Anti-leukemia activity of CD33-specific chimeric cytotoxic T lymphocytes in vitro and in vivo is impaired by addition of the CD28 costimulatory endodomain

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 3052-3052
Author(s):  
A. Dutour ◽  
D. Lee ◽  
S. Napier ◽  
E. Yvon ◽  
H. Finney ◽  
...  
Keyword(s):  
T Cells ◽  
Cytotoxic T Cells ◽  
Surface Protein ◽  
Receptor Expression ◽  
Target Cells ◽  
Chimeric Receptor ◽  
Specific Lysis ◽  
Significant Difference

3052 Background: EBV-specific cytotoxic T cells (EBV-CTLs) expand and have long-term activity in vivo due to the sustained costimulation provided by the EBV-infected cells produced by this persistent virus. We exploited this phenomenon and redirected EBV-CTLs against CD33, a surface protein expressed on the malignant blasts of acute myeloid leukemia (AML) cells. Methods: EBV-CTLs generated from six EBV-seropositive donors were transduced using a retroviral vector encoding CD33 specific chimeric receptor (cR). We evaluated whether the high and sustained activity shown against native EBV+ target cells can be extended to the CD33+ EBV- targets of the chimeric receptor and whether the addition of CD28 signaling domain improved the receptor activity. Results: cRCD33-EBV-CTL maintained killed EBV-LCL and CD33+ targets (specific lysis respectively of 30% and 35% at E:T ratio 25:1). They produced Th-1, Th-2 and Tc cytokines on exposure to CD33+ targets. Addition of the CD28 intracellular domain did not increase cytotoxicity to CD33+ targets. Preincubation of CD33+ cells with the CD33-blocking MoAb resulted in up to 40% inhibition of lysis and up to 60% inhibition of cytokine release by cRCD33-EBV-CTLs confirming the specificity of the TCR interactions with CD33. NOD-SCID mice bearing a human CD33+ AML were injected with EBV-CTLs ×4 weekly starting 5 days after tumor inoculation. Significant tumor reduction was only observed in mice treated with the cRCD33-EBV-CTLs (p<0.05). Immunohistologic analysis showed the presence of a majority of CD8+ human T cells in the tumors of treated mice. Incorporation of the CD28 endodomain resulted in less tumor-infiltrating T cells in mice treated with cRCD33CD28-EBV-CTLs. There was no significant difference in the chemokines receptor expression on cRCD33CD28-EBV-CTLs but their rate of apoptosis was 16 % higher (p<0.05) than the one of cRCD33-EBV-CTLs. Conclusions: EBV- CTL expressing the CD33 chimeric receptor are functional in vitro and in vivo in mice. CD28 signaling may have a deleterious role for the activity of chimeric receptors in vivo. No significant financial relationships to disclose.

scholarly journals The influenza A virus nucleoprotein gene controls the induction of both subtype specific and cross-reactive cytotoxic T cells.

1984 ◽  
Vol 160 (2) ◽  
pp. 552-563 ◽  
Author(s):  
A R Townsend ◽  
J J Skehel
Keyword(s):  
T Cells ◽  
Influenza A ◽  
Cytotoxic T Cells ◽  
Target Cells ◽  
Spleen Cells ◽  
Influenza A Viruses ◽  
Group A ◽  
Selection Of

Using genetically typed recombinant influenza A viruses that differ only in their genes for nucleoprotein, we have demonstrated that repeated stimulation in vitro of C57BL/6 spleen cells primed in vivo with E61-13-H17 (H3N2) virus results in the selection of a population of cytotoxic T lymphocytes (CTL) whose recognition of infected target cells maps to the gene for nucleoprotein of the 1968 virus. Influenza A viruses isolated between 1934 and 1979 fall into two groups defined by their ability to sensitize target cells for lysis by these CTL: 1934-1943 form one group (A/PR/8/34 related) and 1946-1979 form the second group (A/HK/8/68 related). These findings complement and extend our previous results with an isolated CTL clone with specificity for the 1934 nucleoprotein (27, 28). It is also shown that the same spleen cells derived from mice primed with E61-13-H17 virus in vivo, but maintained in identical conditions by stimulation with X31 virus (which differs from the former only in the origin of its gene for NP) in vitro, results in the selection of CTL that cross-react on target cells infected with A/PR/8/1934 (H1N1) or A/Aichi/1968 (H3N2). These results show that the influenza A virus gene for NP can play a role in selecting CTL with different specificities and implicate the NP molecule as a candidate for a target structure recognized by both subtype-directed and cross-reactive influenza A-specific cytotoxic T cells.

scholarly journals Antiviral protection by virus-immune cytotoxic T cells: infected target cells are lysed before infectious virus progeny is assembled.

