Modified T cells as therapeutic agents

Abstract Immunotherapy is now a well-established modality in the treatment of cancer. Although several platforms to redirect the immune response exist, the use of genetically modified T cells has garnered particular attention in recent years. This is due, in large part, to their success in the treatment of B-cell malignancies. Adoptively transferred T cells have also demonstrated efficacy in the treatment of systemic viral infections that occur following hematopoietic cell transplantation prior to immune reconstitution. Here we discuss the techniques that enable redirection of T lymphocytes to treat cancer or infection and the current indications for these therapies.

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Development of Effective Immunotherapy for B-Cell Non-Hodgkin’s Lymphoma with CD19-Specific Cytotoxic T Cells.

Blood ◽

10.1182/blood.v104.11.3277.3277 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3277-3277

Author(s):

Keichiro Mihara ◽

Kazuyoshi Yanagihara ◽

Chihaya Imai ◽

Akiro Kimura ◽

Dario Campana

Keyword(s):

Immune Response ◽

T Cells ◽

T Lymphocytes ◽

B Cell ◽

Hodgkin’S Lymphoma ◽

Cell Killing ◽

Hodgkin's Lymphoma ◽

Single Chain ◽

Non Hodgkin’S Lymphoma ◽

Non Hodgkin's Lymphoma

Abstract Less than 60% of patients with B-cell non-Hodgkin’s lymphoma (B-NHL) can be cured with contemporary therapy. Using artificial receptors it is possible to redirect the specificity of immune cells to tumor-associated antigens, a strategy that holds great potential as a novel cancer therapy. Since B-NHL cells invariably express CD19, we transduced human peripheral blood T lymphocytes with a recently developed receptor (anti-CD19-BB-ζ), which consists of the single-chain variable domain (scFv) of an anti-CD19 monoclonal antibody, the hinge and transmembrane domains of CD8α, and the signaling domains of CD3ζ and 4-1BB. CD3ζ delivers the primary stimulus upon receptor engagement, while 4-1BB delivers co-stimulatory signals that are crucial for T-cell cytotoxicity. It has been shown that elicitation of 4-1BB signaling enhances the immune response to tumors in vivo, even when an immune response cannot be induced by CD28 stimulation. Retroviral transduction led to anti-CD19-BB-ζ expression in T cells with high efficiency: median percent of transduced cells was 60.3% (range, 25.7%–83.4%; n = 9). T lymphocytes expressing anti-CD19-BB-ζ expanded more vigorously that T cells transduced with receptors lacking 4-1BB and exerted powerful cytotoxicity against the CD19+ B-NHL cell lines Raji, Daudi, RL, and HT in vitro: at a 0.5: 1 effector: target ratio, mean (± SD) cell specific lymphoma cell killing was 96.6% ± 4.6% after 5–7 days of culture (4 experiments in each cell line). Transduced T cells were also effective against freshly isolated cells from patients with diffuse large, follicular large, Burkitt, and mantle cell lymphoma cultured on bone marrow-derived mesenchymal cells: in 10 samples, cell killing was 93.6% ± 5.7% at a 0.5: 1 ratio after 5–7 days of culture. Sensitivity to anti-CD19-BB-ζ-mediated killing was observed regardless of high Bcl-2 expression. T cells expressing anti-CD19-BB-ζ were also effective in a xenograft model of NHL, in which NOD/SCID mice were inoculated subcutaneously with lymphoma cells (1 x 107). Subsequent inoculation of T cells (2 x 106) transduced with anti-CD19-BB-ζ receptors significantly suppressed tumor growth, whereas inoculation of T cells transduced with empty control vector had no effect (3 mice for each treatment). These results provide a rationale for clinical testing of autologous T cells modified with anti-CD19-BB-ζ receptors in patients with aggressive or relapsed B-NHLs refractory to conventional therapy.

