ArticlePublisher preview availableLiterature Review

The effect of cyclophosphamide on the immune system: implications for clinical cancer therapy

DOI:10.1007/s00280-016-3152-1
Authors:
Martina Ahlmann
Martina Ahlmann
Martina Ahlmann
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Georg Hempel at University of Münster
Georg Hempel
Read publisher preview
Download citation
To read the full-text of this research, you can request a copy directly from the authors.

Abstract and Figures

Cyclophosphamide is an alkylating agent belonging to the group of oxazaphosporines. As cyclophosphamide is in clinical use for more than 40 years, there is a lot of experience using this drug for the treatment of cancer and as an immunosuppressive agent for the treatment of autoimmune and immune-mediated diseases. Besides antimitotic and antireplicative effects, cyclophosphamide has immunosuppressive as well as immunomodulatory properties. Cyclophosphamide shows selectivity for T cells and is therefore now frequently used in tumour vaccination protocols and to control post-transplant allo-reactivity in haplo-identical unmanipulated bone marrow after transplantation. The schedule of administration is of special importance for the immunological effect: while cyclophosphamide can be used in high-dose therapy for the complete eradication of haematopoietic cells, lower doses of cyclophosphamide are relatively selective for T cells. Of special interest is the fact that a single administration of low-dose cyclophosphamide is able to selectively suppress regulatory T cells (Tregs). This effect can be used to counteract immunosuppression in cancer. However, cyclophosphamide can also increase the number of myeloid-derived suppressor cells. Combination of cyclophosphamide with other immunomodulatory agents could be a promising approach to treat different forms of advanced cancer.
Figures - available from: Cancer Chemotherapy and Pharmacology
This content is subject to copyright. Terms and conditions apply.
  • A preview of this full-text is provided by Springer Nature.
  • Learn more
Preview content only
Content available from Cancer Chemotherapy and Pharmacology
This content is subject to copyright. Terms and conditions apply.
1 3
Cancer Chemother Pharmacol (2016) 78:661–671
DOI 10.1007/s00280-016-3152-1
REVIEW ARTICLE
The effect of cyclophosphamide on the immune system:
implications for clinical cancer therapy
Martina Ahlmann1 · Georg Hempel2
Received: 8 June 2015 / Accepted: 2 September 2016 / Published online: 19 September 2016
© Springer-Verlag Berlin Heidelberg 2016
agents could be a promising approach to treat different
forms of advanced cancer.
Keywords T cells · Oxazaphosphorines · Immune
reactivation · Immunosuppression
Introduction
Cyclophosphamide is one of the oldest anticancer drugs. It
was discovered as early as 1958 and introduced into cancer
therapy in 1959 [1]. It remains a mainstay in the therapy of
haematological malignancies including lymphoma and leu-
kaemia as well as of various epithelial tumours including
breast, ovarian and small-cell lung carcinomas [2]. In Ger-
many, the drug is approved for the therapy of ALL, CLL,
Hodgkin and non-Hodgkin lymphoma, plasmocytoma, breast
cancer, ovarian carcinoma, small-cell lung cancer, Ewing-,
osteo- and rhabdomyosarcoma and neuroblastoma [3].
The drug is applied in many conditioning regimens
before bone marrow transplantation for haematological
malignancies (AML, MDS) as well as for aplastic anaemia.
Besides cancer chemotherapy, other approved indications
are life-threatening events in autoimmune and immune-
mediated diseases as lupus nephritis, Wegener’s granulo-
matosis and multiple sclerosis.
Important side effects are leuco- and thrombocytope-
nia, anaemia, cardio- and bladder toxicity. To avoid haem-
orrhagic cystitis, MESNA (sodium 2-sulfanylethanesul-
fonate) must be administered before cyclophosphamide
administration to neutralise the toxic metabolite acrolein
in the urine. In addition, nephrotoxicity, cardiotoxicity
and liver toxicity occur. All these effects are highly dose
dependent. Cardiotoxicity is dose-limiting in high-dose
schedules of cyclophosphamide [4].
