Cellular Engineering of HSV-tk Transduced, Expanded T Lymphocytes for Graft-versus-Host Disease Management

Burger, Scott R.; Kadidlo, Diane; Basso, Lisa; Bostrom, Nancy; Orchard, Paul J.

doi:10.1159/000072461

Cited by 4 publications

(6 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Table 1 summarizes the HLA-A2.1 affinity/stability (mean F SE, n = 3) of the CD19 and CD20 peptides. Among the peptides tested, the CD19 150-158 (KLMSPKLYV) and CD20 [188][189][190][191][192][193][194][195][196] (SLFLGILSV) peptides displayed the highest HLA-A2.1 binding (CD19 150-158 , FI = 2.66 F 0.36; CD20 [188][189][190][191][192][193][194][195][196] , FI = 2.73 F 0.25), which was close to the affinity of the HLA-A2.1-specific control influenza virus protein matrix peptide [58][59][60][61][62][63][64][65][66] (FI = 3.03 F 0.55). The remaining peptides, CD19 296-304 (TLAYLIFCL), CD20 127-135 (AISGMILSI), CD20 147-155 (LKMESLNFI), CD20 151-159 (SLNFIRAHT), and CD20 154-162 (FIRAHTPYI) displayed minimal binding to HLA-A2.1 as shown by low FI values.…”

Section: Resultsmentioning

confidence: 93%

“…Influenza virus protein matrix peptide [58][59][60][61][62][63][64][65][66] (GILGFVFTL) and MAGE-3 peptide 271-279 (FLWGPRALV) was used as HLA-A2.1-specific peptide controls. All peptides including CD19 and CD20 peptides were synthesized (Biosynthesis, Lewisville, TX) by standard fmoc (9-fluorenylmethyl-oxycarbonyl) chemistry, purified to >90% using reverse-phase chromatography, and validated by mass-spectrometry for molecular weight.…”

Section: Synthetic Peptidementioning

confidence: 99%

See 1 more Smart Citation

Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Bae

Martinson

Klingemann

2005

Clinical Cancer Research

View full text Add to dashboard Cite

The purpose of these studies was to develop immunogenic peptides derived from the CD19 and CD20 self-antigens for the induction of antigen-specific CTLs against B-cell malignancies. A total of seven peptides were designed and examined for their HLA-A2.1 affinity and immunogenicity. Of these peptides, we identified two highly immunogenic HLA-A2.1-specific peptides, CD19 150-158 (KLMSPKLYV) and CD20 [188][189][190][191][192][193][194][195][196] (SLFLGILSV), which were capable of inducing peptide-specific CTLs. The CTLs displayed HLA-A2.1-restricted and antigen-specific cytotoxicity against Burkitt's lymphoma, chronic B cell leukemia, and multiple myeloma cell lines. The CD19 or CD20 peptide -specific CTL cytotoxicity was confirmed using HLA-A2.1 + + T2 cells presenting the appropriate peptide. No cytotoxic activity was observed against T2 cells presenting the irrelevant MAGE-3 peptide or T2 cells alone. In addition, the CTLs displayed a significant (P < 0.05) increase in cell proliferation and IFN-g secretion (>830 ng/mL) following restimulation with HLA-A2.1 + + /CD19 + + /CD20 + + tumor cells. The CTLs also displayed a distinct phenotype consisting of a high percentage of CD69 + + /CD45RO + + and a low percentage of CD45RA + + /CCR7 + + CD4 + + or CD8 + + T cells characteristic of effector memory cell population. Cyclic guanosine 3V ,5V -monophosphate culture conditions using serum-free AIM-V medium containing human AB serum, recombinant human interleukin 2 (Proleukin) and CD3/CD28 Dynabeads were developed resulting in a 35-fold expansion of CD20 peptidespecific CTLs. The expanded CD20-CTLs retained their cytotoxic activity (28-49%) against the Burkitt's lymphoma cell line. In conclusion, we report here on the identification of novel immunogenic CD19 150-158 (KLMSPKLYV) and CD20 [188][189][190][191][192][193][194][195][196] (SLFLGILSV) peptides that have immunotherapeutic potentials as peptide vaccines or targeted T-cell therapies for treating B-cell malignancies.

