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改性柑橘果膠MCP激活T輔助/誘導細胞，毒性T-細胞，B細胞，自然吞噬（NK）細胞與誘導的自然殺傷細胞對抗K562慢性髓系白血病細胞

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

《BMC補充與替代醫(yī)學》2011，11:59

摘要

背景：改性柑橘果膠（MCP）能被人體有效的吸收利用，具有顯著的抗癌效果。本文對于MCP激活人血淋巴細胞亞群如T、B和NK細胞活性的效果進行了驗證。

方法：經(jīng)MCP處理的人血樣品和特定抗體混合進行培養(yǎng)，采用三色流式細胞技術進行分析。分離的正常淋巴細胞與不同濃度的MCP混合，以驗證激活后NK細胞的活性。PKH26標記的相對期K562白血病細胞與淋巴細胞混合培養(yǎng)4小時。上述的混合物用FITC標記的活化型Caspase3抗體進行染色，用二色流式細胞技術進行分析。對于PKH26和FITC呈現(xiàn)陽性的K562細胞作為NK細胞誘導死亡的細胞進行計數(shù)。用高效陰離子交換色譜-脈沖安培檢測法分析MCP的單糖分析。

結果：MCP對于細胞毒性T細胞和B細胞呈現(xiàn)劑量依賴性激活，而對于NK細胞呈現(xiàn)顯著的劑量依賴性激活效應。MCP激活的NK細胞能有效殺傷癌細胞。對于MCP成分分析表明，MCP由含有4,5-不飽和非還原性端的寡聚半乳糖醛酸組成。

結論：MCP對于人血細胞呈現(xiàn)免疫調(diào)節(jié)的功能，包括激活NK細胞對抗K562白血病細胞。不飽和性寡聚半乳糖醛酸可能是MCP中免疫調(diào)節(jié)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.