Abstract Cyclophosphamide is an alkylating agent
belonging to the group of oxazaphosporines. As cyclophos-
phamide is in clinical use for more than 40 years, there is a
lot of experience using this drug for the treatment of can-
cer and as an immunosuppressive agent for the treatment of
autoimmune and immune-mediated diseases. Besides anti-
mitotic and antireplicative effects, cyclophosphamide has
immunosuppressive as well as immunomodulatory proper-
ties. Cyclophosphamide shows selectivity for T cells and is
therefore now frequently used in tumour vaccination proto-
cols and to control post-transplant allo-reactivity in haplo-
identical unmanipulated bone marrow after transplantation.
The schedule of administration is of special importance for
the immunological effect: while cyclophosphamide can be
used in high-dose therapy for the complete eradication of
haematopoietic cells, lower doses of cyclophosphamide are
relatively selective for T cells. Of special interest is the fact
that a single administration of low-dose cyclophosphamide
is able to selectively suppress regulatory T cells (Tregs).
This effect can be used to counteract immunosuppression
in cancer. However, cyclophosphamide can also increase
the number of myeloid-derived suppressor cells. Combina-
tion of cyclophosphamide with other immunomodulatory
* Georg Hempel
georg.hempel@uni-muenster.de
Martina Ahlmann
martina.ahlmann@ukmuenster.de
1 Pädiatrische Hämatologie und Onkologie, Klinik für
Kinder- und Jugendmedizin, Universitätsklinikum Münster,
Albert-Schweitzer-Campus 1, A1, Münster, Germany
2 PharmaCampus, Klinische Pharmazie, Westfälische
Wilhelms-Universität Münster, Corrensstraße 48, 48149,
Münster, Germany
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
... Glyceraldehyde 3-phosphate dehydrogenase (GAPDH; forward: 5'-ACCTTCACTCCTCCATCTT-3' , reverse: 5'-AGGTCACAGACACGGTTG-3') and 5.8S rRNA (forward: 5'-AACTCTTAGCGGTGGATCA-3' , Cyclophosphamide is widely used as an antineoplastic drug because of antireplicative and antimitotic properties (Emadi et al., 2009). Moreover, cyclophosphamide is also used in the treatment of organ transplants and autoimmune diseases through immunosuppressive effect (Ahlmann and Hempel, 2016). A previous study has shown that administration of cyclophosphamide reduced the proliferation of B and T cells, suppressed the activity of cytotoxic T lymphocyte and NK cells, and stimulated the imbalance of type 1 T helper/type 2 T helper (Th1/Th2) cells, thus resulting in sudden immunosuppression in tumor cells (Noh et al., 2019). ...
Norcantharidin alleviates cyclophosphamide-induced immunosuppression via circBCL2L1/miR-30c-3-3p/TRAF6 axis
Article
Full-text available
  • Jun 2022
Cyclophosphamide is a widely used antitumor drug, with induced adverse effects, such as intestinal mucosal injury and immunosuppression. Norcantharidin possesses anticancer activity through enhancement of antitumor immunity. We investigated the role of norcantharidin in cyclophosphamide-induced immunosuppression. Mice were treated with cyclophosphamide, and exposed to norcantharidin. Enzyme-linked-immunosorbent serologic assay was performed to assess the levels of immunoglobulin and cytokines in serum, and the splenic T lymphocytes were analyzed by immunohistochemistry. Incubation with norcantharidin increased the serum levels of immunoglobulin G (IgG), interleukin (IL)-12, interferon-gamma (IFN-γ), and IL-6, and enhanced the percentage of CD4+ and CD8+ T lymphocytes in cyclophosphamide-induced mice. Expression of circBCL2L1 was down-regulated in the spleen of cyclophosphamide-induced mice, while up-regulated by norcantharidin incuba-tion. Norcantharidin attenuated cyclophosphamide-induced up-regulation of miR-30c-3-3p and down-regulation of tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) in mice. Over-expression of circBCL2L1 increased serum levels of immunoglobulin and cytokines, and enhanced the percentage of splenic CD4+ and CD8+ T lymphocytes in cyclophosphamide-induced mice. Moreover, over-expression of circBCL2L1 increased TRAF6 in cyclophosphamide-induced mice through down-regulation of miR-30c-3-3p. Knockdown of TRAF6 attenuated norcantharidin-induced increase of serum levels of IgG, IL-12, IFN-γ, and IL-6, and up-regulation of CD4+ and CD8+ T lymphocytes in cyclophosphamide-induced mice. Norcantharidin exhibited protective effect against cyclophosphamide-induced immunosuppression in mice through regulation of circBCL2L1/miR-30c-3-3p/TRAF6 axis.
