Allogeneic, Off-the-Shelf, Sars-Cov-2-Specific T Cells to Treat High-Risk Patients with COVID-19

Seventy-one patients with hematologic malignancies received bone marrow from a histocompatible sibling (n = 48) or a partially matched relative (n = 23) that had been depleted of CD5+ T cells with either an anti-CD5 mooclonal antibody (MoAb) plus complement (anti-Leu1 + C) or an anti- CD5 MoAb conjugated to ricin A chain (ST1 immunotoxin [ST1-IT]). These patients received intensive chemoradiotherapy consisting of cytosine arabinoside, cyclophosphamide, and fractionated total body irradiation. Both anti-Leu1 + C and ST1-IT ex vivo treatments effectively depleted bone marrow of T cells (97% and 95%, respectively). Overall, primary and late graft failure each occurred in 4% of evaluable patients. The diagnosis of myelodysplasia was a significant risk factor for graft failure (P less than .001), and if myelodysplastic patients were excluded, there were no graft failures in major histocompatibility complex (MHC)-matched patients and 2 of 23 (8.7%) in MHC-mismatched patients. The actuarial risk of grade 2 to 4 acute graft-versus-host disease (GVHD) was 23% in MHC-matched patients and 50% in MHC- mismatched patients. In MHC-matched patients, acute GVHD tended to be mild and treatable with corticosteroids. Chronic GVHD was observed in 6 of 36 (17%) MHC-matched patients and none of 11 MHC-mismatched patients. There were no deaths attributable to GVHD in the MHC-matched group. Epstein-Barr virus-associated lymphoproliferative disorders were observed in 3 of 23 MHC-mismatched patients. The actuarial event-free survival was 38% in the MHC-matched patients versus 21% in the MHC- mismatched patients. However, if outcome is analyzed by risk of relapse, low-risk patients had a 62% actuarial survival compared with 11% in high-risk patients. These data indicate that the use of anti-CD5 MoAbs can effectively control GVHD in histocompatible patients, and that additional strategies are required in MHC-mismatched and high-risk patients.

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CMV-Seropositive Patients Allografted with a CMV-Seronegative Donor Show Delayed or Missing CMV-Specific T-Cell Immune Response and Are at Higher Risk for CMV-Viremia and CMV-Disease.

Blood ◽

10.1182/blood.v106.11.3249.3249 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3249-3249

Author(s):

Susanne Ganepola ◽

Chiara Gentilini ◽

Urte Hilbers ◽

Goetz Hartung ◽

Thoralf Lange ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

High Risk ◽

Cell Response ◽

T Cell Response ◽

First Year ◽

Seronegative Donor ◽

High Risk Patients ◽

Risk Patients ◽

Cmv Disease

Abstract The human cytomegalovirus (CMV) is an important cause for mortality and morbidity after allogeneic stem cell transplantation (allo SCT) especially in patients without a CMV-specific T cell response. CMV seropositive patients allografted with a CMV seronegative donor do not rise up specific CMV-T cell immunity posttransplant and are therefore at great risk for CMV viremia and disease. In a prospective manner, we analyzed blood samples of 38 HLA-A2+ patients with different haematological malignancies for CMV specific T cells. Methods: Frequency of T cells with reactivity against the pp65-peptide (NLVPMVATV) was assessed by ELISPOT assay in 38 patients at defined time points after allo SCT. In patients with high CMV specific T cell frequencies, the T cell phenotype was determined by flow cytometry. Surveillance of CMV viremia was carried out by routine PCR-technique or pp65 Ag staining. Results: In high-risk patients (donor-/recipient+) viremia was observed in 7/9 patients, 3/7 developed clinical severe CMV-disease. In this group, only one patient presented a weak CMV-specific T cell response in the first year after transplantation, although CMV-specific T cells were demonstrated in 5/9 patients before transplantation. In contrast, 64% (11/17) of the CMV seropositive patients having had a CMV seropositive donor showed CMV-specific T cells around day 30. Their T cell frequencies remained stable at a relative high level during the whole time of our investigation period and no CMV disease was observed. CMV seronegative patients allografted with either a CMV seronegative or seropositive donor also remained disease-free. CMV specific T cells were detectable in 2/7 patients of the d+/r− group. FACS analysis revealed that most of the responding cells were of the activated effector phenotype CD8+/CD45RA+/HLA-DR+ with a low expression of CCR7 and CD27. Conclusion: CMV-seropositive patients receiving graft from a seronegative donor are at a high risk and therefore prone for CMV-viremia and -disease. The development of CMV-specific T cells i.e. immune reconstitution in high risk patients remains delayed or is completely missing during the first year post transplant. In contrast, seropositive recipients grafted with a seropositive donor develop a durable T cell response within 3–4 weeks and show no viremia at all. New strategies as donor vaccination and adoptive T cell transfer are warranted to prevent CMV-disease in CMV seropositive patients for whom only a seronegative donor can be found.

