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1.
Bone Marrow Transplant ; 40(6): 585-92, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17637687

ABSTRACT

Donor lymphocyte infusion (DLI) after allogeneic SCT induces complete remissions in approximately 80% of patients with relapsed CML in chronic phase, but some patients do not respond to DLI. We studied absolute numbers of dendritic cell (DC) subsets and chimerism in T cells and two subsets of blood DCs (myeloid DCs (MDCs) and plasmacytoid DCs (PDCs)) in relation to DLI-induced alloreactivity. Based on T cell and DC chimerism, we identified three groups. Four patients were completely donor chimeric in T cells and DC subsets. These patients had an early stage of relapse, and three of the four patients attained complete molecular remission (CMolR) without significant GVHD. Six patients were completely donor in T cells and mixed chimeric in DC subsets. All patients entered CMolR, but this was associated with GVHD in four and cytopenia in three patients. Five patients had mixed chimerism in T cells and complete recipient chimerism in MDC; only two patients entered CMolR. Our data suggest that the combination of donor T cells and mixed chimerism in DC subsets induces a potent graft-versus-leukemia (GVL) effect in association with GVHD. DLI in patients with an early relapse and donor chimerism in both T cells and DC subsets results in GVL reactivity without GVHD.


Subject(s)
Dendritic Cells/immunology , Graft vs Leukemia Effect/immunology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/immunology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy , Lymphocyte Transfusion , T-Lymphocytes/immunology , Transplantation Chimera/immunology , Adult , Aged , Blood Donors , Female , Graft vs Host Disease/immunology , Humans , Lymphocyte Count , Male , Middle Aged , Recurrence , Remission Induction
2.
Leukemia ; 17(3): 621-9, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12646953

ABSTRACT

Analysis of changes in recipient and donor hemopoietic cell origin is extremely useful to monitor the effect of stem cell transplantation (SCT) and sequential adoptive immunotherapy by donor lymphocyte infusions (DLI). We developed a sensitive and accurate method to quantify the percentage of recipient and donor cells by real-time PCR using single nucleotide polymorphisms (SNPs) as markers. Allele-specific PCR of seven SNPs resulted in specific markers for donor or recipient in 97% of HLA-identical sibling pairs. Both, recipient- and donor-derived hemopoietic cells can be simultaneously analyzed in 67% sibling pairs. We expect this can be increased to approximately 99% by developing three additional SNP-PCR. Serial dilution of SNP-positive DNA into either SNP-negative DNA or water revealed a detection limit of 0.1-0.01% depending on the amount of input DNA and start C(t) of the used SNP-PCR. Application of our real-time SNP-PCR method for a CML patient treated by allogeneic SCT and DLI demonstrated its feasibility to follow donor T-cell chimerism and early detection of residual and recurrent autologous hemopoiesis in response to treatment. This detailed monitoring of the genetic origin of hemopoietic cells, in particular immune effector cells and target cells after SCT and DLI, may substantially contribute to understanding of the mechanisms that play a role in the success of treatment.


Subject(s)
Blood Cells , Hematopoietic Stem Cell Transplantation/standards , Polymerase Chain Reaction/methods , Polymorphism, Single Nucleotide , Transplantation Chimera , Alleles , Blood Cells/cytology , Feasibility Studies , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy , Lymphocyte Transfusion , Myeloid Cells/cytology , Polymerase Chain Reaction/standards , Reproducibility of Results , Sensitivity and Specificity , T-Lymphocytes/cytology , Transplantation, Homologous/standards
4.
Leukemia ; 15(9): 1339-46, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11516094

ABSTRACT

In this prospective study we analyzed pre-emptive donor leukocyte infusions (DLI) in 82 consecutive patients transplanted with partially T cell-depleted grafts for acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, refractory anemia with excess of blasts, refractory anemia with excess of blasts in transformation and multiple myeloma. Donors were HLA-identical siblings. Patients without significant acute (>grade 1) and/or chronic GVHD were scheduled to be treated with DLI (35 patients) and 31 actually received DLI. Patients who developed acute GVHD >grade 1 and/or chronic GVHD were not scheduled to receive DLI and served as a comparison group (47 patients). The median interval between BMT and DLI was 22 weeks. The first six patients received 0.7 x 10(8) CD3+ cells/kg body weight (b.w.). Five out of these six patients developed acute GVHD (grade 1: n = 2, grade 3: n = 2 and grade 4: n= 1) which was more frequent and more severe than we had anticipated. In the next 25 patients the number of T lymphocytes was diminished to 0.1 x 10(8) CD3+ cells/kg b.w. which resulted in less frequent and less severe GVHD. Eight patients in this group developed acute GVHD (grade 1: n = 4, grade 2: n = 4) and three patients had limited chronic GVHD. Patients in the DLI group needed more time to establish complete donor chimerism confirmed by a higher number of mixed chimeras at 6 months after BMT. The projected 3-year probability of disease-free survival was 77% for the 35 patients intended to treat with DLI and 45% for the patients of the comparison group (P = 0.024). Relapse rate at 36 months after transplantation was 18% in the patients who were intended to treat with DLI and 44% in the comparison group (P = 0.026). We conclude that pre-emptive DLI is feasible and generates favorable relapse rates in patients who are at high risk for relapse. Furthermore, the incidence and severity of GVHD disease after DLI is dependent on the number of CD3+ cells infused.