... Cyclophosphamide (CP), an alkylating drug, is frequently used as an immunosuppressant during organ transplantation and is used against many types of cancers, autoimmune disorders, and other conditions [1,2]. However, the therapeutic use of CP is restricted due to the side effects it produces, such as nephrotoxicity, cardiotoxicity, and hepatotoxicity [3]. ...
Anti-inflammatory and anti-apoptotic potential of beta-glucan on chemotherapy-induced nephrotoxicity in rats: β-glucan on cyclophosphamide-induced nephrotoxicity
Article
Full-text available
  • Jan 2023
Background/Aim: Cyclophosphamide (CP) is an anti-cancer agent that mediates nephrotoxicity. Beta (β)-glucan has restorative effects on kidney toxicities through its antioxidant potential; however, the effects of β-glucan on CP-induced renal injury remain unknown. In an experimental nephrotoxicity model using rats, we sought to examine the potential protective action of β-glucan on kidney histomorphology, apoptosis, and TNF-α expression. Methods: Male albino Wistar rats were divided equally into four groups: control, CP, β-glucan, and CP+β-glucan. The kidney tissues of the rats were examined for TNF-α and caspase-3 immunostaining to evaluate inflammation and apoptosis, respectively. Hematoxylin and eosin (H&E) and periodic acid–Schiff (PAS) staining were used for histopathological analyses. Results: The CP group showed severe histopathological damage in the renal tissues of rats. In the renal tissue of the CP group, immunoreactivities for TNF-α (1.25 [0.079] and caspase-3 (1.506 [0.143] were also higher than the control group (0.117 [0.006] and 0.116 [0.002], respectively; P<0.001). In the CP+β-glucan group, the histopathological changes significantly improved. Conclusion: Beta-glucan has therapeutic potential against CP-induced nephrotoxicity in rat kidney.
... For example, low dose cisplatin and oxaliplatin increase the number of circulating CD4+ and CD8+ T cells, while high dose regimens decrease the size of lymphocyte in a mouse model of colon cancer (132). Metronomic low dose cyclophosphamide enhance anti-tumor immune response by selectively reducing the amount of Treg cells in tumor patients (133), while high dose cyclophosphamide completely eradicates the hemopoietic cell (134). In addition, a novel administration mode called mediumdose intermittent chemotherapy provokes a striking response depending on the activation of a sustaining anti-tumor immune response (66,135). ...
The application basis of immuno-checkpoint inhibitors combined with chemotherapy in cancer treatment
Article
Full-text available
  • Jan 2023
Immuno-checkpoint inhibitors (ICIs) bring a promising prospect for patients with cancers, which restrains the growth of tumor cells by enhancing anti-tumor activity. Nevertheless, not all patients benefit from the administration of ICIs monotherapy. The partial response or resistance to ICIs is mainly due to the complex and heterogenous tumor microenvironment (TME). The combined therapy is necessary for improving the efficacy of tumor treatment. Chemotherapy is reported not only to kill tumor cells directly, but also to stimulate effective anti-tumor immune responses. Several combined therapies of ICIs and chemotherapeutic agents have been approved for the first-line treatment of cancers, including PD-1/PD-L1 inhibitors. This review summarizes the potential mechanisms of the combined therapy of ICIs and chemotherapeutic agents in inducing immunogenic cell death (ICD) and reprogramming TME, and elucidates the possible anti-tumor effects of combined therapy from the perspective of metabolic reprogramming and microbiome reprogramming.