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Quantification of tumor infiltrating Foxp3+ regulatory T cells enables the identification of high-risk patients for developing synchronous cancers over upper aerodigestive tract

Oral Oncology ◽

10.1016/j.oraloncology.2015.04.015 ◽

2015 ◽

Vol 51 (7) ◽

pp. 698-703 ◽

Cited By ~ 13

Author(s):

Wen-Lun Wang ◽

Wei-Lun Chang ◽

Hsiao-Bai Yang ◽

I-Wei Chang ◽

Ching-Tai Lee ◽

...

Keyword(s):

T Cells ◽

High Risk ◽

Regulatory T Cells ◽

Aerodigestive Tract ◽

Upper Aerodigestive Tract ◽

High Risk Patients ◽

Risk Patients ◽

Synchronous Cancers

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Selective depletion of bone marrow T lymphocytes with anti-CD5 monoclonal antibodies: effective prophylaxis for graft-versus-host disease in patients with hematologic malignancies

Blood ◽

10.1182/blood.v78.8.2139.2139 ◽

1991 ◽

Vol 78 (8) ◽

pp. 2139-2149 ◽

Cited By ~ 83

Author(s):

JH Antin ◽

BE Bierer ◽

BR Smith ◽

J Ferrara ◽

EC Guinan ◽

...

Keyword(s):

Bone Marrow ◽

T Cells ◽

High Risk ◽

Graft Versus Host Disease ◽

Graft Failure ◽

Hematologic Malignancies ◽

Host Disease ◽

Graft Versus Host ◽

High Risk Patients ◽

Risk Patients

Abstract Seventy-one patients with hematologic malignancies received bone marrow from a histocompatible sibling (n = 48) or a partially matched relative (n = 23) that had been depleted of CD5+ T cells with either an anti-CD5 mooclonal antibody (MoAb) plus complement (anti-Leu1 + C) or an anti- CD5 MoAb conjugated to ricin A chain (ST1 immunotoxin [ST1-IT]). These patients received intensive chemoradiotherapy consisting of cytosine arabinoside, cyclophosphamide, and fractionated total body irradiation. Both anti-Leu1 + C and ST1-IT ex vivo treatments effectively depleted bone marrow of T cells (97% and 95%, respectively). Overall, primary and late graft failure each occurred in 4% of evaluable patients. The diagnosis of myelodysplasia was a significant risk factor for graft failure (P less than .001), and if myelodysplastic patients were excluded, there were no graft failures in major histocompatibility complex (MHC)-matched patients and 2 of 23 (8.7%) in MHC-mismatched patients. The actuarial risk of grade 2 to 4 acute graft-versus-host disease (GVHD) was 23% in MHC-matched patients and 50% in MHC- mismatched patients. In MHC-matched patients, acute GVHD tended to be mild and treatable with corticosteroids. Chronic GVHD was observed in 6 of 36 (17%) MHC-matched patients and none of 11 MHC-mismatched patients. There were no deaths attributable to GVHD in the MHC-matched group. Epstein-Barr virus-associated lymphoproliferative disorders were observed in 3 of 23 MHC-mismatched patients. The actuarial event-free survival was 38% in the MHC-matched patients versus 21% in the MHC- mismatched patients. However, if outcome is analyzed by risk of relapse, low-risk patients had a 62% actuarial survival compared with 11% in high-risk patients. These data indicate that the use of anti-CD5 MoAbs can effectively control GVHD in histocompatible patients, and that additional strategies are required in MHC-mismatched and high-risk patients.

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Mucorales-specific T cells emerge in the course of invasive mucormycosis and may be used as a surrogate diagnostic marker in high-risk patients

Blood ◽

10.1182/blood-2011-07-366526 ◽

2011 ◽

Vol 118 (20) ◽

pp. 5416-5419 ◽

Cited By ~ 48

Author(s):

Leonardo Potenza ◽

Daniela Vallerini ◽

Patrizia Barozzi ◽

Giovanni Riva ◽

Fabio Forghieri ◽

...