show abstract

Section: Resultsmentioning

confidence: 93%

Section: Synthetic Peptidementioning

confidence: 99%

Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Bae

Martinson

Klingemann

2005

Clinical Cancer Research

View full text Add to dashboard Cite

show abstract

“…This strategy has been used in the clinic, and several small trials have provided proof of this concept, with results showing reduced GVHD after the administration of ganciclovir [5,13] in the setting of either donor lymphocyte infusion [5,[14][15][16][17] or T-cell add-back to T-cell-depleted marrow at the time of the initial transplantation [13]. However, lower than expected rates of GVHD and GVL were observed in these trials, suggesting that the ex vivo manipulations required to produce the T cells diminished their alloreactivity, possibly because of alterations in the composition of the lymphocyte subsets [10], prolonged culture duration and antibiotic selection [18][19][20][21], or the detrimental effects of exposure to high-dose interleukin (IL)-2 [7,18,22]. Nevertheless, the use of suicidal lymphocytes remains a promising strategy, and investigators in both the United States and Europe [23] have second-generation clinical trials already accruing patients or near to opening.…”

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

“…The suicidal lymphocyte strategy attempts to do this by rendering the GVHD-initiating T cells susceptible to controlled killing by the ex vivo introduction of a suicide gene [4][5][6]. Should these T cells then initiate GVHD, a prodrug is administered, and the suicide gene-bearing T cells are induced into suicide, thereby stopping the GVHD.Numerous groups, including ours, have demonstrated the ability of retroviruses to stably transduce the herpes simplex virus thymidine kinase (HSV-TK) gene into T lymphocytes ex vivo [6][7][8][9][10]. The resultant HSV-TK-transduced lymphocytes convert the prodrug ganciclovir (GCV) to the toxic metabolite GCV-triphosphate [11], which interferes with DNA and RNA transcription, culminating in cell death [12].…”

mentioning

confidence: 99%

“…Numerous groups, including ours, have demonstrated the ability of retroviruses to stably transduce the herpes simplex virus thymidine kinase (HSV-TK) gene into T lymphocytes ex vivo [6][7][8][9][10]. The resultant HSV-TK-transduced lymphocytes convert the prodrug ganciclovir (GCV) to the toxic metabolite GCV-triphosphate [11], which interferes with DNA and RNA transcription, culminating in cell death [12].…”

mentioning

confidence: 99%

See 1 more Smart Citation

Control of graft-versus-host disease with maintenance of the graft-versus-leukemia effect in a murine allogeneic transplant model using retrovirally transduced murine suicidal lymphocytes

Kornblau

Aycox

Stephens

et al. 2007

Experimental Hematology

View full text Add to dashboard Cite

Objective-Limited clinical trials have validated the hypothesis of controlling graft-versus-host disease (GVHD) arising from stem cell transplant utilizing suicidal T-lymphocytes that have been transduced to express the HSV-TK gene. However, clinical utility has been limited by diminished T-cell function arising from the production process. To evaluate strategies for harnessing the graftversus-leukemia (GVL) effect while improving the safety and function of suicidal lymphocytes, we have developed techniques to produce fully functional, retrovirally transduced, HSV-TKpositive murine T cells (TK + TC).Methods-Utilizing a murine major histocompatibility complex-matched transplant model, we evaluated the ability of TK + TC to generate a GVL effect and the ability to control GVHD in experiments where we varied the dose of TK + TC, ganciclovir (GCV) dose, the start of GCV administration (day 4, 7, 10, 13, 15, or 19) posttransplantation, and the GCV administration route (osmotic pump versus intraperitoneal).Results-At TK + TC doses in excess of the standard lethal dose (SLD) of unmanipulated T-cells, GCV administration completely (2 × SLD) and partially (4 × SLD) controlled GVHD. Additionally, GVHD remained reversible despite delaying administration of GCV for a week after GVHD developed. Importantly, GVHD was controlled with a 1-log but not 2-log reduction in GCV dose, and this "partial suicide" preserved more circulating TK + TC compared with standarddose GCV. Survival of leukemia-positive mice receiving TK + TC and GCV was significantly increased compared with control cohorts not receiving GCV or transplanted with unmanipulated T cells, thereby demonstrating a GVL effect. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptConclusion-Retrovirally transduced suicidal lymphocytes generate a potent GVL effect while simultaneously enabling control of GVHD, which results in improved leukemia and GVHD-free survival.The risk of graft-versus-host disease (GVHD) in patients with leukemia who undergo allogeneic bone marrow transplantation necessitates finding a suitably matched donor so that these patients can benefit from the graft-versus-leukemia (GVL) effect [1,2]. However, the inability to identify such donors is a barrier to transplantation for most leukemia patients. Although both prophylactic therapeutic strategies and strategies to manipulate the graft are used to prevent GVHD, as many as 40% of patients who receive HLA-matched transplants develop GVHD that requires systemic therapy [2,3]. The interventions required to treat GVHD further compromise an already impaired immune system, and fatal infections are common during GVHD therapy. Consequently, the risks associated with GVHD decrease the curative potential of allogeneic transplantation by either preventing patients from undergoing transplantation in the first place or complicating the course of those who do.Selectively removing the GVHD-initiating T cells after grafts are established and T cells have been activated might control GVHD...