1977 ◽  
Vol 145 (3) ◽  
pp. 644-651 ◽  
Author(s):  
R M Zinkernagel ◽  
A Althage
Keyword(s):  
T Cells ◽  
Adoptive Transfer ◽  
Infectious Virus ◽  
Cytotoxic T Cells ◽  
Target Cells ◽  
Virus Infections ◽  
Virus Growth ◽  
Virus Progeny

Virus-immune cytotoxic T cells can inhibit effectively growth of vaccinia virus in acutely infected target cells in vitro by destroying infected target cells before infectious virus progeny is assembled. Together with the fact that virus-specific T cells are demonstrable after 3 days, very early during infection, and with strong circumstantial evidence from adoptive transfer models in vivo, these data suggest that in some virus infections T cells may in fact act cytolytically in vivo to prevent virus growth and spread and be an important early antiviral effector mechanism.

scholarly journals Joint recognition by cytotoxic T cells of inactivated Sendai virus and products of the major histocompatibility complex.

1977 ◽  
Vol 145 (3) ◽  
pp. 523-539 ◽  
Author(s):  
J W Schrader ◽  
G M Edelman
Keyword(s):  
T Cells ◽  
Sendai Virus ◽  
Cytotoxic T Cells ◽  
Secondary Response ◽  
Cytotoxic T Cell ◽  
Target Cells ◽  
Murine Spleen ◽  
Virion Antigen

Cytotoxic T cells specific for Sendai virus were generated by culturing murine spleen cells in vitro together with UV-inactivated Sendai virus. In vivo immunization of donor mice with UV-inactivated Sendai virus resulted in an in vitro secondary response of increased magnitude. Cytotoxic activity was demonstrated in a short-term 51Cr-release assay, using syngeneic tumor cells which had been coated with inactivated Sendai virus by incubation at 4 degrees C for 30 min. The lysis of Sendai virus-coated target cells was restricted by the H-2 haplotype of the target cells, suggesting that the H-2 genes of the target cell contributed to the specificity of the lysis. Kinetic experiments showed that susceptibility to lysis by cytotoxic T cells specific for Sendai virus appeared within 30 min after coating target cells with inactivated virus. Furthermore, there was no detectable synthesis of new proteins in cells treated with UV-inactivated Sendai virus. For these reasons, we suggest that neither viral replication nor the synthesis of new proteins are necessary for the production of the antigen recognized by cytotoxic cells specific for Sendai virus. We infer that the virus-specific component on the target cells is probably a preformed virion antigen adsorbed onto or integrated into the cell membrane. These results imply that, if the cytotoxic T cell recognizes a single antigenic determinant specified both by viral and H-2 genes, this determinant is formed by the physical association of H-2 and Sendai virus antigens rather than by their alteration during the processes of synthesis.

scholarly journals Involvement of fusion activity of ultraviolet light-inactivated sendai virus in formation of target antigens recognized by cytotoxic T cells

1978 ◽  
Vol 148 (1) ◽  
pp. 276-287 ◽  
Author(s):  
K Sugamura ◽  
K Shimizu ◽  
FH Back
Keyword(s):  
T Cells ◽  
Ultraviolet Light ◽  
Sendai Virus ◽  
Cytotoxic T Cells ◽  
Target Antigen ◽  
Target Cells ◽  
Fusion Activity ◽  
Ethanol Treatment ◽  
Specific Lysis

Mice inoculated with ultraviolet light-inactivated Sendai virus mount a cell- mediated immune response to the virus. Cytotoxic T cells specific for Sendai virus can be obtained by in vitro secondary stimulation of primed spleen cells with syngeneic stimulator cells coated with UV-inactivated Sendai virus. Neither in vivo nor in vitro stimulation alone is sufficient to generate specific cytotoxic T cells. Sharing of the H-2 haplotype between cytotoxic T cells and target cells is required for the Sendai virus-specific lysis to occur. The fusion (F) glycoprotein of Sendai virus has been implicated in target antigen formation (20). Ethanol treatment of Sendai virus causes complete inactivation of the cell-fusion and hemolytic activities of the envelope, but does not affect the antigenicity of the F glycoprotein; furthermore, hemagglutinin and neuraminidase activities of the envelope HANA glycoprotein are also left intact after ethanol treatment. Target cells can be prepared by coating them with various numbers of UV-inactivated Sendai virus that have been treated with ethanol or, as a control, phosphate-buffered saline (PBS). The amount of virus adsorbed to target cells during the cytotoxicity reaction time using either ethanol-treated or untreated (PBS "treated") virions is essentially identical, but target cells coated with ethanol-treated Sendai virus fail to serve as targets for cytotoxic T cells. These results indicate that fusion activity of the Sendai virus envelope is essential to the formation of the target antigen and that virus adsorption to cell surfaces without fusion of the envelope with cell membranes is not sufficient to allow killing by virus-specific cytotoxic T cells.

scholarly journals Cytotoxic T cells distinguish between trinitrophenyl- and dinitrophenyl-modified syngeneic cells.

