DC vaccine & Anti-tumor “ 生命之树 ” 课题组： 孔琰 唐跃 田亚楠 孔琰 唐跃 田亚楠 王超 王京 王欠欠 王超 王京 王欠欠

The tumor induced immunity DC vaccine Induce in vitro & proliferation Classification DC vaccine The function of DC in anti-tumor Reinfuse of DC Our opinion

The function of DC in anti-tumor Mechanism of DC in anti-tumor Mechanism of DC in anti-tumor Inhibit formation of tumor vessel NO,TNF etc Migration proliferation Tumor Ag DC T CTL Tolerance Cell apoptosis 、 lysis 、 CK

Influence to DC from CKs produced by tumor VEGFC （血管内皮生长因子） —— Depress the differentiation of pre-DC to mature DC IL-10—— Inhibit the expression of co-stimulatory molecules TGF-ß—— Inhibit the activation of specific CTL DC-SIGN

DC vaccine Definition: the DC derive from peripheral blood mononuclear cells(PBMCs) are stimulated by peptide antigen,tumor lysates,whole-cell prepration RNA or DNA of tumor.And the reinfuse the DC into the patient as vaccine.

HOW TO GAIN DC ? HOW TO GAIN DC ? Peripheral blood mononuclear cells are proper for clinical experiment. GM-CSF / IL-4 CD14+ DC CD34+ DC GM-CSF / TNF-α DC vaccine

HOW TO GAIN DC ? HOW TO GAIN DC ? Peripheral blood mononuclear cells are proper for clinical experiment. GM-CSF / IL-4 CD14+ DC CD34+ DC GM-CSF / TNF-α DC vaccine

Ag pep loaded DC subcellular vaccine 4 Classification DC vaccine Peptide-pulsed DC vaccine 1 Tumor cell lysate-sensitized DC 2 DC-tumor cell hybrids 3 Genetically modified DC vaccine 5

1 Peptide-pulsed DC vaccine The tumor peptide in HCC is mainly alpha- fetoprotein(AFP)

Introduction Introduction AFP It has no affection to DC even at high concention. Glypican—3HbsAg

1 Peptide-pulsed DC vaccine Advantages Targeting, avoid unnecessary immune stimulation. higher concentration of tumor Ag stimulate the activation of T cell efficiently Disadvantages There is little confirmed; Specific recognization Little monoclonal tumor Ag

2 Tumor cell lysate-sensitized DC Advantages ： Advantages ： tumor cell Ag prepared by direct supersonic breakdown or repeated freezing&melting etc. is useful ， no desire for Ag recognizing epitope tumor cell Ag prepared by direct supersonic breakdown or repeated freezing&melting etc. is useful ， no desire for Ag recognizing epitope Disadvantages: Disadvantages: hard to supply large amount of fluid tissue ； hard to supply large amount of fluid tissue ； tumor lysate include normal self-Ag,which easily induce self-immune disease after immunization 。 tumor lysate include normal self-Ag,which easily induce self-immune disease after immunization 。

2 Tumor cell lysate-sensitized DC Current Situation ： Current Situation ： Ladhams 、 Iwashita Ladhams 、 Iwashita Ali Ali Hayashi Hayashi

＋ DCTumor cell 3 DC-tumor cell hybrids DC-Tumor hybrids

＋ DCTumor cell 3 DC-tumor cell hybrids DC-Tumor hybrids

Hybrid cell Tumor cell Unrelated tumor cell Tumor cell

Advantages Advantages Easily abtaned from biopsy 、 operation,HCC tissue specimen from ； advanced skill to obtain pre-DC via cell culture ； hybrid cell can present HCC Ag in the form of MHC pep complex ； hybrid cells in vivo can stimulate to form lasing & steady Ag Easily abtaned from biopsy 、 operation,HCC tissue specimen from ； advanced skill to obtain pre-DC via cell culture ； hybrid cell can present HCC Ag in the form of MHC pep complex ； hybrid cells in vivo can stimulate to form lasing & steady Ag Disadvantages: Selection of hybrided cells 3 DC-tumor cell hybrids

4 Antigenic peptide loaded DC subcellular vaccine

DC subcellular structure & other DC hybrids inhense expression of DC

5 Genetically modified DC vaccine Antigen gene (HCC HBsAg AFP) Antigen gene (HCC HBsAg AFP) Cytokine gene(GM-CSF IL-12) Cytokine gene(GM-CSF IL-12) Chemotactic gene (lymphotactin Lpin) Chemotactic gene (lymphotactin Lpin)

5 Genetically modified DC vaccine Advantages ： AFP gene modefied DC vaccine can express AFP gene modefied DC vaccine can express completely endogenously & continuously completely endogenously & continuously reduce autoimmune diseases reduce autoimmune diseases Disadvantages: The technology is not developed The technology is not developed

HOT SPOTS IL-18 & Survivin( 存活素 )

Other methods to improve the effect of DC vaccine NK cell& DC cell interaction Heat shock protein （ HSP ） Mannosalization of Ag

Other methods to improve the effect of DC vaccine NK cell& DC cell interaction Heat shock protein （ HSP ） Mannosalization of Ag

Reinfuse of DC vaccine subcutaneous venousLymph node ?

Ag-loaded DC Produce selective T cell chemotactic factors Migrate to lymph node &spleen Self or other APC secrete IL-12 CD8 + Tcell Secrete CKs & cause damage CD4 + Th0Th1IFN-γ TNF-α GM-CSF IL-3

FORWARD Until now many cancers can be treated, such as: B cell lymphoma cancer Prostate cancer Malignant melanoma

曹雪涛 ( 左 ) 正在进行大肠癌 DC 疫苗研究

Development and maturation of DC

Difference between immature DC and mature DC

Characteristics of DC ： 1. the most potent APC 1. the most potent APC 2.the only cell can active naïve T cell to elicit primary immune response. 2.the only cell can active naïve T cell to elicit primary immune response. 3.play an significant role in anti-tumor 3.play an significant role in anti-tumor

NK cell& DC cell interaction DC attaches at the surface of NK cell and stimulates related receptors,so these two cells can communicate in the form of neural synapse.