITC-conjugated goat anti-mouse IgG. The dotted line shows unstained NIH-3T3/pL2neoSR{alpha}IIISFA-1 cells. Immunoprecipitation (B) and immunoblotting (C) of CD151-transfected NIH-3T3 cells using SFA1.4F11 mAb. NIH-3T3/pL2neoSR{alpha}III (negative control; lane 1) and NIH-3T3/pL2neoSR{alpha}IIISFA-1 (lane 2) cells were surface labeled with 125I, lysed in 0.5% Nonidet P-40, and then immunoprecipitated with SFA1.4F11. The precipitate was fractionated by SDS-PAGE and analyzed by radiography and immunoblotting using mouse serum immunized against the purified histidine-tagged CD151 protein.

 
Expression of CD151 Ag on various cells

The surface expression of the CD151 Ag by various hemopoietic and nonhemopoietic cell lines was examined by flow cytometric analysis using SFA1.4F11 (Fig. 2Go). Strong expression of CD151 Ag was demonstrated on PHA-stimulated CD4+ T cells and two HTLV-I-transformed T cell lines, SF-HT and HUT 102. The other two HTLV-1-transformed T cell lines, MT-2 and MT-4, also showed strong expression of CD151 (data not shown). CD151 Ag was expressed at low levels on CD4+ T cells, T lymphocytes, B lymphocytes, granulocytes, and four other lymphoid cell lines, Jurkat, MOLT-4, SF-EB, and HH-EB, whereas monocytes, U937 cells, and all human nonhemopoietic cell lines used in our previous study (18) showed significant expression of CD151. These results are in good agreement with those by obtained Northern blot analysis (18).



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  		FIGURE 2. Flow cytometric analysis of the expression of CD151 Ag on various hemopoietic and nonhemopoietic cell lines. Cells were incubated with anti-CD151 mAb SFA1.4F11 (solid lines) or IgG1 mouse mAb (dotted lines) and stained with FITC-conjugated goat anti-mouse IgG.

 
Identification of CD151-associated proteins

To examine whether CD151 is associated on the cell surface with other molecules, 125I surface-labeled HUT 102 cells were extracted with the mild detergent CHAPS, and CD151 immunoprecipitates were analyzed by SDS-PAGE. As shown in Figure 3GoA, anti-CD151 mAb SFA1.4F11 immunoprecipitated the CD151 Ag itself (29 kDa). Additional protein bands of 160 and 130 kDa coprecipitated with CD151 molecule. These additional proteins were also obtained from HUT 102 lysates prepared using the milder detergents 1% Tween-20 and 1% Brij 58, but not from 0.5% Nonidet P-40 lysate (data not shown). In addition, pretreatment of cells with EDTA did not alter the pattern of precipitation of these additional proteins (data not shown). These higher molecular mass proteins (160 and 130 kDa) that coprecipitated from HUT 102 cells closely resembled the very late Ag integrins. Next, we reprecipitated the anti-CD151 immunoprecipitate using anti-integrin ({alpha}2, {alpha}3, {alpha}4, {alpha}5, {alpha}6, {alpha}v, and ß1) mAbs. As shown in Figure 3GoA, 160- and 130-kDa bands were reprecipitated by the anti-{alpha}5 and anti-ß1 Abs but not by the other anti-integrin Abs. Conversely, the CD151 molecule was also reprecipitated from an anti-{alpha}5 immunoprecipitate prepared from HUT 102 cells (Fig. 3GoB). However, CD151 was not reprecipitated from an anti-{alpha}4 immunoprecipitate using a combination of immunoprecipitation and immunoblotting (data not shown). As shown in Figure 3GoC, {alpha}5ß1 integrin was observed in anti-CD151 immunoprecipitates prepared from the other three HTLV-1-infected T cell lines, MT-2, MT-4, and SF-HT. These findings show that the CD151 molecule associated preferentially with {alpha}5ß1 integrin in HTLV-1-infected T cell lines. In addition, unknown additional protein bands coprecipitated with the CD151 molecule: a 90-kDa band from the MT-2 lysate and 45-kDa bands from the MT-2 and SF-HT lysates. These additional proteins were either absent or faintly present in anti-CD151 immunoprecipitates prepared from HUT 102 and MT-4 cells and were obtained from lysates prepared using the milder detergents, 1% CHAPS, 1% Tween-20, and 1% Brij 58, but not from the 0.5% Nonidet P-40 lysate (data not shown).



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  		FIGURE 3. Coprecipitation of CD151 Ag and {alpha}5ß1 integrin in HTLV-1-infected T cells. A, HUT 102 cells were surface labeled with 125I, lysed with 1% CHAPS, and then immunoprecipitated with anti-CD151 mAb SFA1.4F11. The anti-CD151 precipitate was eluted using 0.5% Nonidet P-40 and reprecipitated with anti-integrin ({alpha}2, {alpha}3, {alpha}4, {alpha}5, {alpha}6, {alpha}v, and ß1) mAbs. B, 125I surface-labeled HUT 102 cells were immunoprecipitated with anti-{alpha}5 mAb. The anti-{alpha}5 precipitate was reprecipitated with anti-CD151 mAb. C, The lysates prepared from four HTLV-1-infected T cell lines, HUT 102, MT-2, MT-4, and SF-HT, were immunoprecipitated with anti-CD151 mAb.