Subject(s)
Graft vs Leukemia Effect , Hematopoietic Stem Cell Transplantation , Leukemia/therapy , Transplantation Conditioning/methods , Adolescent , Adult , CD3 Complex/analysis , Chimera , Disease-Free Survival , Female , Graft vs Host Disease/prevention & control , Humans , Leukocytes/immunology , Male , Middle Aged , Prospective Studies , Tissue Donors
5.
Bone Marrow Transplant ; 27(10): 1087-93, 2001 May.
Article in English | MEDLINE | ID: mdl-11438826

ABSTRACT

Cytotoxic T lymphocytes (CTL) are thought to play an important role in the graft-versus-leukemia (GVL) response. Unfortunately, GVL reactivity is often associated with life-threatening graft-versus-host disease (GVHD). Characterization of CTL that selectively attack leukemic cells but not normal cells may lead to the development of adjuvant immunotherapy that separates GVL from GVHD. Here, we describe TCR gamma delta (V gamma 9/V delta 1) CTL, isolated from the peripheral blood of an AML patient after stem cell transplantation (SCT), that very efficiently lysed freshly isolated acute myeloid leukemia (AML) cells and AML cell lines. Interestingly, HLA-matched non-malignant hematopoietic cells were not killed. We revealed that the killer cell-inhibitory receptor (KIR) p58.2 (CD158b) specific for group 2 HLA-C molecules negatively regulates the cytotoxic effector function displayed by these TCR gamma delta CTL. First, an antibody against HLA-C enhances lysis of non-malignant cells. Secondly, stable transfection of HLA-Cw*0304 into the class I-negative cell line 721.221 inhibited lysis. Finally, engagement of p58.2 by antibodies immobilized on Fc gamma R-expressing murine P815 cells inhibits CD3- and TCR gamma delta-directed lysis. Compared to non-malignant hematopoietic cells, AML cells express much lower levels of MHC class I molecules making them susceptible to lysis by p58.2(+) TCR gamma delta CTL. Such KIR-regulated CTL reactivity may have a role in the GVL response without affecting normal tissues of the host and leading to GVHD.


Subject(s)
Leukemia, Myeloid/pathology , Receptors, Immunologic/biosynthesis , T-Lymphocytes, Cytotoxic/immunology , Acute Disease , Clone Cells/immunology , Cytotoxicity Tests, Immunologic , HLA-C Antigens/immunology , Humans , Leukemia, Myeloid/immunology , Receptors, Antigen, T-Cell, gamma-delta , Receptors, KIR , Receptors, KIR2DL3 , T-Lymphocytes, Cytotoxic/chemistry , Tumor Cells, Cultured
6.
Leuk Lymphoma ; 32(3-4): 317-25, 1999 Jan.
Article in English | MEDLINE | ID: mdl-10037029

ABSTRACT

Donor leukocyte infusions (DLI) from the original marrow donor have been shown to induce remission in patients with relapse after BMT. We analyzed factors that were associated with remission. Twenty-six patients with a relapse after T cell depleted BMT received DLI. The following pre-DLI factors were analyzed: sex and age of the patients and donors, GVHD after BMT, indication for DLI, percentage of donor T lymphocytes in the patient at the time of DLI, interval between relapse and DLI, and number of T lymphocytes infused. Remission was achieved in 11 of 15 patients (73%) treated for relapsed CML and in one of 11 patients (9%) treated for relapsed AML, ALL or RAEB-t (P = .002). Two of 13 patients (15%) with < or =40% of T lymphocytes from donor origin attained remission compared with 10 of 13 patients (77%) with >40% (P = .002). Two of 13 patients (15%) with an interval of < or =18 months between BMT and first DLI entered remission compared with 10 of 13 patients (77%) with an interval of >18 months (P = .002). Multivariate analysis demonstrated that indication for DLI (CML versus AML/ALL and RAEB-t) and the percentage T lymphocytes from donor origin (< or =40 versus >40) were significantly correlated with remission (P = .03). The occurrence of GVHD post DLI was highly associated with achievement of remission (P = .0001). DLI res ults in remission in a high percentage of patients with relapsed CML after BMT. The percentage of T lymphocytes from donor origin still present in the patient at the time of DLI is highly correlated with achievement of remission.