show abstract

Gamma‐glutamylcysteine synthetase‐based selection strategy for gene therapy of chronic granulomatous disease and graft‐vs.‐host disease

Rappa

Anzanello

Alexeyev

et al. 2007

European J of Haematology

View full text Add to dashboard Cite

Efficient ex vivo/in vivo selection of genetically modified hematopoietic stem/progenitor cells (HPCs) and T lymphocytes could greatly improve several gene therapy strategies. We have previously reported that primary murine HPCs, transduced with a bicistronic retroviral vector, co-expressing the catalytic subunit of gamma-glutamylcysteine synthetase (gamma-GCSh) and eGFP, could be selected by l-buthionine-S,R-sulfoximine (BSO). Upon ex vivo transduction with a low, defined gene dosage and BSO selection, HPCs were able to repopulate the bone marrow of syngeneic myeloablated hosts, showing multi-lineage expression [Hum Gene Ther, 16 (2005), 711]. We now provide 'proof-of-principle' that the same strategy can be applied to the gene therapy of graft-vs.-host disease (GVHD) subsequent to allogeneic bone marrow transplantation (ABMT), and of chromosome X-associated chronic granulomatous disease (CGD). Transfer of the herpes simplex virus-thymidine kinase (HSV-Tk) 'suicide' gene into donor T lymphocytes is a potential method to control GVHD after ABMT. However, an efficient selection system is required to eliminate non-HSV-Tk-expressing T lymphocytes before administration to the patient. We now report that, upon transduction with a retroviral vector, co-expressing gamma-GCSh and eGFP, and subsequent selection by BSO, over 95% human T lymphocytes were found to express eGFP; moreover, upon transduction with a novel retroviral vector co-expressing gamma-GCSh and HSV-Tk, and subsequent BSO treatment, over 95% of T lymphocytes could be eliminated by ganciclovir. The efficacy of the gamma-GCSh-BSO selection strategy was then tested on an in vitro model of CGD. Upon transduction of gp91 (phox)-deficient PLBKO cells with a novel bicistronic retroviral vector co-expressing human gp91 (phox) and gamma-GCSh, exposure to BSO for 48 h eliminated most non-transduced cells, resulting in selection of gp91 (phox)-expressing cells, and reconstitution of NADPH oxidase activity.

show abstract

Cellular Engineering of HSV-tk Transduced, Expanded T Lymphocytes for Graft-versus-Host Disease Management

Cited by 4 publications

References 35 publications

Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Control of graft-versus-host disease with maintenance of the graft-versus-leukemia effect in a murine allogeneic transplant model using retrovirally transduced murine suicidal lymphocytes

Gamma‐glutamylcysteine synthetase‐based selection strategy for gene therapy of chronic granulomatous disease and graft‐vs.‐host disease

Contact Info

Product

Resources

About