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ORIGINAL ARTICLE
Year : 2015  |  Volume : 52  |  Issue : 4  |  Page : 587-589
 

Effect of dendritic cell vaccine therapy on lymphocyte subpopulation in refractory primary brain tumor


1 Department of Oncology, Institute of Oncology, Tianjin, China
2 Department of Oncology, Institute of Oncology, Tianjin; Department of Operation, Shanghai Claison Biotechnology Co. Limited, Shanghai, China
3 Department of Oncology, Institute of Oncology; Institute of Oncology, Tianjin Union Medicine Centre, Tianjin, China

Date of Web Publication10-Mar-2016

Correspondence Address:
Y Pang
Department of Oncology, Institute of Oncology, Tianjin
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0019-509X.178373

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 » Abstract 

BACKGROUND: Dendritic cell (DC)-based immunotherapy has the potential to induce an antitumor response within the immunologically privileged brain. AIMS: The aim of this study was to evaluate the short-term effect of DC vaccine therapy on lymphocyte subsets in patients with refractory primary brain tumor. MATERIALS AND METHODS: Eighteen cases with refractory primary brain tumor who refused any treatment against tumor within 6 months of the therapy, were referred to one medicine center, from January 2011 to October 2012. All patients received 1 × 107 tumor lysate–pulsed DC vaccinations both intradermal injection and intravenous infusion 3 times/week. RESULTS: There were increases of lymphocytes CD8+ (P = 0.002) and CD56+ (P = 4.207E-10), but no change of lymphocytes CD3+ (P = 0.651). Six patients were positive response of delayed-type hypersensitivity. There were improving of appetite in 14 cases and increasing of physical strength 17 cases. CONCLUSIONS: DC vaccine has the potential for inducing an immune cytotoxic effect directed toward tumor cells.


Keywords: Dendritic cell vaccine, immunotherapy, primary brain tumor


How to cite this article:
Niu J, Chang Y, Lu X, Wu X, Pang Y. Effect of dendritic cell vaccine therapy on lymphocyte subpopulation in refractory primary brain tumor. Indian J Cancer 2015;52:587-9

How to cite this URL:
Niu J, Chang Y, Lu X, Wu X, Pang Y. Effect of dendritic cell vaccine therapy on lymphocyte subpopulation in refractory primary brain tumor. Indian J Cancer [serial online] 2015 [cited 2019 Dec 15];52:587-9. Available from: http://www.indianjcancer.com/text.asp?2015/52/4/587/178373





 » Introduction Top


It is believed that dendritic cell (DC) vaccines may play an adjuvant role in primary brain tumor by consolidating the responses to conventional therapy.[1] This study demonstrates the feasibility and safety of DC vaccine for primary brain tumor.


 » Materials and Methods Top


A total of 18 cases with refractory primary brain tumor in this study who refused any treatment against tumor within 6 months of the therapy, were referred to one Immunotherapy Medicine Center, from January 2011 to October 2012. The study protocol was approved by the hospital's Ethical Committees and class III medical techniques of “treatment with autologous immune cells (T-cells, NK cells)” in accordance with the policy by the Minister of Health of China. All patients provided informed consent before treatment.

The patient tumor tissue specimens were removed from the vivid primary brain tumor portion during operation, and immediately placed in phosphate buffer solution (PBS) (5/18 patients were available in this study), or human cell lines U251 (for the remaining 13/18 unresectable primary brain tumor patients) were then mechanically dissociated. Tumor cells were dispersed to create a single-cell suspension. Cells were crushed by ultrasound and large particles were removed by centrifuged with a 600 g for 30 min. The supernatants were collected as tumor lysate for sensitizing DCs and delayed-type hypersensitivity (DTH) test.

Dendritic cells were prepared by the method described in many papers.[2] Briefly, a concentrated 100 ml leukocyte fraction was generated using the Fresenius Kabi system through a 1-h restricted peripheral blood leukapheresis processing 3–4 L of blood with each collection. The leukapheresis product was enriched for monocytes. Peripheral blood mononuclear cells were then purified and cultured in the presence of granulocyte-macrophage colony-stimulating factor, interleukin-4, tumor necrosis factor, etc., at 37°C for 7 days. Then, cells were harvested and used as DCs for pulsing with the tumor lysate. DCs cultured on the 8th day were harvested and viable cells were enumerated and administered. DCs were washed 3 times with PBS and resuspended in PBS for immediate intraderminal or intravenous vaccination; the remaining cells were frozen for the further vaccination.

Dendritic cells cultured on the 1st and 8th days were collected and resuspended in PBS containing 1% bovine serum albumin and 0.1% sodium azide, and stained with fluorescein isothiocyanate labeled mouse antihuman CD14, CD83 and CD86, and the phycoerythrin labeled mouse antihuman monoclonal antibodies against human leukocyte antigen-DR (HLA2-DR) monoclonal antibodies for 30 min at 4°C. Stained cells were washed and analyzed by flow cytometry (FCM).

The release criteria of DCs were: Viability >70%, no organisms on gram stain, cultured samples with no bacterial growth, and endotoxin levels <5 EU/kg. 1 × 107 cells were resuspended in 4 ml NS for two syringes, and injected into the bilateral subaxillary region of patients by the mean of 24 points intraderminal injection. Concurrently, 1 × 107 cells were resuspended in 100 ml normal saline, and infused into the patients intravenously. Patients were vaccinated once a week for 3 weeks. All patients were monitored within 3 months after the first vaccination in order to evaluate the short-term effect of DC vaccine therapy in primary brain tumor. The immune responses to the DC vaccination were analyzed by comparing the immune phenotype changes in peripheral blood of patients and DTH reactions. All toxicities involved in DC vaccine therapy were carefully monitored.