Keyword(s):

T Cells ◽

High Risk ◽

Immune Responses ◽

Diagnostic Marker ◽

High Risk Patients ◽

Risk Patients ◽

Ifn Γ ◽

Human Immune

Abstract Mucorales-specific T cells were investigated in 28 hematologic patients during the course of their treatment. Three developed proven invasive mucormycosis (IM), 17 had infections of known origin but other than IM, and 8 never had fever during the period of observation. Mucorales-specific T cells could be detected only in patients with IM, both at diagnosis and throughout the entire course of the IM, but neither before nor for long after resolution of the infection. Such T cells predominantly produced IL-4, IFN-γ, IL-10, and to a lesser extent IL-17 and belonged to either CD4+ or CD8+ subsets. The specific T cells that produced IFN-γ were able to directly induce damage to Mucorales hyphae. None of the 25 patients without IM had Mucorales-specific T cells. Specific T cells contribute to human immune responses against fungi of the order Mucorales and could be evaluated as a surrogate diagnostic marker of IM.

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A Two Step Approach to Haploidentical Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) for High Risk Patients with Hematologic Disorders.

Blood ◽

10.1182/blood.v110.11.5072.5072 ◽

2007 ◽

Vol 110 (11) ◽

pp. 5072-5072

Author(s):

Neal Flomenberg ◽

Dolores Grosso ◽

Janet Brunner ◽

Matthew Carabasi ◽

Scott Dessain ◽

...

Keyword(s):

T Cells ◽

High Risk ◽

Conditioning Regimen ◽

Additional Patient ◽

Hematopoietic Stem ◽

High Risk Patients ◽

Cd 34 ◽

Risk Patients ◽

Relapsed Disease ◽

Donor Lymphocytes

Abstract Post transplant cyclophosphamide (CTX) has been utilized to induce donor host tolerance in allogeneic HSCT. While promising, the long term risk of MDS and AML from exposure of the graft to CTX is unknown and the optimal T cell dose to deliver using this approach remains undefined. To address these issues, nine high-risk patients, median age 55 (range 19–73) underwent haploidentical HSCT from related donors for various hematopoietic disorders using a two step approach. Seven of 9 (77%) patients had evidence of disease at the time of HSCT. In the GVH direction, 7 of 9 were 4/8 matches with their donors at HLA-A, B, C and DRB1; the other 2 patients were 5 /8 and 6 /8 matches. Seven of the 9 patients were given a myeloablative conditioning regimen of TBI (1.5 Gy) BID on days −9 through −6. Two received a nonmyeloablative conditioning regimen of fludarabine and ARA-C on days-11 through −8 and one 2 Gy fraction of TBI on day −6. The patients received 2x108/kg of their donor’s CD3+ lymphocytes after completion of TBI on day −6, followed by rest days on day −5 and −4. On days −3 and −2, CTX was given at a dose of 60 mg/kg/day for the purpose of tolerization of the donor lymphocytes. On day 0, donor CD 34+ cells were infused. When possible, the number of non-tolerized T-cells in the CD 34+ product was limited to ≤5x104/kg. Tacrolimus and CellCept were used as GVHD prophylaxis. All of the patients developed fevers between DLI and CTX, with some patients also developing mild diarrhea and a rash, which in one case was biopsy-positive for GVHD. All patients engrafted. One patient developed grade III GVHD, two had grade II GVHD, and the rest had grade 0 or I GVHD. GVHD was easily controlled with corticosteroids which could be rapidly tapered in all but one patient. Four patients have died, two of relapsed disease (d+67, d+91) and two of infectious complications (d+101, d+42). An additional patient is alive, but with evidence of relapsed disease at d + 42. Four patients are well and in CR at days +250, +173, +144, and +118 days post HSCT. Three of the 4 have CD3/CD4 counts of greater than 170 cells/ml. This 2 step transplant procedure promotes the establishment of donor-recipient tolerance by in vivo CTX exposure, but avoids any effects of G-CSF on donor lymphocytes, allows delivery of a precise dose of T cells to all patients, and eliminates CTX exposure of the hematopoietic precursors. Patients who lack matched-sibling donors can be successfully transplanted on this protocol with little significant GVHD. Infection and relapse remain problems, but likely can be reduced through transplantation of patients earlier in their disease course. Patient Data Age Disease Transplant Type DLI Dose/kg x10e8 CD 34 Dose/kg x10e6 T Cells/kg in CD34 Product x10e4 GVHD Grade Outcome 19 AML - PIF Ablat 2.0 1.4 0.26 0 Died-Relapse 55 CML & AML - PIF Ablat 2.0 3.6 5.0 0 NED 26 ALL - CR2 Ablat 2.0 2.2 6.9 II NED 44 AML - CR2 Ablat 1.8 3.8 4.4 II NED 32 SAA Ablat 2.0 1.7 0.13 III Died-Infection 59 AML - Resist Relapse 1 Ablat 2.0 4.1 0.39 0 Died-Relapse 60 Biph Leuk - Resist Relapse 1 Ablat 2.0 2.7 0.82 I Died-Infection 67 AML - PIF Non-Ablat 2.0 4.3 4.5 II NED 73 AML - PIF Non-Ablat 2.0 4.4 0.059 0 Alive-Relapsed