 
Effect of anti-CD151 mAb on adhesion of HTLV-1- infected T cells to extracellular matrix proteins

To examine whether anti-CD151 mAb affects the adhesion of HTLV-1-infected T cells to extracellular matrix proteins, we performed in vitro adhesion assays using fibronectin, laminin, collagen type I, or collagen type IV. As shown in Figure 4Go, the anti-CD151 mAb, SFA1.4F11, inhibited adhesion to fibronectin by 40 to 57% for all four HTLV-1-infected T cell lines, but did not have any effect on adhesion to laminin, collagen type I, or collagen type IV. The other anti-CD151 mAb, SFA1.2B4, which reacts with an epitope different from that recognized by SFA1.4F11, was tested, and inhibition of adhesion to fibronectin by SFA1.2B4 was lower (by 85–90% adhesion) than that caused by SFA1.4F11 (data not shown).



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  		FIGURE 4. Inhibition of HTLV-1-infected T cell adhesion to extracellular matrix proteins by anti-CD151 mAb. Four kinds of extracellular matrix proteins, fibronectin, collagen type I, collagen type IV and laminin, were coated on 96-well plates as described in Materials and Methods. The adhesion of HTLV-1-infected T cells to extracellular matrix proteins was measured in the presence of anti-CD151 mAb SFA1.4F11 at the saturating concentration of 100 µg/ml. The percentage of the control value was compared with adhesion in the absence of Ab. All experiments were conducted in triplicate. Statistical analysis was performed using Student’s t test: * indicates p < 0.01.

 
Decreased adhesion of antisense CD151 oligonucleotide-treated HTLV-1-infected T cells to fibronectin

To demonstrate the physiologic significance of the interaction between CD151 and {alpha}5ß1 integrin, we decided to decrease the expression level of CD151 to determine whether this would affect integrin function. The down-regulation of expression of CD151 was achieved using an antisense oligonucleotide designed to inhibit the initiation of CD151 protein synthesis. Antisense oligonucleotide-treated, nonsense oligonucleotide-treated, and control untreated cells showed no difference in cell viability, indicating that the oligonucleotides were not toxic to the cells (data not shown). As shown in Figure 5GoA, addition of the antisense CD151 oligonucleotide to HUT 102 cell cultures resulted in significant inhibition (>60%) of CD151 expression. Treatment with nonsense oligonucleotide at the same concentration did not result in down-regulation. Moreover, addition of antisense or nonsense CD151 oligonucleotides did not have any effect on the expression level of {alpha}5ß1 integrin on HUT 102 cells (data not shown).



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  		FIGURE 5. A, Flow cytometric analysis of the CD151 expression on antisense CD151 oligonucleotide-treated HTLV-1-infected T cells. After HUT 102 cells were treated for 24 h with 5 µM nonsense (clear histogram) or antisense (solid histogram) oligonucleotides, cells were incubated with anti-CD151 mAb SFA1.4F11 and stained with FITC-conjugated goat anti-mouse IgG. The dotted line shows unstained HUT 102 cells. B, Decreased adhesion of the antisense CD151 oligonucleotide-treated HTLV-1-infected T cells to fibronectin. After HUT 102 cells were treated for 24 h with 5 µM nonsense or antisense oligonucleotides, in vitro adhesion assays were performed. All experiments were conducted in triplicate. Statistical analysis was performed using Student’s t test: * indicates p < 0.01.

 
The effect of the antisense CD151 oligonucleotide on adhesion to fibronectin was examined. As shown in Figure 5GoB, antisense oligonucleotide-treated HUT 102 cells showed significant inhibition to 75% of adhesion to fibronectin, whereas there was very little difference in adhesion between the nonsense oligonucleotide-treated and untreated control cells. The other three HTLV-1-infected T cell lines, MT-2, MT-4, and SF-HT, also inhibited the adhesion to fibronectin significantly after antisense-oligonucleotide treatment (data not shown). These findings indicate that the CD151 molecule was associated with the {alpha}5ß1 integrin molecule and that it enhanced the {alpha}5ß1 integrin-mediated adhesion to fibronectin.

Differential expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells in patients of leukemia-type or lymphoma-type ATL

We analyzed the expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on circulating ATL cells isolated from the peripheral blood of 14 patients with leukemia-type ATL or on ATL cells obtained from the lymph nodes of 10 lymphoma-type ATL patients. ATL cells were enriched by the negative immunoselection. Since ATL cells had the phenotype CD4+CD45RO+, the expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells was compared with that on CD4+CD45RO+ T cells purified from control samples. The integrin {alpha}4 chain associates with either ß1- or ß7-chains to form a heterodimeric glycoproteins {alpha}4ß1 and {alpha}4ß7. In all samples, the proportion of ß1 integrin-expressed cells in ATL and that of control CD4+CD45RO+ T cells that expressed {alpha}4 integrin were >90% (data not shown). Therefore, the expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells was assessed by the MFI for staining of anti-CD151 mAb (SFA1.4F11), anti-{alpha}4 integrin mAb (HP2/1), and anti-{alpha}5 integrin mAb (SAM1). As shown in Figure 6Go, there were no significant differences in the expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin between circulating ATL cells from leukemia-type ATL patients and control CD4+CD45RO+ T cells (MFI: leukemia (n = 14), 3.69 ± 1.70; control PBMC (n = 10), 3.21 ± 1.56; vs control lymph node (n = 8), 3.38 ± 1.30; 14.11 ± 7.40 vs 10.72 ± 4.47 vs 12.11 ± 5.34; 11.09 ± 5.90 vs 9.36 ± 4.20 vs 11.01 ± 5.27, respectively). On the other hand, the expression levels of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells from the lymph nodes of lymphoma-type ATL patients were significantly higher than those on circulating ATL cells from leukemia-type ATL patients (MFI: lymphoma (n = 10), 9.55 ± 5.96; vs leukemia (n = 14), 3.69 ± 1.70 (p < 0.05); 25.47 ± 11.42 vs 14.11 ± 7.40 (p < 0.05); 20.16 ± 11.62 vs 11.09 ± 5.90 (p < 0.05), respectively). This suggests that increased expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin may act to retain ATL cells in the lymph nodes in some lymphoma-type ATL patients.



