Dendritic Cell Production from Allogeneic Donor Cd34+ Stem Cells and Mononuclear Cells; Cancer Vaccine

Abstract

Aim: Active immunotherapy provides better recognition of tumor-related antigens by immune system of patient, enhanced immune system and elimination of malignant cells. This modality employs therapeutic potential of donor specific and tumor specific immune responses. Active immunotherapy targets immunosuppressive and tolerogenic mechanisms suppressed by tumor cells. T lymphocytes and antigen-presenting cells (dendritic cells) are two cell lineages that play crucial role in the battle of organism against cancer. Close similarity between cancer cells and normal cell structure is the most important reason of escape from defense cells, namely T lymphocytes. Stimulation and enhancement of T lymphocytes against cancer cells comprise principal part of therapy. Material and Method: To generate allogeneic dendritic cells, leukemic stem cells were isolated from bone marrow samples from patients with acute leukemia. Lysate was prepared from leukemic stem cells identified by flow cytometer. Stem cells and mononuclear cells (1x10>6/kg) obtained from sibling donors by apheresis were separated to produce Dendritic Cells. For Dendritic Cell transformation, GM-CSF and IL-4 were added to media where leukemic stem cell lysate from patient and mononuclear cells from sibling donor were treated. From samples taken from the culture medium, after 48, 72 and 96 hours, dendritic cell surface markers (CD 80, CD 83 and CD 86) was assessed by flow cytometry. CD3, CD14, CD19, CD56, CD66b-negative, and HLA-DR, CD86 positive are indicative of immature DC. HLA-DR, CD 80 and CD83 positive are indicative of mature DC. Results: Mononuclear cells were detected by 27 % among allogeneic hematopoietic cell groups harvested by apheresis. After culture under GMP conditions, mononuclear cell rate was found to be 24% on hours 96 and 120. It was seen that 88% of mononuclear cells transformed to mature dendritic cells after 96 hours culture. Conclusion: In cancer patients, minimal residual disease can be eliminated by active tumor vaccine after reducing tumor burden by standard methods. Tumor vaccine obtained from allogeneic sibling donor can be used in lieu of autologous tumor vaccine and it is thought to be more effective. Allogeneic dendritic cells produced at 37¡C in CO2 media under GMP conditions can be used in tumor immunotherapy. More effective method would have been used by employing dendritic cells against cancer stem cells rather than cancer cells itself. Figure Mature dendritic cells (CD83+) Figure. Mature dendritic cells (CD83+) Disclosures No relevant conflicts of interest to declare.

Modern Methods of Obtaining Immune Dendritic Cells With Anti-Tumor Potential

“…[9] зрілі ДК позиціонують як клітини з фенотипом CD11c + , CD40 + , CD80 + , CD83 + , CD86 + , CCR7 + та HLA-DR + . На поверхні зрілих ДК відсутні такі маркери, як CD3, CD14, CD19, CD56 та CD66b, характерні для T-клітин, моноцитів, В-клітин, природних кілерів і гранулоцитів відповідно [133].…”

Comparitive Study of Dendritic Cell Vaccine Preparation With Presence and Absence of Malpighia Emarginata Fruit Extract Using Tumor Rna Transfection Method: A Promising Approach for Prostate Cancer

“…Although DCs are general presence in most tissues, their total number is low. An ex vivo derivation of dendritic cells improves, and multiple prelusion cells can be used to prepare dendritic cells, such as nonproliferative and proliferative in this nonproliferative CD14+ monocytes from Peripheral blood and the proliferative CD34+ cells from bone marrow and umbilical blood [36,37]. CD14+ monocytes develop ~10% of peripheral blood mononuclear cells, and the dendritic cells derived from peripheral blood monocytes (MoDCs) have been broadly studied and applied.…”

Antigen transfer and its effect on vaccine-induced immune amplification and tolerance

“…The adoptive transfer of ex vivo-activated DCs has been widely recognized with good biosafety and sufficient efficacy in clinical anti-tumor therapy [263]. Briefly, CD34 + bone marrow progenitors or CD14 + peripheral blood monocytes are isolated and stimulated with granulocyte-macrophage colony stimulating factor (GM-CSF) and IL-4 in vitro to generate mo-DCs [264,265]. Then, the resultant mo-DCs are pulsed with 1) TAAs and/or TSAs [266,267]; 2) tumor whole cell lysates [268]; or 3) tumorderived EVs [269,270], and concurrently stimulated with adjuvants for maturation [271].…”