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Immune Reconstitution after Haploidentical Hematopoietic Cell Transplantation: Impact of Reduced Intensity Conditioning and CD3/CD19-Depleted Grafts

Blood ◽

10.1182/blood.v112.11.1175.1175 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1175-1175

Author(s):

Birgit Federmann ◽

Matthias Haegele ◽

Christoph Faul ◽

Wichard Vogel ◽

Lothar Kanz ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Nk Cells ◽

Cell Transplantation ◽

Immune Reconstitution ◽

Hematopoietic Cell Transplantation ◽

Nk Cell ◽

Reduced Intensity Conditioning ◽

Haploidentical Hematopoietic Cell Transplantation ◽

Reduced Intensity

Abstract Haploidentical hematopoietic cell transplantation (HHCT) using CD3/CD19 depleted grafts may lead to faster engraftment and immune reconstitution since grafts contain also graft-facilitating-cells, CD34− progenitors, NK cells, and dendritic cells. Reduced intensity conditioning may also have a positive impact on immune reconstitution following HHCT. 26 adults received CD3/CD19 depleted HHCT after RIC (150–200 mg/m2 fludarabine, 10mg/kg thiothepa, 120 mg/m2 melphalan and 5mg/day OKT-3 (day −5 to +14)) at our institution between 2005–2008. We prospectively evaluated engraftment and immune reconstitution. B-, NK-, T- and T-cell subsets (CD3/8, CD4/8, CD4/45RA/RO), TCR-Vβ repertoire and NK-cell receptors (NKP30, NKP44, NKP46, NKG2D, CD158a/b/e, CD85j, NKG2A, CD161) were analyzed by FACS. Grafts contained 8.8×106 CD34+ (range, 4.3–18.0 ×106), 2.9×104 CD3+ (range, 1.2–9.2×104) and 3.6×107 CD56+ (range, 0.02–23.0 ×107) cells/kg. Engraftment was rapid with a median time to >500 granulocytes/μl of 11 days (range, 9–15) and a median time to >20 000 platelets/μl of 11 days (range, 8–23). Full chimerism was reached on day 14 (median; range, 6–26). NK-cell engraftment was rapid, reaching normal values on day 20 (median of 247 CD16+CD56+CD3− cells/μl (range, 1–886)) with NK cells comprising up to 70% of lymphocytes. B-cell reconstitution was delayed with 81 (range, 0–280) and 335 (range, 11–452) CD19+20+ cells/μl on days 150 and 400, respectively. T-cell reconstitution was impaired with 49 (range, 0–586) and 364 (range, 35–536) CD3+ cells/μl on day 60 and day 150, respectively. We observed an increase of CD3+CD8+ cells in contrast to CD3+CD4+ cells early after HHCT with a median of 24 (range, 0–399) vs 16 (range, 0–257) and 159 (range, 1–402) vs 96 (range, 18–289) cells/μl on day 50 and day 200, respectively. CD4+CD45RA+ T cells increased slowly while CD4+CD45RO+ T cells reconstituted faster with a median of 61 CD4+CD45RO+ cells/μl (range, 0–310) vs 24 CD4+CD45RA+ (range, 0 to 152) on day 100. Within the CD4+CD25+ regulatory T cells there was a slow regeneration with median of 14 CD4+CD25+ cells/μl (range, 0–96) on day 100 and 28 CD4+CD25+ cells/μl (range, 19–160) on day 200. CD14+CD45+ monocytes did not reach normal values within the time of observation with 7 CD14+CD45+ cells/μl (range, 0–21) on day 120 and 7 CD14+CD45+ cells (range, 2–381) on day 400. TCR-Vβ repertoire and NK-cell receptor reconstitution was analyzed so far in 7 and 8 patients, respectively. We found a skewed T-cell repertoire with oligoclonal T-cell expansions to day 100 and normalization after day 200. An increased natural cytotoxicity receptor (NKP30, NKP44, NKP46) and NKG2A, but decreased NKG2D and KIR-expression was observed on NK-cells until day 100. In conclusion, T- and B-cell reconstitution is delayed after HHCT using CD3/CD19 depleted grafts and RIC. However, T-cell reconstitution is faster compared to data published with CD34 selected grafts and myeloablative conditioning. A fast NK-cell reconstitution early after HHCT was observed. Thus a combination of reduced intensity conditioning with CD3/CD19 depleted grafts appears to accelerate the immune recovery after haploidentical stem cell transplantation.