Subject(s)
Blood Donors , Bone Marrow Transplantation , Leukocyte Transfusion , Lymphocyte Depletion , Neoplasm Recurrence, Local , T-Lymphocytes/cytology , Adult , Anemia, Refractory, with Excess of Blasts/therapy , Bone Marrow Transplantation/adverse effects , Female , Graft vs Host Disease/etiology , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy , Leukemia, Myeloid, Acute/therapy , Leukocyte Transfusion/adverse effects , Lymphocyte Count , Male , Middle Aged , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Recurrence , Remission Induction
7.
J Exp Med ; 189(2): 301-8, 1999 Jan 18.
Article in English | MEDLINE | ID: mdl-9892612

ABSTRACT

Human minor histocompatibility antigens (mHags) play an important role in the induction of cytotoxic T lymphocyte (CTL) reactivity against leukemia after human histocompatibility leukocyte antigen (HLA)-identical allogeneic bone marrow transplantation (BMT). As most mHags are not leukemia specific but are also expressed by normal tissues, antileukemia reactivity is often associated with life-threatening graft-versus-host disease (GVHD). Here, we describe a novel mHag, HB-1, that elicits donor-derived CTL reactivity in a B cell acute lymphoblastic leukemia (B-ALL) patient treated by HLA-matched BMT. We identified the gene encoding the antigenic peptide recognized by HB-1-specific CTLs. Interestingly, expression of the HB-1 gene was only observed in B-ALL cells and Epstein-Barr virus-transformed B cells. The HB-1 gene-encoded peptide EEKRGSLHVW is recognized by the CTL in association with HLA-B44. Further analysis reveals that a polymorphism in the HB-1 gene generates a single amino acid exchange from His to Tyr at position 8 within this peptide. This amino acid substitution is critical for recognition by HB-1-specific CTLs. The restricted expression of the polymorphic HB-1 Ag by B-ALL cells and the ability to generate HB-1-specific CTLs in vitro using peptide-loaded dendritic cells offer novel opportunities to specifically target the immune system against B-ALL without the risk of evoking GVHD.


Subject(s)
Burkitt Lymphoma/immunology , HLA Antigens/immunology , HLA-B Antigens/genetics , T-Lymphocytes, Cytotoxic/immunology , Amino Acid Sequence , Base Sequence , Bone Marrow Transplantation/immunology , Cell Line , Clone Cells/immunology , Cloning, Molecular , DNA, Complementary/genetics , Female , Gene Expression Regulation/genetics , Gene Expression Regulation/immunology , Graft vs Host Disease/immunology , Humans , Male , Minor Histocompatibility Antigens , Molecular Sequence Data , Pedigree , Peptide Fragments/immunology , Polymorphism, Genetic/genetics , Sequence Analysis
8.
J Immunol ; 158(2): 560-5, 1997 Jan 15.
Article in English | MEDLINE | ID: mdl-8992968

ABSTRACT

CTL directed against minor histocompatibility Ags (mHag) play a major role in antileukemia reactivity after HLA-identical bone marrow transplantation. Some of these mHag are restricted to hemopoietic cells, others show a broad tissue expression. Therefore, antileukemia reactivity is often associated with graft-vs-host disease. Here, we report the identification of a B cell leukemia-associated mHag, HB-1, recognized by a CD8+ CTL clone derived from peripheral blood of an acute lymphoblastic B cell leukemia patient who has been treated by HLA-matched bone marrow transplantation. Interestingly, the CTL clone that recognizes HB-1 exhibits specific cytotoxicity toward leukemic as well as EBV-transformed B cells, but not against untransformed B cells. Moreover, the CTL clone does not lyse PHA-stimulated T cell blasts, monocytes, and fibroblasts, indicating that HB-1 is mainly expressed by transformed B cells. Further analysis reveals that HB-1 is restricted by HLA-B44 (both B*4402 and B*4403) and that 28% of HLA-B44-positive individuals express HB-1. These findings demonstrate that leukemia-associated mHag with a restricted tissue distribution, such as HB-1, elicit CTL reactivity in vivo. These Ags are of potential use in immunotherapy against leukemia because they generate antileukemia reactivity that is not associated with graft-vs-host disease.