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  		FIGURE 6. Differential expression of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells in leukemia-type or lymphoma-type ATL patients. Freshly isolated ATL cells and CD4+CD45RO+ T cells were purified in the negative immunoselection using a multiple mAb mixture as described in Materials and Methods. Flow cytometric analysis of CD4+CD45RO+ T cells and ATL cells obtained from the peripheral blood of 10 normal volunteers (A, closed circles), the lymph nodes of eight HTLV-1-seronegative individuals (B, closed squares), the peripheral blood of 14 leukemia-type ATL patients (C, open circles, patients 1–14), and the lymph nodes of 10 lymphoma-type ATL patients (D, open circles, patients 15–24) was performed with FITC-conjugated anti-CD4, PE-anti-CD151, PE-anti-{alpha}4, PE-anti-{alpha}5, or PE-nonbinding control IgG1 mouse mAbs. Cells were electronically gated based upon CD4 expression. Each point shows the MFI for CD151, {alpha}4, or {alpha}5 staining, which was corrected by subtracting MFI obtained with the nonbinding control IgG1 mouse mAb. The bars indicate the mean ± SD of each group; the statistical analysis was performed using Student’s t test.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
In this report, we demonstrate for the first time a specific association between CD151 and {alpha}5ß1 integrin in HTLV-1-infected T cells, and we show that the CD151 molecule enhanced {alpha}5ß1 integrin-mediated adhesion to fibronectin. This interaction is likely to be meaningful for the following reasons: 1) {alpha}5ß1 integrin was reprecipitated from anti-CD151 immunoprecipitates prepared from HTLV-1-infected T cell lines; 2) conversely, the CD151 molecule was observed in anti-{alpha}5 immunoprecipitates but not in anti-{alpha}4 immunoprecipitates; 3) the anti-CD151 mAb inhibited the adhesion to fibronectin of HTLV-1-infected T cell lines, but had no effect on their adhesion to laminin, collagen type I, or collagen type IV; and 4) antisense CD151 oligonucleotide-treated HTLV-1-infected T cells showed significantly inhibited of adhesion to fibronectin. Moreover, we also found that the expression levels of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin were higher on ATL cells from the lymph nodes of lymphoma-type ATL patients than those on circulating ATL cells from leukemia-type ATL patients, suggesting that the expression levels of these molecules may affect the retention of ATL cells in lymph nodes.

The TM4SF has 19 members, each containing four highly conserved transmembrane domains, a number of cysteine residues, and a major extracellular region between the third and fourth transmembrane domains. Although the biologic functions of the members of the TM4SF are poorly understood, several studies of their functions using specific mAbs have been undertaken, and the results obtained suggest a role for this superfamily in signal transduction pathways and the regulation of cell activation, development, proliferation, motility, and adhesion (reviewed in Refs. 24–26). Moreover, TM4SF members have been shown to form noncovalent associations with each other, integrins, and coreceptor molecules, such as those involved in adhesion and signal transduction. CD81, CD9, CD53, CD63, and CD82 have all been found in association with certain integrins in various types of human cells. All these molecules associate with the ß1 integrins {alpha}3ß1, {alpha}4ß1, and {alpha}6ß1 (37, 38, 39, 40, 41, 42, 43, 44, 45). These associations appear to be important for cell-cell adhesion and migration. CD81 forms part of a signaling complex with CD21, CD19, and Leu 13 on B cells (46, 47). CD81 also associates with CD82 as well as with CD4 and CD8 coreceptors on T cells (48). The interaction of CD81/CD82 with CD4 is strongly inhibited by p56lck binding to CD4. In the present study, human SFA-1, which was up-regulated by HTLV-I infection and trans-activated by Tax, was associated with {alpha}5ß1 integrin in HTLV-1-infected T cells, and it enhanced the {alpha}5ß1 integrin-mediated adhesion to fibronectin. PETA-3, which was originally identified as a human platelet surface glycoprotein, was detected using a mAb, 14A2.H1. Although PETA-3 was present in low abundance on the platelet surface, 14A2.H1 stimulated platelet aggregation and mediator release. In addition, this Ab showed synergy with subthreshold concentrations of other agonists, ADP, adrenaline, collagen, and serotonin, in mediating platelet activation. Sincock et al. (49) examined the distribution of PETA-3 (CD151), CD9, CD63, {alpha}5ß1, and the integrin ß1-chain in normal human tissues by the indirect immunoperoxidase and alkaline phosphatase-anti-alkaline phosphatase techniques. CD151 showed a broad distribution and was expressed by endothelium, epithelium, Schwann cells, dendritic cells, and skeletal, smooth, and cardiac muscle. They showed colocalization of CD151 with CD9, CD63, {alpha}5ß1, and ß1 in particular tissues, demonstrating that CD151/integrin complexes may occur. CD151 formed a complex with {alpha}5ß1 integrin, but not with CD9 or CD63, on HTLV-1-infected T cells (H. Hasegawa, et al., unpublished observations). Their finding and our results suggest that CD151 associates strongly with {alpha}5ß1 integrin in particular cells and tissues.