The cell surface phenotypes of peripheral blood T lymphocytes D3+, CD8+ and CD56+ expression level were detected before and after DC vaccine therapy with FCM assay. DTH was tested with intradermal administration of 40 μg/0.1 mL tuSmor lysate into the forearm 1 week after the end of vaccination. A positive skin test reaction was defined as >2 mm diameter in duration after 48 h.

Data were analyzed with Statistical Analysis System (version 8.1). Means are reported with a standard deviation The pre- and post-vaccination data were compared using Student's t-test. P < 0.05 was determined statistically significant.


 » Results Top


There were an increase of cell surface phenotypes of CD83+, CD86+, HLA2DR and decrease that of CD14+ in immune phenotype of matured DCs after 7 days cultured by the method of FCM. This difference was significant (data not shown). There were 18 cases in this study, 10 men and 8 women with an age range from 4 to 72 years (average 51.8 years), showed in [Table 1]. There were five cases of high-grade (Grade III–IV) brain tumors (three cases of Grade III astrocytoma and two glioblastoma multiforme) and 13 cases of low grade (Grade I–II) brain tumors (seven cases of Grades I–II astrocytoma, two ependymomas, one pituitary tumor, two meningiomas and one craniopharyngioma, respectively). Of 18 cases, 13 had no proper treatment against tumor and five patients had undergone a repetitive surgery, radiotherapy, and chemotherapy. All 18 cases received no cancer directed treatment within 6 months of DC vaccination.
Table 1: Characteristics of patients and clinical responses of DC vaccine therapy in 18 patients
(s, n=18)


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Six patients complained of recurrent headache lasting for 2–5 days that were relieved by intracranial pressure lowering agent - mannitol. Four patients developed generalized seizures lasting for 10 min to 3 h after vaccination, which resolved with antiepileptic medication. One patient with severe and two with mild erythema at the injection site were observed at the end of vaccination. Fever was seen in 11 cases received DCs vaccine, which relieved with 5 mg dexamethasone injection and/or physical cooling. Of 11 cases with fever, eight were mild (37–39°C), and three were severe (>39°C). There were no other serious adverse events observed.

There were increases of lymphocytes CD8+ (45.25 ± 4.82 vs. 39.75 ± 5.17, P = 0.002) and CD56+ (31.90 ± 4.85 vs. 19. 48 ± 3. 69, P = 4.207E-10), but no change of lymphocytes CD3+ (64.98 ± 7.64 vs. 66.28 ± 9.35, P = 0.651), showed in [Table 2]. To test the cell-mediated cytotoxicity response, DTH skin test was performed 1 week after the vaccination. Of 18 patients, six were positive, and 12 were DTH-negative.
Table 2: Analysis of immune function in 18 patients (s, n=18)


Click here to view



 » Discussion Top


In this studies, the cell surface phenotypes of peripheral blood T-lymphocytes CD8+ (45.25 ± 4.82 vs. 39.75 ± 5.17, P = 0.002) and CD56+ (31.90 ± 4.85 vs. 19. 48 ± 3. 69, P = 4.207E-10) increased significantly after DC vaccine therapy. Six out of 18 patients were DTH positive. Since a compromised immune function is a common feature of advanced malignancy. Increases of the cell surface phenotypes of peripheral blood T-lymphocytes CD8+ and CD56+ and positive DTH response means DCs vaccine has affected the patients' immune system. Because DTH test is simple and cheap, it could be as a therapeutically effective marker to predict the clinical response of primary brain tumor patients under the DCs vaccination.[3]

Our results indicated that DC vaccine showed a promising approach for inducing an immune cytotoxic effect directed toward tumor cells and an effective clinical response in patients with primary brain tumor. The protocols should be determined more in detail. The development of methods for combining the use of the DCs vaccination with other treatments will enhance the clinical usefulness of DCs-based immunotherapy for primary brain tumor.[4],[5]

 
 » References Top

1.
Ardon H, Verbinnen B, Maes W, Beez T, Van Gool S, De Vleeschouwer S. Technical advancement in regulatory T cell isolation and characterization using CD127 expression in patients with malignant glioma treated with autologous dendritic cell vaccination. J Immunol Methods 2010;352:169-73.  Back to cited text no. 1
    
2.
Zhang SN, Choi IK, Huang JH, Yoo JY, Choi KJ, Yun CO. Optimizing DC vaccination by combination with oncolytic adenovirus coexpressing IL-12 and GM-CSF. Mol Ther 2011;19:1558-68.  Back to cited text no. 2
    
3.
Wolchok JD, Hoos A, O'Day S, Weber JS, Hamid O, Lebbé C, et al. Guidelines for the evaluation of immune therapy activity in solid tumors: Immune-related response criteria. Clin Cancer Res 2009;15:7412-20.  Back to cited text no. 3
    
4.
Steinman RM. Decisions about dendritic cells: Past, present, and future. Annu Rev Immunol 2012;30:1-22.  Back to cited text no. 4
    
5.
Okada H, Kalinski P, Ueda R, Hoji A, Kohanbash G, Donegan TE, et al. Induction of CD8+T-cell responses against novel glioma-associated antigen peptides and clinical activity by vaccinations with {alpha}-type 1 polarized dendritic cells and polyinosinic-polycytidylic acid stabilized by lysine and carboxymethylcellulose in patients with recurrent malignant glioma. J Clin Oncol 2011;29:330-6.  Back to cited text no. 5
    



 
 
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  [Table 1], [Table 2]



 

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