1977 ◽  
Vol 146 (2) ◽  
pp. 600-605 ◽  
Author(s):  
J Forman
Keyword(s):  
T Cells ◽  
Cytotoxic Effect ◽  
Cytotoxic T Cells ◽  
Cytotoxic T Cell ◽  
Target Cells ◽  
Spleen Cells ◽  
Competition Assay ◽  
Effector Cells

Spleen cells sensitized against trinitrophenyl (TNP)-modified stimulator cells displayed a cytotoxic effect against syngeneic TNP-modified but not dinitrophenyl (DNP)-modified target cells. The same finding was observed in the opposite direction; that is, effector cells sensitized against DNP-modified stimulator cells did not cross kill TNP-modified targets. The specificity of the anti-TNP effector cells was confirmed in a cold target competition assay. Presensitization in vivo with hapten-modified cells followed by rechallenge and testing in vitro did not alter the specificity of the response between the haptens. These data indicate that the receptor(s) on the cytotoxic T cell can distinguish between two closely related haptenic molecules.

Cord blood comprises antigen-experienced T cells specific for maternal minor histocompatibility antigen HA-1

Blood ◽  
2005 ◽  
Vol 105 (4) ◽  
pp. 1823-1827 ◽  
Author(s):  
Bregje Mommaas ◽  
Janine A. Stegehuis-Kamp ◽  
Astrid G. van Halteren ◽  
Michel Kester ◽  
Jürgen Enczmann ◽  
...  
Keyword(s):  
T Cells ◽  
Cord Blood ◽  
Ex Vivo ◽  
Cytotoxic T Cells ◽  
Cord Blood Transplantation ◽  
Human Leukocyte ◽  
Leukemic Cells ◽  
Target Cells ◽  
Graft Versus Leukemia

AbstractUmbilical cord blood transplantation is applied as treatment for mainly pediatric patients with hematologic malignancies. The clinical results show a relatively low incidence of graft-versus-host disease and leukemia relapse. Since maternal cells traffic into the fetus during pregnancy, we questioned whether cord blood has the potential to generate cytotoxic T cells specific for the hematopoietic minor histocompatibility (H) antigen HA-1 that would support the graft-versus-leukemia effect. Here, we demonstrate the feasibility of ex vivo generation of minor H antigen HA-1-specific T cells from cord blood cells. Moreover, we observed pre-existing HA-1-specific T cells in cord blood samples. Both the circulating and the ex vivo-generated HA-1-specific T cells show specific and hematopoietic restricted lysis of human leukocyte antigen-A2pos/HA-1pos (HLA-A2pos/HA-1pos) target cells, including leukemic cells. The cord blood-derived HA-1-specific cytotoxic T cells are from child origin. Thus, the so-called naive cord blood can comprise cytotoxic T cells directed at the maternal minor H antigen HA-1. The apparent immunization status of cord blood may well contribute to the in vivo graft-versus-leukemia activity after transplantation. Moreover, since the fetus cannot be primed against Y chromosome-encoded minor H antigens, cord blood is an attractive stem cell source for male patients. (Blood. 2005;105:1823-1827)

scholarly journals Participation of the H-2 antigens of tumor cells in their lysis by syngeneic T cells.

1976 ◽  
Vol 143 (3) ◽  
pp. 601-614 ◽  
Author(s):  
J W Schrader ◽  
G M Edelman
Keyword(s):  
T Cells ◽  
Tumor Cell ◽  
Tumor Cells ◽  
Target Cell ◽  
Cytotoxic T Cells ◽  
Hybrid Origin ◽  
Target Cells ◽  
Cytotoxic Lymphocytes ◽  
Effector Cells

Cytotoxic T lymphocytes were generated in vitro against H-2 compatible or syngeneic tumor cells. In vitro cytotoxic activity was inhibited by specific anti-H2 sera, suggesting that H-2 antigens are involved in cell lysis. Two observations directly demonstrated the participation of the H-2 antigens on the tumor cells in their lysis by H-2-compatible T cells. First, coating of the H-2 antigens on the target tumor cell reduced the number of cells lysed on subsequent exposure to cytotoxic T cells. Second, when cytotoxic T cells were activated against an H-2 compatible tumor and assayed against an H-2-incompatible tumor, anti-H-2 serum that could bind to the target cell, but not to the cytotoxic lymphocyte, inhibited lysis. H-2 antigens were also shown to be present on the cytotoxic lymphocytes. Specific antisera reacting with these H-2 antigens, but not those of the target cell, failed to inhibit lysis when small numbers of effector cells were assayed against H-2-incompatible target cells or when effector cells of F1-hybrid origin and bearing two H-2 haplotypes were assayed against a tumor cell of one of the parental strains. These findings suggest that it is the H-2 antigens on the tumor cell and not those on the cytotoxic lymphocytes that are important in cell-mediated lysis of H-2-compatible tumor cells.