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DISCORDANT RESPONSE OF CD4+ T LYMPHOCYTES TO ANTIRETROVIRAL THERAPY

HIV Infection and Immunosuppressive Disorders ◽

10.22328/2077-9828-2019-11-1-16-30 ◽

2019 ◽

Vol 11 (1) ◽

pp. 16-30 ◽

Cited By ~ 2

Author(s):

K. V. Shmagel

Keyword(s):

Immune Response ◽

T Cells ◽

Viral Load ◽

Immune System ◽

Antiretroviral Therapy ◽

T Lymphocytes ◽

Lymphocyte Proliferation ◽

Immune Reconstitution ◽

Cell Counts ◽

Increased Risk

Antiretroviral therapy (ART) in HIV infected patients generally results in the suppression of viral replication and reconstitution of CD4+ T lymphocytes cell counts. In some patients (about 20%), however, a disturbance in regeneration of immune competent cells with a background of low viral load occurs. The term «immunological nonresponders» has been used to describe this phenomenon. Discordant immune response to antiviral therapy may be caused by increasing of depletion and reducing of production of CD4+ T cells. However, mechanisms for low immune reconstitution are not currently well understood. «Immunological nonresponders» exhibit booster lymphocyte proliferation, increased immune activation and reducing of CD4+ T lymphocytes survival time in comparison with patients with concordant response to the therapy. Their immune system is characterized by more pronounced aging and exhaustion. This leads to early and frequent manifestation of AIDSrelated diseases. Besides, immunological nonresponders have an increased risk of non-AIDS-related diseases due to pronounced systemic inflammation. The objective of the present review was to highlight the important problem that is rather common on аntiretroviral therapy and to enlist the specialists to the solving of this issue.

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Method of Mobilization: Implication on Cell Subsets in the Graft and Immune Reconstitution Post Autologous Hematopoietic Cell Transplantation (AHCT)

Blood ◽

10.1182/blood.v124.21.1132.1132 ◽

2014 ◽

Vol 124 (21) ◽

pp. 1132-1132

Author(s):

Melhem M. Solh ◽

Rathmann Kristin ◽

Sauvi chang-Fong ◽

Jeremiah Oyer ◽

Wesam B. Ahmed ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Transplantation ◽

Peripheral Blood ◽

Immune Reconstitution ◽

Hematopoietic Cell Transplantation ◽

Hematopoietic Cell ◽

P Value ◽

Autologous Hematopoietic Cell Transplantation ◽

Graft Composition

Abstract Method of Mobilization: Implication on Cell Subsets in The Graft and Immune Reconstitution post Autologous Hematopoietic Cell Transplantation (AHCT) The optimal mobilization method for either myeloma or lymphoma patients undergoing AHCT is still debatable and strategies for graft collection vary between different institutions. Plerixafor, a CXCR4 antagonist is used for peripheral blood stem cell mobilization in multiple myeloma and non-Hodgkins lymphoma patients requiring AHCT. The effect of plerixafor on graft composition has scarce data that are based mostly on cryopreserved samples. Moreover; the effect of plerixafor on immune reconstitution and hematologic recovery post AHCT has not been well evaluated. The goal of our study was to compare graft composition, hematologic and immune reconstitution recovery among patients mobilized with plerixafor plus G-CSF to those mobilized