Subject(s)
Antigens, Neoplasm/analysis , Antigens, Neoplasm/immunology , Burkitt Lymphoma/immunology , Minor Histocompatibility Antigens/analysis , Minor Histocompatibility Antigens/immunology , T-Lymphocytes, Cytotoxic/immunology , Adult , Burkitt Lymphoma/metabolism , Cell Line, Transformed , Cytotoxicity, Immunologic/immunology , Female , HLA-B Antigens/genetics , HLA-B44 Antigen , Herpesvirus 4, Human/pathogenicity , Humans , Minor Histocompatibility Antigens/biosynthesis , Organ Specificity/immunology , Pedigree
9.
Ned Tijdschr Geneeskd ; 140(42): 2087-91, 1996 Oct 19.
Article in Dutch | MEDLINE | ID: mdl-8965951

ABSTRACT

OBJECTIVE: Evaluation of induction of complete remission with infusion of lymphocytes from the original bone marrow donor in patients with leukaemia which relapsed after allogeneic bone marrow transplantation. SETTING: Division of Haematology, University Hospital Nijmegen and the Red Cross Blood Bank Nijmegen, the Netherlands. DESIGN: Prospective, non-randomized study. METHODS: Twenty-eight patients who relapsed after allogeneic bone marrow transplantation were treated with infusion of lymphocytes from the original bone marrow donor. Lymphocytes were collected by means of leukapheresis. Follow-up was done by frequent checks at the outpatient clinic. RESULTS: Eleven of 15 (73%) patients with relapsed chronic myeloid leukaemia (CML) and only one of 13 patients (8%) with a relapse of acute leukaemia went into complete remission (p < 0.001). Entering complete remission was always preceded by acute or chronic graft-versus-host disease (GVHD). The development of acute and/or chronic GVHD was significantly associated with the origin of T-lymphocytes in the blood of the recipient at the time of infusion. If the T-lymphocytes came mostly from the patient himself, the infusion remained usually without effect. If the T-lymphocytes came mostly from the donor, the patients went into complete remission. CONCLUSION: Patients with a relapse of leukaemia after allogeneic bone marrow transplantation may enter complete remission after infusion of lymphocytes from the original marrow donor. This form of immunotherapy can be successful especially in patients with a relapsed CML with a relatively low percentage of autologous T-lymphocytes at the time of infusion.


Subject(s)
Leukemia, Myeloid, Acute/therapy , Lymphocyte Transfusion/methods , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Adolescent , Adult , Bone Marrow Transplantation , Cytogenetics/methods , Female , Humans , Immunotherapy, Adoptive/methods , Male , Middle Aged , Prospective Studies , Recurrence , Transplantation, Homologous
10.
Bone Marrow Transplant ; 18(2): 339-45, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8864444

ABSTRACT

After lymphocyte-depleted BMT, CD8+ T cells have been expanded to or above normal levels in 45% of the recipients within 3 months. The mechanisms underlying proliferation of donor-derived CD8+ T cells after BMT are still unclear. We investigated whether these CD8+ T cells proliferate in response to specific antigens by determination of TCR clonality and whether these cells exert specific cytotoxicity. PCR analysis of TCR-gamma gene rearrangements showed a marked clonal predominance in CD8+ T cells of recipients with a high number of these cells. Strong association between expansion of CD8+ T cells and CMV infection suggests involvement of CMV antigens. Therefore, we examined CMV-specific cytotoxicity of freshly isolated CD8+ T cells of two BMT recipients with clonal expansion after the onset of CMV infection. CD8+ T cells exerted HLA-restricted cytotoxicity directed against CMV-infected fibroblasts indicating that CMV stimulates proliferation. The majority of CD8+ T cells in these recipients expressed CD57. We demonstrated that TCR clonality was irrespective of CD57 expression. Both CD8+CD57+ and CD8+CD57- T cells showed significant HLA-restricted CMV-specific cytotoxicity. These studies strongly suggest that CMV antigens can induce expansion of clonal CD8+ T cells after BMT.