The {alpha}5ß1 integrin has been identified as the classical receptor for fibronectin. At least eight integrins have been reported to bind to fibronectin: {alpha}3ß1, {alpha}4ß1, {alpha}5ß1, {alpha}vß1, {alpha}IIbß3, {alpha}vß3, {alpha}vß6, and {alpha}4ß7 (50). Of these integrins, lymphocytes interact with fibronectin via two main receptors, {alpha}5ß1 and {alpha}4ß1 integrins. The interaction of {alpha}5ß1 integrin with fibronectin results in augmented cell adhesion, migration, differentiation, and signal transduction and has been implicated in important events in early embryogenesis, such as gastrulation (51, 52, 53, 54, 55). The {alpha}5ß1 integrin-mediated adhesion to fibronectin causes localization of the receptor to focal contacts and results in stable cell-substratum adhesion (56). The capacity of {alpha}5ß1 integrin to bind fibronectin is regulated by divalent cations and GM3 ganglioside. The recognition of fibronectin by {alpha}5ß1 integrin is enhanced by Mn2+ and is inhibited by Ca2+ (57). GM3 ganglioside, within an optimal concentration range, enhances the ability of {alpha}5ß1 integrin to adhere to fibronectin (58). Down-regulation of CD151 expression on HUT 102 cells caused by antisense oligonucleotide resulted in significant inhibition of adhesion to fibronectin. Some members of the TM4SF function as adaptor proteins that organize the relative positions of other cell surface molecules and modulate their function. For example, CD81 forms a complex consisting of CD19, CD21, and Leu 13 on B cells, and this complex lowers the threshold for signal transduction through the B cell Ag receptor complex (46, 47). This result indicates that CD81 facilitates interactions with integrins, resulting in cell adhesion, and that CD81 stabilizes the CD21/CD19 interaction. CD151 may also affect the accumulation of {alpha}5ß1 integrin at focal contacts or the maintenance of the appropriate conformation of {alpha}5ß1 integrin for optimal adhesion to fibronectin.

It has been reported that the expression of {alpha}4ß1 integrin and {alpha}5ß1 integrin on HTLV-1-infected lymphocytes and of {alpha}5ß1 integrin on T lymphocytes of patients with HIV-1 infection is increased and that alteration of expression of these integrins might contribute to the abnormal proliferation of T cells in ATL or to the abnormal localization of activated or infected T cells (59, 60). Fibronectin is widely distributed in plasma and connective tissue subendothelial matrexes and stroma and is produced by various cells, such as endothelial, mesenchymal, and epithelial cells. In lymph nodes, fibronectin is mainly produced by stromal cells and is present in all areas except the mantle zone (61). Fibronectin is especially abundant in the interfollicular compartments, which are rich in T cells. The follicular dendritic cells in the germinal center express VCAM-1. {alpha}4ß1 integrin expressed at increased levels on B lymphoma cells has been reported to play a role in the formation of follicular non-Hodgkin’s lymphoma by binding to VCAM-1 expressed on follicular dendritic cells (62, 63). On the other hand, lymphocyte adhesion to high endothelium has been reported to be mediated by L-selectin, {alpha}4ß1 integrin, and {alpha}5ß1 integrin, resulting in contributing to the transmigration of lymphocytes (64, 65). The expression levels of CD151, {alpha}4ß1 integrin, and {alpha}5ß1 integrin on ATL cells from lymph nodes of lymphoma-type patients were higher than those on circulating ATL cells from leukemia-type patients. In ATL, the increased expression of these integrins may contribute to the formation of a lymphoma through adhesion of extracellular matrix and dendritic cells, rather than contributing to transmigration. The fibronectin-rich microenvironment in the lymph node can also serve as a site of ATL cell adhesion and support for ATL cell proliferation. Adhesion studies using ATL cells and HUVECs revealed that E-selectin-mediated adhesion was a major pathway for the adherence of ATL cells to HUVECs, whereas the {alpha}4ß1 integrin/VCAM-1 pathway was partly involved in adhesion in some cases (31). Three pairs of adhesion molecules, L-selectin/peripheral lymph node addressin, cutaneous lymphocyte-associated Ag/E-selectin, and {alpha}4ß7 integrin/mucosal VCAM-1 have been reported to be associated with preferential recirculation of ATL cells to peripheral lymph nodes, skin, and gastrointestinal mucosa, respectively (66). The circulating ATL cells from patients with lymph node, skin, and gut involvement had higher expression levels of L-selectin, cutaneous lymphocyte-associated Ag, and {alpha}4ß7 integrin, respectively, compared with those from patients without involvement of these organs. These findings together with our results suggest that these adhesion molecules, rather than {alpha}4ß1 integrin and {alpha}5ß1 integrin, play an important role in the transmigration and organ infiltration of ATL cells.

In this report, we showed that CD151 associated with {alpha}5ß1 integrin and regulated adhesion of HTLV-1-infected T cells and ATL cells to fibronectin. Since CD151 is widely expressed on various cells and tissues, however, CD151 may have other unknown biologic functions in other cells and tissues. It is necessary to study the biologic functions of CD151 further in other cells and tissues.


   Footnotes
 
1 This work was supported in part by a grant-in-aid from the Osaka Cancer Research Foundation and Scientific Research, and the Ministry of Education, Science, and Culture of Japan. Back

2 Address correspondence and reprint requests to Dr. Hitoshi Hasegawa, First Department of Internal Medicine, Ehime University School of Medicine, Shigenobu, Ehime 791-02, Japan. E-mail address: Back

3 Abbreviations used in this paper: HTLV-1, human T cell leukemia virus type 1; ATL, adult T cell leukemia; TM4SF, transmembrane 4 superfamily; PE, phycoerythrin; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; MFI, mean fluorescence intensity. Back