Lyophilized Equine Platelet-Rich Plasma (L-GF equina ) Antagonize the Reproductive Toxicity and Oxidative Stress Induced by Cyclophosphamide in Female Rats
Preprint
Full-text available
  • Jan 2023
Background: The antineoplastic agent Cyclophosphamide (CP) induces reproductive toxicity. New strategies for protecting ovarian tissue damage in women with chemotherapy-induced reproductive toxicity are essential. This study was designed to evaluate the possible protective effect of combined treatment with L-GFequina on CP-induced reproductive toxicity in the mature female rat. Methodology: Forty mature female rats were assigned into four groups: First group, control: rats were intraperitoneally injected (IP) with 200 μl sterile saline solution on days 1 and 10; Group 2 (CP): were IP injected with 75 mg/kg on days 1 and 10 to induce POI); Group 3 (CP + L-GFequina): as in group 2 + IP injected with 200 μl rehydrated L-GFequina half-hour after CP injection on day 1 and 10); Group 4 (L-GFequina): rats were IP injected with 200 μl L-GFequina on day 1 and 10). Blood samples were collected for complete blood picture and determinations of nitric oxide and malondialdehyde. Animals were sacrificed on Day-21, genitalis was dissected, weighted and fixed in 10% formalin for histopathological and morphometric evaluation. Results: On day 21 of the experiment, body weight, ovarian parameters (Ovarian weight, uterine weight, the number of ovarian follicles, and corpora lutea (CL) were determined, and histopathological changes, blood profile, as well as antioxidant activity assessment, were performed. CP significantly suppresses ovarian and uterine functions and increased MAD, NO levels, RBCs, hemoglobin, WBCs and platelet count compared to the control group ( P < 0.05). While, in CP + L-GFequina group, gross, histomorphometric parameters, blood, and biochemical markers were similar to that in the control. IP injection of L-GFequina alone significantly (P<0.05) increased body weight, and ovarian and uterine morphometry compared with the control. Conclusion: co-administration of L-GFequina with CP might protect the reproductive organs in rats through its high antioxidant capacity.
Chemotherapy Induced Cardiotoxicity: A State of the Art Review on General Mechanisms, Prevention, Treatment and Recent Advances in Novel Therapeutics
Article
As medicine advances to employ sophisticated anticancer agents to treat a vast array of oncological conditions, it is worth considering side effects associated with several chemotherapeutics. One adverse effect observed with several classes of chemotherapy agents is cardiotoxicity which leads to reduced ejection fraction (EF), cardiac arrhythmias, hypertension and Ischemia/myocardial infarction that can significantly impact quality of life and patient outcomes. Research into possible mechanisms has elucidated several mechanisms, such as ROS generation, calcium overload and apoptosis. However, there is a relative scarcity of literature detailing the relationship between exact mechanism of cardiotoxicity for each anticancer agent and observed clinical effects. This review comprehensively describes cardiotoxicity associated with various classes of anticancer agents and possible mechanisms. Further research exploring possible mechanisms for cardiotoxicity observed with anticancer agents could provide valuable insight into susceptibility for developing symptoms and management guidelines. Chemotherapeutics are associated with several side effects. Several classes of chemotherapy agents cause cardiotoxicity leading to a reduced ejection fraction (EF), cardiac arrhythmias, hypertension, and Ischemia/myocardial infarction. Research into possible mechanisms has elucidated several mechanisms, such as ROS generation, calcium overload, and apoptosis. However, there is a relative scarcity of literature detailing the relationship between the exact mechanism of cardiotoxicity for each anticancer agent and observed clinical effects. This review describes cardiotoxicity associated with various classes of anticancer agents and possible mechanisms. Further research exploring mechanisms for cardiotoxicity observed with anticancer agents could provide insight that will guide management.
Immunotherapy for Triple-Negative Breast Cancer: Combination Strategies to Improve Outcome
Article
Full-text available
  • Jan 2023
Due to the absence of hormone receptor (both estrogen receptors and progesterone receptors) along with human epidermal growth factor receptor 2 (HER-2) amplification, the treatment of triple-negative breast cancer (TNBC) cannot benefit from endocrine or anti-HER-2 therapy. For a long time, chemotherapy was the only systemic treatment for TNBC. Due to the lack of effective treatment options, the prognosis for TNBC is extremely poor. The successful application of immune checkpoint inhibitors (ICIs) launched the era of immunotherapy in TNBC. However, the current findings show modest efficacy of programmed cell death- (ligand) 1 (PD-(L)1) inhibitors monotherapy and only a small proportion of patients can benefit from this approach. Based on the basic principles of immunotherapy and the characteristics of the tumor immune microenvironment (TIME) in TNBC, immune combination therapy is expected to further enhance the efficacy and expand the beneficiary population of patients. Given the diversity of drugs that can be combined, it is important to select effective biomarkers to identify the target population. Moreover, the side effects associated with the combination of multiple drugs should also be considered.