with G-CSF alone. Methods: 49 patients eligible for AHCT were enrolled on a single arm prospective trial at a single transplant center. All patients were mobilized with G-CSF 10µg/kg/day for 4 consecutive days. A peripheral blood CD34 level of <20/µl on day 4 was used as a cutoff to use plerixafor 0.24mg/kg in addition to G-CSf on 9pm of the fourth day. Peripheral blood collection was started on day 5 and was continued till the target dose is achieved or a minimum CD 34+ cell dose of >2x106 cells/Kg was obtained after 3 collection days. Samples from the freshly collected graft and patients' peripheral blood on days +30 and +60 were analyzed by flow cytometry (BD FACSCanto II) . A single platform assay was used (Beckman-Coulter Stem kit) via a ISHAGE protocol. The antibody cocktail contained the following pre-conjugated monoclonal antibodies: CD56-PE (Miltenyi Biotech, Auburn, CA), CD3-APC, CD16-FITC, (Beckman Coulter, Brea, CA), CD19-PE-CY7 (BD Biosciences, San Jose, CA). Data were acquired using BD FACSCanto II (BD Biosciences) and analyzed with the FACSDiva software (BD Biosciences) to quantify CD3+ T cells, CD3+ CD56+ NK-like T cells, CD56+ CD16+ and CD56+ CD16- NK cells as well as CD19+ B cells. Results: 49 patients with a median age of 58 years (range 21-75) were mobilized with either G-CSF alone (N=16) or plerixafor +G-CSF (G+P)(N=33).The median number of collection days was 1.42 and 1.81 (p=0.2) and the median collected CD34+ dose was 8.28x106/kg and 5.24x106 /kg (p=022) in the G+P and G-CSF alone groups respectively. Both groups had similar times to neutrophil and platelet engraftment. The graft analysis showed a white blood count of 309x109/l and 262x109/l (p=0.38), median percentage of CD34+ cells of 0.75% and 0.73% (p=0.81), percentage of CD3+ T cells of 25.6% and 22% (p=0.6) in the G+P and G-CSF alone groups resepectively. Both groups had similar proportions of CD3+, CD4+,CD8+, NK, NKT and iNKT cells in the mobilized grafts. Peripheral blood samples at day +30 and day +60 were analyzed for T cell markers and hematologic recovery (table 1). There was no significant difference between absolute lymphocyte counts, NK cell counts, T cells and absolute neutrophil count. Conclusion: Plerixafor when combined with G-CSF helps in achieving mobilization goals in patients predicted to be poor mobilizers based on peripheral CD34 levels. The addition of plerixafor doesn't not seem to affect T cell composition of the graft and yields similar hematologic and immune recovery when compared to mobilization with G-CSF alone. Table 1: Immune Reconstitution at Day 30 and Day 60 post Autologous Transplantation Treatment Group G-CSF (N=16) Plerixafor + G-CSF (N=33) P-value G-CSF (N=16) Plerixafor + G-CSF (N=33) P-value Day 30 Day 60 WBC 5.08 5.41 0.873 4.94 5.38 0.654 HGB 10.86 11.19 0.353 11.22 11.17 0.757 HCT 32.35 33.66 0.321 33.36 33.53 0.565 PLT 119.88 161.42 0.068 166.94 173.73 0.949 Abs Lymph 1.09 1.44 0.296 1.41 1.50 0.974 % NK 26.14 30.38 0.277 11.53 20.09 0.095 Abs NK 0.31 0.35 0.186 0.17 0.21 0.470 % T cell 67 60 0.183 76.15 67.39 0.340 Abs T cell 0.72 0.96 0.717 1.35 .82 0.095 NKT%* 5.28 3.33 8.25 3.38 B cell % 2.38 1.52 0.922 2.63 5.58 0.424 Abs. Neut count 2.99 2.64 0.488 2.85 3.01 0.848 Disclosures No relevant conflicts of interest to declare.