Subject(s)
Bone Marrow Transplantation , Cytomegalovirus/immunology , T-Lymphocytes, Cytotoxic/immunology , CD57 Antigens/physiology , Cytotoxicity, Immunologic , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor , Humans
11.
Br J Haematol ; 90(2): 300-7, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7540855

ABSTRACT

Peripheral blood lymphocytes of 46 recipients of lymphocyte-depleted bone marrow allografts were phenotypically analysed over a period of 1 year. We investigated the repopulation of lymphocyte subpopulations and their relation with clinical parameters such as graft-versus-host disease (GVHD), graft-versus-leukaemia and cytomegalovirus (CMV) infection. The number of repopulated T cells varied strongly between the blood samples of the recipients. In 45% of the recipients the number of T cells recovered to or above normal levels within 3 months after bone marrow transplantation (BMT), whereas the other recipients remained below normal up to 1 year after BMT. In recipients with a high repopulation, the CD8+ T-cell subset contributed more to this high repopulation than the CD4+ T-cell subset. We showed that the majority of T cells of these recipients expressed the alpha beta T-cell receptor, CD8, CD57 and CD11b. HLA-DR was also highly expressed reflecting the activation stage of T cells in these recipients. BMT recipients with a high repopulation of CD8+ T cells showed a lower incidence of leukaemic relapse than recipients with a low repopulation. The 3-year probability of relapse was 19% versus 64% (P = 0.03), respectively. The relative high number of CD8+ T cells at 3 months after BMT was not associated with the incidence of GVHD. In contrast, occurrence of CMV infection after BMT was significantly higher in these recipients. Our results indicate that CD8+ T cells, predominantly CD57+, of BMT recipients with an expansion of these cells represent an in vivo activated cell population. This CD8+ T-cell population may consist partially of cytotoxic cells with anti-leukaemic activity as suggested by a low relapse rate. The signal for the strong expansion of these CD8+CD57+ T cells after BMT is still unclear, but association with CMV infection suggests that viral antigens are involved.


Subject(s)
Antigens, CD/immunology , Antigens, Differentiation, T-Lymphocyte/immunology , Bone Marrow Transplantation , CD8-Positive T-Lymphocytes/immunology , Leukemia/therapy , T-Lymphocyte Subsets/immunology , Adolescent , Adult , Anemia/immunology , CD57 Antigens , Cytomegalovirus Infections/immunology , Female , Graft vs Host Disease/immunology , Humans , Leukemia/immunology , Lymphoma/immunology , Male , Middle Aged , Recurrence
12.
J Immunol ; 151(2): 767-76, 1993 Jul 15.
Article in English | MEDLINE | ID: mdl-7687622

ABSTRACT

Episialin (MUC1) is a mucin-like glycoprotein abundantly expressed on most carcinoma cells. As a result of its extended and rigid structure, it reduces intercellular adhesion. We investigated whether this antiadhesion function allows tumor cells expressing high levels of episialin to escape from immune recognition. To test this hypothesis, we transfected episialin-negative (episialin-) melanoma cells (A375) with the MUC1 cDNA-encoding episialin. The results demonstrated that episialin-positive (episialin+) melanoma cells were significantly less susceptible to lysis than episialin- melanoma cells by both alloantigen or rIL-2-stimulated cytotoxic effector cells. In addition, cold target inhibition experiments with episialin+ and episialin- cells clearly demonstrated preferential lysis of episialin- cells. Furthermore, antibody blocking studies showed that lysis of episialin+, but not of episialin-, melanoma cells was predominantly dependent on the leukocyte function-associated Ag-1/intracellular adhesion molecule adhesion route, suggesting that episialin+ target cells adhere less efficiently to effector cells than episialin- target cells. This notion was supported by the observation that conjugate formation of the effector cells with episialin+ target cells was significantly impaired. From these results we conclude that over-expression of episialin as found on many tumor cells may indeed affect efficient lysis by cytotoxic lymphocytes and thus may contribute to escape from immune surveillance.


Subject(s)
Antigens, Neoplasm/physiology , Cell Adhesion , Cytotoxicity, Immunologic , Membrane Glycoproteins/physiology , T-Lymphocytes, Cytotoxic/immunology , Antigens, CD/physiology , CD58 Antigens , Cell Adhesion Molecules/physiology , Cells, Cultured , Humans , Intercellular Adhesion Molecule-1 , Melanoma/immunology , Mucin-1 , Transfection , Tumor Cells, Cultured
13.
Int J Cancer ; 54(2): 315-21, 1993 May 08.
Article in English | MEDLINE | ID: mdl-8387465