IgA nephropathy: an overview of drug treatments in clinical trials
Article
  • Jan 2023
Introduction IgA nephropathy (IgAN) is the commonest primary glomerulonephritis worldwide and may progress to end-stage kidney disease (ESKD) within a 10–20 year period. Its slowly progressive course has made clinical trials challenging to perform, however the acceptance of proteinuria reduction as a surrogate end point has significantly improved the feasibility of conducting clinical trials in IgAN, with several novel and repurposed therapies currently undergoing assessment. Already, interim results are demonstrating value to some of these, offering great hope to those with IgAN. Areas covered This review explores the rationale, candidates, clinical precedents, and trial status of therapies that are currently or have recently been evaluated for efficacy in IgAN. All IgAN trials registered with the U.S. National Library of Medicine; ClinicalTrials.gov were reviewed. Expert opinion For the first time, effective treatment options beyond supportive care are becoming available for those with IgAN. This is the culmination of commendable international efforts and signifies a new era for those with IgAN. As more therapies become available, future challenges will revolve around deciding which treatments are most appropriate for individual patients, which is likely to push IgAN into the realm of precision medicine.
Decreased FOXP3+ and GARP+ Tregs to neoadjuvant chemotherapy associated with favorable prognosis in advanced gastric cancer
Article
Full-text available
  • Jun 2016
Neoadjuvant chemotherapy (NACT) has been an increasingly used therapeutic strategy to improve the outcome of advanced gastric cancer (GC) over the past few decades. Lymphocytic infiltration has been reported to be associated with response to NACT, but the immune cell subpopulation and its prognosis contributing to response in GC have not been clarified yet. In the current study, the tumor infiltration of FOXP3+ and GARP+ regulatory T-cells (Tregs, marked by FOXP3 and GARP) response to NACT in advanced GC and their correlation with prognosis were evaluated. The infiltration of FOXP3+ and GARP+ Tregs in 102 patients with advanced GC who were treated with or without NACT was measured using immunohistochemical method. The infiltration of FOXP3+ and GARP+ Tregs was significantly decreased in the NACT group than in the non-NACT group (P=0.023 and P=0.012, respectively) and significantly associated with tumor, node, metastasis stage (P=0.019 and P=0.011, respectively). There was no significant difference in patient's overall survival between the NACT and non-NACT groups (P=0.166); however, patients in the NACT group with decreased infiltration of FOXP3+ and GARP+ Tregs had longer overall survival (P=0.002 and P<0.001, respectively). Univariate and multivariate analyses indicated that the infiltration of GARP+ Tregs and lymph node metastasis were independent prognostic factors (P=0.038 and P=0.013, respectively). The results demonstrated that NACT could decrease the infiltration of FOXP3+ and GARP+ Tregs, and that the infiltration of GARP+ Tregs may serve as a new prognostic factor of human GC response to NACT.