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Haploidentical TCRab Depleted Hematopoietic Cell Transplantation Leads To Rapid Immune Reconstitution and Promising Clinical Outcome In Adults

Blood ◽

10.1182/blood.v122.21.5475.5475 ◽

2013 ◽

Vol 122 (21) ◽

pp. 5475-5475

Author(s):

Sebastian P. Haen ◽

Michael Schumm ◽

Wichard Vogel ◽

Christoph Faul ◽

Rupert Handgretinger ◽

...

Keyword(s):

T Cells ◽

Body Weight ◽

Clinical Outcome ◽

Cell Transplantation ◽

Immune Reconstitution ◽

Hematopoietic Cell Transplantation ◽

Reduced Intensity Conditioning ◽

Virus Pneumonia ◽

Reduced Intensity ◽

Observation Period

Abstract Immunomagnetic TCRab-depletion and reduced intensity conditioning (RIC) for haploidentical hematopoietic cell transplantation (HHCT) has been reported to result in improved immune reconstitution and clinical outcome in children. We here report the first clinical results in 10 adult patients. HHCT using TCRab-depleted grafts was performed in 7 men and 3 women (median age at transplantation 35 years, range 21-68 years). Patients were treated for ALL (n=1), AML (n=5), CLL (n=1), NHL (n=2) or SAA (n=1). Disease status prior transplantation was CR in two patients, minimal residual disease but cytologic CR in 1 patient, conditioning out of aplasia after salvage chemotherapie in 4 patients, and active disease in 3 patients. Haploidentical donors were parents (n=3), siblings (n=4) and children (n=3). Seven patients were treated with HHCT due to relapse after first (n=6) or second (n=1) allogeneic HCT. In 3 patients, HHCT was performed as primary transplantation. Conditioning regimen was myeloablative (12 Gy TBI/high dose cyclophosphamide) in 1 patient and reduced intensity conditioning (fludarabine/clofarabine, thiotepa, melphalan) in the remaining 9 patients. All conditioning regimens included ATG and mycophenolate mofetil as postgrafting immunosuppression until day +30. Patients received a median of 6.18 x 106 (range 3.4-20.4 x106) CD34+,5.92 x 106 (range 2.26-14.84) CD3+and 36.77 x 106 (range 15.75-51.84) CD56+ cells per kilogram body weight. TCRab-depletion of the grafts was a median of 4.5 logs with 0.38 104 TCRab+ CD3+ (range 0.06-1.17) cells per kilogram body weight in the graft. Median engraftment of neutrophils (> 500/µl) and thrombocytes (> 25.000/µl) occurred on days 12 (range day 8-15) and 14 (range day 10-23), respectively, with one patient not achieving thrombocyte engraftment until death. Patients were followed for a median of 276 days after HHCT (range 41-723 days). To date, 6 patients remain alive and in CR (60%). Four patients died due to viral infections (n=4, 40%) at a median of 92 days after HHCT (range 41-100 days) comprising adenovirus viremia, varicella zoster virus encephalitis, respiratory syncytial virus pneumonia and herpes simplex virus pneumonia. One patient also suffered from severe graft versus host disease (GvHD) of the liver. Acute GvHD ≥II occurred in a total of 5 patients with development of chronic GvHD not being observed. None of the patients experienced relapse of the underlying hematologic disease, the non-relapse mortality was 40%. Median overall and disease free survival were not reached during the observation period. Immune reconstitution was studied by flow cytometry at least every week for a median follow-up of 122 days after transplantation (range 25-558 days). Detailed information on immune reconstitution data is provided in the table. Median T cell (CD3+) engraftment (> 200/µl) occurred on day 42 (range 21-397 days). Two patients did not reach > 200/µl CD3+ cells until death. Engraftment of CD4+ T cells > 200/µl was only achieved in 3 patients during the observation period. Our data indicate that in comparison to approaches applying CD34 selected or CD3/CD19 depleted grafts, depletion of TCRab T cells in HHCT might lead to more rapid immune reconstitution. A prospective study evaluating the role of TCRab depletion in HHCT in adults is presently ongoing. Table Peripheral cell counts of leukocyte subsets. NK cells T cells T helper cells Cytotoxic T cells B cells CD56+ CD16+ CD3- CD3+ CD3+ CD4+ CD3+ CD8+ CD19+ Day 30 (median [/µl]) 435 28 3 12 1 range [/µl] 113-642 5-1,655 0-292 2-509 0-212 Day 60 (median [/µl]) 433 457 54 342 213 range [/µl] 188-504 1-1,367 0-175 0-368 0-556 Day 100 (median [/µl]) 319 74 9 39 82 range [/µl] 118-1,223 0-2,292 0-229 0-1,289 19-344 Peak (median [/µl]) 466 416 186 221 309 range [/µl] 134-2,650 5-4,529 3-787 2-4,029 1-3,065 Day Peak (median [day]) 52 62 59 62 63 range [days] 19-337 25-397 8-558 25-344 19-344 Disclosures: No relevant conflicts of interest to declare.

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An Unconventional View of T Cell Reconstitution After Allogeneic Hematopoietic Cell Transplantation

Frontiers in Oncology ◽

10.3389/fonc.2020.608923 ◽

2021 ◽

Vol 10 ◽

Author(s):

Hana Andrlová ◽

Marcel R. M. van den Brink ◽

Kate A. Markey

Keyword(s):

T Cells ◽

Cell Transplantation ◽

Immune Reconstitution ◽

Hematopoietic Cell Transplantation ◽

Hematopoietic Cell ◽

Allogeneic Hematopoietic Cell Transplantation ◽

Intestinal Microbiome ◽

Post Transplantation ◽

Curative Intent

Allogeneic hematopoietic cell transplantation (allo-HCT) is performed as curative-intent therapy for hematologic malignancies and non-malignant hematologic, immunological and metabolic disorders, however, its broader implementation is limited by high rates of transplantation-related complications and a 2-year mortality that approaches 50%. Robust reconstitution of a functioning innate and adaptive immune system is a critical contributor to good long-term patient outcomes, primarily to prevent and overcome post-transplantation infectious complications and ensure adequate graft-versus-leukemia effects. There is increasing evidence that unconventional T cells may have an important immunomodulatory role after allo-HCT, which may be at least partially dependent on the post-transplantation intestinal microbiome. Here we discuss the role of immune reconstitution in allo-HCT outcome, focusing on unconventional T cells, specifically mucosal-associated invariant T (MAIT) cells, γδ (gd) T cells, and invariant NK T (iNKT) cells. We provide an overview of the mechanistic preclinical and associative clinical studies that have been performed. We also discuss the emerging role of the intestinal microbiome with regard to hematopoietic function and overall immune reconstitution.

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