Received for publication January 29, 1998. Accepted for publication May 11, 1998.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Osame, M., K. Usuku, S. Izumo, N. Ijichi, H. Amatani, A. Igata, M. Matsumoto, M. Tara. 1986. HTLV-1 associated myelopathy, a new clinical entity. Lancet i:1031.
  2. Vernant, J. C., G. Buisson, J. Magdeleine, J. De Thore, A. Jouannelle, C. Neisson-Vernant, N. Monplaisir. 1988. T-lymphocyte alveolitis, tropical spastic paresis, and Sjogren syndrome. Lancet i:177.
  3. Nishioka, N., I. Maruyama, K. Sato, I. Kitajima, Y. Nakajima, M. Osame. 1989. Chronic inflammatory arthropathy associated with HTLV-1. Lancet 1:441.[Medline]
  4. Mochizuki, M., K. Yamaguchi, K. Takatsuki, T. Watanabe, S. Mori, K. Tajima. 1992. HTLV-1 and uveitis. Lancet 339:1110.[Medline]
  5. Fujii, M., P. Sassone-Corsi, I. M. Verma. 1988. c-fos promoter transactivation by the tax1 protein of human T-cell leukemia virus type 1. Proc. Natl. Acad. Sci. USA 85:8526.[Abstract/Free Full Text]
  6. Inoue, J.-K., M. Seiki, T. Taniguchi, S. Tsuru, M. Yoshida. 1986. Induction of interleukin-2 receptor gene expression by p40 encoded by human T-cell leukemia virus type 1. EMBO J. 5:2883.[Medline]
  7. Sjekevitz, M., M. B. Feinberg, N. Holbrook, F. Wong-Staal, W. C. Greene. 1987. Activation of interleukin-2 and interleukin-2 receptor (tac) promoter expression by the trans-activator (tat) gene product of human T cell leukemia virus type 1. Proc. Natl. Acad. Sci. USA 84:5389.[Abstract/Free Full Text]
  8. Miyatake, S., M. Seiki, M. Yoshida, K.-I. Arai. 1988. T-cell activation signals and human T-cell leukemia virus type 1-encoded p40 protein activate the mouse granulocyte-macrophage colony-stimulating factor gene through a common DNA element. Mol. Cell. Biol. 8:5581.[Abstract/Free Full Text]
  9. Tschachler, E., M. Robert-Guroff, R. C. Gallo, M. S. Reitz. 1989. Human T-lymphotropic-virus-1-infected T cells constitutively express lymphotoxin in vitro. Blood 73:194.[Abstract/Free Full Text]
  10. Kim, S.-J., J. H. Kehrl, J. Burton, C. L. Tendler, K.-T. Jeang, D. Danielpour, C. Thevenin, K. Y. Kim, M. B. Sporn, A. B. Roberts. 1990. Transactivation of the transforming growth factor ß1 (TGF-ß1) gene by human T lymphotropic virus type 1 Tax: a potential mechanism for the increased production of TGF-ß1 in adult T cell leukemia. J. Exp. Med. 172:121.[Abstract/Free Full Text]
  11. Alexandre, C., B. Verrier. 1991. Four regulatory elements in the human c-fos promoter mediate transactivation by HTLV-1 Tax protein. Oncogene 6:543.[Medline]
  12. Alexandre, C., P. Charnay, B. Verrier. 1991. Transactivation of Krox-20 and Krox-24 promoters by the HTLV-1 Tax protein through common regulatory elements. Oncogene 6:1852.
  13. Liliebaum, A., M. Duc Dodon, C. Alexandre, L. Gazzolo, D. Paulin. 1990. Effect of human T-cell leukemia virus type 1 Tax protein on activation of the human vimentin gene. J. Virol. 64:256.[Abstract/Free Full Text]
  14. Jeang, K.-T., S. G. Widen, IV O. J. Semmes, S. H. Wilson. 1990. HTLV-1 transactivator protein, Tax, is a transrepressor of the human ß polymerase gene. Science 247:1082.[Abstract/Free Full Text]
  15. Uittenbogaard, M. N., H. A. Giebler, D. Reisman, J. K. Nyborg. 1995. Transcriptional repression of p53 by human T-cell leukemia virus type 1 Tax protein. J. Biol. Chem. 270:28503.[Abstract/Free Full Text]
  16. Feigenbaum, L., K. Fujita, F. S. Collins, G. Jay. 1996. Repression of the NF1 gene by Tax may explain the development of neurofibromas in human T-lymphotropic virus type 1 transgenic mice. J. Virol. 70:3280.[Abstract]
  17. Lemasson, I., V. Robert-Hebmann, S. Hamaia, M. D. Dodon, L. Gazzolo, C. Devaux. 1997. Transrepression of lck gene expression of human T-cell leukemia virus type 1-encoded p40tax. J. Virol. 71:1975.[Abstract]
  18. Hasegawa, H., Y. Utsunomiya, K. Kishimoto, K. Yanagisawa, S. Fujita. 1996. SFA-1, a novel cellular gene induced by human T-cell leukemia virus type 1, is a member of the transmembrane 4 superfamily. J. Virol. 70:3258.[Abstract]
  19. Hasegawa, H., K. Kishimoto, K. Yanagisawa, H. Terasaki, M. Shimadzu, S. Fujita. 1997. Assignment of SFA-1 (PETA-3), a member of the transmembrane 4 superfamily, to human chromosome 11p15.5 by fluorescence in situ hybridization. Genomics 40:193.[Medline]
  20. Hasegawa, H., H. Watanabe, T. Nomura, Y. Utsunomiya, K. Yanagisawa, S. Fujita. 1997. Molecular cloning and expression of mouse homologue of SFA-1/PETA-3 (CD151), a member of the transmembrane 4 superfamily. Biochim. Biophys. Acta 1353:125.[Medline]
  21. Fitter, S., T. J. Tetaz, M. C. Berndt, L. K. Ashman. 1995. Molecular cloning of cDNA encoding a novel platelet-endothelial cell tetra-span antigen, PETA-3. Blood 86:1348.[Abstract/Free Full Text]
  22. Ashman, L. K., G. W. Aylett, P. A. Mehrabani, L. J. Bendall, S. Niutta, A. C. Cambareri, S. R. Cole, M. C. Berndt. 1991. The murine monoclonal antibody, 14A2.H1, identifies a novel platelet surface antigen. Br. J. Haematol. 79:263.[Medline]
  23. Roberts, J. J., S. E. Rodgers, J. Drury, L. K. Ashman, J. V. Lloyd. 1995. Platelet activation induced by a murine monoclonal antibody directed against a novel tetra-span antigen. Br. J. Haematol. 89:853.[Medline]
  24. Maecker, H. T., S. C. Todd, S. Levy. 1997. The tetraspanin superfamily: molecular facilitators. FASEB J. 11:428.[Abstract]
  25. Hemler, M. E., A. B. Mannion, F. Berditchevski. 1996. Association of TM4SF proteins with integrins: relevance to cancer. Biochim. Biophys. Acta 1287:67.[Medline]
  26. Wright, M. D., M. G. Tomlinson. 1994. The ins and outs of the transmembrane 4 superfamily. Immunol. Today 15:588.[Medline]
  27. Hasegawa, H., Y. Utsunomiya, K. Kishimoto, Y. Tange, M. Yasukawa, S. Fujita. 1996. SFA-2, a novel bZIP transcription factor induced by human T-cell leukemia virus type 1, is highly expressed in mature lymphocytes. Biochem. Biophys. Res. Commun. 222:164.[Medline]
  28. Hasegawa, H., Y. Satake, Y. Kobayashi. 1990. Effect of cytokines on Japanese encephalitis virus production by human monocytes. Microbiol. Immunol. 34:459.[Medline]
  29. Hasegawa, H., Y. Utsunomiya, M. Yasukawa, K. Yanagisawa, S. Fujita. 1994. Induction of G protein-coupled peptide receptor EBI 1 by human herpesvirus 6 and 7 infection in CD4+ T cells. J. Virol. 68:5326.[Abstract/Free Full Text]
  30. Shimoyama, M., and members of The Lymphoma Study Group (1984–87). 1991. Diagnostic criteria and classification of clinical subtypes of adult T-cell leukemia-lymphoma. Br. J. Haematol. 79:428.
  31. Ishikawa, T., A. Imura, K. Tanaka, H. Shirane, M. Okuma, T. Uchiyama. 1993. E-selectin and vascular cell adhesion molecule-1 mediate adult T-cell leukemia cell adhesion to endothelial cells. Blood 82:1590.[Abstract/Free Full Text]
  32. Takebe, Y., M. Seiki, J. Fujisawa, P. Hoy, K. Yokota, K. Arai, M. Yoshida, N. Arai. 1988. The SR{alpha} promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol. Cell. Biol. 8:466.[Abstract/Free Full Text]
  33. Hasegawa, H., M. Yoshida, T. Shiosaka, S. Fujita, Y. Kobayashi. 1992. Mutations in the envelope protein of Japanese encephalitis virus affect entry into cultured cells and virulence in mice. Virology 191:158.[Medline]
  34. Hasegawa, H., M. Yoshida, Y. Kobayashi, S. Fujita. 1995. Antigenic analysis of Japanese encephalitis viruses in Asia by using monoclonal antibodies. Vaccine 13:1713.[Medline]