Glutathione S Transferases Polymorphisms Are Independent Prognostic Factors in Lupus Nephritis Treated with Cyclophosphamide
Article
Full-text available
Objective To investigate association between genetic polymorphisms of GST, CYP and renal outcome or occurrence of adverse drug reactions (ADRs) in lupus nephritis (LN) treated with cyclophosphamide (CYC). CYC, as a pro-drug, requires bioactivation through multiple hepatic cytochrome P450s and glutathione S transferases (GST). Methods We carried out a multicentric retrospective study including 70 patients with proliferative LN treated with CYC. Patients were genotyped for polymorphisms of the CYP2B6, CYP2C19, GSTP1, GSTM1 and GSTT1 genes. Complete remission (CR) was defined as proteinuria ≤0.33g/day and serum creatinine ≤124 µmol/l. Partial remission (PR) was defined as proteinuria ≤1.5g/day with a 50% decrease of the baseline proteinuria value and serum creatinine no greater than 25% above baseline. Results Most patients were women (84%) and 77% were Caucasian. The mean age at LN diagnosis was 41 ± 10 years. The frequency of patients carrying the GST null genotype GSTT1-, GSTM1-, and the Ile→105Val GSTP1 genotype were respectively 38%, 60% and 44%. In multivariate analysis, the Ile→105Val GSTP1 genotype was an independent factor of poor renal outcome (achievement of CR or PR) (OR = 5.01 95% CI [1.02–24.51]) and the sole factor that influenced occurrence of ADRs was the GSTM1 null genotype (OR = 3.34 95% CI [1.064–10.58]). No association between polymorphisms of cytochrome P450s gene and efficacy or ADRs was observed. Conclusion This study suggests that GST polymorphisms highly impact renal outcome and occurrence of ADRs related to CYC in LN patients.
Dendritic cells combined with doxorubicin induces immunogenic cell death and exhibits antitumor effects for osteosarcoma
Article
Full-text available
  • Feb 2016
The effects of dendritic cells (DCs) with low dose doxorubicin on the enhancement of the systemic immune response, including the effects on calreticulin (CRT) expression, heat shock protein 70 (HSP70) on the cell surface expression, and the enhancement of high mobility group box 1 (HMGB1) release from cancer cells, remain unclear. The present study investigated whether the combination of DCs and doxorubicin (ADM) induces immune cell death, and leads to tumor growth inhibition in a murine osteosarcoma model. To evaluate immune response activation in vivo, 4 groups of mice were established: i) untreated mice, ii) DC-treated mice, iii) ADM-treated mice, and iv) DC and ADM-treated mice. Immunological cell death and CRT, HSP70, and HMGB1 expression levels were higher in doxorubicin-treated cells than those in untreated or those treated with DCs alone. NF-κB expression was higher in the DCs after ligand activation using CRT, HSP70, or HMGB1 in vitro. Mice treated with DCs and ADM displayed an increased number of CD8(+) T-lymphocytes within metastatic tumors and inhibition of metastatic growth. The expression of CRT and the release of HMGB1 from tumor tissues were increased in the ADM-treated groups. Treatment with DCs and ADM resulted in the highest serum interferon-γ levels. Combining ADM, which can induce immunogenic cell death, with DCs enhanced the systemic immune response. The findings of the present study provide further support for the continued development of antitumor agents that induce cell death and the immune response to target osteosarcoma.
Increased circulating myeloid-derived suppressor cells correlate with clinical cancer stage, metastatic tumor burden, and doxorubicin-cyclophosphamide chemotherapy
Article
Full-text available
  • Jan 2009
Abnormal accumulation of myeloid-derived suppressor cells (MDSC) is an important mechanism of tumor immune evasion. Cyclophosphamide (CTX) has also been shown in non-tumor bearing animals to cause transient surges in MDSC. Knowledge of MDSC is primarily based on preclinical work, and to date only few published studies have involved cancer patients. The goal of this study was to test the hypothesis that circulating MDSC levels correlate with clinical cancer stage, CTX-based chemo-therapy, and metastatic tumor burden. Whole blood was collected from 106 newly diagnosed solid tumor patients (stages I–IV). Percentages of circulating MDSC (Lin ¡/Lo , HLA DR¡, CD33 + CD11b +) were determined prior to initiation of systemic therapy. In 17 early stage breast cancer patients receiving doxorubicin–cyclophosphamide chemo-therapy every 14 days (ddAC) blood was collected on day 1 of each cycle. Circulating MDSC were signiWcantly increased in cancer patients of all stages relative to healthy volunteers. A signiWcant correlation between circulating MDSC and clinical cancer stage was also observed. Moreover , among stage IV patients, those with extensive meta-static tumor burden had the highest percent and absolute number of MDSC. SigniWcant increases in circulating MDSC were observed with ddAC when compared with pretreatment levels. Circulating MDSC levels correlate with clinical cancer stage, ddAC, and metastatic tumor burden. This information must be incorporated into the design of future trials exploring immune-based therapeutic strategies. Pharmacologic modulation of MDSC should also be tested in future clinical trials.