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Tandom Autologous Transplantation and Combined Infusion of CD19 and Bcma-Specific Chimeric Antigen Receptor T Cells for High Risk MM: Initial Safety and Efficacy Report from a Clinical Pilot Study

Blood ◽

10.1182/blood-2018-99-117964 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 1009-1009 ◽

Cited By ~ 28

Author(s):

Xiaolan Shi ◽

Lingzhi Yan ◽

Jingjing Shang ◽

Su Qu ◽

Liqing Kang ◽

...

Keyword(s):

Flow Cytometry ◽

T Cells ◽

High Risk ◽

Autologous Transplantation ◽

Cytokine Release ◽

Cytokine Release Syndrome ◽

Safety And Efficacy ◽

High Risk Patients ◽

Risk Patients ◽

Cd19 Expression

Abstract Background: Multiple myeloma (MM) is an incurable plasma cell malignancies despite the advent of numerously new drugs. Survival was poor particularly for high risk patients such as R-ISS stage III. The preliminary data from our center showed that the median PFS after auto-HSCT was 24 months for patients with R-ISS III stage, while 17 months for patients that achieved PR or less after induction. Chimeric antigen receptor (CAR)-transduced T cells is a promising strategy for cancer immunotherapy. Our previous study showed good response for RRMM patients after CD19 and BCMA-specific CART therapy without severe CRS and other deadly side effects. To improve the survival of high risk patients, this study was designed to observe the safety and efficacy of combined infusion of CD19 and BCMA-specific CART cells after autologous transplantation (SZ-MM-CART02 study, NCT 03455972). Methods:18-65y NDMM in R-ISS stage III, or who only achieved PR or less after 4 cycles of PAD triplet induction were enrolled with serum creatinine (Cr) <2.0 mg/dL, and adequate hepatic, cardiac and pulmonary function. BCMA and CD19 expression on MM cells were analyzed by flow cytometry. Lymphocytes were collected from PBSCs and cultured with an anti-CD3 monoclonal antibody to activate T-cell proliferation. The cells were transduced with recombinant lentiviral verctors which respectively contained the anti-BCMA or anti-CD19 single chain variable fragment (scFv), the cytoplasmic portion of the OX40 and CD28 costimulatory moiety, and the CD3z T-cell activation domain. This is the new third generation CAR technique applied in clinic. BUCY were used as conditioning, followed by infusion of autologous stem cells. Median time to engraftment were 10 days for nutrophils. CART-19 (1×107/kg on d0) and CART-BCMA cells as split-dose (40% on d1 and 60% on d2) were infused derectly on d14 to d20 after autologous transplantation. Levels of CAR-transduced cells are measured by qPCR. The cytokine release syndrome (CRS) was graded according to the UPen cytokine release syndrome grading system. Neurotoxic side effects and other toxicities were assessed according to the CTCAE v 4.03. Plasma levels of IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma, and IL-17A proteins were determined with a cytokine kit. Imids alone were given as maintenance therapy. Responses were assessed by IMWG criteria. 10-color flow cytometry was used to monitor MRD regularly after CART treatment. The median of follow-up was 3 (2∼11) months. Results: To date, 9 patients have completed the CART cells infusion (cohort 1). All cases expressed BCMA >50% without CD19 expression on MM cells. CRS occurred in 9 patients (100%) grade1 or 2 associated with fever(n=9), fatigue (n=9), elevated IL-6 and CRP (n=9), elevated ALT (n=1, grade 1). Two patients needed to use low-dose vascular active drugs (pts 04 and 07). Other toxicities to date included coagulopathy (n=6, grade 1 for 4 pts and grade 2 for 2 pts), elevated troponin T (n=4, grade 1), and atrial flutter (n=1). There was no serious CRS or neurologic complications occurred in this group of patients. The ORR was 100% with all patients were monitored for a period of more than 2 months, which may be eventually further improved. There were 2 CR, 1 VGPR, 4 PR, 2 SD after induction; 3 CR, 2 VGPR, 4 PR after APBSCT; 3 CR, 6 VGPR after CART therapy. MRD negativity in BM increased from 37.5% after transplantation to 66.7% after CART therapy latest. Four patients (pts 02, 03, 06 and 07) obtained partial PR after transplantation, and got VGPR after CART cells infusion. We found dramatic in vivo CART expansion that median of peak value of CART copies was 1059.54 folds (ranged from 536.90 to 10997.93 folds) which was 100 folds to that with RRMM patients in our previous study. Conclusions: Tandom autologous transplantation and combined infusion of CART-19 and CART-BCMA cells could be another choice of consolidation treatment for high risk MM patients. Toxicities to date including CRS and organ function impairment seemed to be mild and reversable. It is worthy of further study to compare DFS, OS between single autologous transplantation and tandom transplantation with CART therapy. Immune environment in high risk patients with multiple myelomaI remodelled by auto-HSCT may contribute to more rapid expansion of CART cells than that in RRMM patients, suggesting that the extent of CART expansion depends more than tumor burden. Table Table. Disclosures No relevant conflicts of interest to declare.