ABSTRACT

We compared integrin-mediated adhesion to extracellular matrix (ECM) components of cultured human melanocytes and 6 human melanoma cell lines with different metastatic capacities in nude mice. Cultured melanocytes and most melanoma cell lines adhered strongly to fibronectin (FN), whereas only highly metastatic cell lines adhered to laminin (LM), collagen type I (COI) and type IV (COIV). Adhesion to LM and CO could be blocked by anti-alpha 6 and anti-alpha 2 monoclonal antibodies (MAbs) respectively. This observation is consistent with the finding that expression of LM receptor alpha 6 beta 1 and LM/CO receptor alpha 2 beta 1 was low on melanocytes and non- or poorly metastatic cell lines, whereas these integrins were strongly expressed on highly metastatic cell lines. In addition, immunoprecipitation from [35S]-methionine-labeled cells demonstrated increased synthesis of alpha 6, alpha 2 and beta 1 in highly metastatic cell lines and immunohistochemistry showed expression of alpha 6 beta 1 and alpha 2 beta 1 only in xenograft lesions from highly metastatic cell lines. Furthermore, the observation that adhesion of melanocytes and non- or poorly metastatic cell lines could be stimulated with anti beta 1 MAbs demonstrates that these receptors, on these cells, are expressed in an inactive state. Our results suggest that alpha 2 beta 1 and alpha 6 beta 1 play a role in human melanoma metastasis in nude mice and demonstrate that interactions of these integrins with their ligands can be regulated at the level of surface expression and activation state of the receptor.


Subject(s)
Collagen/metabolism , Integrins/metabolism , Laminin/metabolism , Melanoma/pathology , Neoplasm Metastasis , Receptors, Cell Surface/metabolism , Receptors, Laminin/metabolism , Animals , Antibodies, Monoclonal/immunology , Cell Adhesion , Extracellular Matrix/metabolism , Flow Cytometry , Humans , Immunoenzyme Techniques , Mice , Mice, Nude , Neoplasm Transplantation , Receptors, Collagen , Transplantation, Heterologous , Tumor Cells, Cultured
14.
J Exp Med ; 177(1): 185-90, 1993 Jan 01.
Article in English | MEDLINE | ID: mdl-7678112

ABSTRACT

Lymphocyte function-associated antigen 1/intercellular adhesion molecule 1 (LFA-1/ICAM-1)-and very late antigen 4/vascular cell adhesion molecule 1 (VLA-4/VCAM-1)-mediated adhesion of T lymphocytes to endothelial cells (EC) can be regulated by increased expression of ICAM-1 and VCAM-1 upon cytokine treatment of EC, or by activation of the integrin molecules LFA-1 and VLA-4 on T cells. Here, we provide evidence that preferential usage of LFA-1 over VLA-4 is yet another mechanism to control T cell adhesion. We observed that binding of activated T lymphocytes, as opposed to resting T cells, to EC is essentially mediated through LFA-1 and not through VLA-4. VLA-4-mediated adhesion of T cells to EC is only found when LFA-1 is not expressed or not functional, as observed for several T cell leukemia cell lines. These results suggest that LFA-1-mediated adhesion dominates and may downregulate VLA-4-mediated adhesion through an unidentified mechanism.


Subject(s)
Endothelium, Vascular/physiology , Lymphocyte Activation , Lymphocyte Function-Associated Antigen-1/physiology , Receptors, Very Late Antigen/physiology , T-Lymphocytes/physiology , Animals , Cell Adhesion , Cell Adhesion Molecules/physiology , Humans , Intercellular Adhesion Molecule-1 , Mice , Vascular Cell Adhesion Molecule-1
15.
Cancer Immunol Immunother ; 36(3): 210-3, 1993.
Article in English | MEDLINE | ID: mdl-8439983

ABSTRACT

A patient with renal cell cancer developed acute renal failure due to biopsy-proven acute tubulo-interstitial nephritis (AIN) in the 6th week of continuous infusion of 9 x 10(6) IU m-2 day-1 recombinant interleukin-2 (rIL-2). We investigated whether the AIN was the result of a cellular cytotoxic reaction induced by the rIL-2 treatment. The cytolytic activity of cryopreserved peripheral blood lymphocytes (PBL), isolated before and at the end of the rIL-2 treatment (at the time of AIN), was studied after 5 days of culture with or without rIL-2 or anti-CD28 and immobilized anti-CD3 antibodies. The PBL isolated before and at the end of the rIL-2 treatment showed cytolytic activity towards a number of allogeneic targets. However, only the PBL isolated at the end of the rIL-2 treatment showed, when stimulated with rIL-2 in vitro, significant cytolytic activity against an autologous renal cell line cultured from the AIN biopsy specimen and against an allogeneic renal cell cancer cell line. These PBL displayed no enhanced killing capacity towards autologous PBL and the melanoma cell line M14. These observations suggest that the AIN may be the result of a cytotoxic lymphocyte-mediated reaction induced by the rIL-2 treatment.