scholarly journals Modulation of the CCR5 Receptor/Ligand Axis by Seminal Plasma and the Utility of In Vitro versus In Vivo Models

2019 ◽  
Vol 93 (11) ◽  
Author(s):  
Jennifer A. Juno ◽  
Kathleen M. Wragg ◽  
Anne B. Kristensen ◽  
Wen Shi Lee ◽  
Kevin J. Selva ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Seminal Plasma ◽  
Target Cells ◽  
Ccr5 Expression ◽  
Ccr5 Receptor ◽  
The Impact ◽  
Hiv 1

ABSTRACT Sexual HIV-1 transmission occurs primarily in the presence of semen. Although data from macaque studies suggest that CCR5+ CD4+ T cells are initial targets for HIV-1 infection, the impact of semen on T cell CCR5 expression and ligand production remains inconclusive. To determine if semen modulates the lymphocyte CCR5 receptor/ligand axis, primary human T cell CCR5 expression and natural killer (NK) cell anti-HIV-1 antibody-dependent beta chemokine production was assessed following seminal plasma (SP) exposure. Purified T cells produce sufficient quantities of RANTES to result in a significant decline in CCR5bright T cell frequency following 16 h of SP exposure (P = 0.03). Meanwhile, NK cells retain the capacity to produce limited amounts of MIP-1α/MIP-1β in response to anti-HIV-1 antibody-dependent stimulation (median, 9.5% MIP-1α+ and/or MIP-1β+), despite the immunosuppressive nature of SP. Although these in vitro experiments suggest that SP-induced CCR5 ligand production results in the loss of surface CCR5 expression on CD4+ T cells, the in vivo implications are unclear. We therefore vaginally exposed five pigtail macaques to SP and found that such exposure resulted in an increase in CCR5+ HIV-1 target cells in three of the animals. The in vivo data support a growing body of evidence suggesting that semen exposure recruits target cells to the vagina that are highly susceptible to HIV-1 infection, which has important implications for HIV-1 transmission and vaccine design. IMPORTANCE The majority of HIV-1 vaccine studies do not take into consideration the impact that semen exposure might have on the mucosal immune system. In this study, we demonstrate that seminal plasma (SP) exposure can alter CCR5 expression on T cells. Importantly, in vitro studies of T cells in culture cannot replicate the conditions under which immune cells might be recruited to the genital mucosa in vivo, leading to potentially erroneous conclusions about the impact of semen on mucosal HIV-1 susceptibility.

Stimulation of cytotoxic T cells against idiotype immunoglobulin of malignant lymphoma with protein-pulsed or idiotype-transduced dendritic cells

Blood ◽  
2000 ◽  
Vol 95 (4) ◽  
pp. 1342-1349 ◽  
Author(s):  
Frank Osterroth ◽  
Annette Garbe ◽  
Paul Fisch ◽  
Hendrik Veelken
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
Light Chain ◽  
Cytotoxic T Cells ◽  
Killer Cell ◽  
Cell Activity ◽  
Expression Vectors ◽  
Target Cells ◽  
Fab Fragment ◽  
Specific Lysis

Because of their hypervariable regions and somatic mutations, the antigen receptor molecules of lymphomas (idiotypes) are tumor-specific antigens and attractive targets for antilymphoma immunotherapy. For the optimal induction of human idiotype-specific cytotoxic T cells (CTL), idiotype was presented to CD8+ peripheral blood mononuclear cells by monocyte-derived autologous dendritic cells (DC) after the endocytosis of idiotype protein or by idiotype-expressing DC. Recombinant idiotype was obtained as a functionally folded Fab fragment by periplasmic expression in Escherichia coli. Idiotype-expressing DC were generated by transduction with recombinant Semliki forest virus vectors encompassing heavy- or light-chain idiotype genes. Autologous lymphoblastoid cell lines stably transfected with Epstein-Barr virus-based idiotype expression vectors were used as target cells to detect idiotype-specific lysis. CTL stimulated with idiotype-loaded DC showed strong specific, CD8-mediated, and major histocompatibility complex (MHC) class I-restricted cytotoxicity against autologous heavy- and light-chain idiotype. In contrast, stimulation with idiotype-transduced DC resulted in only moderate natural killer cell activity. These data confirm the existence of idiotype-specific CTL in patients with lymphoma, define a “good manufacturing practice”-compatible protocol for the generation of these cells without the requirement of viable lymphoma cells, and favor the processing of exogenous antigen over DC transduction for the induction of MHC I-restricted CTL against idiotypes with unknown antigenicity.

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