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Identification of tumour immune microenvironment-related alternative splicing events for the prognostication of pancreatic adenocarcinoma

BMC Cancer ◽

10.1186/s12885-021-08962-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Bo Chen ◽

Tuo Deng ◽

Liming Deng ◽

Haitao Yu ◽

Bangjie He ◽

...

Keyword(s):

T Cells ◽

Alternative Splicing ◽

High Risk ◽

Pancreatic Adenocarcinoma ◽

Gene Set Enrichment Analysis ◽

Prognostic Signature ◽

Immune Microenvironment ◽

High Risk Patients ◽

Risk Patients ◽

Alternative Splicing Events

Abstract Purpose Pancreatic adenocarcinoma (PAAD) is characterized by low antitumour immune cell infiltration in an immunosuppressive microenvironment. This study aimed to systematically explore the impact on prognostic alternative splicing events (ASs) of tumour immune microenvironment (TIME) in PAAD. Methods The ESTIMATE algorithm was implemented to compute the stromal/immune-related scores of each PAAD patient, followed by Kaplan–Meier (KM) survival analysis of patients with different scores grouped by X-tile software. TIME-related differentially expressed ASs (DEASs) were determined and evaluated through functional annotation analysis. In addition, Cox analyses were implemented to construct a TIME-related signature and an AS clinical nomogram. Moreover, comprehensive analyses, including gene set enrichment analysis (GSEA), immune infiltration, immune checkpoint gene expression, and tumour mutation were performed between the two risk groups to understand the potential mechanisms. Finally, Cytoscape was implemented to illuminate the AS-splicing factor (SF) regulatory network. Results A total of 437 TIME-related DEASs significantly related to PAAD tumorigenesis and the formation of the TIME were identified. Additionally, a robust TIME-related prognostic signature based on seven DEASs was generated, and an AS clinical nomogram combining the signature and four clinical predictors also exhibited prominent discrimination by ROC (0.762 ~ 0.804) and calibration curves. More importantly, the fractions of CD8 T cells, regulatory T cells and activated memory CD4 T cells were lower, and the expression of four immune checkpoints—PD-L1, CD47, CD276, and PVR—was obviously higher in high-risk patients. Finally, functional analysis and tumour mutations revealed that aberrant immune signatures and activated carcinogenic pathways in high-risk patients may be the cause of the poor prognosis. Conclusion We extracted a list of DEASs associated with the TIME through the ESTIMATE algorithm and constructed a prognostic signature on the basis of seven DEASs to predict the prognosis of PAAD patients, which may guide advanced decision-making for personalized precision intervention.

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