Subject(s)
Carcinoma, Renal Cell/drug therapy , Interleukin-2/adverse effects , Kidney Neoplasms/drug therapy , Nephritis, Interstitial/chemically induced , T-Lymphocytes, Cytotoxic/physiology , Acute Kidney Injury/chemically induced , Acute Kidney Injury/etiology , Humans , Immunophenotyping , Interleukin-2/therapeutic use , Lymphocyte Activation/drug effects , Lymphocyte Activation/physiology , Lymphocytes/drug effects , Lymphocytes/physiology , Lymphocytes, Tumor-Infiltrating/drug effects , Lymphocytes, Tumor-Infiltrating/physiology , Male , Melanoma/immunology , Middle Aged , Nephritis, Interstitial/etiology , Recombinant Proteins/adverse effects , Recombinant Proteins/therapeutic use , T-Lymphocytes, Cytotoxic/drug effects , Tumor Cells, Cultured
16.
Eur J Immunol ; 22(6): 1467-75, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1376259

ABSTRACT

We investigated the capacity of T lymphocytes from a leukocyte adhesion-deficient (LAD) patient to respond to alloantigen. Leukocytes of this patient completely lacked LFA-1 surface expression due to the absence of mRNA coding for the LFA-1 beta chain. Despite the absence of LFA-1, T lymphocytes obtained from this patient, cultured with allogeneic stimulator cells (lymphoblastoid B cells JY), were capable of lysing JY cells. Furthermore, two T cell clones (one CD4+ and one CD8+), generated from this lymphocyte culture, specifically lysed the allogeneic lymphoblastoid JY cells. The cytolytic capacity of LFA-1-negative T lymphocytes and T cell clones was comparable to that of control LFA-1-positive T cells with allospecificity against JY. Detailed analysis of the CD4 positive and LFA-1-negative T cell clone demonstrated that it specifically recognized HLA-DQ. Antibody inhibition studies showed that the CTL/target cell interaction was mediated through the CD2/LFA-3 adhesion pathway. LFA-1 expressed by the target cells did not participate in the CTL/target cell conjugate formation and contributed only minimally to the cytotoxic activity. Moreover, when allogeneic LFA-1-deficient B cells, bearing the appropriate HLA-DQ alloantigen, were used as target cells, significant levels of specific cytotoxicity were measured, further excluding a role for LFA-1 in this interaction. The adhesion molecules, VLA-4, CD44 and L-selectin (LECAM1) were not involved. These results demonstrate that LFA-1-negative T lymphocytes can exert allospecific cytotoxicity and that CTL/target cell contact is mediated through the CD2/LFA-3 route. This observation may explain in part why in LAD patients viral infections, cleared largely by T cells, are less frequently observed than bacterial infections, in which phagocytic cells play a major role.


Subject(s)
Antigens, Differentiation, T-Lymphocyte/physiology , Antigens, Surface/physiology , Cytotoxicity, Immunologic/immunology , Membrane Glycoproteins/physiology , Receptors, Immunologic/physiology , T-Lymphocytes/immunology , Antibodies, Monoclonal , Antigens, CD/analysis , B-Lymphocytes/immunology , Blotting, Northern , CD2 Antigens , CD4-Positive T-Lymphocytes/immunology , CD58 Antigens , Cell Adhesion/immunology , Cell Adhesion Molecules , Cell Division , Clone Cells , Flow Cytometry , Granzymes , HLA-DQ Antigens/physiology , Humans , Immunity, Cellular , Immunophenotyping , Intercellular Adhesion Molecule-1 , Lymphocyte Function-Associated Antigen-1/physiology , RNA/analysis , Serine Endopeptidases/biosynthesis , T-Lymphocytes/physiology , T-Lymphocytes, Cytotoxic/immunology
17.
J Cell Biol ; 117(2): 461-70, 1992 Apr.
Article in English | MEDLINE | ID: mdl-1560035

ABSTRACT

Investigating the regulation of very late antigen (VLA)-mediated functions, we found that TS2/16, a mAb directed against the beta chain of the VLA group of integrins, can induce binding of resting peripheral blood lymphocytes, cloned T lymphocytes, and Epstein Barr virus-transformed B cells to extracellular matrix components, fibronectin, laminin, and collagen, but not to fibrinogen. The antibody stimulates VLA-4-, VLA-5-, and VLA-6-mediated binding. Furthermore, it induces VLA-4-mediated binding to vascular cell adhesion molecule-1 expressed by rTNF-alpha-stimulated endothelial cells, but it does not stimulate homotypic aggregation of cells as described for a number of anti-VLA-4 alpha antibodies (Bednarczyk, J.L., and B. W. McIntyre. 1990. J. Immunol. 144: 777-784; Campanero, M. R., R. Pulido, M. A. Ursa, M. Rodríguez-Moya, M. O. de Landázuri, and F. Sánchez-Madrid. 1990. J. Cell Biol. 110:2157-2165). Therefore, the stimulating activity of this anti-beta 1 antibody clearly contrasts with that of the anti-VLA-4 alpha antibodies, which induce homotypic cell aggregation, but not binding of cells to extracellular matrix components or endothelial cells, indicating that TS2/16 may generate different signals. The observation that also F(ab')2 or Fab fragments of this anti-beta 1 antibody stimulate binding to extracellular matrix components and endothelial cells excludes the possibility that binding requires receptor crosslinking, or is Fc receptor mediated. Induction of this adhesion is cation and energy dependent and requires an intact cytoskeleton. Although changes in the conformation of VLA integrins induced by this antibody may regulate their functional activity, the dependence on metabolic energy indicates that intracellular processes may also play a role.