A randomized phase II clinical trial of personalized peptide vaccination with metronomic low-dose cyclophosphamide in patients with metastatic castration-resistant prostate cancer
Article
Full-text available
This study investigated the effect of metronomic cyclophosphamide (CPA) in combination with personalized peptide vaccination (PPV) on regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSC), and whether it could improve the antitumor effect of PPV. Seventy patients with metastatic castration-resistant prostate cancer were randomly assigned (1:1) to receive PPV plus oral low-dose CPA (50 mg/day), or PPV alone. PPV treatment used a maximum of four peptides chosen from 31 pooled peptides according to human leukocyte antigen types and antigen-specific humoral immune responses before PPV, for 8 subcutaneous weekly injections. Peptide-specific cytotoxic T lymphocyte (CTL) and immunoglobulin G responses were measured before and after PPV. The incidence of grade 3 or 4 hematologic adverse events was higher in the PPV plus CPA arm than in the PPV alone arm. Decrease in Treg and increase in MDSC were more pronounced in PPV plus CPA treatment than in PPV alone (p = 0.036 and p = 0.048, respectively). There was no correlation between the changes in Treg or MDSC and CTL response. There was no difference in positive immune responses between the two arms, although overall survival in patients with positive immune responses was longer than in those with negative immune responses (p = 0.001). Significant differences in neither progression-free survival nor overall survival were observed between the two arms. Low-dose CPA showed no change in the antitumor effect of PPV, possibly due to the simultaneous decrease in Treg and increase in MDSC, in patients under PPV.
Enhancement of adoptive T Cell transfer with single low dose pretreatment of doxorubicin or paclitaxel in mice
Article
Full-text available
  • Dec 2015
Ex vivo expansion of CD8+ T-cells has been a hindrance for the success of adoptive T cell transfer in clinic. Currently, preconditioning with chemotherapy is used to modulate the patient immunity before ACT, however, the tumor microenvironment beneficial for transferring T cells may also be damaged. Here preconditioning with single low dose of doxorubicin or paclitaxel combined with fewer CD8+ T-cells was investigated to verify whether the same therapeutic efficacy of ACT could be achieved. An E.G7/OT1 animal model that involved adoptive transfer of OVA-specific CD8+ T-cells transduced with a granzyme B promoter-driven firefly luciferase and tomato fluorescent fusion reporter gene was used to evaluate this strategy. The result showed that CD8+ T-cells were activated and sustained longer in mice pretreated with one low-dose Dox or Tax. Enhanced therapeutic efficacy was found in Dox or Tax combined with 2x106 CD8+ T-cells and achieved the same level of tumor growth inhibition as that of 5x106 CD8+ T-cells group. Notably, reduced numbers of Tregs and myeloid derived suppressor cells were shown in combination groups. By contrast, the number of tumor-infiltrating cytotoxic T lymphocytes and IL-12 were increased. The NF-κB activity and immunosuppressive factors such as TGF-β, IDO, CCL2, VEGF, CCL22, COX-2 and IL-10 were suppressed. This study demonstrates that preconditioning with single low dose Dox or Tax and combined with two fifth of the original CD8+ T-cells could improve the tumor microenvironment via suppression of NF-κB and its related immunosuppressors, and activate more CD8+ T-cells which also stay longer.