  35. Sonnenberg, A., C. J. T. Linders, P. W. Modderman, C. H. Damsky, M. Aumailley, R. Timpl. 1990. Integrin recognition of different cell binding fragments of laminin (P1, E3, E8) and evidence that {alpha}6ß1 but not {alpha}6ß4 functions as a major laminin receptor for fragment E8. J. Cell. Biol. 110:2145.[Abstract/Free Full Text]
  36. Camichael, J., W. G. Degraff, A. F. Gazdar, J. D. Minna, J. B. Mitchell. 1987. Evaluation of a tetrazolium-based semi-automated colorimetric assay: assessment of radiosensitivity. Cancer Res. 47:943.[Abstract/Free Full Text]
  37. Berditchevski, F., M. H. Zutter, M. E. Hemler. 1996. Characterization of novel complexes on the cell surface between integrins and proteins with 4 transmembrane domains (TM4 proteins). Mol. Biol. Cell 7:193.[Abstract]
  38. Radford, K. J., R. F. Thorne, P. Hersey. 1996. CD63 associates with transmembrane 4 superfamily members, CD9 and CD81, and with ß1 integrins in human melanoma. Biochem. Biophys. Res. Commun. 222:13.[Medline]
  39. Berditchevski, F., G. Bazzoni, M. E. Hemler. 1995. Specific association of CD63 with the VLA-3 and VLA-6 integrins. J. Biol. Chem. 270:17784.[Abstract/Free Full Text]
  40. Rubinstein, E., F. Le Naour, M. Billard, M. Prenant, C. Boucheix. 1994. CD9 antigen is an accessory subunit of the VLA integrin complexes. Eur. J. Immunol. 24:3005.[Medline]
  41. Nakamura, K., R. Iwamoto, E. Mekata. 1995. Membrane-anchored heparin-binding EGF-like growth factor (HB-ECF) and diphtheria toxin receptor-associated protein (DRAP27)/CD9 form a complex with integrin {alpha}3ß1 at cell-cell contact sites. J. Cell Biol. 129:1691.[Abstract/Free Full Text]
  42. Rubinstein, E., F. Le Naour, C. Lagaudriere-Gesbert, M. Billard, H. Conjeaud, C. Boucheix. 1996. CD9, CD63, CD81, and CD82 are components of a surface tetraspan network connected to HLA-DR and VLA integrins. Eur. J. Immunol. 26:2657.[Medline]
  43. Behr, S., F. Schriever. 1995. Engaging CD19 or target of an antiproliferative antibody 1 on human B lymphocytes induces binding of B cells to the interfollicular stroma of human tonsils via integrin {alpha}4/ß1 and fibronectin. J. Exp. Med. 182:1191.[Abstract/Free Full Text]
  44. Hadjiargyrou, M., Z. Kaprielian, N. Kato, P. H. Patterson. 1996. Association of the tetraspan protein CD9 with integrins on the surface of S-16 Schwann cells. J. Neurochem. 67:2505.[Medline]
  45. Mannion, B. A., F. Berditchevski, S.-K. Kraeft, L. B. Chen, M. E. Hemler. 1996. Transmembrane-4 superfamily proteins CD81 (TAPA-1), CD82, CD63, and CD53 specifically associate with integrin {alpha}4ß1 (CD49d/CD29). J. Immunol. 157:2039.[Abstract]
  46. Matsumoto, A. K., D. R. Martin, R. H. Carter, L. B. Klickstein, J. M. Ahearn, D. T. Fearon. 1993. Functional dissection of the CD21/CD19/TAPA-1/Leu-13 complex of B lymphocytes. J. Exp. Med. 178:1407.[Abstract/Free Full Text]
  47. Bradbury, L. E., G. S. Kansas, S. Levy, R. L. Evans, T. F. Tedder. 1992. The CD19/CD21 signal transducing complex of human B lymphocytes includes the target of antiproliferative antibody-1 and Leu-13 molecules. J. Immunol. 149:2841.[Abstract]
  48. Imai, T., M. Kakizaki, M. Nishimura, O. Yoshie. 1995. Molecular analysis of the association of CD4 with two members of the transmembrane 4 superfamily, CD81 and CD82. J. Immunol. 155:1229.[Abstract]
  49. Sincock, P. M., G. Mayrhofer, L. K. Ashman. 1997. Localization of the transmembrane 4 superfamily (TM4SF) member PETA-3 (CD151) in normal human tissues: comparison with CD9, CD63, and {alpha}5ß1 integrin. J. Histochem. Cytochem. 45:515.[Abstract/Free Full Text]
  50. Akiyama, S. K., K. Olden, K. M. Yamada. 1995. Fibronectin and integrins in invasion and metastasis. Cancer Metastasis Rev. 14:173.[Medline]
  51. Plantefaber, L. C., R. O. Hynes. 1989. Changes in integrin receptors on oncogenically transformed cells. Cell 56:281.[Medline]
  52. Schreiner, C. L., J. S. Bauer, Y. N. Danilov, S. Hussein, M. M. Sczekan, R. L. Juliano. 1989. Isolation and characterization of Chinese hamster ovary cell variants deficient in the expression of fibronectin receptor. J. Cell Biol. 109:3157.[Abstract/Free Full Text]
  53. Giancotti, F., E. Ruoslahti. 1990. Elevated levels of {alpha}5/ß1 fibronectin receptor suppress the transformed phenotype in CHO cells. Cell 60:849.[Medline]
  54. Zutter, M. M., G. Mazoujian, S. A. Santoro. 1990. Decreased expression of integrin adhesive protein receptors in adenocarcinoma of the breast. Am. J. Pathol. 137:863.[Abstract]
  55. Gui, G. P. H., J. R. Puddefoot, G. P. Vinson, C. A. Wells, R. Carpenter. 1997. Altered cell-matrix contact: a prerequisite for breast cancer metastasis?. Br. J. Cancer 75:623.[Medline]
  56. Tawil, N., P. Wilson, S. Carbonetto. 1993. Integrins in point contacts mediate cell spreading: factors that regulate integrin accumulation in point contacts vs. focal contacts. J. Cell Biol. 120:261.[Abstract/Free Full Text]
  57. Gailit, J., E. Rouslahti. 1988. Regulation of the fibronectin receptor affinity by divalent cations. J. Biol. Chem. 263:12927.[Abstract/Free Full Text]
  58. Zheng, M., T. Tsuruoka, T. Tsuji, S. Hakomori. 1992. Regulatory role of GM3 ganglioside in integrin function, as evidenced by its effect on function of {alpha}5ß1-liposomes: a preliminary note. Biochem. Biophys. Res. Commun. 186:1397.[Medline]
  59. Dhawan, S., B. S. Weeks, F. Abbasi, H. R. Gralnick, A. L. Natkins, M. E. Klotmon, K. M. Yamada, P. E. Klotman. 1993. Increased expression of {alpha}4ß1 and {alpha}5ß1 integrins on HTLV-I-infected lymphocytes. Virology 197:778.[Medline]
  60. Torre, D., G. Ferrario, M. Issi, A. Pugliese, F. Speranza. 1996. Expression of the {alpha}5ß1 fibronectin receptor on T lymphocytes of patients with HIV-1 infection. J. Clin. Pathol. 49:733.[Abstract/Free Full Text]
  61. Castanos-Velez, E., P. Biberfeld, M. Patarroyo. 1995. Extracellular matrix proteins and integrin receptors in reactive and non-reactive lymph nodes. Immunology 86:270.[Medline]
  62. Koopman, G., H. K. Parmentier, H. J. Schuurman, W. Newman, C. J. Meijer, S. T. Pals. 1991. Adhesion of human B cells to follicular dendritic cells involves both the lymphocyte function-associated antigen 1/intercellular adhesion molecule 1 and very late antigen 4/vascular cell adhesion molecule 1 pathway. J. Exp. Med. 173:1297.[Abstract/Free Full Text]
  63. Freedman, A. S., J. M. Munro, C. Morimoto, B. W. McIntyre, K. Rhynhart, N. Lee, L. M. Nadler. 1992. Follicular non-Hodgkin’s lymphoma cell adhesion to normal germinal centers and neoplastic follicles involves very late antigen-4 and vascular cell adhesion molecule-1. Blood 79:206.[Abstract/Free Full Text]
  64. Lasky, L. A., M. S. Singer, D. Dowbenko, Y. Imai, W. J. Henzel, C. Grimley, C. Fennie, N. Gillett, S. R. Watson, S. D. Rosen. 1992. An endothelial ligand for L-selectin is a novel mucin-like molecule. Cell 69:927.[Medline]
  65. Szekanecz, Z., M. J. Humphries, A. Ager. 1992. Lymphocyte adhesion to high endothelium is mediated by two ß1 integrin receptors for fibronectin, {alpha}4ß1 and {alpha}5ß1. J. Cell Sci. 101:885.[Abstract/Free Full Text]
  66. Tanaka, Y., A. Wake, K. J. Horgan, S. Murakami, M. Aso, K. Saito, S. Oda, I. Morimoto, H. Uno, H. Kikuchi, Y. Izumi, S. Eto. 1997. Distinct phenotype of leukemic T cells with various tissue tropisms. J. Immunol. 158:3822.[Abstract]