Subject(s)
B-Lymphocytes/physiology , Endothelium/physiology , Extracellular Matrix/physiology , Receptors, Very Late Antigen/physiology , T-Lymphocytes/physiology , Antibodies, Monoclonal/immunology , Cell Adhesion , Cell Aggregation , Cell Line , Cell Line, Transformed , Cells, Cultured , Collagen/metabolism , Fibrinogen/metabolism , Fibronectins/metabolism , Humans , Laminin/metabolism , Receptors, Very Late Antigen/immunology , Tetradecanoylphorbol Acetate/pharmacology
18.
J Immunol ; 148(4): 1093-101, 1992 Feb 15.
Article in English | MEDLINE | ID: mdl-1371131

ABSTRACT

Patients with the leukocyte adhesion deficiency (LAD) syndrome have a genetic defect in the common beta 2-chain (CD18) of the leukocyte integrins. This defect can result in the absence of cell surface expression of all three members of the leukocyte integrins. We investigated the capacity of T cell clones obtained from the blood of an LAD patient and of normal T cell clones to adhere to human umbilical vein endothelial cells (EC). Adhesion of the number of LAD T cells to unstimulated EC was approximately half of that of leukocyte function-associated antigen (LFA)-1+ T cells. Stimulation of EC with human rTNF-alpha resulted in an average 2- and 2.5-fold increase in adhesion of LFA-1+ and LFA-1- cells, respectively. This effect was maximal after 24 h and lasted for 48 to 72 h. The involvement of surface structures known to participate in cell adhesion (integrins, CD44) was tested by blocking studies with mAb directed against these structures. Adhesion of LFA-1+ T cells to unstimulated EC was inhibited (average inhibition of 58%) with mAb to CD11a or CD18. Considerably less inhibition of adhesion occurred with mAb to CD11a or CD18 (average inhibition, 20%) when LFA-1+ T cells were incubated with rTNF-alpha-stimulated EC. The adhesion of LFA-1- T cells to EC stimulated with rTNF-alpha, but not to unstimulated EC, was inhibited (average inhibition, 56%) by incubation with a mAb directed to very late antigen (VLA)-4 (CDw49d). In contrast to LAD T cell clones and the LFA-1+ T cell line Jurkat, mAb to VLA-4 did not inhibit adhesion of normal LFA-1+ T cell clones to EC, whether or not the EC had been stimulated with rTNF-alpha. We conclude that the adhesion molecule pair LFA-1/intercellular adhesion molecule (ICAM)-1 plays a major role in the adhesion of LFA-1+ T cell clones derived from normal individuals to unstimulated EC. Adhesion of LFA-1-T cells to TNF-alpha-stimulated EC is mediated by VLA-4/vascular cell adhesion molecule (VCAM)-1 interactions. Since we were unable to reduce significantly the adhesion of cultured normal LFA-1+ T cells to 24 h with TNF-alpha-stimulated endothelium with antibodies that block LFA-1/ICAM-1 or VLA-4/VCAM-1 interactions, and lectin adhesion molecule-1 and endothelial leukocyte adhesion molecule-1 appeared not to be implicated, other as yet undefined cell surface structures are likely to participate in T cell/EC interactions.


Subject(s)
Cell Adhesion , Endothelium, Vascular/physiology , Lymphocyte Function-Associated Antigen-1/physiology , Receptors, Very Late Antigen/physiology , T-Lymphocytes/physiology , Cell Adhesion Molecules/analysis , Cell Adhesion Molecules/physiology , Cells, Cultured , Clone Cells , Humans , Intercellular Adhesion Molecule-1 , Lymphocyte Function-Associated Antigen-1/analysis , Receptors, Lymphocyte Homing/physiology , T-Lymphocytes/immunology , Tetradecanoylphorbol Acetate/pharmacology , Tumor Necrosis Factor-alpha/pharmacology , Vascular Cell Adhesion Molecule-1
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