Nitric oxide-producing CD11b+Ly-6G(Gr-1)+CD31(ER-MP12)+cells in the spleen of cyclophosphamide–treated mice: implications for T-cell responses in immunosuppressed mice
Article
  • Jan 2000
During recovery from intensive chemotherapy with cyclophosphamide (CTX), mice suffer a severe but transitory impairment in spleen cell proliferation to T-cell mitogens (Con A or anti-CD3 plus IL-2). Although CTX treatment reduced spleen T-cell cellularity, this cannot fully account for T-cell unresponsiveness. The results showed that CTX induces the colonization of spleen by an immature myeloid CD11b+Ly-6G+CD31+ population. Its presence closely correlated with the maximum inhibition of T-cell proliferation. Moreover, this suppressive activity was dependent on nitric oxide (NO) production in cultures since (1) higher amounts of nitric oxide and inducible nitric oxide synthase (iNOS) mRNA were produced in CTX spleen cells (CTX-SC) than in control splenocyte cultures and (2) NOS inhibitors greatly improved the proliferation of T lymphocytes. Nitric oxide production and suppressive activity were also dependent on endogenous interferon-γ (IFN-γ) production since anti–IFN-γ abrogated both effects. Finally, iNOS protein expression was restricted to a heterogeneous population of CD31+cells in which CD11b+Ly-6G+ cells were required to suppress T-cell proliferation. These results indicated that CTX might also cause immunosuppression by a mechanism involving the presence of immature myeloid cells with suppressor activity. This may have implications in clinical praxis since inappropriate immunotherapies in patients treated with intensive chemotherapy could lead to deleterious T-cell responses. (Blood. 2000;95:212-220)
Relationship of glutathione S-transferase genotypes with side-effects of pulsed cyclophsphamide therapy in patients with systemic lupus erythematosus
Article
  • Jul 2006
b>Aims Cyclophosphamide (CTX) is an established treatment of severe systemic lupus erythematosus (SLE). Cytotoxic CTX metabolites are mainly detoxified by multiple glutathione S-transferases (GSTs). However, data are lacking on the relationship between the short-term side-effects of CTX therapy and GST genotypes. In the present study, the effects of common GSTM1 , GSTT1 , and GSTP1 genetic mutations on the severity of myelosuppression, gastrointestinal (GI) toxicity, and infection incidences induced by pulsed CTX therapy were evaluated in patients SLE. Methods DNA was extracted from peripheral leucocytes in patients with confirmed SLE diagnosis ( n = 102). GSTM1 and GSTT1 null mutations were analyzed by a polymerase chain reaction (PCR)-multiplex procedure, whereas the GSTP1 codon 105 polymorphism (Ile→Val) was analyzed by a PCR-restriction fragment length polymorphism (RFLP) assay. Results Our study demonstrated that SLE patients carrying the genotypes with GSTP1 codon 105 mutation [ GSTP1 *-105I/V (heterozygote) and GSTP1 *-105 V/V (homozygote)] had an increased risk of myelotoxicity when treated with pulsed high-dose CTX therapy (Odds ratio (OR) 5.00, 95% confidence interval (CI) 1.96, 12.76); especially in patients younger than 30 years (OR 7.50, 95% CI 2.14, 26.24), or in patients treated with a total CTX dose greater than 1.0 g (OR 12.88, 95% CI 3.16, 52.57). Similarly, patients with these genotypes ( GSTP1 *I/V and GSTP1 *V/V) also had an increased risk of GI toxicity when treated with an initial pulsed high-dose CTX regimen (OR 3.33, 95% CI 1.03, 10.79). However, GSTM1 and GSTT1 null mutations did not significantly alter the risks of these short-term side-effects of pulsed high-dose CTX therapy in SLE patients. Conclusions The GSTP1 codon 105 polymorphism, but not GSTM1 or GSTT1 null mutations, significantly increased the risks of short-term side-effects of pulsed high-dose CTX therapy in SLE patients. Because of the lack of selective substrates for a GST enzyme phenotyping study, timely detection of this mutation on codon 105 may assist in optimizing pulsed high-dose CTX therapy in SLE patients.<br /
Immunological Effects of Conventional Chemotherapy and Targeted Anticancer Agents
Article
The tremendous clinical success of checkpoint blockers illustrates the potential of reestablishing latent immunosurveillance for cancer therapy. Although largely neglected in the clinical practice, accumulating evidence indicates that the efficacy of conventional and targeted anticancer agents does not only involve direct cytostatic/cytotoxic effects, but also relies on the (re)activation of tumor-targeting immune responses. Chemotherapy can promote such responses by increasing the immunogenicity of malignant cells, or by inhibiting immunosuppressive circuitries that are established by developing neoplasms. These immunological "side" effects of chemotherapy are desirable, and their in-depth comprehension will facilitate the design of novel combinatorial regimens with improved clinical efficacy.