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PNAS, February 8, 2005; 102(6): 1939 - 1944.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Epidemiol. Biomarkers Prev.Home page
J. Ang, M. Lijovic, L. K. Ashman, K. Kan, and A. G. Frauman
CD151 Protein Expression Predicts the Clinical Outcome of Low-Grade Primary Prostate Cancer Better than Histologic Grading: A New Prognostic Indicator?
Cancer Epidemiol. Biomarkers Prev., November 1, 2004; 13(11): 1717 - 1721.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
Y. Imaizumi, H. Murota, S. Kanda, Y. Hishikawa, T. Koji, T. Taguchi, Y. Tanaka, Y. Yamada, S. Ikeda, T. Kohno, et al.
Expression of the c-Met Proto-Oncogene and Its Possible Involvement in Liver Invasion in Adult T-cell Leukemia
Clin. Cancer Res., January 1, 2003; 9(1): 181 - 187.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
M. Sasaki, H. Hasegawa, M. Kohno, A. Inoue, M. R. Ito, and S. Fujita
Antagonist of Secondary Lymphoid-Tissue Chemokine (CCR Ligand 21) Prevents the Development of Chronic Graft-Versus-Host Disease in Mice
J. Immunol., January 1, 2003; 170(1): 588 - 596.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
N. Anzai, Y. Lee, B.-S. Youn, S. Fukuda, Y.-J. Kim, C. Mantel, M. Akashi, and H. E. Broxmeyer
c-kit associated with the transmembrane 4 superfamily proteins constitutes a functionally distinct subunit in human hematopoietic progenitors
Blood, May 29, 2002; 99(12): 4413 - 4421.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
L. M. T. Sterk, C. A. W. Geuijen, J. G. van den Berg, N. Claessen, J. J. Weening, and A. Sonnenberg
Association of the tetraspanin CD151 with the laminin-binding integrins {alpha}3{beta}1, {alpha}6{beta}1, {alpha}6{beta}4 and {alpha}7{beta}1 in cells in culture and in vivo
J. Cell Sci., March 15, 2002; 115(6): 1161 - 1173.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
T. Tokuhara, H. Hasegawa, N. Hattori, H. Ishida, T. Taki, S. Tachibana, S. Sasaki, and M. Miyake
Clinical Significance of CD151 Gene Expression in Non-Small Cell Lung Cancer
Clin. Cancer Res., December 1, 2001; 7(12): 4109 - 4114.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
F. Berditchevski
Complexes of tetraspanins with integrins: more than meets the eye
J. Cell Sci., January 12, 2001; 114(23): 4143 - 4151.
[Abstract] [Full Text] [PDF]

