改性柑橘果胶MCP激活T辅助/诱导细胞，毒性T-细胞，B细胞，自然吞噬（NK）细胞与诱导的自然杀伤细胞对抗K562慢性髓系白血病细胞

Cheppail Ramachandran, Barry J Wilk, Arland Hotchkiss, Hoa Chau, Isaac Eliaz and Steven J Melnick

《BMC补充与替代医学》2011，11:59

摘要

背景：改性柑橘果胶（MCP）能被人体有效的吸收利用，具有显著的抗癌效果。本文对于MCP激活人血淋巴细胞亚群如T、B和NK细胞活性的效果进行了验证。

方法：经MCP处理的人血样品和特定抗体混合进行培养，采用三色流式细胞技术进行分析。分离的正常淋巴细胞与不同浓度的MCP混合，以验证激活后NK细胞的活性。PKH26标记的相对期K562白血病细胞与淋巴细胞混合培养4小时。上述的混合物用FITC标记的活化型Caspase3抗体进行染色，用二色流式细胞技术进行分析。对于PKH26和FITC呈现阳性的K562细胞作为NK细胞诱导死亡的细胞进行计数。用高效阴离子交换色谱-脉冲安培检测法分析MCP的单糖分析。

结果：MCP对于细胞毒性T细胞和B细胞呈现剂量依赖性激活，而对于NK细胞呈现显著的剂量依赖性激活效应。MCP激活的NK细胞能有效杀伤癌细胞。对于MCP成分分析表明，MCP由含有4,5-不饱和非还原性端的寡聚半乳糖醛酸组成。

结论：MCP对于人血细胞呈现免疫调节的功能，包括激活NK细胞对抗K562白血病细胞。不饱和性寡聚半乳糖醛酸可能是MCP中免疫调节主要成分。

Activation of Human T-Helper/Inducer Cell, T-Cytotoxic Cell, B-Cell, and Natural Killer (NK)-Cells and induction of Natural Killer Cell Activity against K562 Chronic Myeloid Leukemia Cells with Modified Citrus Pectin

Cheppail Ramachandran, Barry J Wilk, Arland Hotchkiss, Hoa Chau, Isaac Eliaz and Steven J Melnick

MC Complementary and Alternative Medicine 2011,11:59   

Abstract 

Background:Modified citrus pectin (MCP) is known for its anti-cancer effects and its ability to be absorbed and circulated in the human body. In this report we tested the ability of MCP to induce the activation of human blood lymphocyte subsets like T, B and NK-cells. 

Methods:MCP treated human blood samples were incubated with specific antibody combinations and analyzed in a flow cytometer using a 3-color protocol. To test functionality of the activated NK-cells, isolated normal lymphocytes were treated with increasing concentrations of MCP. Log-phase PKH26-labeled K562 leukemic cells were added to the lymphocytes and incubated for 4 h. The mixture was stained with FITC-labeled active form of caspase 3 antibody and analyzed by a 2-color flow cytometry protocol. The percentage of K562 cells positive for PKH26 and FITC were calculated as the dead cells induced by NK-cells. Monosaccharide analysis of the MCP was 

performed by high-performance anion-exchange chromatography with pulse amperometric detection (HPAECPAD).

Results:MCP activated T-cytotoxic cells and B-cell in a dose-dependent manner, and induced significant dosedependent activation of NK-cells. MCP-activated NK-cells demonstrated functionality in inducing cancer cell death. MCP consisted of oligogalacturonic acids with some containing 4,5-unsaturated non-reducing ends.

Conclusions:MCP has immunostimulatory properties in human blood samples, including the activation of functional NK cells against K562 leukemic cells in culture. Unsaturated oligogalacturonic acids appear to be the immunostimulatory carbohydrates in MCP.