Home page
 JCBHome page
L. M.Th. Sterk, C. A.W. Geuijen, L. C.J.M. Oomen, J. Calafat, H. Janssen, and A. Sonnenberg
The Tetraspan Molecule Cd151, a Novel Constituent of Hemidesmosomes, Associates with the Integrin {alpha}6{beta}4 and May Regulate the Spatial Organization of Hemidesmosomes
J. Cell Biol., May 15, 2000; 149(4): 969 - 982.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
C. Stipp and M. Hemler
Transmembrane-4-superfamily proteins CD151 and CD81 associate with alpha 3 beta 1 integrin, and selectively contribute to alpha 3 beta 1-dependent neurite outgrowth
J. Cell Sci., January 6, 2000; 113(11): 1871 - 1882.
[Abstract] [PDF]

Home page
 BloodHome page
H. Hasegawa, T. Nomura, M. Kohno, N. Tateishi, Y. Suzuki, N. Maeda, R. Fujisawa, O. Yoshie, and S. Fujita
Increased chemokine receptor CCR7/EBI1 expression enhances the infiltration of lymphoid organs by adult T-cell leukemia cells
Blood, January 1, 2000; 95(1): 30 - 38.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. S. Stipp, T. V. Kolesnikova, and M. E. Hemler
EWI-2 Is a Major CD9 and CD81 Partner and Member of a Novel Ig Protein Subfamily
J. Biol. Chem., October 26, 2001; 276(44): 40545 - 40554.
[Abstract] [Full Text] [PDF]

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